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EP4661911A1 - Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof - Google Patents

Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof

Info

Publication number
EP4661911A1
EP4661911A1 EP24704907.5A EP24704907A EP4661911A1 EP 4661911 A1 EP4661911 A1 EP 4661911A1 EP 24704907 A EP24704907 A EP 24704907A EP 4661911 A1 EP4661911 A1 EP 4661911A1
Authority
EP
European Patent Office
Prior art keywords
saccharide
another embodiment
amount
glycoconjugate
activated
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP24704907.5A
Other languages
German (de)
French (fr)
Inventor
Catherine ALEX
Jin-Hwan Kim
Justin Keith Moran
Suddham Singh
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Pfizer Corp Belgium
Pfizer Corp SRL
Pfizer Inc
Original Assignee
Pfizer Corp Belgium
Pfizer Corp SRL
Pfizer Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Pfizer Corp Belgium, Pfizer Corp SRL, Pfizer Inc filed Critical Pfizer Corp Belgium
Publication of EP4661911A1 publication Critical patent/EP4661911A1/en
Pending legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/646—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/09—Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
    • A61K39/092—Streptococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/6415—Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6037—Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]

Definitions

  • PC72949A Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof Field of the Invention
  • the present invention relates to new conjugated capsular saccharide antigens (glycoconjugates), immunogenic compositions comprising said glycoconjugates and uses thereof.
  • Immunogenic compositions of the present invention will typically comprise glycoconjugates, wherein the saccharides are derived from bacterial capsular polysaccharide antigens, in particular a capsular polysaccharide derived from pathogenic bacteria.
  • the invention also relates to vaccination of human subjects, in particular infants and elderly, against infections using said glycoconjugates.
  • Gram-positive and Gram-negative bacteria may produce an extracellular compartment, the capsule, which covers the bacterial cell and often prevents the reaction of underlying cell surface antigens with their homologous antibodies.
  • Capsules are found in several bacteria of medical importance, especially in extraintestinal and invasive strains.
  • Bacterial capsules are largely composed of polysaccharides. They form a gelatinous mass around the cell. Capsular polysaccharides are important immunogens and has led to them being an important component in the design of vaccines. They have proved useful in eliciting immune responses especially when linked to carrier proteins.
  • Conjugates have been successfully generated using various cross-linking or coupling reagents, such as homobifunctional, heterobifunctional, or zero-length crosslinkers.
  • Many methods are currently available for coupling immunogenic molecules, such as saccharides, proteins, and peptides, to peptide or protein carriers. Most methods create amine, amide, urethane, isothiourea, or disulfide bonds, or in some cases thioethers.
  • a disadvantage to the use of cross-linking or coupling reagents which introduce reactive sites into the side chains of reactive amino acid molecules on carrier and/or immunogenic molecules is that the reactive sites, if not neutralized, are free to react with any unwanted molecule either in vitro (thus potentially adversely affecting the functionality or stability of the conjugates) or in vivo (thus posing a potential risk of adverse events in persons or animals immunized with the preparations).
  • Such excess reactive sites can be reacted or “capped”, so as to inactivate these sites, utilizing various known chemical reactions, but these reactions may be otherwise disruptive to the functionality of the conjugates.
  • FIG.1 shows an example of a scheme for the preparation of glycoconjugates of the invention prepared using click chemistry and using propargylamine as alkyne linker.
  • CP Carrier Protein
  • CDI 1,1’-carbonyldiimidazole.
  • glycoconjugates of the invention The present invention is directed in part to conjugated bacterial capsular saccharide antigens (also named glycoconjugates).
  • conjugated bacterial capsular saccharide antigens also named glycoconjugates.
  • the term 'glycoconjugate' indicates a capsular saccharide (in particular a bacterial capsular saccharide) linked covalently to a carrier protein.
  • saccharide of the invention may indicate polysaccharide or oligosaccharide and includes both.
  • saccharide of the invention may be oligosaccharides. Oligosaccharides have a low number of repeat units (typically 5-15 repeat units) and are typically derived synthetically or by hydrolysis of polysaccharides.
  • all of the saccharides of the present invention and in the immunogenic compositions of the present invention are polysaccharides. High molecular weight polysaccharides are able to induce certain antibody immune responses due to the epitopes present on the antigenic surface.
  • the saccharide is a polysaccharide.
  • the saccharide used in the present invention is a bacterial capsular saccharide (also named ‘capsular saccharide’ herein).
  • Capsules are found in several bacteria of medical importance. Bacterial capsules are largely composed of polysaccharides. Capsular saccharides are prepared by standard techniques known to those of ordinary skill in the art.
  • the saccharide is a S. pneumoniae capsular polysaccharide.
  • the capsular saccharide used in the present invention is a synthetic carbohydrate.
  • the source of bacterial capsular saccharide according to this invention can be bacterial cells.
  • Bacterial strains which can be used as source of capsular saccharide may be obtained from established culture collections (such as for example from the American Type Culture Collection (ATCC, Manassas, VA USA) or the Streptococcal Reference Laboratory (Centers for Disease Control and Prevention, Atlanta, GA USA)) or clinical specimens.
  • Bacterial capsular saccharides can be obtained directly from bacteria using isolation procedures known to one of ordinary skill in the art (see for example methods disclosed in US2006/0228380, US2006/0228381, US2007/0184071, US2007/0184072, US2007/0231340, and US2008/0102498 and WO2008/118752). They can also be produced using synthetic protocols known to the man skilled in the art.
  • the bacterial capsular saccharide is obtained directly from bacteria, the bacterial cells can be grown in a medium. Following fermentation of bacterial cells that produce the capsular saccharide, the bacterial cells can be lysed to produce a cell lysate.
  • the capsular saccharide may then be isolated from the cell lysate using purification techniques known in the art, including the use of centrifugation, depth filtration, precipitation, ultra-filtration, treatment with activate carbon, diafiltration and/or column chromatography (see, for example, US2006/0228380, US2006/0228381 and WO2008/118752).
  • the purified capsular saccharide can then be used for the preparation of immunogenic conjugates.
  • the isolated capsular saccharide obtained by purification can be characterized by different parameters including, for example the weight average molecular weight (Mw).
  • Mw weight average molecular weight
  • the molecular weight of the polysaccharide can be measured by Size Exclusion Chromatography (SEC) combined with Multiangle Laser Light Scattering detector (MALLS).
  • the capsular saccharide used in the method of making or part of the glycoconjugate of the present invention is a capsular saccharide from a pathogenic bacteria.
  • the capsular saccharide used in the present invention is a capsular saccharide from a pathogenic Streptococcus, a pathogenic Staphylococcus, a pathogenic Enterococcus, a pathogenic Bacillus, a pathogenic Corynebacterium, a pathogenic Listeria, a pathogenic Erysipelothrix, a pathogenic Clostridium, a pathogenic Haemophilus, a pathogenic Neisseria or a pathogenic Escherichia.
  • the capsular saccharide used in the present invention is a capsular saccharide from a pathogenic Streptococcus, a pathogenic Neisseria or a pathogenic Escherichia.
  • the capsular saccharide used in the present invention is a capsular saccharide from Aeromonas hydrophila and other species (spp.); Bacillus anthracis; Bacillus cereus; Botulinum neurotoxin producing species of Clostridium; Brucella abortus; Brucella melitensis; Brucella suis; Burkholderia mallei (formally Pseudomonas mallei); Burkholderia pseudomallei (formerly Pseudomonas pseudomallei); Campylobacter jejuni; Chlamydia psittaci; Chlamydia trachomatis, Clostridium botulinum; Clostridioides difficile (formally Clostridium difficile); Clostridium perfringens;
  • the capsular saccharide used in the present invention is a capsular saccharide from Enterococcus faecalis, Escherichia coli, Staphylococcus aureus or Streptococcus.
  • the capsular saccharide used in the present invention is a capsular saccharide from Haemophilus influenzae, Neisseria meningitidis, S. pneumoniae, S. pyogenes, S. agalactiae, Group C & G Streptococci or Escherichia coli. More preferably, the capsular saccharide used in the present invention is a capsular saccharide from Neisseria meningitidis, S. pneumoniae, S.
  • the capsular saccharide used in the present invention is a capsular saccharide from S. pneumoniae or S. agalactiae. Even more preferably, the capsular saccharide used in the present invention is a capsular saccharide from S. pneumoniae. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Staphylococcus aureus. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Staphylococcus aureus type 5 or Staphylococcus aureus type 8.
  • the capsular saccharide used in the present invention is a capsular saccharide from Enterococcus faecalis. In yet a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from is Haemophilus influenzae type b. In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Neisseria meningitidis. In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup A (MenA), N. meningitidis serogroup W135 (MenW135), N.
  • the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup A (MenA). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup W135 (MenW135). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup Y (MenY).
  • the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup C (MenC). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup X (MenX). In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Escherichia coli. In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Enterococcus faecalis.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus agalactiae (Group B streptococcus (GBS)).
  • the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III, IV, V, VI, VII or VIII.
  • the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III or V.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus agalactiae (Group B streptococcus (GBS)).
  • the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III, IV, V, VI, VII, VIII or IX.
  • the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III, IV or V.
  • the capsular saccharide used in the present invention is a capsular saccharide from Escherichia coli.
  • the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli part of the Enterovirulent Escherichia coli group (EEC Group) such as Escherichia coli - enterotoxigenic (ETEC), Escherichia coli - enteropathogenic (EPEC), Escherichia coli - O157:H7 enterohemorrhagic (EHEC), or Escherichia coli - enteroinvasive (EIEC).
  • ETEC Escherichia coli - enterotoxigenic
  • EPEC Escherichia coli - enteropathogenic
  • EHEC Escherichia coli - O157:H7 enterohemorrhagic
  • EIEC Escherichia coli - enteroinvasive
  • the capsular saccharide used in the present invention is a capsular saccharide from an Uropathogenic Escherichia coli (UPEC).
  • the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype selected from the group consisting of serotypes O157:H7, O26:H11, O111:H- and O103:H2.
  • the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype selected from the group consisting of serotypes O6:K2:H1 and O18:K1:H7.
  • the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype selected from the group consisting of serotypes O45:K1, O17:K52:H18, O19:H34 and O7:K1.
  • the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O104:H4.
  • the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O1:K12:H7.
  • the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O127:H6. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O139:H28. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O128:H2. In a preferred embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae.
  • the capsular saccharide used in the present invention is a capsular saccharide from a Streptococcus pneumoniae serotype selected from the group consisting of serotypes 1, 2, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9V, 9N, 10A, 10B, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A, 23B, 23F, 24B, 24F, 27, 29, 31, 33B, 33F, 34, 35B, 35F, 38, 72 and 73.
  • a Streptococcus pneumoniae serotype selected from the group consisting of serotypes 1, 2, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9V, 9N, 10A, 10B, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A,
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 1. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 2. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 3. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 4. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 5.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 6A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 6B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 7C. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 7F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 8.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 9V. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 9N. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 10A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 10B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 11A.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 12F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 14. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 15A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 15B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 15C.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 16F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 17F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 18C. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 19A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 19F.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 20. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 21. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 22A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 22F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 23A.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 23B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 23F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 24B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 24F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 27.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 29. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 31. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 33B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 33F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 34.
  • the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 35B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 35F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 38. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 72. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 73.
  • the capsular saccharide used in the present invention (purified before further treatment) has a weight average molecular weight between 50 kDa and 5000 kDa. In a preferred embodiment, the capsular saccharide used in the present invention has a weight average molecular weight between 500 kDa and 5000 kDa. In another preferred embodiment, the capsular saccharide used in the present invention has a weight average molecular weight between 1000 kDa and 5000 kDa. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure.
  • sizing of the saccharide to a target molecular weight range is performed prior to the conjugation to a carrier protein.
  • the size of the purified capsular saccharide is reduced while preserving critical features of the structure of the polysaccharide. Mechanical or chemical sizing maybe employed.
  • the size of the purified capsular saccharide is reduced by chemical hydrolysis. Chemical hydrolysis maybe conducted using a mild acid (e.g., acetic acid, formic acid, propanoic acid). In an embodiment, chemical hydrolysis is conducted using formic acid. In an embodiment, chemical hydrolysis is conducted using propanoic acid.
  • chemical hydrolysis is conducted using acetic acid.
  • Chemical hydrolysis may also be conducted using a diluted strong acid (such as diluted hydrochloric acid, diluted sulfuric acid, diluted phosphoric acid, diluted nitric acid or diluted perchloric acid).
  • chemical hydrolysis is conducted using diluted hydrochloric acid.
  • chemical hydrolysis is conducted using diluted sulfuric acid.
  • chemical hydrolysis is conducted using diluted phosphoric acid.
  • chemical hydrolysis is conducted using diluted nitric acid.
  • chemical hydrolysis is conducted using diluted perchloric acid.
  • the size of the purified capsular saccharide can also be reduced by mechanical homogenization.
  • the size of the purified capsular saccharide is reduced by high pressure homogenization.
  • High pressure homogenization achieves high shear rates by pumping the process stream through a flow path with sufficiently small dimensions.
  • the shear rate is increased by using a larger applied homogenization pressure, and exposure time can be increased by recirculating the feed stream through the homogenizer.
  • the high-pressure homogenization process can be appropriate for reducing the size of the purified capsular saccharide while preserving the structural features of the saccharide.
  • the isolated capsular saccharide is sized to a weight average molecular weight between 10 kDa and 1000 kDa.
  • the isolated capsular saccharide is sized to a weight average molecular weight between 50 kDa and 500 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 50 kDa and 400 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 50 kDa and 250 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 250 kDa and 1000 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 250 kDa and 500 kDa.
  • the isolated capsular saccharide is sized to a weight average molecular weight between 250 kDa and 400 kDa. In a preferred embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 200 kDa and 800 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 250 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 300 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 350 kDa.
  • the isolated capsular saccharide is sized to a weight average molecular weight of about 400 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 450 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 500 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 550 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 600 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 700 kDa.
  • the isolated capsular saccharide is sized to a weight average molecular weight of about 800 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 900 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 1000 kDa. In an embodiment, the isolated capsular saccharide is not sized.
  • the isolated capsular saccharide described above may be activated (e.g., chemically activated) to make them capable of reacting (e.g., with a linker) and then incorporated into glycoconjugates, as further described herein.
  • the term 'glycoconjugate' indicates a saccharide covalently linked to a carrier protein.
  • covalent conjugation of saccharides to carriers enhances the immunogenicity of saccharides as it converts them from T-independent antigens to T-dependent antigens, thus allowing priming for immunological memory. Conjugation is particularly useful for pediatric vaccines.
  • 1.2 Capsular saccharide glycoconjugates of the invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said polysaccharide before conjugation is between 10 kDa and 2,000 kDa.
  • the weight average molecular weight (Mw) of the saccharide before conjugation refers to the Mw before the activation of the saccharide (i.e., after an eventual sizing step but before reacting the saccharide with an activating agent).
  • Mw of the saccharide is not substantially modified by the activation step and the Mw of the saccharide incorporated in the conjugate is similar to the Mw of the saccharide as measured before activation.
  • the saccharide is activated with a carbonic acid derivative (e.g., CDI or CDT) in combination with an alkyne linker (see sections 1.3 below).
  • the saccharide is activated with CDI in combination with an alkyne linker (see sections 1.3 below). In an embodiment, the saccharide is activated with CDT in combination with an alkyne linker (see sections 1.3 below).
  • the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 50 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 500 kDa.
  • the weight average molecular weight (Mw) is between 50 kDa and 250 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 200 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 150 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 100 kDa.
  • the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 75 kDa and 1,000 kDa.
  • the weight average molecular weight (Mw) is between 75 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 500 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 250 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 200 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 150 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 100 kDa.
  • the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 100 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 500 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 250 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 200 kDa.
  • the weight average molecular weight (Mw) is between 100 kDa and 150 kDa.
  • the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 150 kDa and 1,000 kDa.
  • the weight average molecular weight (Mw) is between 150 kDa and 750 kDa.
  • the weight average molecular weight (Mw) is between 150 kDa and 500 kDa.
  • the weight average molecular weight (Mw) is between 150 kDa and 250 kDa.
  • the weight average molecular weight (Mw) is between 150 kDa and 200 kDa.
  • the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 200 kDa and 1,000 kDa.
  • the weight average molecular weight (Mw) is between 200 kDa and 750 kDa.
  • the weight average molecular weight (Mw) is between 200 kDa and 500 kDa.
  • the weight average molecular weight (Mw) is between 200 kDa and 300 kDa.
  • the weight average molecular weight (Mw) is between 200 kDa and 250 kDa.
  • the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 500 kDa and 1,000 kDa.
  • the weight average molecular weight (Mw) is between 500 kDa and 750 kDa.
  • the weight average molecular weight (Mw) is between 500 kDa and 700 kDa.
  • the weight average molecular weight (Mw) is between 500 kDa and 600 kDa.
  • the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is about 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is about 750 kDa. In an embodiment, the weight average molecular weight (Mw) is about 700 kDa. In an embodiment, the weight average molecular weight (Mw) is about 600 kDa. In an embodiment, the weight average molecular weight (Mw) is about 500 kDa. In an embodiment, the weight average molecular weight (Mw) is about 400 kDa.
  • the weight average molecular weight (Mw) is about 300 kDa. In an embodiment, the weight average molecular weight (Mw) is about 200 kDa. In an embodiment, the weight average molecular weight (Mw) is about 150 kDa. In an embodiment, the weight average molecular weight (Mw) is about 125 kDa. In an embodiment, the weight average molecular weight (Mw) is about 100 kDa. In some embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 250 kDa and 20,000 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 15,000 kDa. In yet other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 7,500 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 2,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 1,500 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 1,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 750 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 7,500 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 2,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 1,500 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 1,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 5,000 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 2,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 1,500 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 10,000 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 4,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 3,000 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 3,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,250 kDa and 3,500 kDa. In preferred embodiment, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 2,500 kDa. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 10,000 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 9,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 8,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 6,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 5,000 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 4,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 3,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 3,250 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 3,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 2,500 kDa.
  • the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 1,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 1,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 750 kDa.
  • Another way to characterize the glycoconjugates of the invention is by the number of lysine residues in the carrier protein (e.g., CRM 197 or SCP) that become conjugated to the saccharide which can be characterized as a range of conjugated lysines (degree of conjugation).
  • the evidence for lysine modification of the carrier protein, due to covalent linkages to the saccharides, can be obtained by amino acid analysis using routine methods known to those of skill in the art. Conjugation results in a reduction in the number of lysine residues recovered compared to the carrier protein starting material used to generate the conjugate materials.
  • the degree of conjugation of the glycoconjugate of the invention is between 2 and 15.
  • the degree of conjugation of the glycoconjugate of the invention is between 2 and 13. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 10. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 8. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 6. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 5. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 4. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 13.
  • the degree of conjugation of the glycoconjugate of the invention is between 3 and 10. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 8. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 6. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 5. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 4. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 5 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 5 and 10. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 8 and 15.
  • the degree of conjugation of the glycoconjugate of the invention is between 8 and 12. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 10 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 10 and 12. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 2. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 3. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 4. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 5. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 6.
  • the degree of conjugation of the glycoconjugate of the invention is about 7. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 8. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 9. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 10, about 11. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 12. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 13. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 14. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 15.
  • the degree of conjugation of the glycoconjugate of the invention is between 4 and 7.
  • the carrier protein is CRM 197 .
  • the carrier protein is SCP.
  • the glycoconjugates of the invention may also be characterized by the ratio (weight/weight) of saccharide to carrier protein.
  • the ratio of saccharide to carrier protein in the glycoconjugate (w/w) is between 0.5 and 3.0.
  • the saccharide to carrier protein ratio (w/w) is between 0.5 and 2.0.
  • the saccharide to carrier protein ratio (w/w) is between 0.5 and
  • the saccharide to carrier protein ratio (w/w) is between 0.8 and 1.2.
  • the saccharide to carrier protein ratio (w/w) is between 0.5 and 1.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is between 1.0 and 1.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is between 1.0 and 2.0. In further embodiments, the saccharide to carrier protein ratio (w/w) is between 0.8 and 1.2. In a preferred embodiment, the ratio of saccharide to carrier protein in the conjugate is between 0.9 and 1.1. In an embodiment, the saccharide to carrier protein ratio (w/w) is about 0.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 0.6.
  • the saccharide to carrier protein ratio (w/w) is about 0.7. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 0.8. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 0.9. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.1. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.2. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.3. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.4.
  • the saccharide to carrier protein ratio (w/w) is about 1.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.6. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.7. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.8. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.9. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.1. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.2.
  • the saccharide to carrier protein ratio is about 2.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.8. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 3.0. In some such embodiments, the carrier protein is CRM 197 . In other such embodiments, the carrier protein is SCP.
  • the glycoconjugates of the invention may also be characterized by the number of covalent linkages between the carrier protein and the saccharide as a function of repeat units of the saccharide. In one embodiment, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 4 saccharide repeat units of the saccharide.
  • the covalent linkage between the carrier protein and the saccharide occurs at least once in every 10 saccharide repeat units of the saccharide. In another embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 15 saccharide repeat units of the saccharide. In a further embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 25 saccharide repeat units of the saccharide. In a further embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 50 saccharide repeat units of the saccharide. In yet a further embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 100 saccharide repeat units of the saccharide.
  • the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 5 to 10 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 2 to 7 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 6 to 11 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 9 to 14 saccharide repeat units of the saccharide.
  • the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 10 to 20 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 4 to 25 saccharide repeat units of the saccharide.
  • the carrier protein is CRM 197 . In other embodiments, the carrier protein is SCP. In some embodiments, the carrier protein is CRM 197 and the covalent linkage between the CRM 197 and the saccharide occurs at least once in every 4, 10, 15 or 25 saccharide repeat units of the saccharide.
  • the carrier protein is SCP and the covalent linkage between the SCP and the saccharide occurs at least once in every 4, 10, 15 or 25 saccharide repeat units of the saccharide.
  • the glycoconjugates and immunogenic compositions of the invention may contain free saccharide that is not covalently conjugated to the carrier protein but is nevertheless present in the glycoconjugate composition.
  • the free saccharide may be noncovalently associated with (i.e., noncovalently bound to, adsorbed to, or entrapped in or with) the glycoconjugate.
  • the glycoconjugate comprises less than about 50% of free saccharide compared to the total amount of said saccharide.
  • the glycoconjugate comprises less than about 40% of free saccharide compared to the total amount of said saccharide. In a yet preferred embodiment, the glycoconjugate comprises less than about 25% of free saccharide compared to the total amount of said saccharide. In an even preferred embodiment, the glycoconjugate comprises less than about 20% of free saccharide compared to the total amount of said saccharide. In a yet preferred embodiment, the glycoconjugate comprises less than about 15% of free saccharide compared to the total amount of said saccharide.
  • the glycoconjugates may also be characterized by their molecular size distribution (K d ). Size exclusion chromatography media (CL-4B) can be used to determine the relative molecular size distribution of the conjugate.
  • At least 30% of the glycoconjugate of the invention has a K d below or equal to 0.3 in a CL-4B column.
  • at least 40% of the glycoconjugate of the invention has a K d below or equal to 0.3 in a CL-4B column.
  • at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, or 85% of the glycoconjugate of the invention has a K d below or equal to 0.3 in a CL-4B column.
  • at least 60% of the glycoconjugate of the invention has a K d below or equal to 0.3 in a CL-4B column.
  • glycoconjugate of the invention has a K d below or equal to 0.3 in a CL-4B column. In a preferred embodiment, between 65% and 80% of the glycoconjugate of the invention has a K d below or equal to 0.3 in a CL-4B column.
  • Capsular saccharide glycoconjugates of the invention prepared using click chemistry The glycoconjugates of the present invention are prepared using click chemistry. The invention also relates to a method of making a glycoconjugate, as disclosed herein.
  • click chemistry comprises three steps, (a) reacting an isolated capsular saccharide with a carbonic acid derivative and an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide (activation of the saccharide), (b) reacting a carrier protein with an agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group where the NHS moiety reacts with the amino groups to form an amide linkage thereby obtaining an azido functionalized carrier protein (activation of the carrier protein), (c) reacting the activated alkynyl saccharide of step (a) with the activated azido-carrier protein of step (b) by Cu +1 mediated azide-alkyne cycloaddition reaction to form a glycoconjugate.
  • NPS N-Hydroxysuccinimide
  • the saccharide is said to be activated and is referred to herein as “activated saccharide” or “activated alkynyl saccharide”.
  • the carrier is said to be activated and is referred to as “activated carrier” or “activated azido-carrier”.
  • sizing of the saccharide to a target molecular weight (MW) range can be performed before the activation (a), sizing of the saccharide to a target molecular weight (MW) range can be performed. Therefore, in an embodiment, the isolated saccharide is sized before activation with a carbonic acid derivative and an alkyne linker. In an embodiment, the isolated saccharide is sized to any of the target molecular weight (MW) range defined above.
  • said carbonic acid derivative is selected from the group consisting of 1,1’-carbonyldiimidazole (CDI), 1,1’-carbonyl-di-(1,2,4-triazole) (CDT), disuccinimidyl carbonate (DSC) and N-hydroxysuccinimidyl chloroformate.
  • said carbonic acid derivative is 1,1’-carbonyldiimidazole (CDI).
  • said carbonic acid derivative is 1,1'-Carbonyl-di-(1,2,4-triazole) (CDT).
  • said carbonic acid derivative is disuccinimidyl carbonate (DSC).
  • said carbonic acid derivative is N-hydroxysuccinimidyl chloroformate.
  • said carbonic acid derivative is 1,1’-carbonyldiimidazole (CDI) or 1,1'- Carbonyl-di-(1,2,4-triazole) (CDT).
  • said carbonic acid derivative is 1,1’- carbonyldiimidazole (CDI).
  • said alkyne linker is a compound of formula (I), H 2 N X (I) wherein X is selected from the group consisting of CH 2 , CH 2 (CH 2 ) n , (CH 2 CH 2 O) m CH 2 CH 2 , NHCO(CH 2 ) n , NHCO(CH 2 CH 2 O) m CH 2 CH 2 , OCH 2 (CH 2 ) n and O(CH 2 CH 2 O) m CH 2 CH 2 ; where n is selected from 1 to 10 and m is selected from 1 to 4.
  • said alkyne linker is a compound of formula (I), wherein X is CH 2 .
  • said alkyne linker is a compound of formula (I), wherein X is CH 2 (CH 2 ) n , and n is selected from 1 to 10. In an embodiment, n is selected from 1 to 5. In an embodiment, n is selected from 1 to 4. In an embodiment, n is selected from 1 to 3. In an embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In yet another embodiment, n is 3. In yet a further embodiment, n is 4. In yet a further embodiment, n is 5. In yet a further embodiment, n is 6. In yet a further embodiment, n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9.
  • n is 10.
  • said alkyne linker is a compound of formula (I), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , wherein m is selected from 1 to 4.
  • m is selected from 1 to 3.
  • m is selected from 1 to 2.
  • m is 1.
  • m is 2.
  • m is 3.
  • m is 4.
  • said alkyne linker is a compound of formula (I), wherein X is NHCO(CH 2 ) n , and n is selected from 1 to 10. In an embodiment, n is selected from 1 to 5.
  • n is selected from 1 to 4. In an embodiment, n is selected from 1 to 3. In an embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In yet another embodiment, n is 3. In yet a further embodiment, n is 4. In yet a further embodiment, n is 5. In yet a further embodiment, n is 6. In yet a further embodiment, n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9. In yet a further embodiment, n is 10. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4.
  • m is selected from 1 to 3. In an embodiment, m is selected from 1 to 2. In a particular embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet a further embodiment, m is 4. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is OCH 2 (CH 2 ) n , and n is selected from 1 to 10. In an embodiment, n is selected from 1 to 5. In an embodiment, n is selected from 1 to 4. In an embodiment, n is selected from 1 to 3. In an embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In yet another embodiment, n is 3.
  • n is 4. In yet a further embodiment, n is 5. In yet a further embodiment, n is 6. In yet a further embodiment, n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9. In yet a further embodiment, n is 10.
  • said alkyne linker is a compound of formula (I), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4. In an embodiment, m is selected from 1 to 3. In an embodiment, m is selected from 1 to 2. In a particular embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet a further embodiment, m is 4.
  • said alkyne linker is a compound of formula (II), Hence in a preferred embodiment, said alkyne linker is propargylamine.
  • n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9. In yet a further embodiment, n is 10.
  • m is selected from 0 to 3. In an embodiment, m is selected from 0 to 2. In an embodiment, m is selected from 0 to 1. In a particular embodiment, m is 0. In a particular embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3.
  • step a) comprises reacting the saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative-activated saccharide with an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-10 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-10 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 2-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 3-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 5-10 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-5 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-5 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 2-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 3-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-3 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-3 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-3 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 2-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-2 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-2 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-1 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-1 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-0.5 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-0.5 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-0.4 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-0.4 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-0.4 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-0.3 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.01 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.05 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.08 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.3 molar equivalent to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 2 molar equivalents to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 3 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 4 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 5 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 8 molar equivalents to the amount of saccharide present in the reaction mixture.
  • step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 10 molar equivalents to the amount of saccharide present in the reaction mixture.
  • the isolated saccharide is reacted with a carbonic acid derivative in an aprotic solvent.
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylsulphoxide (DMSO) or dimethylformamide (DMF).
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylformamide (DMF).
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylsulphoxide (DMSO).
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylsulphoxide (DMSO).
  • DMSO dimethylsulphoxide
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylsulphoxide (DMSO) or dimethylformamide (DMF).
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylformamide (DMF).
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylsulphoxide (DMSO).
  • the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylacetamide. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of N-methyl-2- pyrrolidone. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of hexamethylphosphoramide (HMPA). In a preferred embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylsulphoxide (DMSO).
  • DMSO dimethylsulphoxide
  • the isolated saccharide is reacted with a carbonic acid derivative in dimethylsulphoxide (DMSO) or dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in dimethylsulphoxide (DMSO). In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in dimethylacetamide. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in N-methyl-2-pyrrolidone. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in hexamethylphosphoramide (HMPA).
  • HMPA hexamethylphosphoramide
  • the isolated saccharide is reacted with CDI in dimethylsulphoxide (DMSO).
  • DMSO dimethylsulphoxide
  • the isolated saccharide is reacted with CDI in anhydrous DMSO. It has been surprisingly found that reacting the isolated saccharide with CDI in an environment with a moisture level of about 0.1% to 1% (v/v) allows to avoid side reactions. Therefore, in one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.8% (v/v) water.
  • the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.2% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 1% (v/v) water.
  • the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.3% to 0.8% (v/v) water.
  • the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.3% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.3% to 0.4% (v/v) water. Preferably, in one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.2% (v/v) water.
  • the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.6% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.7% (v/v) water.
  • the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.9% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.5% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.2% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.8% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.3% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.3% to 0.5% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising 0.3% to 0.4% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.5% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising about 0.1% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising about 0.2% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising about 0.3% (v/v) water.
  • the isolated saccharide is reacted with CDI in DMSO comprising about 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.6% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.7% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.9% (v/v) water.
  • the free carbonic acid derivative is then quenched by the addition of water before the addition of the alkyne linker.
  • Water can inactivate free CDI. Therefore, in an embodiment, carbonic acid derivative activation is followed by the addition of water.
  • water is added to bring the total water content in the mixture to between about 1% to about 10% (v/v). In an embodiment, water is added to bring the total water content in the mixture to between about 1.2% to about 8% (v/v). In an embodiment, water is added to bring the total water content in the mixture to between about 1.5% to about 5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to between about 1.5% to about 3% (v/v).
  • water is added to bring the total water content in the mixture to between about 1.5% to about 2.5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1 % (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1.2% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1.4% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1.5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 2% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 2.5% (v/v).
  • water is added to bring the total water content in the mixture to about 3% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 7% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 10% (v/v).
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-10 molar equivalent to the amount of polysaccharide Repeat Unit of the activated saccharide (molar equivalent of RU). In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-0.1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-0.1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 4-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 4-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 4-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 5-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 5-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 8-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.01 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.05 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 3 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 4 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide.
  • step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 8 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 10 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide.
  • said carbonic acid derivative is preferably CDI. In another embodiment, said carbonic acid derivative is CDT.
  • the degree of activation of the activated saccharide following step a) is between 0.5 to 50%.
  • the degree of activation of the alkynyl saccharide being defined as the percentage of Repeating Unit linked to an alkyne linker.
  • the degree of activation of the activated saccharide following step a) is between 1 to 30%. In another embodiment the degree of activation of the activated saccharide following step a) is between 2 to 25%. In another embodiment the degree of activation of the activated saccharide following step a) is between 3 to 20%. In another embodiment the degree of activation of the activated saccharide following step a) is between 3 to 15%. In another embodiment the degree of activation of the activated saccharide following step a) is between 4 to 15%. In an embodiment the degree of activation of the activated saccharide following step a) is between 1 to 6%.
  • the degree of activation of the activated saccharide following step a) is between 3 to 6%. In an embodiment the degree of activation of the activated saccharide following step a) is between 10 to 15%. In an embodiment the degree of activation of the activated saccharide following step a) is about 1%. In an embodiment the degree of activation of the activated saccharide following step a) is about 2%. In an embodiment the degree of activation of the activated saccharide following step a) is about 3%. In an embodiment the degree of activation of the activated saccharide following step a) is about 4%. In an embodiment the degree of activation of the activated saccharide following step a) is about 5%.
  • the degree of activation of the activated saccharide following step a) is about 6%. In an embodiment the degree of activation of the activated saccharide following step a) is about 7%. In an embodiment the degree of activation of the activated saccharide following step a) is about 8%. In an embodiment the degree of activation of the activated saccharide following step a) is about 9%. In an embodiment the degree of activation of the activated saccharide following step a) is about 10%. In an embodiment the degree of activation of the activated saccharide following step a) is about 11%. In an embodiment the degree of activation of the activated saccharide following step a) is about 12%.
  • the degree of activation of the activated saccharide following step a) is about 13%. In an embodiment the degree of activation of the activated saccharide following step a) is about 14%. In an embodiment the degree of activation of the activated saccharide following step a) is about 15%. In an embodiment the degree of activation of the activated saccharide following step a) is about 16%. In an embodiment the degree of activation of the activated saccharide following step a) is about 17%. In an embodiment the degree of activation of the activated saccharide following step a) is about 18%. In an embodiment the degree of activation of the activated saccharide following step a) is about 19%.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-10 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide NHS
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-10 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-10 molar equivalents to the lysines on the carrier protein.
  • NHS N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 3-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 5-10 molar equivalents to the lysines on the carrier protein.
  • NHS N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 7.5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-7.5 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide NHS
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-7.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-7.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-7.5 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide NHS
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-7.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 3-7.5 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide NHS
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 3-7.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-5 molar equivalents to the lysines on the carrier protein.
  • NHS N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 3-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-3 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-3 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-3 molar equivalents to the lysines on the carrier protein.
  • NHS N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-2.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-2.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-2 molar equivalents to the lysines on the carrier protein.
  • NHS N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-2 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-1.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-1.5 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide NHS
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-1.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-1.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-1 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-1 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-0.5 molar equivalents to the lysines on the carrier protein.
  • N-Hydroxysuccinimide NHS
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-0.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 7.5 molar equivalents to the lysines on the carrier protein.
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 2.5 molar equivalents to the lysines on the carrier protein.
  • NHS N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 1.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 1 molar equivalent to the lysines on the carrier protein.
  • NHS N-Hydroxysuccinimide
  • step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 0.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 0.1 molar equivalents to the lysines on the carrier protein. In one embodiment the degree of activation of the activated carrier following step b) is between 1 and 50.
  • the degree of activation of the activated carrier being defined as the number of lysine residues in the carrier protein that become linked to the agent bearing an N- Hydroxysuccinimide (NHS) moiety and an azido group.
  • the carrier protein is CRM 197 , which contains 39 lysine residues.
  • the degree of activation of the activated carrier following step b) may be between 1 to 30.
  • the degree of activation of the activated carrier (CRM 197 ) following step b) is between 5 to 20.
  • the degree of activation of the activated carrier (CRM 197 ) following step b) is between 9 to 18.
  • the degree of activation of the activated carrier (CRM197) following step b) is between 8 to 11. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is between 15 to 20. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 5. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 6. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 7. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 8.
  • the degree of activation of the activated carrier (CRM 197 ) following step b) is about 9. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 10. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 11. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 12. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 13. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 14.
  • the degree of activation of the activated carrier (CRM 197 ) following step b) is about 15. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 16. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 17. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 18. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 19. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 20.
  • the degree of activation of the activated carrier (CRM 197 ) following step b) is about 21. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 22. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 23. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 24. In another embodiment the degree of activation of the activated carrier (CRM 197 ) following step b) is about 25. In an embodiment, the carrier protein is SCP or a fragment thereof. In said embodiment the degree of activation of the activated carrier following step b) may be between 1 to 50.
  • the degree of activation of the activated carrier (SCP) following step b) is between 5 to 50. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 7 to 45. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 5 to 15. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 20 to 30. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 30 to 50. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 30 to 40.
  • the degree of activation of the activated carrier (SCP) following step b) is between 10 to 40. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 5. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 7. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 10. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 13. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 15. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 20.
  • the degree of activation of the activated carrier (SCP) following step b) is about 26. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 30. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 35. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 37. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 40. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 45. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 50.
  • the carrier protein is TT or a fragment thereof.
  • the degree of activation of the activated carrier following step b) may be between 1 to 30. In another embodiment the degree of activation of the activated carrier (TT) following step b) is between 5 to 25. In another embodiment the degree of activation of the activated carrier (TT) following step b) is between 7 to 25. In another embodiment the degree of activation of the activated carrier (TT) following step b) is between 10 to 20. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 5. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 7.
  • the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is between 0.6 and 1.5. In a preferred embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is between 0.8 and 1. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 0.5. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 0.6.
  • the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.1. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.2. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.3. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.4.
  • this alkyne group capping agent is 3-azido-1-propanol. Therefore, in an embodiment, following step (c) the process further comprises a step of capping the unreacted alkyne groups remained in the conjugates with an alkyne group capping agent. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.05 to 20 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.1 to 15 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide.
  • the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.5 to 1 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 1 to 5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 1 to 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide.
  • Formula (VIII) is a schematic representation of the preferred glycoconjugates of the invention. It should not be understood that a linkage is present at every repeat unit of the saccharide (the structure in square brackets). Rather, a majority of the saccharide repeating unit remains unmodified and covalent linkages between the carrier protein and the saccharide is for a minority of the saccharide repeat units. Additionally, an individual carrier protein (CP) molecule may be linked to more than one saccharide molecule and an individual saccharide molecule can be linked to more than one individual carrier protein (CP) molecule.
  • the structure in square brackets is a schematic representation of a repeat unit of the saccharide.
  • n’ is selected from 0 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 0 and n’’ is 0. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is à and n’’ is 1. In a particular embodiment, n’ is 1 and n’’ is 1.
  • n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5.
  • n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 0 and n’’ is 6. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH 2 (CH 2 ) n’ , where n’ is selected from 0 to 10 and wherein X' is CH 2 O(CH 2 ) n’’ CH 2 where n’’ is selected from 0 to 10.
  • n’ is selected from 0 to 5 and n’’ is selected from 0 to 10.
  • n’ is selected from 0 to 5 and n’’ is selected from 0 to 5.
  • n’ is selected from 0 to 3 and n’’ is selected from 0 to 3.
  • n’ is selected from 0 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 0 and n’’ is 0. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is à and n’’ is 1. In a particular embodiment, n’ is 1 and n’’ is 1.
  • n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 0 and n’’ is 2. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2.
  • n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 0 and n’’ is 4.
  • n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5.
  • n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 0 and n’’ is 6. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6.
  • n’ is selected from 0 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 0 to 3, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 0 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In an embodiment, n’ is selected from 0 to 1, m’ is selected from 0 to 1 and n’’ is selected from 0 to 1. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 0. In another embodiment, n’ is 0, m’ is 1 and n’’ is 0.
  • n’ is 0, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 0. In another embodiment, n’ is 1, m’ is 1 and n’’ is 0. In another embodiment, n’ is 1, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In another embodiment, n’ is 2, m’ is 0 and n’’ is 0. In another embodiment, n’ is 2, m’ is 1 and n’’ is 0.
  • n’ is 2, m’ is 2 and n’’ is 0. In another embodiment, n’ is 2, m’ is 3 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 0.
  • n’ is 4, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 0.In yet a further embodiment, n’ is 5, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 0. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 1.
  • n’ is 0, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 1. In another embodiment, n’ is 2, m’ is 0 and n’’ is 1. In another embodiment, n’ is 2, m’ is 1 and n’’ is 1.
  • n’ is 2, m’ is 2 and n’’ is 1. In another embodiment, n’ is 2, m’ is 3 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 1.
  • n’ is 4, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 1. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 2. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 2.
  • n’ is 0, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 2. In another embodiment, n’ is 2, m’ is 0 and n’’ is 2. In another embodiment, n’ is 2, m’ is 1 and n’’ is 2.
  • n’ is 2, m’ is 2 and n’’ is 2. In another embodiment, n’ is 2, m’ is 3 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 2.
  • n’ is 4, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 2. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 3.
  • n’ is 0, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 3. In another embodiment, n’ is 2, m’ is 0 and n’’ is 3. In another embodiment, n’ is 2, m’ is 1 and n’’ is 3.
  • n’ is 2, m’ is 2 and n’’ is 3. In another embodiment, n’ is 2, m’ is 3 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 3.
  • n’ is 4, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 3. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 4. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 4.
  • n’ is 0, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 4. In another embodiment, n’ is 2, m’ is 0 and n’’ is 4. In another embodiment, n’ is 2, m’ is 1 and n’’ is 4.
  • n’ is 2, m’ is 2 and n’’ is 4. In another embodiment, n’ is 2, m’ is 3 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 4.
  • n’ is 4, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 4. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 5.
  • n’ is 0, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 5. In another embodiment, n’ is 2, m’ is 0 and n’’ is 5. In another embodiment, n’ is 2, m’ is 1 and n’’ is 5.
  • n’ is 2, m’ is 2 and n’’ is 5. In another embodiment, n’ is 2, m’ is 3 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 5.
  • n’ is 4, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 5.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 (CH 2 ) n” , where n’’ is selected from 0 to 10.
  • X is (CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 (CH 2 ) n” , where n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 3.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2.
  • m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5.
  • m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 O(CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10.
  • X is (CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 O(CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 3.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2.
  • m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5.
  • m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is (CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 O(CH 2 CH 2 O) m’ (CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4.
  • m is selected from 1 to 3
  • m’ is selected from 0 to 4
  • n’’ is selected from 0 to 10.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 4 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 2
  • n’’ is selected from 0 to 3.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 2
  • n’’ is selected from 0 to 1.
  • m is 1, m’ is 0 and n’’ is 0.
  • m is 1, m’ is 1 and n’’ is 0.
  • m is 1, m’ is 2 and n’’ is 0.
  • m is 1, m’ is 3 and n’’ is 0.
  • m is 2, m’ is 0 and n’’ is 0. In another embodiment, m is 2, m’ is 1 and n’’ is 0. In another embodiment, m is 2, m’ is 2 and n’’ is 0. In another embodiment, m is 2, m’ is 3 and n’’ is 0. In yet another embodiment, m is 3, m’ is 0 and n’’ is 0. In yet another embodiment, m is 3, m’ is 1 and n’’ is 0. In yet another embodiment, m is 3, m’ is 2 and n’’ is 0. In yet another embodiment, m is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, m is 4, m’ is 0 and n” is 0.
  • m is 4, m’ is 1 and n” is 0. In yet a further embodiment, m is 4, m’ is 2 and n” is 0. In yet a further embodiment, m is 4, m’ is 3 and n” is 0. In a particular embodiment, m is 1, m’ is 0 and n’’ is 1. In a particular embodiment, m is 1, m’ is 1 and n’’ is 1. In a particular embodiment, m is 1, m’ is 2 and n’’ is 1. In a particular embodiment, m is 1, m’ is 3 and n’’ is 1. In another embodiment, m is 2, m’ is 0 and n’’ is 1.
  • m is 2, m’ is 1 and n’’ is 1. In another embodiment, m is 2, m’ is 2 and n’’ is 1. In another embodiment, m is 2, m’ is 3 and n’’ is 1. In yet another embodiment, m is 3, m’ is 0 and n’’ is 1. In yet another embodiment, m is 3, m’ is 1 and n’’ is 1. In yet another embodiment, m is 3, m’ is 2 and n’’ is 1. In yet another embodiment, m is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, m is 4, m’ is 0 and n” is 1. In yet a further embodiment, m is 4, m’ is 1 and n” is 1.
  • m is 4, m’ is 2 and n” is 1. In yet a further embodiment, m is 4, m’ is 3 and n” is 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 2. In a particular embodiment, m is 1, m’ is 1 and n’’ is 2. In a particular embodiment, m is 1, m’ is 2 and n’’ is 2. In a particular embodiment, m is 1, m’ is 3 and n’’ is 2. In another embodiment, m is 2, m’ is 0 and n’’ is 2. In another embodiment, m is 2, m’ is 1 and n’’ is 2.
  • m is 2, m’ is 2 and n’’ is 2. In another embodiment, m is 2, m’ is 3 and n’’ is 2. In yet another embodiment, m is 3, m’ is 0 and n’’ is 2. In yet another embodiment, m is 3, m’ is 1 and n’’ is 2. In yet another embodiment, m is 3, m’ is 2 and n’’ is 2. In yet another embodiment, m is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, m is 4, m’ is 0 and n” is 2. In yet a further embodiment, m is 4, m’ is 1 and n” is 2. In yet a further embodiment, m is 4, m’ is 2 and n” is 2.
  • m is 4, m’ is 3 and n” is 2. In a particular embodiment, m is 1, m’ is 0 and n’’ is 3. In a particular embodiment, m is 1, m’ is 1 and n’’ is 3. In a particular embodiment, m is 1, m’ is 2 and n’’ is 3. In a particular embodiment, m is 1, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 0 and n’’ is 3. In another embodiment, m is 2, m’ is 1 and n’’ is 3. In another embodiment, m is 2, m’ is 2 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3.
  • m is 3, m’ is 0 and n’’ is 3. In yet another embodiment, m is 3, m’ is 1 and n’’ is 3. In yet another embodiment, m is 3, m’ is 2 and n’’ is 3. In yet another embodiment, m is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, m is 4, m’ is 0 and n” is 3. In yet a further embodiment, m is 4, m’ is 1 and n” is 3. In yet a further embodiment, m is 4, m’ is 2 and n” is 3. In yet a further embodiment, m is 4, m’ is 3 and n” is 3.
  • m is 1, m’ is 0 and n’’ is 4. In a particular embodiment, m is 1, m’ is 1 and n’’ is 4. In a particular embodiment, m is 1, m’ is 2 and n’’ is 4. In a particular embodiment, m is 1, m’ is 3 and n’’ is 4. In another embodiment, m is 2, m’ is 0 and n’’ is 4. In another embodiment, m is 2, m’ is 1 and n’’ is 4. In another embodiment, m is 2, m’ is 2 and n’’ is 4. In another embodiment, m is 2, m’ is 3 and n’’ is 4. In yet another embodiment, m is 3, m’ is 0 and n’’ is 4.
  • m is 3, m’ is 1 and n’’ is 4. In yet another embodiment, m is 3, m’ is 2 and n’’ is 4. In yet another embodiment, m is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, m is 4, m’ is 0 and n” is 4. In yet a further embodiment, m is 4, m’ is 1 and n” is 4. In yet a further embodiment, m is 4, m’ is 2 and n” is 4. In yet a further embodiment, m is 4, m’ is 3 and n” is 4. In a particular embodiment, m is 1, m’ is 0 and n’’ is 5.
  • m is 1, m’ is 1 and n’’ is 5. In a particular embodiment, m is 1, m’ is 2 and n’’ is 5. In a particular embodiment, m is 1, m’ is 3 and n’’ is 5. In another embodiment, m is 2, m’ is 0 and n’’ is 5. In another embodiment, m is 2, m’ is 1 and n’’ is 5. In another embodiment, m is 2, m’ is 2 and n’’ is 5. In another embodiment, m is 2, m’ is 3 and n’’ is 5. In yet another embodiment, m is 3, m’ is 0 and n’’ is 5. In yet another embodiment, m is 3, m’ is 1 and n’’ is 5.
  • m is 3, m’ is 2 and n’’ is 5. In yet another embodiment, m is 3, m’ is 3 and n’’ is 5. In yet another embodiment, m is 4, m’ is 0 and n” is 5. In yet a further embodiment, m is 4, m’ is 1 and n” is 5. In yet a further embodiment, m is 4, m’ is 2 and n” is 5. In yet a further embodiment, m is 4, m’ is 3 and n” is 5.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH 2 ) n’ , where n’ is selected from 1 to 10 and wherein X' is CH 2 (CH 2 ) n” , where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3 and n’’ is selected from 0 to 3.
  • n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1.
  • n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 1 and n’’ is 3.
  • n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4.
  • n’ is 6 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH 2 ) n’ , where n’ is selected from 1 to 10 and wherein X' is CH 2 O(CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10.
  • n’ is selected from 1 to 5 and n’’ is selected from 0 to 10.
  • n’ is selected from 1 to 5 and n’’ is selected from 0 to 5.
  • n’ is selected from 1 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1.
  • n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2.
  • n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4.
  • n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH 2 ) n’ , where n’ is selected from 1 to 10 and wherein X' is CH 2 O(CH 2 CH 2 O) m’ (CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4.
  • n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 0. In another embodiment, n’ is 1, m’ is 1 and n’’ is 0.
  • n’ is 1, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In another embodiment, n’ is 2, m’ is 0 and n’’ is 0. In another embodiment, n’ is 2, m’ is 1 and n’’ is 0. In another embodiment, n’ is 2, m’ is 2 and n’’ is 0. In another embodiment, n’ is 2, m’ is 3 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 0.
  • n’ is 3, m’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 0.
  • n’ is 5, m’ is 1 and n” is 0.In yet a further embodiment, n’ is 5, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 0. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 1. In another embodiment, n’ is 2, m’ is 0 and n’’ is 1.
  • n’ is 2, m’ is 1 and n’’ is 1. In another embodiment, n’ is 2, m’ is 2 and n’’ is 1. In another embodiment, n’ is 2, m’ is 3 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 1.
  • n’ is 1, m’ is 1 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 2. In another embodiment, n’ is 2, m’ is 0 and n’’ is 2. In another embodiment, n’ is 2, m’ is 1 and n’’ is 2. In another embodiment, n’ is 2, m’ is 2 and n’’ is 2. In another embodiment, n’ is 2, m’ is 3 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 2.
  • n’ is 3, m’ is 1 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 2.
  • n’ is 5, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 2. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 3. In another embodiment, n’ is 2, m’ is 0 and n’’ is 3.
  • n’ is 2, m’ is 1 and n’’ is 3. In another embodiment, n’ is 2, m’ is 2 and n’’ is 3. In another embodiment, n’ is 2, m’ is 3 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 3.
  • n’ is 4, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 3. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 4.
  • n’ is 1, m’ is 1 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 4. In another embodiment, n’ is 2, m’ is 0 and n’’ is 4. In another embodiment, n’ is 2, m’ is 1 and n’’ is 4. In another embodiment, n’ is 2, m’ is 2 and n’’ is 4. In another embodiment, n’ is 2, m’ is 3 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 4.
  • n’ is 3, m’ is 1 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 4.
  • n’ is 5, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 4. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 5. In another embodiment, n’ is 2, m’ is 0 and n’’ is 5.
  • n’ is 2, m’ is 1 and n’’ is 5. In another embodiment, n’ is 2, m’ is 2 and n’’ is 5. In another embodiment, n’ is 2, m’ is 3 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 5.
  • n’ is 4, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 5.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 (CH 2 ) n” , where n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 3.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2.
  • m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5.
  • m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 O(CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 3.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2.
  • m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5.
  • m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 O(CH 2 CH 2 O) m’ (CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. In an embodiment, m is selected from 1 to 3, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 4 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 2
  • n’’ is selected from 0 to 3.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 2
  • n’’ is selected from 0 to 1.
  • m is 1, m’ is 0 and n’’ is 0.
  • m is 1, m’ is 1 and n’’ is 0.
  • m is 1, m’ is 2 and n’’ is 0.
  • m is 1, m’ is 3 and n’’ is 0.
  • m is 2, m’ is 0 and n’’ is 0. In another embodiment, m is 2, m’ is 1 and n’’ is 0. In another embodiment, m is 2, m’ is 2 and n’’ is 0. In another embodiment, m is 2, m’ is 3 and n’’ is 0. In yet another embodiment, m is 3, m’ is 0 and n’’ is 0. In yet another embodiment, m is 3, m’ is 1 and n’’ is 0. In yet another embodiment, m is 3, m’ is 2 and n’’ is 0. In yet another embodiment, m is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, m is 4, m’ is 0 and n” is 0.
  • m is 4, m’ is 1 and n” is 0. In yet a further embodiment, m is 4, m’ is 2 and n” is 0. In yet a further embodiment, m is 4, m’ is 3 and n” is 0. In a particular embodiment, m is 1, m’ is 0 and n’’ is 1. In a particular embodiment, m is 1, m’ is 1 and n’’ is 1. In a particular embodiment, m is 1, m’ is 2 and n’’ is 1. In a particular embodiment, m is 1, m’ is 3 and n’’ is 1. In another embodiment, m is 2, m’ is 0 and n’’ is 1.
  • m is 2, m’ is 1 and n’’ is 1. In another embodiment, m is 2, m’ is 2 and n’’ is 1. In another embodiment, m is 2, m’ is 3 and n’’ is 1. In yet another embodiment, m is 3, m’ is 0 and n’’ is 1. In yet another embodiment, m is 3, m’ is 1 and n’’ is 1. In yet another embodiment, m is 3, m’ is 2 and n’’ is 1. In yet another embodiment, m is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, m is 4, m’ is 0 and n” is 1. In yet a further embodiment, m is 4, m’ is 1 and n” is 1.
  • m is 4, m’ is 2 and n” is 1. In yet a further embodiment, m is 4, m’ is 3 and n” is 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 2. In a particular embodiment, m is 1, m’ is 1 and n’’ is 2. In a particular embodiment, m is 1, m’ is 2 and n’’ is 2. In a particular embodiment, m is 1, m’ is 3 and n’’ is 2. In another embodiment, m is 2, m’ is 0 and n’’ is 2. In another embodiment, m is 2, m’ is 1 and n’’ is 2.
  • m is 2, m’ is 2 and n’’ is 2. In another embodiment, m is 2, m’ is 3 and n’’ is 2. In yet another embodiment, m is 3, m’ is 0 and n’’ is 2. In yet another embodiment, m is 3, m’ is 1 and n’’ is 2. In yet another embodiment, m is 3, m’ is 2 and n’’ is 2. In yet another embodiment, m is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, m is 4, m’ is 0 and n” is 2. In yet a further embodiment, m is 4, m’ is 1 and n” is 2. In yet a further embodiment, m is 4, m’ is 2 and n” is 2.
  • m is 4, m’ is 3 and n” is 2. In a particular embodiment, m is 1, m’ is 0 and n’’ is 3. In a particular embodiment, m is 1, m’ is 1 and n’’ is 3. In a particular embodiment, m is 1, m’ is 2 and n’’ is 3. In a particular embodiment, m is 1, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 0 and n’’ is 3. In another embodiment, m is 2, m’ is 1 and n’’ is 3. In another embodiment, m is 2, m’ is 2 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3.
  • m is 3, m’ is 0 and n’’ is 3. In yet another embodiment, m is 3, m’ is 1 and n’’ is 3. In yet another embodiment, m is 3, m’ is 2 and n’’ is 3. In yet another embodiment, m is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, m is 4, m’ is 0 and n” is 3. In yet a further embodiment, m is 4, m’ is 1 and n” is 3. In yet a further embodiment, m is 4, m’ is 2 and n” is 3. In yet a further embodiment, m is 4, m’ is 3 and n” is 3.
  • m is 1, m’ is 0 and n’’ is 4. In a particular embodiment, m is 1, m’ is 1 and n’’ is 4. In a particular embodiment, m is 1, m’ is 2 and n’’ is 4. In a particular embodiment, m is 1, m’ is 3 and n’’ is 4. In another embodiment, m is 2, m’ is 0 and n’’ is 4. In another embodiment, m is 2, m’ is 1 and n’’ is 4. In another embodiment, m is 2, m’ is 2 and n’’ is 4. In another embodiment, m is 2, m’ is 3 and n’’ is 4. In yet another embodiment, m is 3, m’ is 0 and n’’ is 4.
  • m is 3, m’ is 1 and n’’ is 4. In yet another embodiment, m is 3, m’ is 2 and n’’ is 4. In yet another embodiment, m is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, m is 4, m’ is 0 and n” is 4. In yet a further embodiment, m is 4, m’ is 1 and n” is 4. In yet a further embodiment, m is 4, m’ is 2 and n” is 4. In yet a further embodiment, m is 4, m’ is 3 and n” is 4. In a particular embodiment, m is 1, m’ is 0 and n’’ is 5.
  • m is 1, m’ is 1 and n’’ is 5. In a particular embodiment, m is 1, m’ is 2 and n’’ is 5. In a particular embodiment, m is 1, m’ is 3 and n’’ is 5. In another embodiment, m is 2, m’ is 0 and n’’ is 5. In another embodiment, m is 2, m’ is 1 and n’’ is 5. In another embodiment, m is 2, m’ is 2 and n’’ is 5. In another embodiment, m is 2, m’ is 3 and n’’ is 5. In yet another embodiment, m is 3, m’ is 0 and n’’ is 5. In yet another embodiment, m is 3, m’ is 1 and n’’ is 5.
  • m is 3, m’ is 2 and n’’ is 5. In yet another embodiment, m is 3, m’ is 3 and n’’ is 5. In yet another embodiment, m is 4, m’ is 0 and n” is 5. In yet a further embodiment, m is 4, m’ is 1 and n” is 5. In yet a further embodiment, m is 4, m’ is 2 and n” is 5. In yet a further embodiment, m is 4, m’ is 3 and n” is 5.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is OCH 2 (CH 2 ) n’ , where n’ is selected from 1 to 10 and wherein X' is CH 2 (CH 2 ) n” , where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3 and n’’ is selected from 0 to 3.
  • n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1.
  • n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 1 and n’’ is 3.
  • n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4.
  • n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6.
  • n’ is 6 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is OCH 2 (CH 2 ) n’ , where n’ is selected from 1 to 10 and wherein X' is CH 2 O(CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10.
  • n’ is selected from 1 to 5 and n’’ is selected from 0 to 10.
  • n’ is selected from 1 to 5 and n’’ is selected from 0 to 5.
  • n’ is selected from 1 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1.
  • n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2.
  • n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4.
  • n’ is 6 and n” is 4. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6.
  • n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is OCH 2 (CH 2 ) n’ , where n’ is selected from 1 to 10 and wherein X' is CH 2 O(CH 2 CH 2 O) m’ (CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4.
  • n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 0. In another embodiment, n’ is 1, m’ is 1 and n’’ is 0.
  • n’ is 1, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In another embodiment, n’ is 2, m’ is 0 and n’’ is 0. In another embodiment, n’ is 2, m’ is 1 and n’’ is 0. In another embodiment, n’ is 2, m’ is 2 and n’’ is 0. In another embodiment, n’ is 2, m’ is 3 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 0.
  • n’ is 3, m’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 0.
  • n’ is 5, m’ is 1 and n” is 0.In yet a further embodiment, n’ is 5, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 0. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 1. In another embodiment, n’ is 2, m’ is 0 and n’’ is 1.
  • n’ is 2, m’ is 1 and n’’ is 1. In another embodiment, n’ is 2, m’ is 2 and n’’ is 1. In another embodiment, n’ is 2, m’ is 3 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 1.
  • n’ is 4, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 2.
  • n’ is 1, m’ is 1 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 2. In another embodiment, n’ is 2, m’ is 0 and n’’ is 2. In another embodiment, n’ is 2, m’ is 1 and n’’ is 2. In another embodiment, n’ is 2, m’ is 2 and n’’ is 2. In another embodiment, n’ is 2, m’ is 3 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 2.
  • n’ is 3, m’ is 1 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 2.
  • n’ is 5, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 2. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 3. In another embodiment, n’ is 2, m’ is 0 and n’’ is 3.
  • n’ is 2, m’ is 1 and n’’ is 3. In another embodiment, n’ is 2, m’ is 2 and n’’ is 3. In another embodiment, n’ is 2, m’ is 3 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 3.
  • n’ is 4, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 3. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 4.
  • n’ is 1, m’ is 1 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 4. In another embodiment, n’ is 2, m’ is 0 and n’’ is 4. In another embodiment, n’ is 2, m’ is 1 and n’’ is 4. In another embodiment, n’ is 2, m’ is 2 and n’’ is 4. In another embodiment, n’ is 2, m’ is 3 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 4.
  • n’ is 3, m’ is 1 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 4.
  • n’ is 5, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 4. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 5. In another embodiment, n’ is 2, m’ is 0 and n’’ is 5.
  • n’ is 2, m’ is 1 and n’’ is 5. In another embodiment, n’ is 2, m’ is 2 and n’’ is 5. In another embodiment, n’ is 2, m’ is 3 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 5.
  • n’ is 4, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 5.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 (CH 2 ) n” , where n’’ is selected from 0 to 10.
  • X is O(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 (CH 2 ) n” , where n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 3.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2.
  • m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5.
  • m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH2O(CH2)n’’CH2, where n’’ is selected from 0 to 10.
  • X is O(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH2O(CH2)n’’CH2, where n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 3 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 3.
  • m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2.
  • m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5.
  • m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6.
  • the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is O(CH 2 CH 2 O) m CH 2 CH 2 , where m is selected from 1 to 4 and wherein X' is CH 2 O(CH 2 CH 2 O) m’ (CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4.
  • m is selected from 1 to 3
  • m’ is selected from 0 to 4 and n’’ is selected from 0 to 10.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 4 and n’’ is selected from 0 to 5.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 2
  • n’’ is selected from 0 to 3.
  • m is selected from 1 to 2
  • m’ is selected from 0 to 2
  • n’’ is selected from 0 to 1.
  • m is 1, m’ is 0 and n’’ is 0.
  • m is 1, m’ is 1 and n’’ is 0.
  • m is 1, m’ is 2 and n’’ is 0.
  • m is 1, m’ is 3 and n’’ is 0.
  • m is 2, m’ is 0 and n’’ is 0. In another embodiment, m is 2, m’ is 1 and n’’ is 0. In another embodiment, m is 2, m’ is 2 and n’’ is 0. In another embodiment, m is 2, m’ is 3 and n’’ is 0. In yet another embodiment, m is 3, m’ is 0 and n’’ is 0. In yet another embodiment, m is 3, m’ is 1 and n’’ is 0. In yet another embodiment, m is 3, m’ is 2 and n’’ is 0. In yet another embodiment, m is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, m is 4, m’ is 0 and n” is 0.
  • m is 4, m’ is 1 and n” is 0. In yet a further embodiment, m is 4, m’ is 2 and n” is 0. In yet a further embodiment, m is 4, m’ is 3 and n” is 0. In a particular embodiment, m is 1, m’ is 0 and n’’ is 1. In a particular embodiment, m is 1, m’ is 1 and n’’ is 1. In a particular embodiment, m is 1, m’ is 2 and n’’ is 1. In a particular embodiment, m is 1, m’ is 3 and n’’ is 1. In another embodiment, m is 2, m’ is 0 and n’’ is 1.
  • m is 2, m’ is 1 and n’’ is 1. In another embodiment, m is 2, m’ is 2 and n’’ is 1. In another embodiment, m is 2, m’ is 3 and n’’ is 1. In yet another embodiment, m is 3, m’ is 0 and n’’ is 1. In yet another embodiment, m is 3, m’ is 1 and n’’ is 1. In yet another embodiment, m is 3, m’ is 2 and n’’ is 1. In yet another embodiment, m is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, m is 4, m’ is 0 and n” is 1. In yet a further embodiment, m is 4, m’ is 1 and n” is 1.
  • m is 4, m’ is 2 and n” is 1. In yet a further embodiment, m is 4, m’ is 3 and n” is 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 2. In a particular embodiment, m is 1, m’ is 1 and n’’ is 2. In a particular embodiment, m is 1, m’ is 2 and n’’ is 2. In a particular embodiment, m is 1, m’ is 3 and n’’ is 2. In another embodiment, m is 2, m’ is 0 and n’’ is 2. In another embodiment, m is 2, m’ is 1 and n’’ is 2.
  • m is 2, m’ is 2 and n’’ is 2. In another embodiment, m is 2, m’ is 3 and n’’ is 2. In yet another embodiment, m is 3, m’ is 0 and n’’ is 2. In yet another embodiment, m is 3, m’ is 1 and n’’ is 2. In yet another embodiment, m is 3, m’ is 2 and n’’ is 2. In yet another embodiment, m is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, m is 4, m’ is 0 and n” is 2. In yet a further embodiment, m is 4, m’ is 1 and n” is 2. In yet a further embodiment, m is 4, m’ is 2 and n” is 2.
  • m is 4, m’ is 3 and n” is 2. In a particular embodiment, m is 1, m’ is 0 and n’’ is 3. In a particular embodiment, m is 1, m’ is 1 and n’’ is 3. In a particular embodiment, m is 1, m’ is 2 and n’’ is 3. In a particular embodiment, m is 1, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 0 and n’’ is 3. In another embodiment, m is 2, m’ is 1 and n’’ is 3. In another embodiment, m is 2, m’ is 2 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3.
  • m is 3, m’ is 0 and n’’ is 3. In yet another embodiment, m is 3, m’ is 1 and n’’ is 3. In yet another embodiment, m is 3, m’ is 2 and n’’ is 3. In yet another embodiment, m is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, m is 4, m’ is 0 and n” is 3. In yet a further embodiment, m is 4, m’ is 1 and n” is 3. In yet a further embodiment, m is 4, m’ is 2 and n” is 3. In yet a further embodiment, m is 4, m’ is 3 and n” is 3.
  • m is 1, m’ is 0 and n’’ is 4. In a particular embodiment, m is 1, m’ is 1 and n’’ is 4. In a particular embodiment, m is 1, m’ is 2 and n’’ is 4. In a particular embodiment, m is 1, m’ is 3 and n’’ is 4. In another embodiment, m is 2, m’ is 0 and n’’ is 4. In another embodiment, m is 2, m’ is 1 and n’’ is 4. In another embodiment, m is 2, m’ is 2 and n’’ is 4. In another embodiment, m is 2, m’ is 3 and n’’ is 4. In yet another embodiment, m is 3, m’ is 0 and n’’ is 4.
  • m is 3, m’ is 1 and n’’ is 4. In yet another embodiment, m is 3, m’ is 2 and n’’ is 4. In yet another embodiment, m is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, m is 4, m’ is 0 and n” is 4. In yet a further embodiment, m is 4, m’ is 1 and n” is 4. In yet a further embodiment, m is 4, m’ is 2 and n” is 4. In yet a further embodiment, m is 4, m’ is 3 and n” is 4. In a particular embodiment, m is 1, m’ is 0 and n’’ is 5.
  • m is 1, m’ is 1 and n’’ is 5. In a particular embodiment, m is 1, m’ is 2 and n’’ is 5. In a particular embodiment, m is 1, m’ is 3 and n’’ is 5. In another embodiment, m is 2, m’ is 0 and n’’ is 5. In another embodiment, m is 2, m’ is 1 and n’’ is 5. In another embodiment, m is 2, m’ is 2 and n’’ is 5. In another embodiment, m is 2, m’ is 3 and n’’ is 5. In yet another embodiment, m is 3, m’ is 0 and n’’ is 5. In yet another embodiment, m is 3, m’ is 1 and n’’ is 5.
  • m is 3, m’ is 2 and n’’ is 5. In yet another embodiment, m is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, m is 4, m’ is 0 and n” is 5. In yet a further embodiment, m is 4, m’ is 1 and n” is 5. In yet a further embodiment, m is 4, m’ is 2 and n” is 5. In yet a further embodiment, m is 4, m’ is 3 and n” is 5. 1.5 Carrier protein of the glycoconjugates of the invention A component of the glycoconjugate is a carrier protein to which the saccharide is conjugated.
  • the carrier protein of the glycoconjugate of the invention is selected in the group consisting of: DT (Diphtheria toxoid), TT (tetanus toxoid) or fragment C of TT, CRM 197 (a nontoxic but antigenically identical variant of diphtheria toxin), other DT mutants (such as CRM 176 , CRM 228 , CRM 45 (Uchida et al. (1973) J. Biol.
  • Patent No.5,843,711 pneumococcal pneumolysin (ply) (Kuo et al. (1995) Infect lmmun 63:2706-2713) including ply detoxified in some fashion, for example dPLY-GMBS (WO 2004/081515, WO 2006/032499) or dPLY-formol, PhtX, including PhtA, PhtB, PhtD, PhtE (sequences of PhtA, PhtB, PhtD or PhtE are disclosed in WO 00/37105 and WO 00/39299) and fusions of Pht proteins, for example PhtDE fusions, PhtBE fusions, Pht A-E (WO 01/98334, WO 03/054007, WO 2009/000826), OMPC (meningococcal outer membrane protein), which is usually extracted from Neisseria meningitidis serogroup B (EP0372501), PorB (from N.
  • PD Hemophilus influenzae protein D
  • PD Hemophilus influenzae protein D
  • synthetic peptides EP0378881, EP0427347
  • heat shock proteins WO 93/17712, WO 94/03208
  • pertussis proteins WO 98/58668, EP0471177
  • cytokines lymphokines
  • growth factors or hormones WO 91/01146
  • artificial proteins comprising multiple human CD4+ T cell epitopes from various pathogen derived antigens (Falugi et al. (2001) Eur J Immunol 31:3816-3824) such as N19 protein (Baraldoi et al.
  • Other proteins such as ovalbumin, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA) or purified protein derivative of tuberculin (PPD) also can be used as carrier proteins.
  • suitable carrier proteins include inactivated bacterial toxins such as cholera toxoid (e.g., as described in WO 2004/083251), Escherichia coli LT, E. coli ST, and exotoxin A from P. aeruginosa.
  • Another suitable carrier protein is a C5a peptidase from Streptococcus (SCP).
  • the carrier protein of the glycoconjugate of the invention is TT, DT, DT mutants (such as CRM 197 ) or a C5a peptidase from Streptococcus (SCP).
  • the carrier protein of the glycoconjugate of the invention is DT (Diphtheria toxoid).
  • the carrier protein of the glycoconjugate of the invention is TT (tetanus toxoid).
  • the carrier protein of the glycoconjugate of the invention is PD (H. influenzae protein D; see, e.g., EP0594610 B).
  • the carrier protein of the glycoconjugate of the invention is CRM 197 or a C5a peptidase from Streptococcus (SCP).
  • the carrier protein of the glycoconjugate of the invention is CRM 197 .
  • the CRM 197 protein is a nontoxic form of diphtheria toxin but is immunologically indistinguishable from the diphtheria toxin.
  • CRM 197 is produced by Corynebacterium diphtheriae infected by the nontoxigenic phage ⁇ 197 tox- created by nitrosoguanidine mutagenesis of the toxigenic corynephage beta (Uchida et al. (1971) Nature New Biology 233:8-11).
  • the CRM 197 protein has the same molecular weight as the diphtheria toxin but differs therefrom by a single base change (guanine to adenine) in the structural gene. This single base change causes an amino acid substitution (glutamic acid for glycine) in the mature protein and eliminates the toxic properties of diphtheria toxin.
  • the CRM 197 protein is a safe and effective T-cell dependent carrier for saccharides. Further details about CRM 197 and production thereof can be found, e.g., in U.S. Patent No.5,614,382.
  • the carrier protein of the glycoconjugate of the invention is the A chain of CRM 197 (see CN103495161).
  • the carrier protein of the glycoconjugate of the invention is the A chain of CRM 197 obtained via expression by genetically recombinant E. coli (see CN103495161).
  • the carrier protein of the glycoconjugate of the invention is SCP (Streptococcal C5a Peptidase).
  • Streptococcus pyogenes group A Streptococcus, GAS
  • Streptococcus agalactiae group B Streptococcus, GBS
  • GAS necrotizing fasciitis
  • GBS neonatal sepsis
  • the scp genes from GAS and GBS encode a polypeptide containing between 1,134 and 1,181 amino acids (Brown et al., PNAS, 2005, vol.102, no.51 pages 18391–18396).
  • the first 31 residues are the export signal presequence and are removed upon passing through the cytoplasmic membrane.
  • the next 68 residues serve as a pro-sequence and must be removed to produce active SCP.
  • the next 10 residues can be removed without loss of protease activity.
  • Lys-1034 are four consecutive 17-residue motifs followed by a cell sorting and cell-wall attachment signal.
  • This combined signal is composed of a 20-residue hydrophilic sequence containing an LPTTND sequence, a 17-residue hydrophobic sequence, and a short basic carboxyl terminus.
  • SCP can be divided in domains (see figure 1B of Brown et al., PNAS, 2005, vol.102, no. 51 pages 18391–18396).
  • These domains are the Pre/Pro domain (which comprises the export signal presequence (commonly the first 31 residues) and the pro-sequence (commonly the next 68 residues)), the protease domain (which is splitted in two part (protease part 1 commonly residues 89–333/334 and protease domain part 2 and commonly residues 467/468–583/584), the protease-associated domain (PA domain) (commonly residues 333/334–467/468), three fibronectin type III (Fn) domains (Fn1, commonly residues 583/584–712/713; Fn2, commonly residues 712/713–928/929/930; commonly Fn3, residues 929/930-1029/1030/1031) and a cell wall anchor domain (commonly residues 1029/1030/1031 to the C-terminus).
  • the protease domain which is splitted in two part (protease part 1 commonly residues 89–333/334 and protea
  • the carrier protein of the glycoconjugate of the invention is an SCP from GBS (SCPB).
  • SCPB GBS
  • An example of SCPB is provided at SEQ ID.NO: 3 of WO97/26008. See also SEQ ID NO: 3 of WO00/34487.
  • the carrier protein of the glycoconjugate of the invention is an SCP from GAS (SCPA). Examples of SCPA can be found at SEQ ID.No.1 and SEQ ID.No.2 of WO97/26008. See also SEQ ID NO: 1, 2 and 23 of WO00/34487.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP from GBS (SCPB). In another preferred embodiments, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP from GAS (SCPA). In an embodiment, the carrier protein of the glycoconjugate of the invention is a fragment of an SCP. In an embodiment, the carrier protein of the glycoconjugate of the invention is a fragment of an SCPA. Preferably, the carrier protein of the glycoconjugate of the invention is a fragment of an SCPB.
  • the carrier protein of the glycoconjugate of the invention is a fragment of an SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain.
  • the carrier protein of the glycoconjugate of the invention is a fragment of an SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP which comprises the protease domain, the protease- associated domain (PA domain) and two of the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP.
  • said enzymatically inactive fragment of SCP comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCPA.
  • said enzymatically inactive fragment of an SCPA comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB.
  • said enzymatically inactive fragment of SCPB comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain.
  • the enzymatic activity of SCP is inactivated by replacing at least one amino acid of the wild type sequence.
  • said replacement is selected from the group consisting of D130A, H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence.
  • said replacement of at least one amino acid is in the protease domain.
  • said replacement of at least one amino acid is in part 1 of the protease domain.
  • said replacement of at least one amino acid is in part 2 of the protease domain.
  • said replacement is selected from the group consisting of D130A, H193A, N295A and S512A.
  • said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain.
  • said replacement is selected from the group consisting of D130A, H193A, N295A and S512A.
  • said replacement is D130A.
  • said replacement is H193A.
  • said replacement is N295A.
  • said replacement is S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence.
  • said replacement of at least one amino acid is in the protease domain.
  • said replacement of at least one amino acid is in part 1 of the protease domain.
  • said replacement of at least one amino acid is in part 2 of the protease domain.
  • said replacement is selected from the group consisting of D130A, H193A, N295A and S512A.
  • said replacement is D130A.
  • said replacement is H193A.
  • said replacement is N295A.
  • said replacement is S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence.
  • said replacement of at least one amino acid is in the protease domain.
  • said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence.
  • said replacement of at least one amino acid is in the protease domain.
  • said replacement of at least one amino acid is in part 1 of the protease domain.
  • said replacement of at least one amino acid is in part 2 of the protease domain.
  • said replacement is selected from the group consisting of D130A, H193A, N295A and S512A.
  • said replacement is D130A.
  • said replacement is H193A.
  • said replacement is N295A.
  • said replacement is S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence.
  • said replacement of at least one amino acid is in the protease domain.
  • said replacement of at least one amino acid is in part 1 of the protease domain.
  • said replacement of at least one amino acid is in part 2 of the protease domain.
  • said replacement is selected from the group consisting of D130A, H193A, N295A and S512A.
  • said replacement is D130A.
  • said replacement is H193A.
  • said replacement is N295A.
  • said replacement is S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence.
  • said replacement of at least one amino acid is in the protease domain.
  • said replacement of at least one amino acid is in part 1 of the protease domain.
  • said replacement of at least one amino acid is in part 2 of the protease domain.
  • said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A.
  • the enzymatic activity of SCP is inactivated by replacing at least two amino acids of the wild type sequence. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A.
  • said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. Therefore, in an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acid is in part 1 of the protease domain.
  • said replacement of at least two amino acid is in part 2 of the protease domain.
  • said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least two amino acids replacements are D130A and H193A.
  • said at least two amino acids replacements are D130A and N295A.
  • said at least two amino acids replacements are D130A and S512A.
  • said at least two amino acids replacements are H193A and N295A.
  • said at least two amino acids replacements are H193A and S512A.
  • said at least two amino acids replacements are N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence.
  • said replacement of at least two amino acids is in the protease domain.
  • said replacement of at least two amino acids is in part 1 of the protease domain.
  • said replacement of at least two amino acid is in part 2 of the protease domain.
  • said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence.
  • said replacement of at least two amino acids is in the protease domain.
  • said replacement of at least two amino acids is in part 1 of the protease domain.
  • said replacement of at least two amino acid is in part 2 of the protease domain.
  • said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least two amino acids replacements are D130A and H193A.
  • said at least two amino acids replacements are D130A and N295A.
  • said at least two amino acids replacements are D130A and S512A.
  • said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence.
  • said replacement of at least two amino acids is in the protease domain.
  • said replacement of at least two amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least two amino acid is in part 2 of the protease domain.
  • said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence.
  • said replacement of at least two amino acids is in the protease domain.
  • said replacement of at least two amino acids is in part 1 of the protease domain.
  • said replacement of at least two amino acid is in part 2 of the protease domain.
  • said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence.
  • said replacement of at least two amino acids is in the protease domain.
  • said replacement of at least two amino acids is in part 1 of the protease domain.
  • said replacement of at least one amino acid is in part 2 of the protease domain.
  • said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence.
  • said replacement of at least two amino acids is in the protease domain.
  • said replacement of at least two amino acids is in part 1 of the protease domain.
  • said replacement of at least two amino acids is in part 2 of the protease domain.
  • said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A.
  • the enzymatic activity of SCP is inactivated by replacing at least three amino acids of the wild type sequence.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least three amino acids replacements are D130A, H193A and N295A.
  • said at least three amino acids replacements are D130A, H193A and S512A.
  • said at least three amino acids replacements are D130A, N295A and S512A.
  • said at least three amino acids replacements are H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence.
  • said replacement of at least three amino acids is in the protease domain.
  • said replacement of at least three amino acid is in part 1 of the protease domain.
  • said replacement of at least three amino acid is in part 2 of the protease domain.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least three amino acids replacements are D130A, H193A and N295A.
  • said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence.
  • said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain.
  • said replacement of at least three amino acid is in part 2 of the protease domain.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least three amino acids replacements are D130A, H193A and N295A.
  • said at least three amino acids replacements are D130A, H193A and S512A.
  • said at least three amino acids replacements are D130A, N295A and S512A.
  • said at least three amino acids replacements are H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence.
  • said replacement of at least three amino acids is in the protease domain.
  • said replacement of at least three amino acids is in part 1 of the protease domain.
  • said replacement of at least three amino acid is in part 2 of the protease domain.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least three amino acids replacements are D130A, H193A and N295A.
  • said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence.
  • said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain.
  • said replacement of at least three amino acid is in part 2 of the protease domain.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A.
  • said at least three amino acids replacements are D130A, H193A and N295A.
  • said at least three amino acids replacements are D130A, H193A and S512A.
  • said at least three amino acids replacements are D130A, N295A and S512A.
  • said at least three amino acids replacements are H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence.
  • said replacement of at least three amino acids is in the protease domain.
  • said replacement of at least three amino acids is in part 1 of the protease domain.
  • said replacement of at least three amino acid is in part 2 of the protease domain.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence.
  • said replacement of at least three amino acids is in the protease domain.
  • said replacement of at least three amino acids is in part 1 of the protease domain.
  • said replacement of at least three amino acids is in part 2 of the protease domain.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence.
  • said replacement of at least three amino acids is in the protease domain.
  • said replacement of at least three amino acids is in part 1 of the protease domain.
  • said replacement of at least three amino acids is in part 2 of the protease domain.
  • said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the enzymatic activity of SCP is inactivated by replacing at least four amino acids of the wild type sequence.
  • said at least four amino acids replacements are D130A, H193A, N295A and S512A. Therefore, in an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least four amino acid is in part 2 of the protease domain.
  • said at least four amino acids replacements are D130A, H193A, N295A and S512A
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence.
  • said replacement of at least four amino acids is in the protease domain.
  • said replacement of at least four amino acids is in part 1 of the protease domain.
  • said replacement of at least four amino acid is in part 2 of the protease domain.
  • said at least four amino acids replacements are D130A, H193A, N295A and S512A
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence.
  • said replacement of at least four amino acids is in the protease domain.
  • said replacement of at least four amino acids is in part 1 of the protease domain.
  • said replacement of at least four amino acid is in part 2 of the protease domain.
  • said at least four amino acids replacements are D130A, H193A, N295A and S512A
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence.
  • said replacement of at least four amino acids is in the protease domain.
  • said replacement of at least four amino acids is in part 1 of the protease domain.
  • said replacement of at least four amino acid is in part 2 of the protease domain.
  • said at least four amino acids replacements are D130A, H193A, N295A and S512A
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence.
  • said replacement of at least four amino acids is in the protease domain.
  • said replacement of at least four amino acids is in part 1 of the protease domain.
  • said replacement of at least four amino acid is in part 2 of the protease domain.
  • said at least four amino acids replacements are D130A, H193A, N295A and S512A
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence.
  • said replacement of at least four amino acids is in the protease domain.
  • said replacement of at least four amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence.
  • said replacement of at least four amino acids is in the protease domain.
  • said replacement of at least four amino acids is in part 1 of the protease domain.
  • said replacement of at least four amino acids is in part 2 of the protease domain.
  • said at least four amino acids replacements are D130A, H193A, N295A and S512A
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which consists of SEQ ID NO: 41.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which consists of SEQ ID NO: 42.
  • SEQ ID NO: 41 MAKTADTPATSKATIRDLNDPSQVKTLQEKAGKGAGTVVAVIAAGFDKNH EAWRLTDKAKARYQSKEDLEKAKKEHGITYGEWVNDKVAYYHDYSKDGKT AVDQEHGTHVSGILSGNAPSETKEPYRLEGAMPEAQLLLMRVEIVNGLAD YARNYAQAIRDAINLGAKVINMSFGNAALAYANLPDETKKAFDYAKSKGV SIVTSAGNDSSFGGKTRLPLADHPDYGVVGTPAAADSTLTVASYSPDKQL TETVTVKTADQQDKEMPVLSTNRFEPNKAYDYAYANRGTKEDDFKDVKGK IALIERGDIDFKDKIAKAKKAGAVGVLIYDNQDKGFPIELPNVDQMPAAF ISRKDGLLLKDNPQKTITFNATPKVLPTASGTKLSRFSSWGLTADGNIKP DIAAPGQDILSSVANNKYAKLSGTAMSAPLVAGI
  • SEQ ID NO: 42 AKTADTPATSKATIRDLNDPSQVKTLQEKAGKGAGTVVAVIAAGFDKNH EAWRLTDKAKARYQSKEDLEKAKKEHGITYGEWVNDKVAYYHDYSKDGKT AVDQEHGTHVSGILSGNAPSETKEPYRLEGAMPEAQLLLMRVEIVNGLAD YARNYAQAIRDAINLGAKVINMSFGNAALAYANLPDETKKAFDYAKSKGV SIVTSAGNDSSFGGKTRLPLADHPDYGVVGTPAAADSTLTVASYSPDKQL TETVTVKTADQQDKEMPVLSTNRFEPNKAYDYAYANRGTKEDDFKDVKGK IALIERGDIDFKDKIAKAKKAGAVGVLIYDNQDKGFPIELPNVDQMPAAF ISRKDGLLLKDNPQKTITFNATPKVLPTASGTKLSRFSSWGLTADGNIKP DIAAPGQDILSSVANNKYAKLSGTAMSAPLVAGI
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 90% identity with SEQ ID NO: 41.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity with SEQ ID NO: 41.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99% identity with SEQ ID NO: 41.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.5% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.8% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.85% identity with SEQ ID NO: 41.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 90% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99% identity with SEQ ID NO: 42.
  • the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.5% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.8% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.85% identity with SEQ ID NO: 42.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention (as disclosed at section 1 above). In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising from 1 to 25 different glycoconjugates. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising from 26 to 35 different glycoconjugates. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising from 1 to 25 glycoconjugates from different serotypes of S. pneumoniae (1 to 25 pneumococcal conjugates).
  • the invention relates to an immunogenic composition comprising glycoconjugates from 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 different serotypes of S. pneumoniae.
  • the immunogenic composition comprises glycoconjugates from 16 or 20 different serotypes of S. pneumoniae.
  • the immunogenic composition is a 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20-valent pneumococcal conjugate compositions.
  • the immunogenic composition is a 14, 15, 16, 17, 18 or 19-valent pneumococcal conjugate compositions.
  • the immunogenic composition is a 16-valent pneumococcal conjugate composition.
  • the immunogenic composition is a 19- valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 20-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 21, 22, 23, 24 or 25-valent pneumococcal conjugate compositions. In an embodiment the immunogenic composition is a 21- valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 25-valent pneumococcal conjugate composition.
  • the immunogenic composition is a 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35-valent pneumococcal conjugate compositions. In an embodiment the immunogenic composition is a 26-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 27-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 28-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 29-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 30-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 31-valent pneumococcal conjugate composition.
  • the immunogenic composition is a 32- valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 33-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 34-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 35-valent pneumococcal conjugate composition. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F. In an embodiment the immunogenic composition is an 11-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition
  • a glycoconjugate of the invention comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F.
  • the immunogenic composition is a 13-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and further comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F.
  • the immunogenic composition is a 15-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F.
  • the immunogenic composition is a 20-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and further comprising glycoconjugates from S.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 2, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 22,F, 23F and 33F.
  • the immunogenic composition is a 24-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F and 33F.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 24F and 33F.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and further comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F and 33F.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F, 24F and 33F.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F and 33F.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 24F and 33F.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F and 33F.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 33F and 35B.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F and 33F.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 24F and 33F.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F and 33F.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 33F and 35B.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 33F and 35B.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F and 33F and 35B.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F and 35B.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 24-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F and 33F.
  • the immunogenic composition is a 24-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 24F, 33F and 35B.
  • the immunogenic composition is a 24-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 24-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F and 35B.
  • the immunogenic composition is a 24-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F and 35B.
  • the immunogenic composition is a 25-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention selected from the group consisting of glycoconjugates from S.
  • the invention relates to an immunogenic composition
  • an immunogenic composition comprising a glycoconjugate of the invention selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition
  • a glycoconjugate of the invention and comprising twenty one glycoconjugates selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38.
  • the immunogenic composition is a 22-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising at least one glycoconjugate selected from the group consisting of glycoconjugates from S.
  • the invention relates to an immunogenic composition
  • a glycoconjugate of the invention and comprising twenty two glycoconjugates selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising twenty three glycoconjugates selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38.
  • the immunogenic composition is a 24-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition
  • an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 2, 9N, 15A, 17F, 20, 23A, 23B, 24F and 35B.
  • the immunogenic composition is a 10-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 2, 9N, 15A, 17F, 19A, 19F, 20, 23A, 23B, 24F and 35B.
  • the immunogenic composition is a 12-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition
  • a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38.
  • the immunogenic composition is a 23-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S.
  • the immunogenic composition is a 25-valent pneumococcal conjugate compositions.
  • the saccharides are each individually conjugated to different molecules of the protein carrier (each molecule of protein carrier only having one type of saccharide conjugated to it).
  • the capsular saccharides are said to be individually conjugated to the carrier protein.
  • all the glycoconjugates of the above immunogenic compositions are individually conjugated to the carrier protein.
  • the glycoconjugate of the invention is conjugated to CRM 197 .
  • the glycoconjugate of the invention is conjugated to SCP.
  • the glycoconjugate from S. pneumoniae serotype 22F is conjugated to CRM 197 .
  • the glycoconjugate from S. pneumoniae serotype 33F is conjugated to CRM 197 .
  • the glycoconjugate from S. pneumoniae serotype 15B is conjugated to CRM 197 .
  • pneumoniae serotype 12F is conjugated to CRM 197 .
  • the glycoconjugate from S. pneumoniae serotype 10A is conjugated to CRM 197 .
  • the glycoconjugate from S. pneumoniae serotype 11A is conjugated to CRM 197 .
  • the glycoconjugate from S. pneumoniae serotype 8 is conjugated to CRM 197 .
  • the glycoconjugates from S. pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F and 23F are conjugated to CRM 197 .
  • the glycoconjugates from S. pneumoniae serotypes 1, 5 and 7F are conjugated to CRM 197 .
  • the glycoconjugates from S. pneumoniae serotypes 6A and 19A are conjugated to CRM 197 .
  • the glycoconjugates of any of the above immunogenic compositions are all individually conjugated to CRM 197 .
  • the glycoconjugate of the invention is conjugated to SCP and the other glycoconjugate(s) is/are all individually conjugated to CRM 197 .
  • the glycoconjugate from S. pneumoniae serotypes 1, 4, 5, 6B, 7F, 9V, 14 and/or 23F of any of the above immunogenic compositions are individually conjugated to PD.
  • the glycoconjugate from S. pneumoniae serotype 18C of any of the above immunogenic compositions is conjugated to TT.
  • the glycoconjugate from S. pneumoniae serotype 19F of any of the above immunogenic compositions is conjugated to DT.
  • the glycoconjugates from S. pneumoniae serotypes 1, 4, 5, 6B, 7F, 9V, 14 and/or 23F of any of the above immunogenic compositions are individually conjugated to PD, the glycoconjugate from S.
  • the glycoconjugate from S. pneumoniae serotype 18C is conjugated to TT and the glycoconjugate from S. pneumoniae serotype 19F is conjugated to DT.
  • the above immunogenic compositions comprise from 8 to 25 different serotypes of S. pneumoniae.
  • the above immunogenic compositions comprise from 26 to 35 different serotypes of S. pneumoniae.
  • the glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 18C, 19A, 19F, 23F and 33F of any of the above immunogenic compositions are individually conjugated to CRM197, the glycoconjugates from S.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F, wherein the glycoconjugates from S.
  • the immunogenic composition is a 21-valent pneumococcal conjugate compositions.
  • Compositions of the invention may include a small amount of free carrier.
  • the unconjugated form is preferably no more than 5% of the total amount of the carrier protein in the composition as a whole, and more preferably present at less than 2% by weight.
  • the amount of glycoconjugate(s) in each dose is selected as an amount which induces an immunoprotective response without significant, adverse side effects in typical vaccinees. Such amount will vary depending upon which specific immunogen is employed and how it is presented. The amount of a particular glycoconjugate in an immunogenic composition can be calculated based on total saccharide for that conjugate (conjugated and non-conjugated).
  • a glycoconjugate with 20% free saccharide will have about 80 ⁇ g of conjugated saccharide and about 20 ⁇ g of nonconjugated saccharide in a 100 ⁇ g saccharide dose.
  • the amount of glycoconjugate can vary depending upon the bacteria and bacteria serotype.
  • the saccharide concentration can be determined by the uronic acid assay.
  • the "immunogenic amount" of the different saccharide components in the immunogenic composition may diverge and each may comprise about 0.5 ⁇ g, about 0.75 ⁇ g, about 1 ⁇ g, about 2 ⁇ g, about 3 ⁇ g, about 4 ⁇ g, about 5 ⁇ g, about 6 ⁇ g, about 7 ⁇ g, about 8 ⁇ g, about 9 ⁇ g, about 10 ⁇ g, about 15 ⁇ g, about 20 ⁇ g, about 30 ⁇ g, about 40 ⁇ g, about 50 ⁇ g, about 60 ⁇ g, about 70 ⁇ g, about 80 ⁇ g, about 90 ⁇ g, or about 100 ⁇ g of any particular saccharide antigen.
  • each dose will comprise 0.1 ⁇ g to 100 ⁇ g of saccharide. In an embodiment each dose will comprise 0.1 ⁇ g to 100 ⁇ g of saccharide. In a preferred embodiment each dose will comprise 0.5 ⁇ g to 20 ⁇ g. In a preferred embodiment each dose will comprise 1.0 ⁇ g to 10 ⁇ g. In an even preferred embodiment, each dose will comprise 2.0 ⁇ g to 5.0 ⁇ g of serotype 3 polysaccharide. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure. In an embodiment, each dose will comprise about 0.5 ⁇ g of saccharide. In an embodiment, each dose will comprise about 0.55 ⁇ g of saccharide.
  • each dose will comprise about 0.75 ⁇ g of saccharide. In an embodiment, each dose will comprise about 1.0 ⁇ g of saccharide. In an embodiment, each dose will comprise about 1.1 ⁇ g of saccharide. In an embodiment, each dose will comprise about 1.5 ⁇ g of saccharide. In an embodiment, each dose will comprise about 2.0 ⁇ g of saccharide. In an embodiment, each dose will comprise about 2.2 ⁇ g of saccharide. In an embodiment, each dose will comprise about 2.5 ⁇ g of saccharide. In an embodiment, each dose will comprise about 3.0 ⁇ g of saccharide. In an embodiment, each dose will comprise about 3.5 ⁇ g of saccharide. In an embodiment, each dose will comprise about 4.0 ⁇ g of saccharide.
  • each dose will comprise about 4.4 ⁇ g of saccharide. In an embodiment, each dose will comprise about 5.0 ⁇ g of saccharide. In an embodiment, each dose will comprise about 5.5 ⁇ g of saccharide. In an embodiment, each dose will comprise about 6.0 ⁇ g of saccharide. Generally, each dose will comprise 0.1 ⁇ g to 100 ⁇ g of saccharide for a given bacteria or serotype. In an embodiment each dose will comprise 0.1 ⁇ g to 100 ⁇ g of saccharide for a given bacteria or serotype. In a preferred embodiment each dose will comprise 0.5 ⁇ g to 20 ⁇ g. In a preferred embodiment each dose will comprise 1.0 ⁇ g to 10 ⁇ g.
  • each dose will comprise 2.0 ⁇ g to 5.0 ⁇ g of saccharide for a given bacteria or serotype. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure.
  • each dose will comprise about 0.5 ⁇ g of saccharide for each particular glycoconjugate.
  • each dose will comprise about 0.55 ⁇ g of saccharide for each particular glycoconjugate.
  • each dose will comprise about 0.75 ⁇ g of saccharide for each particular glycoconjugate.
  • each dose will comprise about 1.0 ⁇ g of saccharide for each particular glycoconjugate.
  • each dose will comprise about 1.1 ⁇ g of saccharide for each particular glycoconjugate.
  • each dose will comprise about 1.5 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 2.0 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 2.2 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 2.5 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 3.0 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 3.5 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 4.0 ⁇ g of saccharide for each particular glycoconjugate.
  • each dose will comprise about 4.4 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 5.0 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 5.5 ⁇ g of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 6.0 ⁇ g of saccharide for each particular glycoconjugate.
  • Carrier amount Generally, each dose will comprise 10 ⁇ g to 150 ⁇ g of carrier protein, particularly 15 ⁇ g to 100 ⁇ g of carrier protein, more particularly 25 ⁇ g to 75 ⁇ g of carrier protein, and even more particularly 40 ⁇ g to 60 ⁇ g of carrier protein. In an embodiment, said carrier protein is CRM 197 .
  • each dose will comprise about 10 ⁇ g, about 15 ⁇ g, about 20 ⁇ g, about 25 ⁇ g, about 26 ⁇ g, about 27 ⁇ g, about 28 ⁇ g, about 29 ⁇ g, about 30 ⁇ g, about 31 ⁇ g, about 32 ⁇ g, about 33 ⁇ g, about 34 ⁇ g, about 35 ⁇ g, about 36 ⁇ g, about 37 ⁇ g, about 38 ⁇ g, about 39 ⁇ g, about 40 ⁇ g, about 41 ⁇ g, about 42 ⁇ g, about 43 ⁇ g, about 44 ⁇ g, about 45 ⁇ g, about 46 ⁇ g, about 47 ⁇ g, about 48 ⁇ g, about 49 ⁇ g, about 50 ⁇ g, about 51 ⁇ g, about 52 ⁇ g, about 53 ⁇ g, about 54 ⁇ g, about 55 ⁇ g, about 56 ⁇ g, about 57 ⁇ g, about 58 ⁇ g, about 59 ⁇ g, about 60 ⁇ g
  • each dose will comprise about 25 ⁇ g, about 26 ⁇ g, about 27 ⁇ g, about 28 ⁇ g, about 29 ⁇ g, about 30 ⁇ g, about 31 ⁇ g, about 32 ⁇ g, about 33 ⁇ g, about 34 ⁇ g, about 35 ⁇ g, about 36 ⁇ g, about 37 ⁇ g, about 38 ⁇ g, about 39 ⁇ g, about 40 ⁇ g, about 41 ⁇ g, about 42 ⁇ g, about 43 ⁇ g, about 44 ⁇ g, about 45 ⁇ g, about 46 ⁇ g, about 47 ⁇ g, about 48 ⁇ g, about 49 ⁇ g, about 50 ⁇ g, about 51 ⁇ g, about 52 ⁇ g, about 53 ⁇ g, about 54 ⁇ g, about 55 ⁇ g, about 56 ⁇ g, about 57 ⁇ g, about 58 ⁇ g, about 59 ⁇ g, about 60 ⁇ g, about 61 ⁇ g, about 62 ⁇ g, about 63 ⁇ g, about 64 ⁇ g, about
  • each dose will comprise about 30 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 31 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 32 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 33 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 34 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 45 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 40 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 41 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 42 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 43 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 44 ⁇ g of carrier protein.
  • each dose will comprise about 45 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 48 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 49 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 50 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 51 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 52 ⁇ g of carrier protein. In an embodiment, each dose will comprise about 53 ⁇ g of carrier protein. In an embodiment, said carrier protein is CRM197. In an embodiment, said carrier protein is SCP. 2.4 Further antigens Immunogenic compositions of the invention comprise conjugated saccharide antigen(s) (glycoconjugate(s)).
  • antigen(s) from other pathogen(s), particularly from bacteria and/or viruses are selected from: a diphtheria toxoid (D), a tetanus toxoid (T), a pertussis antigen (P), which is typically acellular (Pa), a hepatitis B virus (HBV) surface antigen (HBsAg), a hepatitis A virus (HAV) antigen, a conjugated Haemophilus influenzae type b capsular saccharide (Hib), inactivated poliovirus vaccine (IPV).
  • D diphtheria toxoid
  • T tetanus toxoid
  • P pertussis antigen
  • P which is typically acellular (Pa)
  • HBV hepatitis B virus
  • HAV hepatitis A virus
  • Hib conjugated Haemophilus influenzae type b capsular saccharide
  • IPV inactivated poliovirus vaccine
  • the immunogenic compositions of the invention comprise D-T-Pa-Hib, D-T-Pa- IPV or D-T-Pa-HBsAg. In an embodiment, the immunogenic compositions of the invention comprise D-T-Pa-HBsAg-IPV or D-T-Pa-HBsAg-Hib. In an embodiment, the immunogenic compositions of the invention comprise D-T-Pa-HBsAg-IPV-Hib.
  • Pertussis antigens Bordetella pertussis causes whooping cough. Pertussis antigens in vaccines are either cellular (whole cell, in the form of inactivated B. pertussis cells) or acellular.
  • cellular pertussis antigens are well documented (e.g., it may be obtained by heat inactivation of phase I culture of B. pertussis).
  • the invention uses acellular antigens.
  • acellular antigens it is preferred to use one, two or (preferably) three of the following antigens: (1) detoxified pertussis toxin (pertussis toxoid, or PT); (2) filamentous hemagglutinin (FHA); (3) pertactin (also known as the 69 kilodalton outer membrane protein).
  • FHA and pertactin may be treated with formaldehyde prior to use according to the invention.
  • PT is preferably detoxified by treatment with formaldehyde and/or glutaraldehyde.
  • Acellular pertussis antigens are preferably adsorbed onto one or more aluminum salt adjuvants. As an alternative, they may be added in an unadsorbed state. Where pertactin is added then it is preferably already adsorbed onto an aluminum hydroxide adjuvant.
  • PT and FHA may be adsorbed onto an aluminum hydroxide adjuvant or an aluminum phosphate. Adsorption of all of PT, FHA and pertactin to aluminum hydroxide is most preferred.
  • Inactivated poliovirus vaccine Poliovirus causes poliomyelitis. Rather than use oral poliovirus vaccine, preferred embodiments of the invention use IPV.
  • Poliomyelitis can be caused by one of three types of poliovirus. The three types are similar and cause identical symptoms, but they are antigenically different and infection by one type does not protect against infection by others. It is therefore preferred to use three poliovirus antigens in the invention: poliovirus Type 1 (e.g., Mahoney strain), poliovirus Type 2 (e.g., MEF- 1 strain), and poliovirus Type 3 (e.g., Saukett strain).
  • the viruses are preferably grown, purified and inactivated individually, and are then combined to give a bulk trivalent mixture for use with the invention.
  • Diphtheria toxoid Corynebacterium diphtheriae causes diphtheria. Diphtheria toxin can be treated (e.g., using formalin or formaldehyde) to remove toxicity while retaining the ability to induce specific anti-toxin antibodies after injection. These diphtheria toxoids are used in diphtheria vaccines. Preferred diphtheria toxoids are those prepared by formaldehyde treatment. The diphtheria toxoid can be obtained by growing C. diphtheriae in growth medium, followed by formaldehyde treatment, ultrafiltration and precipitation. The toxoided material may then be treated by a process comprising sterile filtration and/or dialysis.
  • the diphtheria toxoid is preferably adsorbed onto an aluminum hydroxide adjuvant.
  • Tetanus toxoid Clostridium tetani causes tetanus. Tetanus toxin can be treated to give a protective toxoid.
  • the toxoids are used in tetanus vaccines. Preferred tetanus toxoids are those prepared by formaldehyde treatment.
  • the tetanus toxoid can be obtained by growing C. tetani in growth medium, followed by formaldehyde treatment, ultrafiltration and precipitation. The material may then be treated by a process comprising sterile filtration and/or dialysis.
  • Hepatitis A virus antigens Hepatitis A virus (HAV) is one of the known agents which causes viral hepatitis. A preferred HAV component is based on inactivated virus, and inactivation can be achieved by formalin treatment. Hepatitis B virus (HBV) is one of the known agents which causes viral hepatitis. The major component of the capsid is a protein known as HBV surface antigen or, more commonly, HBsAg, which is typically a 226-amino acid polypeptide with a molecular weight of ⁇ 24 kDa.
  • HBV surface antigen or, more commonly, HBsAg, which is typically a 226-amino acid polypeptide with a molecular weight of ⁇ 24 kDa.
  • HBsAg has been made in two ways: purification of the antigen in particulate form from the plasma of chronic hepatitis B carriers or expression of the protein by recombinant DNA methods (e.g., recombinant expression in yeast cells). Unlike native HBsAg (i.e., as in the plasma-purified product), yeast-expressed HBsAg is generally non-glycosylated, and this is the most preferred form of HBsAg for use with the invention.
  • Haemophilus influenzae type b causes bacterial meningitis.
  • Hib vaccines are typically based on the capsular saccharide antigen, the preparation of which is well documented.
  • the Hib saccharide can be conjugated to a carrier protein in order to enhance its immunogenicity, especially in children.
  • Typical carrier proteins are tetanus toxoid, diphtheria toxoid, CRM 197 , H.influenzae protein D, and an outer membrane protein complex from serogroup B meningococcus.
  • the saccharide moiety of the conjugate may comprise full-length polyribosylribitol phosphate (PRP) as prepared from Hib bacteria, and/or fragments of full-length PRP.
  • Hib conjugates may or may not be adsorbed to an aluminum salt adjuvant.
  • Adjuvant(s) In some embodiments, the immunogenic compositions disclosed herein may further comprise at least one, two or three adjuvants. In some embodiments, the immunogenic compositions disclosed herein may further comprise at least one adjuvant. In some embodiments, the immunogenic compositions disclosed herein may further comprise one adjuvant. In some embodiments, the immunogenic compositions disclosed herein may further comprise two adjuvants.
  • adjuvant refers to a compound or mixture that enhances the immune response to an antigen. Antigens may act primarily as a delivery system, primarily as an immune modulator or have strong features of both. Suitable adjuvants include those suitable for use in mammals, including humans.
  • alum e.g., aluminum phosphate, aluminum sulfate or aluminum hydroxide
  • calcium phosphate e.g., calcium phosphate
  • liposomes e.g., calcium phosphate, liposomes
  • oil-in-water emulsions such as MF59 (4.3% w/v squalene, 0.5% w/v polysorbate 80 (Tween 80), 0.5% w/v sorbitan trioleate (Span 85)
  • water-in-oil emulsions such as Montanide
  • PLG poly(D,L-lactide-co-glycolide)
  • the immunogenic compositions disclosed herein comprise aluminum salts (alum) as adjuvant (e.g., aluminum phosphate, aluminum sulfate or aluminum hydroxide).
  • the immunogenic compositions disclosed herein comprise aluminum phosphate or aluminum hydroxide as adjuvant.
  • the immunogenic compositions disclosed herein comprise aluminum phosphate as adjuvant.
  • adjuvants to enhance effectiveness of the immunogenic compositions as disclosed herein include, but are not limited to: (1) oil-in-water emulsion formulations (with or without other specific immunostimulating agents such as muramyl peptides (see below) or bacterial cell wall components), such as for example (a) SAF, containing 10% Squalene, 0.4% Tween 80, 5% pluronic-blocked polymer L121, and thr-MDP either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion, and (b) RIBITM adjuvant system (RAS), (Ribi Immunochem, Hamilton, MT) containing 2% Squalene, 0.2% Tween 80, and one or more bacterial cell wall components such as monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL + CWS (DETOXTM); (2) saponin adjuvity
  • Muramyl peptides include N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-25 acetyl- normuramyl-L-alanyl-D-isoglutamine (nor-MDP), N-acetylmuramyl-L-alanyl-D-isoglutarninyl-L- alanine-2-(1'-2'-dipalmitoyl-sn-gIycero-3-hydroxyphosphoryloxy)-ethylamine MTP-PE), etc.
  • the immunogenic compositions as disclosed herein comprise a CpG Oligonucleotide as adjuvant.
  • a CpG oligonucleotide as used herein refers to an immunostimulatory CpG oligodeoxynucleotide (CpG ODN), and accordingly these terms are used interchangeably unless otherwise indicated.
  • Immunostimulatory CpG oligodeoxynucleotides contain one or more immunostimulatory CpG motifs that are unmethylated cytosine-guanine dinucleotides, optionally within certain preferred base contexts.
  • the methylation status of the CpG immunostimulatory motif generally refers to the cytosine residue in the dinucleotide.
  • An immunostimulatory oligonucleotide containing at least one unmethylated CpG dinucleotide is an oligonucleotide which contains a 5' unmethylated cytosine linked by a phosphate bond to a 3' guanine, and which activates the immune system through binding to Toll- like receptor 9 (TLR-9).
  • TLR-9 Toll- like receptor 9
  • the immunostimulatory oligonucleotide may contain one or more methylated CpG dinucleotides, which will activate the immune system through TLR9 but not as strongly as if the CpG motif(s) was/were unmethylated.
  • CpG immunostimulatory oligonucleotides may comprise one or more palindromes that in turn may encompass the CpG dinucleotide.
  • CpG oligonucleotides have been described in a number of issued patents, published patent applications, and other publications, including U.S. Patent Nos. 6,194,388; 6,207,646; 6,214,806; 6,218,371; 6,239,116; and 6,339,068.
  • the immunogenic compositions as disclosed herein comprise any of the CpG Oligonucleotide described at page 3, line 22, to page 12, line 36 of WO 2010/125480. Different classes of CpG immunostimulatory oligonucleotides have been identified.
  • the immunogenic compositions as disclosed herein comprise an A class CpG oligonucleotide.
  • the "A class" CpG oligonucleotide of the invention has the following nucleic acid sequence: 5’ GGGGACGACGTCGTGGGGGGG 3’ (SEQ ID NO: 1).
  • A-Class oligonucleotides include: 5’ G*G*G_G_A_C_G_A_C_G_T_C_G_T_G_G*G*G*G*G*G*G*G 3’ (SEQ ID NO: 2); wherein “*” refers to a phosphorothioate bond and “_” refers to a phosphodiester bond.
  • the immunogenic compositions as disclosed herein comprise a B class CpG Oligonucleotide.
  • the CpG oligonucleotide for use in the present invention is a B class CpG oligonucleotide represented by at least the formula: 5' X 1 X 2 CGX 3 X 4 3’, wherein X1, X2, X3, and X4 are nucleotides.
  • X 2 is adenine, guanine, or thymine.
  • X 3 is cytosine, adenine, or thymine.
  • the B class CpG oligonucleotide sequences of the invention are those broadly described above as well as disclosed in WO 96/02555, WO 98/18810 and U.S. Patent Nos.
  • the "B class" CpG oligonucleotide of the invention has the following nucleic acid sequence: 5’ TCGTCGTTTTTCGGTGCTTTT 3’ (SEQ ID NO: 3), or 5’ TCGTCGTTTTTCGGTCGTTTT 3’ (SEQ ID NO: 4), or 5’ TCGTCGTTTTGTCGTTTTGTCGTT 3’ (SEQ ID NO: 5), or 5’ TCGTCGTTTCGTCGTTTTGTCGTT 3’ (SEQ ID NO: 6), or 5’ TCGTCGTTTTGTCGTTTTTTTCGA 3’ (SEQ ID NO: 7).
  • all of the linkages may be all phosphorothioate bonds.
  • one or more of the linkages may be phosphodiester, preferably between the “C” and the “G” of the CpG motif making a semi-soft CpG oligonucleotide.
  • an ethyl-uridine or a halogen may substitute for the 5' T; examples of halogen substitutions include but are not limited to bromo-uridine or iodo-uridine substitutions.
  • B-Class oligonucleotides include: 5’ T*C*G*T*C*G*T*T*T*T*T*C*G*G*T*G*C*T*T*T 3’ (SEQ ID NO: 8), or 5’ T*C*G*T*C*G*T*T*T*T*T*C*G*G*T*C*G*T*T*T 3’ (SEQ ID NO: 9), or 5’ T*C*G*T*C*G*T*T*T*T*T*T*T*T*T*T*T*T*G*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T*T 3’ (SEQ ID NO: 10), or 5’ T*C*G*T*C*G*T*T*T*C*G*T*C*T*T*T*T*T*T
  • the immunogenic compositions as disclosed herein comprise a C class CpG Oligonucleotide.
  • the "C class" CpG oligonucleotides of the invention have the following nucleic acid sequence: 5’ TCGCGTCGTTCGGCGCGCCG 3’ (SEQ ID NO: 13), or 5’ TCGTCGACGTTCGGCGCGCCG 3’ (SEQ ID NO: 14), or 5’ TCGGACGTTCGGCGCGCGCCG 3’ (SEQ ID NO: 15), or 5’ TCGGACGTTCGGCGCGCCG 3’ (SEQ ID NO: 16), or 5’ TCGCGTCGTTCGGCGCGCCG 3’ (SEQ ID NO: 17), or 5’ TCGACGTTCGGCGCGCCG 3’ (SEQ ID NO: 18), or 5’ TCGACGTTCGGCGCGCCG 3’ (SEQ ID NO: 19), or 5’
  • all of the linkages may be all phosphorothioate bonds.
  • one or more of the linkages may be phosphodiester, preferably between the “C” and the “G” of the CpG motif making a semi-soft CpG oligonucleotide.
  • C-Class oligonucleotides include: 5’ T*C_G*C_G*T*C_G*T*T*C_G*G*C*G*C*G*C*G*C*G*C*G 3’ (SEQ ID NO: 26), or 5’ T*C_G*T*C_G*A*C_G*T*T*C_G*G*C*G*C_G*C*G*C*C*G 3’ (SEQ ID NO: 27), or 5’ T*C_G*G*A*C_G*T*T*C_G*G*C*G*C_G*C*C*C*G 3’ (SEQ ID NO: 28), or 5’ T*C_G*G*A*C_G*T*T*C_G*G*C*G*C*C*C*G 3’ (SEQ ID NO: 29), or 5’ T*C_G*C_G*T*C_G*T*C_G*T
  • an ethyl-uridine or a halogen may substitute for the 5' T; examples of halogen substitutions include but are not limited to bromo-uridine or iodo-uridine substitutions.
  • the immunogenic compositions as disclosed herein comprise a P class CpG Oligonucleotide.
  • the CpG oligonucleotide for use in the present invention is a P class CpG oligonucleotide containing a 5' TLR activation domain and at least two palindromic regions, one palindromic region being a 5' palindromic region of at least 6 nucleotides in length and connected to a 3' palindromic region of at least 8 nucleotides in length either directly or through a spacer, wherein the oligonucleotide includes at least one YpR dinucleotide.
  • said oligonucleotide is not T*C_G*T*C_G*A*C_G*T*T*C_G*G*C*G*C_G*C*G*C*G (SEQ ID NO: 27).
  • the P class CpG oligonucleotide includes at least one unmethylated CpG dinucleotide.
  • the TLR activation domain is TCG, TTCG, TTTCG, TYpR, TTYpR, TTTYpR, UCG, UUCG, UUUCG, TTT, or TTTT. In yet another embodiment the TLR activation domain is within the 5' palindromic region.
  • an ethyl-uridine or a halogen may substitute for the 5' T; examples of halogen substitutions include but are not limited to bromo-uridine or iodo-uridine substitutions.
  • a non-limiting example of P-Class oligonucleotides include: 5’ T*C_G*T*C_G*A*C_G*A*T*C_G*G*C*G*C*C_G*C*C*C*G 3’ (SEQ ID NO: 40) wherein “*” refers to a phosphorothioate bond and “_” refers to a phosphodiester bond.
  • all the internucleotide linkages of the CpG oligonucleotides disclosed herein are phosphodiester bonds (“soft” oligonucleotides, as described in WO 2007/026190).
  • CpG oligonucleotides of the invention are rendered resistant to degradation (e.g., are stabilized).
  • a "stabilized oligonucleotide” refers to an oligonucleotide that is relatively resistant to in vivo degradation (e.g., via an exo- or endo-nuclease). Nucleic acid stabilization can be accomplished via backbone modifications.
  • Oligonucleotides having phosphorothioate linkages provide maximal activity and protect the oligonucleotide from degradation by intracellular exo- and endo-nucleases.
  • the immunostimulatory oligonucleotides may have a chimeric backbone, which have combinations of phosphodiester and phosphorothioate linkages.
  • Oligonucleotides of any size greater than 6 nucleotides are capable of inducing an immune response if sufficient immunostimulatory motifs are present, because larger oligonucleotides are degraded inside cells.
  • the CpG oligonucleotides are 6 to 100 nucleotides long, preferentially 8 to 30 nucleotides long.
  • nucleic acids and oligonucleotides of the invention are not plasmids or expression vectors.
  • the CpG oligonucleotide disclosed herein comprise substitutions or modifications, such as in the bases and/or sugars as described at paragraphs 134 to 147 of WO 2007/026190.
  • the immunogenic composition of the invention comprises 0.1 mg to 50 mg of CpG oligonucleotide, preferably from 0.2 mg to 10 mg CpG oligonucleotide, more preferably from 0.3 mg to 5 mg CpG oligonucleotide.. In a particular embodiment of the present invention, the immunogenic composition of the invention comprises from 0.3 mg to 5 mg CpG oligonucleotide. Even preferably, the immunogenic composition of the invention may comprise from 0.5 to 2 mg CpG oligonucleotide. Most preferably, the immunogenic composition of the invention may comprise from 0.75 to 1.5 mg CpG oligonucleotide.
  • Immunogenic compositions of the disclosure may comprise one or more of a buffer, a salt, a divalent cation, a non-ionic detergent, a cryoprotectant such as a sugar, and an anti-oxidant such as a free radical scavenger or chelating agent, or any multiple combinations thereof.
  • the immunogenic compositions of the disclosure comprise a buffer.
  • said buffer has a pKa of about 3.5 to about 7.5.
  • the buffer is phosphate, succinate, histidine or citrate.
  • the buffer is succinate.
  • the buffer is histidine.
  • the buffer is succinate at a final concentration of 1 mM to 10 mM.
  • the surfactant is polysorbate 80.
  • the final concentration of polysorbate 80 in the formulation is at least 0.0001% to 10% polysorbate 80 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 80 in the formulation is at least 0.001% to 1% polysorbate 80 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 80 in the formulation is at least 0.01% to 1% polysorbate 80 weight to weight (w/w). In other embodiments, the final concentration of polysorbate 80 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 80 (w/w).
  • the final concentration of the polysorbate 80 in the formulation is 0.02% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.01% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.03% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.04% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.05% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 1% polysorbate 80 (w/w). In one particular embodiment, the surfactant is polysorbate 20.
  • the final concentration of the polysorbate 20 in the formulation is 0.02% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.01% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.03% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.04% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.05% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 1% polysorbate 20 (w/w). In one particular embodiment, the surfactant is polysorbate 40.
  • the final concentration of polysorbate 60 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 60 (w/w).
  • the final concentration of the polysorbate 60 in the formulation is 1% polysorbate 60 (w/w).
  • the surfactant is polysorbate 65.
  • the final concentration of polysorbate 65 in the formulation is at least 0.0001% to 10% polysorbate 65 weight to weight (w/w).
  • the final concentration of polysorbate 65 in the formulation is at least 0.001% to 1% polysorbate 65 weight to weight (w/w).
  • the immunogenic composition of the disclosure has a pH of 5.5 to 7.5, more preferably a pH of 5.6 to 7.0, even more preferably a pH of 5.8 to 6.0.
  • the present disclosure provides a container filled with any of the immunogenic compositions disclosed herein.
  • the container is selected from the group consisting of a vial, a syringe, a flask, a fermentor, a bioreactor, a bag, a jar, an ampoule, a cartridge and a disposable pen.
  • the container is siliconized.
  • the subject is a mammal, such as a human, non-human primate, cat, sheep, pig, horse, bovine or dog. In one aspect, the subject is a human.
  • the immunogenic compositions described herein may be used in therapeutic or prophylactic methods for preventing, treating or ameliorating a bacterial infection, disease or condition in a subject.
  • the disclosure provides a method of preventing, treating or ameliorating an infection, disease or condition associated with a bacterial infection in a subject, comprising administering to the subject an immunologically effective amount of an immunogenic composition of the disclosure.
  • the subject to be vaccinated can be about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11 or about 12 months of age.
  • the subject to be vaccinated is about 2, about 4 or about 6 months of age.
  • the subject to be vaccinated is less than 2 years of age.
  • the subject to be vaccinated can be about 12 to about 15 months of age.
  • a second, third or fourth dose may be given (see section 8 below).
  • the subject to be vaccinated is a human adult 50 years of age or older, more preferably a human adult 55 years of age or older. In an embodiment, the subject to be vaccinated is a human adult 65 years of age or older, 70 years of age or older, 75 years of age or older or 80 years of age or older. In an embodiment the subject to be vaccinated is an immunocompromised individual, in particular a human.
  • An immunocompromised individual is generally defined as a person who exhibits an attenuated or reduced ability to mount a normal humoral or cellular defense to challenge by infectious agents.
  • the immunocompromised subject to be vaccinated suffers from a disease selected from the groups consisting of: HIV-infection, acquired immunodeficiency syndrome (AIDS), cancer, chronic heart or lung disorders, congestive heart failure, diabetes mellitus, chronic liver disease, alcoholism, cirrhosis, spinal fluid leaks, cardiomyopathy, chronic bronchitis, emphysema, chronic obstructive pulmonary disease (COPD), spleen dysfunction (such as sickle cell disease), lack of spleen function (asplenia), blood malignancy, leukemia, multiple myeloma, Hodgkin’s disease, lymphoma, kidney failure, nephrotic syndrome and asthma.
  • AIDS acquired immunodeficiency syndrome
  • cancer chronic heart or lung disorders
  • congestive heart failure diabetes mellitus
  • chronic liver disease chronic liver disease
  • alcoholism alcoholism
  • cirrhosis chronic obstructive pulmonary disease
  • COPD chronic obstruct
  • the immunocompromised subject to be vaccinated has a white blood cell count (leukocyte count) below 5 x 10 9 cells per liter, or below 4 x 10 9 cells per liter, or below 3 x 10 9 cells per liter, or below 2 x 10 9 cells per liter, or below 1 x 10 9 cells per liter, or below 0.5 x 10 9 cells per liter, or below 0.3 x 10 9 cells per liter, or below 0.1 x 10 9 cells per liter.
  • White blood cell count (leukocyte count) The number of white blood cells (WBC) in the blood. The WBC is usually measured as part of the CBC (complete blood count).
  • White blood cells are the infection-fighting cells in the blood and are distinct from the red (oxygen-carrying) blood cells known as erythrocytes.
  • red blood cells known as erythrocytes.
  • white blood cells include neutrophils (polymorphonuclear leukocytes; PMN), band cells (slightly immature neutrophils), T-type lymphocytes (T-cells), B-type lymphocytes (B-cells), monocytes, eosinophils, and basophils. All the types of white blood cells are reflected in the white blood cell count.
  • the normal range for the white blood cell count is usually between 4,300 and 10,800 cells per cubic millimeter of blood.
  • the immunocompromised subject to be vaccinated suffers from neutropenia.
  • the immunocompromised subject to be vaccinated has a neutrophil count below 2 x 10 9 cells per liter, or below 1 x 10 9 cells per liter, or below 0.5 x 10 9 cells per liter, or below 0.1 x 10 9 cells per liter, or below 0.05 x 10 9 cells per liter.
  • a low white blood cell count or “neutropenia” is a condition characterized by abnormally low levels of neutrophils in the circulating blood.
  • the immunocompromised subject to be vaccinated has a CD4+ cell count below 500/mm 3 , or CD4+ cell count below 300/mm 3 , or CD4+ cell count below 200/mm 3 , CD4+ cell count below 100/mm 3 , CD4+ cell count below 75/mm 3 , or CD4+ cell count below 50/mm 3 .
  • CD4 cell tests are normally reported as the number of cells in mm 3 .
  • any of the immunocompromised subjects disclosed herein is a human male. In an embodiment of the invention, any of the immunocompromised subjects disclosed herein is a human female. 6 Regimen In some cases, as little as one dose of the immunogenic composition according to the invention is needed, but under some circumstances, such as conditions of greater immune deficiency, a second, third or fourth dose may be given. Following an initial vaccination, subjects can receive one or several booster immunizations adequately spaced. In an embodiment, the schedule of vaccination of the immunogenic composition according to the invention is a single dose.
  • said single dose schedule is for healthy persons being at least 2 years of age.
  • the schedule of vaccination of the immunogenic composition according to the invention is a multiple dose schedule.
  • said multiple dose schedule consists of a series of 2 doses separated by an interval of about 1 month to about 2 months.
  • said multiple dose schedule consists of a series of 2 doses separated by an interval of about 1 month, or a series of 2 doses separated by an interval of about 2 months.
  • said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month to about 2 months.
  • said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month, or a series of 3 doses separated by an interval of about 2 months. In another embodiment, said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month to about 2 months followed by a fourth dose about 10 months to about 13 months after the first dose. In another embodiment, said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month followed by a fourth dose about 10 months to about 13 months after the first dose, or a series of 3 doses separated by an interval of about 2 months followed by a fourth dose about 10 months to about 13 months after the first dose.
  • the multiple dose schedule consists of at least one dose (e.g., 1, 2 or 3 doses) in the first year of age followed by at least one toddler dose.
  • the multiple dose schedule consists of a series of 2 or 3 doses separated by an interval of about 1 month to about 2 months (for example 28-56 days between doses), starting at 2 months of age, and followed by a toddler dose at 12-18 months of age.
  • said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month to about 2 months (for example 28-56 days between doses), starting at 2 months of age, and followed by a toddler dose at 12-15 months of age.
  • said multiple dose schedule consists of a series of 2 doses separated by an interval of about 2 months, starting at 2 months of age, and followed by a toddler dose at 12-18 months of age.
  • the multiple dose schedule consists of a 4-dose series of vaccine at 2, 4, 6, and 12-15 months of age.
  • a prime dose is given at day 0 and one or more boosts are given at intervals that range from about 2 to about 24 weeks, preferably with a dosing interval of 4-8 weeks.
  • a prime dose is given at day 0 and a boost is given about 3 months later. 7.
  • the invention also provides the following embodiments as defined in the following numbered paragraphs 1 to 86 1.
  • a method of making a capsular saccharide glycoconjugate comprising the steps of: (a) reacting an isolated capsular saccharide with a carbonic acid derivative and an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide, (b) reacting a carrier protein with an agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group where the NHS moiety reacts with the amino groups to form an amide linkage thereby obtaining an azido functionalized carrier protein, (c) reacting the activated alkynyl saccharide of step (a) with the azido functionalized carrier protein of step (b) by Cu +1 mediated azide-alkyne cycloaddition reaction to form a glycoconjugate.
  • NHS N-Hydroxysuccinimide
  • the isolated saccharide is sized before the activation step (a). 3. The method of paragraph 2 wherein, the isolated capsular saccharide is sized to a weight average molecular weight of between 50 kDa and 500 kDa. 4. The method of any one of paragraphs 1-3 wherein, said carbonic acid derivative is selected from the group consisting of 1,1’-carbonyldiimidazole (CDI), 1,1’-carbonyl-di-(1,2,4-triazole) (CDT), disuccinimidyl carbonate (DSC) and N-hydroxysuccinimidyl chloroformate. 5.
  • CDI 1,1’-carbonyldiimidazole
  • CDT 1,1’-carbonyl-di-(1,2,4-triazole)
  • DSC disuccinimidyl carbonate
  • N-hydroxysuccinimidyl chloroformate 5.
  • said carbonic acid derivative is 1,1’- carbonyldiimidazole (CDI). 6. The method of any one of paragraphs 1-4 wherein, said carbonic acid derivative is 1,1'- Carbonyl-di-(1,2,4-triazole) (CDT). 7.
  • alkyne linker is a compound of formula (I), H 2 N X (I) wherein X is selected from the group consisting of CH 2 , CH 2 (CH 2 ) n , (CH 2 CH 2 O) m CH 2 CH 2 , NHCO(CH 2 ) n , NHCO(CH 2 CH 2 O) m CH 2 CH 2 , OCH 2 (CH 2 ) n and O(CH 2 CH 2 O) m CH 2 CH 2 ; where n is selected from 1 to 10 and m is selected from 1 to 4.
  • said alkyne linker is a compound of formula (II), 9.
  • step (a) comprises reacting the capsular saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative-activated capsular saccharide with an alkyne linker in an aprotic solvent to produce an activated alkynyl capsular saccharide.
  • step (a) comprises reacting the capsular saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative-activated capsular saccharide with an alkyne linker in an aprotic solvent to produce an activated alkynyl capsular saccharide.
  • step (a) comprises reacting the capsular saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative-activated capsular saccharide with an alkyne linker in an aprotic solvent to produce an activated alkynyl capsular saccharide.
  • step (a) comprises reacting the capsular saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative-activated capsular saccharide with an alkyne link
  • step (a) further comprises reacting the carbonic acid derivative-activated saccharide with an amount of alkyne linker that is between 0.01-10 molar equivalent to the amount of saccharide Repeat Unit of the activated saccharide.
  • step (b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-10 molar equivalents to the lysines on the carrier.
  • step (c) is carried out in aqueous buffer in the presence of copper (I) as catalyst and ascorbate as oxidant, wherein the reaction mixture further comprises THPTA (tris(3- hydroxypropyltriazolylmethyl)amine) and aminoguanidine.
  • the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step (c) is between 0.1 and 3. 28.
  • step (c) the method further comprises a step of capping the unreacted azido groups remained in the conjugate with an azido group capping agent.
  • said azido group capping agent is a compound of formula (V), (V) wherein X is (CH 2 ) n wherein n is selected from 1 to 15.
  • said azido group capping agent is propargyl alcohol.
  • the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.05 to 20 molar equivalents to the amount of saccharide repeat unit of the activated saccharide.
  • step (c) the method further comprises a step of capping the unreacted alkyne groups remained in the conjugate with an alkyne group capping agent.
  • said alkyne group capping agent is an agent bearing an azido group.
  • said alkyne group capping agent is a compound of formula (VI), (VI) wherein X is (CH 2 ) n wherein n is selected from 1 to 15.
  • said alkyne group capping agent is 3-azido-1-propanol. 36.
  • a capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII): (VII), wherein X is selected from the group consisting of CH 2 (CH 2 ) n’ , (CH 2 CH 2 O) m CH 2 CH 2 , NHCO(CH 2 ) n’ , NHCO(CH 2 CH 2 O) m CH 2 CH 2 , OCH 2 (CH 2 ) n’ and O(CH 2 CH 2 O) m CH 2 CH 2 ; where n’ is selected from 0 to 10 and m is selected from 1 to 4, and wherein X' is selected from the group consisting of CH 2 (CH 2 ) n”, CH 2 O(CH 2 ) n’’ CH 2 , CH 2 O(CH 2 CH 2 O) m’ (CH 2 ) n’’ CH 2 , where n’’ is selected from 0 to 10 and m’ is selected from
  • a capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH 2 (CH 2 ) n’ , where n’ is 0 and wherein X' is CH 2 (CH 2 ) n” where n’’ is 0. 41.
  • a capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VIII), (VIII). 42.
  • the capsular saccharide glycoconjugate of any one of paragraphs 38 to 41 comprising a capsular saccharide wherein the weight average molecular weight (Mw) of said capsular saccharide before conjugation is between 200 kDa and 750 kDa.
  • the capsular saccharide glycoconjugate of any one of paragraphs 38 to 44 having a weight average molecular weight (Mw) of between 250 kDa and 20,000 kDa.
  • the capsular saccharide glycoconjugate of any one of paragraphs 38 to 44 having a weight average molecular weight (Mw) of between 500 kDa and 5,000 kDa. 47.
  • the capsular saccharide glycoconjugate of any one of paragraphs 38 to 44 having a weight average molecular weight (Mw) of between 750 kDa and 2,500 kDa. 48.
  • Mw weight average molecular weight
  • said carrier protein is an enzymatically inactive fragment of an SCP
  • said enzymatically inactive fragment of SCP comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain.
  • PA domain protease-associated domain
  • Fn fibronectin type III
  • said carrier protein is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence and wherein said replacement is selected from the group consisting of D130A, H193A, N295A and S512A where the numbers indicate the amino acid residue position in the peptidase according to the numbering of SEQ ID NO: 1 of WO00/34487. 61.
  • said carrier protein is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence wherein said at least two amino acids replacements are D130A and S512A where the numbers indicate the amino acid residue position in the peptidase according to the numbering of SEQ ID NO: 1 of WO00/34487. 62.
  • said carrier protein is DT (Diphtheria toxoid).
  • said carrier protein is TT (tetanus toxoid).
  • said carrier protein is PD (H. influenzae protein D).
  • a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68 wherein said capsular saccharide is a capsular saccharide from a pathogenic Streptococcus, a pathogenic Staphylococcus, a pathogenic Enterococcus, a pathogenic Bacillus, a pathogenic Corynebacterium, a pathogenic Listeria, a pathogenic Erysipelothrix, a pathogenic Clostridium, a pathogenic Haemophilus, a pathogenic Neisseria or a pathogenic Escherichia. 71.
  • the method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from is Haemophilus influenzae type b. 79.
  • a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68 wherein said capsular saccharide is a capsular saccharide from a Streptococcus pneumoniae serotype selected from the group consisting of serotypes 1, 2, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9V, 9N, 10A, 10B, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A, 23B, 23F, 24B, 24F, 27, 29, 31, 33B, 33F, 34, 35B, 35F, 38, 72 and 73.
  • a Streptococcus pneumoniae serotype selected from the group consisting of serotypes 1, 2, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9V, 9N, 10A, 10B, 11A, 12
  • An immunogenic composition comprising a capsular saccharide glycoconjugate of any one of paragraphs 38 to 85.
  • the term "about” means within a statistically meaningful range of a value, such as a stated concentration range, time frame, molecular weight, temperature or pH. Such a range can be within an order of magnitude, typically within 20%, more typically within 10%, and even more typically within 5% or within 1% of a given value or range.
  • an “immunogenic amount”, an “immunologically effective amount”, a “therapeutically effective amount”, a “prophylactically effective amount”, or “dose”, each of which is used interchangeably herein, generally refers to the amount of antigen or immunogenic composition sufficient to elicit an immune response, either a cellular (T cell) or humoral (B cell or antibody) response, or both, as measured by standard assays known to one skilled in the art. Any whole number integer within any of the ranges of the present document is contemplated as an embodiment of the disclosure. All references or patent applications cited within this patent specification are incorporated by reference herein. The invention is illustrated in the accompanying examples.
  • EXAMPLE Example 1 Activation of Serotype 3 Capsular Polysaccharide with an alkyne linker Lyophilized S. pneumoniae serotype 3 capsular polysaccharide (300 mg) was reconstituted with anhydrous DMSO (75 mL). The reaction mixture was then warmed to 35°C, and then CDI (100 mg/mL in DMSO), 288 ⁇ L (0.2 MEq) was added. The reaction mixture was stirred at 35°C for 3 hrs.
  • Example 2 Activation of a carrier protein (SCP) with an agent bearing an N- Hydroxysuccinimide (NHS) moiety and an azido group (azidoacetic acid NHS ester (AAN)) To the SCP solution (500 mg, 86 mL) WFI 14 mL and 0.5 M SPB (pH 8.3) 25 mL were added.
  • SCP carrier protein
  • AAN azidoacetic acid NHS ester
  • Azidoacetic acid NHS ester (AAN) (20 mg/mL in DMSO) 2.0 mL (0.5 MEq to lysine on SCP) was added to the reaction mixture dropwise maintaining the reaction temperature at 8 ⁇ 3 oC. After the reaction mixture was stirred for 30 min at 10 oC, purified by UF/DF using 10K MWCO PES membrane (Millipore Pellicon 2 Mini) against 10 mM sodium phosphate buffer in saline (pH 7.0) (30X diavolume). After UF/DF, retentate was taken and then sucrose was added as solid and then filtered through 0.22 um filter and analyzed.
  • AAN Azidoacetic acid NHS ester
  • reaction mixture was stirred for 1.5 hours at 23°C, just before capping step, additional mixture of 5 mM copper sulfate (2.5 mL) and 25 mM THPTA (2.5 mL) was added to the reaction mixture at 23°C and followed by the additional addition of 100 mM aminoguanidine (5 mL) and 100 mM ascorbate (5 mL).
  • 3-Azido-propyl alcohol (20 MEq per alkyne on polysaccharide) was added to cap the unreacted alkyne group on the polysaccharide and stirred for 1 hour at the same temperature, and then propargyl alcohol (40 MEq per alkyne on poly) was added to cap the unreacted azido group on SCP and stirred for 1 hour at 23°C.
  • reaction mixture was purified by first UF/DF using 100K MWCO RC membrane (Millipore Pellicon 2 Mini) against 10 mM EDTA + 10 mM SPB in saline (pH 7.0) (30X diavolume) and followed by 5 mM succinate in saline (pH 6.0) (20X diavolume). After UF/DF, filtered through 0.22 um filter (Millipak 40), and analyzed. Table 1.

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Abstract

The present invention relates to new conjugated capsular saccharide antigens (glycoconjugates), immunogenic compositions comprising said glycoconjugates and uses thereof.

Description

PC72949A Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof Field of the Invention The present invention relates to new conjugated capsular saccharide antigens (glycoconjugates), immunogenic compositions comprising said glycoconjugates and uses thereof. Immunogenic compositions of the present invention will typically comprise glycoconjugates, wherein the saccharides are derived from bacterial capsular polysaccharide antigens, in particular a capsular polysaccharide derived from pathogenic bacteria. The invention also relates to vaccination of human subjects, in particular infants and elderly, against infections using said glycoconjugates. Background of the Invention Both Gram-positive and Gram-negative bacteria may produce an extracellular compartment, the capsule, which covers the bacterial cell and often prevents the reaction of underlying cell surface antigens with their homologous antibodies. Capsules are found in several bacteria of medical importance, especially in extraintestinal and invasive strains. Bacterial capsules are largely composed of polysaccharides. They form a gelatinous mass around the cell. Capsular polysaccharides are important immunogens and has led to them being an important component in the design of vaccines. They have proved useful in eliciting immune responses especially when linked to carrier proteins. The approach to increasing immunogenicity of poorly immunogenic molecules by conjugating these molecules to “carrier” molecules has been utilized successfully for decades (see, e.g., Goebel et al. (1939) J. Exp. Med. 69: 53). For example, many immunogenic compositions have been described in which purified capsular polymers have been conjugated to carrier proteins to create more effective immunogenic compositions by exploiting this “carrier effect.” Schneerson et al. (1984) Infect. Immun.45: 582-591). Conjugation has also been shown to bypass the poor antibody response usually observed in infants when immunized with a free polysaccharide (Anderson et al. (1985) J. Pediatr.107: 346; Insel et al. (1986) J. Exp. Med.158: 294). Conjugates have been successfully generated using various cross-linking or coupling reagents, such as homobifunctional, heterobifunctional, or zero-length crosslinkers. Many methods are currently available for coupling immunogenic molecules, such as saccharides, proteins, and peptides, to peptide or protein carriers. Most methods create amine, amide, urethane, isothiourea, or disulfide bonds, or in some cases thioethers. A disadvantage to the use of cross-linking or coupling reagents which introduce reactive sites into the side chains of reactive amino acid molecules on carrier and/or immunogenic molecules is that the reactive sites, if not neutralized, are free to react with any unwanted molecule either in vitro (thus potentially adversely affecting the functionality or stability of the conjugates) or in vivo (thus posing a potential risk of adverse events in persons or animals immunized with the preparations). Such excess reactive sites can be reacted or “capped”, so as to inactivate these sites, utilizing various known chemical reactions, but these reactions may be otherwise disruptive to the functionality of the conjugates. Thus, there remains a need for new glycoconjugates appropriately capped and methods to prepare said conjugates, such that the functionality is preserved, and the conjugate retains the ability to elicit the desired immune response. The present inventors have found a new and efficacious method to generate glycoconjugates. The method allows for generating conjugates with very low free saccharide and a good yield. Figures FIG.1 shows an example of a scheme for the preparation of glycoconjugates of the invention prepared using click chemistry and using propargylamine as alkyne linker. CP = Carrier Protein, CDI = 1,1’-carbonyldiimidazole. Detailed description of the Invention The present inventors have developed new polysaccharide-carrier protein conjugates and processes for making these conjugates. The method to generate glycoconjugates has been found to allow for producing glycoconjugates with very low free saccharide and a good yield. 1. Glycoconjugates of the invention The present invention is directed in part to conjugated bacterial capsular saccharide antigens (also named glycoconjugates). For the purpose of the invention the term 'glycoconjugate' indicates a capsular saccharide (in particular a bacterial capsular saccharide) linked covalently to a carrier protein. 1.1 Capsular saccharide of the invention The term "saccharide" throughout this specification may indicate polysaccharide or oligosaccharide and includes both. In an embodiment, saccharide of the invention may be oligosaccharides. Oligosaccharides have a low number of repeat units (typically 5-15 repeat units) and are typically derived synthetically or by hydrolysis of polysaccharides. Preferably though, all of the saccharides of the present invention and in the immunogenic compositions of the present invention are polysaccharides. High molecular weight polysaccharides are able to induce certain antibody immune responses due to the epitopes present on the antigenic surface. The isolation and purification of high molecular weight capsular polysaccharides is preferably contemplated for use in the conjugates, compositions and methods of the present invention. Therefore, in a preferred embodiment of the present invention, the saccharide is a polysaccharide. Preferably, the saccharide used in the present invention is a bacterial capsular saccharide (also named ‘capsular saccharide’ herein). Capsules are found in several bacteria of medical importance. Bacterial capsules are largely composed of polysaccharides. Capsular saccharides are prepared by standard techniques known to those of ordinary skill in the art. In a most preferred embodiment of the present invention, the saccharide is a S. pneumoniae capsular polysaccharide. In an embodiment, the capsular saccharide used in the present invention is a synthetic carbohydrate. In a preferred embodiment though, the source of bacterial capsular saccharide according to this invention can be bacterial cells. Bacterial strains which can be used as source of capsular saccharide may be obtained from established culture collections (such as for example from the American Type Culture Collection (ATCC, Manassas, VA USA) or the Streptococcal Reference Laboratory (Centers for Disease Control and Prevention, Atlanta, GA USA)) or clinical specimens. Bacterial capsular saccharides can be obtained directly from bacteria using isolation procedures known to one of ordinary skill in the art (see for example methods disclosed in US2006/0228380, US2006/0228381, US2007/0184071, US2007/0184072, US2007/0231340, and US2008/0102498 and WO2008/118752). They can also be produced using synthetic protocols known to the man skilled in the art. In case the bacterial capsular saccharide is obtained directly from bacteria, the bacterial cells can be grown in a medium. Following fermentation of bacterial cells that produce the capsular saccharide, the bacterial cells can be lysed to produce a cell lysate. The capsular saccharide may then be isolated from the cell lysate using purification techniques known in the art, including the use of centrifugation, depth filtration, precipitation, ultra-filtration, treatment with activate carbon, diafiltration and/or column chromatography (see, for example, US2006/0228380, US2006/0228381 and WO2008/118752). The purified capsular saccharide can then be used for the preparation of immunogenic conjugates. The isolated capsular saccharide obtained by purification can be characterized by different parameters including, for example the weight average molecular weight (Mw). The molecular weight of the polysaccharide can be measured by Size Exclusion Chromatography (SEC) combined with Multiangle Laser Light Scattering detector (MALLS). In an embodiment, the capsular saccharide used in the method of making or part of the glycoconjugate of the present invention is a capsular saccharide from a pathogenic bacteria. Preferably, the capsular saccharide used in the present invention is a capsular saccharide from a pathogenic Streptococcus, a pathogenic Staphylococcus, a pathogenic Enterococcus, a pathogenic Bacillus, a pathogenic Corynebacterium, a pathogenic Listeria, a pathogenic Erysipelothrix, a pathogenic Clostridium, a pathogenic Haemophilus, a pathogenic Neisseria or a pathogenic Escherichia. More preferably, the capsular saccharide used in the present invention is a capsular saccharide from a pathogenic Streptococcus, a pathogenic Neisseria or a pathogenic Escherichia. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Aeromonas hydrophila and other species (spp.); Bacillus anthracis; Bacillus cereus; Botulinum neurotoxin producing species of Clostridium; Brucella abortus; Brucella melitensis; Brucella suis; Burkholderia mallei (formally Pseudomonas mallei); Burkholderia pseudomallei (formerly Pseudomonas pseudomallei); Campylobacter jejuni; Chlamydia psittaci; Chlamydia trachomatis, Clostridium botulinum; Clostridioides difficile (formally Clostridium difficile); Clostridium perfringens; Coccidioides immitis; Coccidioides posadasii; Cowdria ruminantium (Heartwater); Coxiella burnetii; Enterococcus faecalis; Enterovirulent Escherichia coli group (EEC Group) such as Escherichia coli - enterotoxigenic (ETEC), Escherichia coli - enteropathogenic (EPEC), Escherichia coli - O157:H7 enterohemorrhagic (EHEC), and Escherichia coli - enteroinvasive (EIEC); Ehrlichia spp. such as Ehrlichia chajfeensis; Francisella tularensis; Legionella pneumophilia; Liberobacter africanus; Liberobacter asiaticus; Listeria monocytogenes; miscellaneous enterics such as Klebsiella, Enterobacter, Proteus, Citrobacter, Aerobacter, Providencia, and Serratia; Mycobacterium bovis; Mycobacterium tuberculosis; Mycoplasma capricolum; Mycoplasma mycoides ssp mycoides; Peronosclerosporaphilippinensis; Phakopsora pachyrhizi; Plesiomonas shigelloides; Ralstonia solanacearum race 3, biovar 2; Rickettsia prowazekii; Rickettsia rickettsii; Salmonella spp.; Schlerophthora rayssiae var zeae; Shigella spp.; Staphylococcus aureus; Streptococcus; Synchytrium endobioticum; Vibrio cholerae non-01; Vibrio cholerae 01; Vibrio par ahaemo Iy ticus and other Vibrios; Vibrio vulnificus; Xanthomonas oryzae; Xylella fastidiosa (citrus variegated chlorosis strain); Yersinia enterocolitica and Yersinia pseudotuberculosis; or Yersinia pestis. Preferably, the capsular saccharide used in the present invention is a capsular saccharide from Enterococcus faecalis, Escherichia coli, Staphylococcus aureus or Streptococcus. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Haemophilus influenzae, Neisseria meningitidis, S. pneumoniae, S. pyogenes, S. agalactiae, Group C & G Streptococci or Escherichia coli. More preferably, the capsular saccharide used in the present invention is a capsular saccharide from Neisseria meningitidis, S. pneumoniae, S. agalactiae or Escherichia coli. Even more preferably, the capsular saccharide used in the present invention is a capsular saccharide from S. pneumoniae or S. agalactiae. Even more preferably, the capsular saccharide used in the present invention is a capsular saccharide from S. pneumoniae. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Staphylococcus aureus. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Staphylococcus aureus type 5 or Staphylococcus aureus type 8. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Enterococcus faecalis. In yet a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from is Haemophilus influenzae type b. In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Neisseria meningitidis. In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup A (MenA), N. meningitidis serogroup W135 (MenW135), N. meningitidis serogroup Y (MenY), N. meningitidis serogroup X (MenX) or N. meningitidis serogroup C (MenC). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup A (MenA). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup W135 (MenW135). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup Y (MenY). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup C (MenC). In an embodiment the capsular saccharide used in the present invention is a capsular saccharide from N. meningitidis serogroup X (MenX). In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Escherichia coli. In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Enterococcus faecalis. In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus agalactiae (Group B streptococcus (GBS)). In some embodiments, the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III, IV, V, VI, VII or VIII. In some embodiments, the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III or V. In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus agalactiae (Group B streptococcus (GBS)). In some embodiments, the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III, IV, V, VI, VII, VIII or IX. In some embodiments, the capsular saccharide used in the present invention is a capsular saccharide from GBS type Ia, Ib, II, III, IV or V. In a further embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Escherichia coli. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli part of the Enterovirulent Escherichia coli group (EEC Group) such as Escherichia coli - enterotoxigenic (ETEC), Escherichia coli - enteropathogenic (EPEC), Escherichia coli - O157:H7 enterohemorrhagic (EHEC), or Escherichia coli - enteroinvasive (EIEC). In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Uropathogenic Escherichia coli (UPEC). In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype selected from the group consisting of serotypes O157:H7, O26:H11, O111:H- and O103:H2. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype selected from the group consisting of serotypes O6:K2:H1 and O18:K1:H7. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype selected from the group consisting of serotypes O45:K1, O17:K52:H18, O19:H34 and O7:K1. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O104:H4. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O1:K12:H7. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O127:H6. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O139:H28. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from an Escherichia coli serotype O128:H2. In a preferred embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae. Preferably, the capsular saccharide used in the present invention is a capsular saccharide from a Streptococcus pneumoniae serotype selected from the group consisting of serotypes 1, 2, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9V, 9N, 10A, 10B, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A, 23B, 23F, 24B, 24F, 27, 29, 31, 33B, 33F, 34, 35B, 35F, 38, 72 and 73. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 1. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 2. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 3. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 4. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 5. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 6A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 6B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 7C. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 7F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 8. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 9V. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 9N. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 10A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 10B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 11A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 12F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 14. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 15A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 15B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 15C. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 16F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 17F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 18C. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 19A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 19F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 20. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 21. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 22A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 22F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 23A. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 23B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 23F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 24B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 24F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 27. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 29. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 31. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 33B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 33F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 34. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 35B. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 35F. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 38. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 72. In an embodiment, the capsular saccharide used in the present invention is a capsular saccharide from Streptococcus pneumoniae serotype 73. In a preferred embodiment, the capsular saccharide used in the present invention (purified before further treatment) has a weight average molecular weight between 50 kDa and 5000 kDa. In a preferred embodiment, the capsular saccharide used in the present invention has a weight average molecular weight between 500 kDa and 5000 kDa. In another preferred embodiment, the capsular saccharide used in the present invention has a weight average molecular weight between 1000 kDa and 5000 kDa. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure. Preferably, in order to generate glycoconjugates with advantageous filterability characteristics, immunogenicity and/or yields, sizing of the saccharide to a target molecular weight range is performed prior to the conjugation to a carrier protein. Advantageously, the size of the purified capsular saccharide is reduced while preserving critical features of the structure of the polysaccharide. Mechanical or chemical sizing maybe employed. In an embodiment, the size of the purified capsular saccharide is reduced by chemical hydrolysis. Chemical hydrolysis maybe conducted using a mild acid (e.g., acetic acid, formic acid, propanoic acid). In an embodiment, chemical hydrolysis is conducted using formic acid. In an embodiment, chemical hydrolysis is conducted using propanoic acid. In a preferred embodiment, chemical hydrolysis is conducted using acetic acid. Chemical hydrolysis may also be conducted using a diluted strong acid (such as diluted hydrochloric acid, diluted sulfuric acid, diluted phosphoric acid, diluted nitric acid or diluted perchloric acid). In an embodiment, chemical hydrolysis is conducted using diluted hydrochloric acid. In an embodiment, chemical hydrolysis is conducted using diluted sulfuric acid. In an embodiment, chemical hydrolysis is conducted using diluted phosphoric acid. In an embodiment, chemical hydrolysis is conducted using diluted nitric acid. In an embodiment, chemical hydrolysis is conducted using diluted perchloric acid. The size of the purified capsular saccharide can also be reduced by mechanical homogenization. In an embodiment, the size of the purified capsular saccharide is reduced by high pressure homogenization. High pressure homogenization achieves high shear rates by pumping the process stream through a flow path with sufficiently small dimensions. The shear rate is increased by using a larger applied homogenization pressure, and exposure time can be increased by recirculating the feed stream through the homogenizer. The high-pressure homogenization process can be appropriate for reducing the size of the purified capsular saccharide while preserving the structural features of the saccharide. In a preferred embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 10 kDa and 1000 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 50 kDa and 500 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 50 kDa and 400 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 50 kDa and 250 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 250 kDa and 1000 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 250 kDa and 500 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 250 kDa and 400 kDa. In a preferred embodiment, the isolated capsular saccharide is sized to a weight average molecular weight between 200 kDa and 800 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 250 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 300 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 350 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 400 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 450 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 500 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 550 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 600 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 700 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 800 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 900 kDa. In an embodiment, the isolated capsular saccharide is sized to a weight average molecular weight of about 1000 kDa. In an embodiment, the isolated capsular saccharide is not sized. The isolated capsular saccharide described above may be activated (e.g., chemically activated) to make them capable of reacting (e.g., with a linker) and then incorporated into glycoconjugates, as further described herein. For the purposes of the invention the term 'glycoconjugate' indicates a saccharide covalently linked to a carrier protein. In general, covalent conjugation of saccharides to carriers enhances the immunogenicity of saccharides as it converts them from T-independent antigens to T-dependent antigens, thus allowing priming for immunological memory. Conjugation is particularly useful for pediatric vaccines. 1.2 Capsular saccharide glycoconjugates of the invention In some embodiments, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said polysaccharide before conjugation is between 10 kDa and 2,000 kDa. The weight average molecular weight (Mw) of the saccharide before conjugation refers to the Mw before the activation of the saccharide (i.e., after an eventual sizing step but before reacting the saccharide with an activating agent). In the context of the present invention the Mw of the saccharide is not substantially modified by the activation step and the Mw of the saccharide incorporated in the conjugate is similar to the Mw of the saccharide as measured before activation. In an embodiment, the saccharide is activated with a carbonic acid derivative (e.g., CDI or CDT) in combination with an alkyne linker (see sections 1.3 below). In an embodiment, the saccharide is activated with CDI in combination with an alkyne linker (see sections 1.3 below). In an embodiment, the saccharide is activated with CDT in combination with an alkyne linker (see sections 1.3 below). In an embodiment, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 50 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 500 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 250 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 200 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 150 kDa. In an embodiment, the weight average molecular weight (Mw) is between 50 kDa and 100 kDa. In an embodiment, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 75 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 500 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 250 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 200 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 150 kDa. In an embodiment, the weight average molecular weight (Mw) is between 75 kDa and 100 kDa. In an embodiment, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 100 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 500 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 250 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 200 kDa. In an embodiment, the weight average molecular weight (Mw) is between 100 kDa and 150 kDa. In an embodiment, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 150 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 150 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 150 kDa and 500 kDa. In an embodiment, the weight average molecular weight (Mw) is between 150 kDa and 250 kDa. In an embodiment, the weight average molecular weight (Mw) is between 150 kDa and 200 kDa. In an embodiment, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 200 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 200 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 200 kDa and 500 kDa. In an embodiment, the weight average molecular weight (Mw) is between 200 kDa and 300 kDa. In an embodiment, the weight average molecular weight (Mw) is between 200 kDa and 250 kDa. In an embodiment, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is between 500 kDa and 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is between 500 kDa and 750 kDa. In an embodiment, the weight average molecular weight (Mw) is between 500 kDa and 700 kDa. In an embodiment, the weight average molecular weight (Mw) is between 500 kDa and 600 kDa. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure. In an embodiment, the glycoconjugate of the present invention comprises a capsular saccharide wherein the weight average molecular weight (Mw) of said saccharide before conjugation is about 1,000 kDa. In an embodiment, the weight average molecular weight (Mw) is about 750 kDa. In an embodiment, the weight average molecular weight (Mw) is about 700 kDa. In an embodiment, the weight average molecular weight (Mw) is about 600 kDa. In an embodiment, the weight average molecular weight (Mw) is about 500 kDa. In an embodiment, the weight average molecular weight (Mw) is about 400 kDa. In an embodiment, the weight average molecular weight (Mw) is about 300 kDa. In an embodiment, the weight average molecular weight (Mw) is about 200 kDa. In an embodiment, the weight average molecular weight (Mw) is about 150 kDa. In an embodiment, the weight average molecular weight (Mw) is about 125 kDa. In an embodiment, the weight average molecular weight (Mw) is about 100 kDa. In some embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 250 kDa and 20,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 15,000 kDa. In yet other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 2,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 1,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 1,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 500 kDa and 750 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 2,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 1,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 750 kDa and 1,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 2,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 1,500 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 4,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 3,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,000 kDa and 3,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 2,250 kDa and 3,500 kDa. In preferred embodiment, the glycoconjugate of the invention has a weight average molecular weight (Mw) of between 1,000 kDa and 2,500 kDa. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 10,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 9,000 kDa. In other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 8,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 7,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 6,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 5,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 4,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 3,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 3,250 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 3,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 2,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 2,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 1,500 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 1,000 kDa. In still other embodiments, the glycoconjugate of the invention has a weight average molecular weight (Mw) of about 750 kDa. Another way to characterize the glycoconjugates of the invention is by the number of lysine residues in the carrier protein (e.g., CRM197 or SCP) that become conjugated to the saccharide which can be characterized as a range of conjugated lysines (degree of conjugation). The evidence for lysine modification of the carrier protein, due to covalent linkages to the saccharides, can be obtained by amino acid analysis using routine methods known to those of skill in the art. Conjugation results in a reduction in the number of lysine residues recovered compared to the carrier protein starting material used to generate the conjugate materials. In a preferred embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 13. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 10. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 8. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 6. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 5. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 2 and 4. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 13. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 10. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 8. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 6. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 5. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 3 and 4. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 5 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 5 and 10. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 8 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 8 and 12. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 10 and 15. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is between 10 and 12. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 2. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 3. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 4. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 5. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 6. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 7. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 8. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 9. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 10, about 11. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 12. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 13. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 14. In an embodiment, the degree of conjugation of the glycoconjugate of the invention is about 15. In a preferred embodiment, the degree of conjugation of the glycoconjugate of the invention is between 4 and 7. In some such embodiments, the carrier protein is CRM197. In other such embodiments, the carrier protein is SCP. The glycoconjugates of the invention may also be characterized by the ratio (weight/weight) of saccharide to carrier protein. In some embodiments, the ratio of saccharide to carrier protein in the glycoconjugate (w/w) is between 0.5 and 3.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is between 0.5 and 2.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is between 0.5 and In other embodiments, the saccharide to carrier protein ratio (w/w) is between 0.8 and 1.2. In other embodiments, the saccharide to carrier protein ratio (w/w) is between 0.5 and 1.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is between 1.0 and 1.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is between 1.0 and 2.0. In further embodiments, the saccharide to carrier protein ratio (w/w) is between 0.8 and 1.2. In a preferred embodiment, the ratio of saccharide to carrier protein in the conjugate is between 0.9 and 1.1. In an embodiment, the saccharide to carrier protein ratio (w/w) is about 0.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 0.6. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 0.7. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 0.8. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 0.9. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.1. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.2. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.3. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.4. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.6. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.7. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.8. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 1.9. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.0. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.1. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.2. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.5. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 2.8. In other embodiments, the saccharide to carrier protein ratio (w/w) is about 3.0. In some such embodiments, the carrier protein is CRM197. In other such embodiments, the carrier protein is SCP. The glycoconjugates of the invention may also be characterized by the number of covalent linkages between the carrier protein and the saccharide as a function of repeat units of the saccharide. In one embodiment, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 4 saccharide repeat units of the saccharide. In another embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 10 saccharide repeat units of the saccharide. In another embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 15 saccharide repeat units of the saccharide. In a further embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 25 saccharide repeat units of the saccharide. In a further embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 50 saccharide repeat units of the saccharide. In yet a further embodiment, the covalent linkage between the carrier protein and the saccharide occurs at least once in every 100 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 5 to 10 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 2 to 7 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 6 to 11 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 9 to 14 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 10 to 20 saccharide repeat units of the saccharide. In other embodiments, the glycoconjugate of the invention comprises at least one covalent linkage between the carrier protein and the saccharide for every 4 to 25 saccharide repeat units of the saccharide. In frequent embodiments, the carrier protein is CRM197. In other embodiments, the carrier protein is SCP. In some embodiments, the carrier protein is CRM197 and the covalent linkage between the CRM197 and the saccharide occurs at least once in every 4, 10, 15 or 25 saccharide repeat units of the saccharide. In other embodiments, the carrier protein is SCP and the covalent linkage between the SCP and the saccharide occurs at least once in every 4, 10, 15 or 25 saccharide repeat units of the saccharide. The glycoconjugates and immunogenic compositions of the invention may contain free saccharide that is not covalently conjugated to the carrier protein but is nevertheless present in the glycoconjugate composition. The free saccharide may be noncovalently associated with (i.e., noncovalently bound to, adsorbed to, or entrapped in or with) the glycoconjugate. In a preferred embodiment, the glycoconjugate comprises less than about 50% of free saccharide compared to the total amount of said saccharide. In a preferred embodiment the glycoconjugate comprises less than about 40% of free saccharide compared to the total amount of said saccharide. In a yet preferred embodiment, the glycoconjugate comprises less than about 25% of free saccharide compared to the total amount of said saccharide. In an even preferred embodiment, the glycoconjugate comprises less than about 20% of free saccharide compared to the total amount of said saccharide. In a yet preferred embodiment, the glycoconjugate comprises less than about 15% of free saccharide compared to the total amount of said saccharide. The glycoconjugates may also be characterized by their molecular size distribution (Kd). Size exclusion chromatography media (CL-4B) can be used to determine the relative molecular size distribution of the conjugate. Size Exclusion Chromatography (SEC) is used in gravity fed columns to profile the molecular size distribution of conjugates. Large molecules excluded from the pores in the media elute more quickly than small molecules. Fraction collectors are used to collect the column eluate. The fractions are tested colorimetrically by saccharide assay. For the determination of Kd, columns are calibrated to establish the fraction at which molecules are fully excluded (V0), (Kd=0), and the fraction representing the maximum retention (Vi), (Kd=1). The fraction at which a specified sample attribute is reached (Ve), is related to Kd by the expression, Kd = (Ve - V0)/ (Vi - V0). In a preferred embodiment, at least 30% of the glycoconjugate of the invention has a Kd below or equal to 0.3 in a CL-4B column. In a preferred embodiment, at least 40% of the glycoconjugate of the invention has a Kd below or equal to 0.3 in a CL-4B column. In a preferred embodiment, at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, or 85% of the glycoconjugate of the invention has a Kd below or equal to 0.3 in a CL-4B column. In a preferred embodiment, at least 60% of the glycoconjugate of the invention has a Kd below or equal to 0.3 in a CL-4B column. In a preferred embodiment, between 50% and 80% of the glycoconjugate of the invention has a Kd below or equal to 0.3 in a CL-4B column. In a preferred embodiment, between 65% and 80% of the glycoconjugate of the invention has a Kd below or equal to 0.3 in a CL-4B column. 1.3 Capsular saccharide glycoconjugates of the invention prepared using click chemistry The glycoconjugates of the present invention are prepared using click chemistry. The invention also relates to a method of making a glycoconjugate, as disclosed herein. According to the present invention, click chemistry comprises three steps, (a) reacting an isolated capsular saccharide with a carbonic acid derivative and an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide (activation of the saccharide), (b) reacting a carrier protein with an agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group where the NHS moiety reacts with the amino groups to form an amide linkage thereby obtaining an azido functionalized carrier protein (activation of the carrier protein), (c) reacting the activated alkynyl saccharide of step (a) with the activated azido-carrier protein of step (b) by Cu+1 mediated azide-alkyne cycloaddition reaction to form a glycoconjugate. Following step (a) the saccharide is said to be activated and is referred to herein as “activated saccharide” or “activated alkynyl saccharide”. Following step (b) the carrier is said to be activated and is referred to as “activated carrier” or “activated azido-carrier”. As mentioned above, before the activation (a), sizing of the saccharide to a target molecular weight (MW) range can be performed. Therefore, in an embodiment, the isolated saccharide is sized before activation with a carbonic acid derivative and an alkyne linker. In an embodiment, the isolated saccharide is sized to any of the target molecular weight (MW) range defined above. In an embodiment, said carbonic acid derivative is selected from the group consisting of 1,1’-carbonyldiimidazole (CDI), 1,1’-carbonyl-di-(1,2,4-triazole) (CDT), disuccinimidyl carbonate (DSC) and N-hydroxysuccinimidyl chloroformate. In an embodiment, said carbonic acid derivative is 1,1’-carbonyldiimidazole (CDI). In another embodiment, said carbonic acid derivative is 1,1'-Carbonyl-di-(1,2,4-triazole) (CDT). In another embodiment, said carbonic acid derivative is disuccinimidyl carbonate (DSC). In yet a further embodiment, said carbonic acid derivative is N-hydroxysuccinimidyl chloroformate. In an embodiment, said carbonic acid derivative is 1,1’-carbonyldiimidazole (CDI) or 1,1'- Carbonyl-di-(1,2,4-triazole) (CDT). Preferably, said carbonic acid derivative is 1,1’- carbonyldiimidazole (CDI). In an embodiment, said alkyne linker is a compound of formula (I), H2N X (I) wherein X is selected from the group consisting of CH2, CH2(CH2)n, (CH2CH2O)mCH2CH2, NHCO(CH2)n, NHCO(CH2CH2O)mCH2CH2, OCH2(CH2)n and O(CH2CH2O)mCH2CH2; where n is selected from 1 to 10 and m is selected from 1 to 4. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is CH2. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is CH2(CH2)n, and n is selected from 1 to 10. In an embodiment, n is selected from 1 to 5. In an embodiment, n is selected from 1 to 4. In an embodiment, n is selected from 1 to 3. In an embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In yet another embodiment, n is 3. In yet a further embodiment, n is 4. In yet a further embodiment, n is 5. In yet a further embodiment, n is 6. In yet a further embodiment, n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9. In yet a further embodiment, n is 10. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is (CH2CH2O)mCH2CH2, wherein m is selected from 1 to 4. In an embodiment, m is selected from 1 to 3. In an embodiment, m is selected from 1 to 2. In a particular embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet a further embodiment, m is 4. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is NHCO(CH2)n, and n is selected from 1 to 10. In an embodiment, n is selected from 1 to 5. In an embodiment, n is selected from 1 to 4. In an embodiment, n is selected from 1 to 3. In an embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In yet another embodiment, n is 3. In yet a further embodiment, n is 4. In yet a further embodiment, n is 5. In yet a further embodiment, n is 6. In yet a further embodiment, n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9. In yet a further embodiment, n is 10. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is NHCO(CH2CH2O)mCH2CH2, where m is selected from 1 to 4. In an embodiment, m is selected from 1 to 3. In an embodiment, m is selected from 1 to 2. In a particular embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet a further embodiment, m is 4. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is OCH2(CH2)n, and n is selected from 1 to 10. In an embodiment, n is selected from 1 to 5. In an embodiment, n is selected from 1 to 4. In an embodiment, n is selected from 1 to 3. In an embodiment, n is selected from 1 to 2. In a particular embodiment, n is 1. In another embodiment, n is 2. In yet another embodiment, n is 3. In yet a further embodiment, n is 4. In yet a further embodiment, n is 5. In yet a further embodiment, n is 6. In yet a further embodiment, n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9. In yet a further embodiment, n is 10. In an embodiment, said alkyne linker is a compound of formula (I), wherein X is O(CH2CH2O)mCH2CH2, where m is selected from 1 to 4. In an embodiment, m is selected from 1 to 3. In an embodiment, m is selected from 1 to 2. In a particular embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet a further embodiment, m is 4. In an embodiment, said alkyne linker is a compound of formula (II), Hence in a preferred embodiment, said alkyne linker is propargylamine. In an embodiment, said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (III), where X is selected from the group consisting of (CH2)nCH2C=O and (CH2CH2O)mCH2CH2=O where n is selected from 0 to 10 and m is selected from 0 to 4. In an embodiment, said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (III), wherein X is (CH2)nCH2C=O, where n is selected from 0 to 10. In an embodiment, n is selected from 0 to 5. In an embodiment, n is selected from 0 to 4. In an embodiment, n is selected from 0 to 3. In an embodiment, n is selected from 0 to 2. In a particular embodiment, n is 0. In a particular embodiment, n is 1. In another embodiment, n is 2. In yet another embodiment, n is 3. In yet a further embodiment, n is 4. In yet a further embodiment, n is 5. In yet a further embodiment, n is 6. In yet a further embodiment, n is 7. In yet a further embodiment, n is 8. In yet a further embodiment, n is 9. In yet a further embodiment, n is 10. In an embodiment, said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (III), wherein X is (CH2CH2O)mCH2CH2=O, where m is selected from 0 to 4. In an embodiment, m is selected from 0 to 3. In an embodiment, m is selected from 0 to 2. In an embodiment, m is selected from 0 to 1. In a particular embodiment, m is 0. In a particular embodiment, m is 1. In another embodiment, m is 2. In yet another embodiment, m is 3. In yet a further embodiment, m is 4. In an embodiment, said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (IV): Hence in an embodiment, said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is (2,5-Dioxopyrrolidin-1-yl) 2-azidoacetate. In an embodiment, step a) comprises reacting the saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative-activated saccharide with an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 2-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 3-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 5-10 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 2-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 3-5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 2-3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 1-2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.5-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.8-1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.4-0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.3-0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.01-0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.05-0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.1-0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative that is between 0.2-0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.01 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.05 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.08 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.2 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.3 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.4 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 0.5 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 1 molar equivalent to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 2 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 3 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 4 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 5 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 8 molar equivalents to the amount of saccharide present in the reaction mixture. In one embodiment step a) comprises reacting the saccharide with an amount of carbonic acid derivative of about 10 molar equivalents to the amount of saccharide present in the reaction mixture. In an embodiment, at step a) the isolated saccharide is reacted with a carbonic acid derivative in an aprotic solvent. In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylsulphoxide (DMSO) or dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylsulphoxide (DMSO). In a preferred embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution comprising dimethylsulphoxide (DMSO). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylsulphoxide (DMSO) or dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylsulphoxide (DMSO). In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylacetamide. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of N-methyl-2- pyrrolidone. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of hexamethylphosphoramide (HMPA). In a preferred embodiment the isolated saccharide is reacted with a carbonic acid derivative in a solution consisting essentially of dimethylsulphoxide (DMSO). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in dimethylsulphoxide (DMSO) or dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in dimethylformamide (DMF). In one embodiment the isolated saccharide is reacted with a carbonic acid derivative in dimethylsulphoxide (DMSO). In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in dimethylacetamide. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in N-methyl-2-pyrrolidone. In an embodiment, the isolated saccharide is reacted with a carbonic acid derivative in hexamethylphosphoramide (HMPA). In a preferred embodiment the isolated saccharide is reacted with CDI in dimethylsulphoxide (DMSO). In an embodiment the isolated saccharide is reacted with CDI in anhydrous DMSO. It has been surprisingly found that reacting the isolated saccharide with CDI in an environment with a moisture level of about 0.1% to 1% (v/v) allows to avoid side reactions. Therefore, in one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.2% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.2% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.3% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.3% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.3% to 0.4% (v/v) water. Preferably, in one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.2% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.6% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.7% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in an aprotic solvent comprising about 0.9% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.2% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.2% to 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.3% to 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.3% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.3% to 0.4% (v/v) water. Preferably, in one embodiment the isolated saccharide is reacted with CDI in DMSO comprising 0.1% to 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.1% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.2% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.3% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.4% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.5% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.6% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.7% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.8% (v/v) water. In one embodiment the isolated saccharide is reacted with CDI in DMSO comprising about 0.9% (v/v) water. In one embodiment the free carbonic acid derivative is then quenched by the addition of water before the addition of the alkyne linker. Water can inactivate free CDI. Therefore, in an embodiment, carbonic acid derivative activation is followed by the addition of water. In an embodiment, water is added to bring the total water content in the mixture to between about 1% to about 10% (v/v). In an embodiment, water is added to bring the total water content in the mixture to between about 1.2% to about 8% (v/v). In an embodiment, water is added to bring the total water content in the mixture to between about 1.5% to about 5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to between about 1.5% to about 3% (v/v). In an embodiment, water is added to bring the total water content in the mixture to between about 1.5% to about 2.5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1 % (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1.2% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1.4% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 1.5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 2% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 2.5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 3% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 5% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 7% (v/v). In an embodiment, water is added to bring the total water content in the mixture to about 10% (v/v). Once the saccharide has been reacted with carbonic acid derivative and following an eventual quenching of carbonic acid derivative with water, the carbonic acid derivative-activated saccharide is reacted with an alkyne linker. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-10 molar equivalent to the amount of polysaccharide Repeat Unit of the activated saccharide (molar equivalent of RU). In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.01-0.1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.05-0.1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.1-0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 0.5-1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 1-2 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 2-3 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 3-4 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 4-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 4-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 4-5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 5-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 5-8 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is between 8-10 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.01 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.05 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 3 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 4 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 8 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In one embodiment step a) further comprises reacting the carbonic acid derivative- activated saccharide with an amount of alkyne linker that is about 10 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In the above embodiments, said carbonic acid derivative is preferably CDI. In another embodiment, said carbonic acid derivative is CDT. In one embodiment the degree of activation of the activated saccharide following step a) is between 0.5 to 50%. The degree of activation of the alkynyl saccharide being defined as the percentage of Repeating Unit linked to an alkyne linker. In one embodiment the degree of activation of the activated saccharide following step a) is between 1 to 30%. In another embodiment the degree of activation of the activated saccharide following step a) is between 2 to 25%. In another embodiment the degree of activation of the activated saccharide following step a) is between 3 to 20%. In another embodiment the degree of activation of the activated saccharide following step a) is between 3 to 15%. In another embodiment the degree of activation of the activated saccharide following step a) is between 4 to 15%. In an embodiment the degree of activation of the activated saccharide following step a) is between 1 to 6%. In an embodiment the degree of activation of the activated saccharide following step a) is between 3 to 6%. In an embodiment the degree of activation of the activated saccharide following step a) is between 10 to 15%. In an embodiment the degree of activation of the activated saccharide following step a) is about 1%. In an embodiment the degree of activation of the activated saccharide following step a) is about 2%. In an embodiment the degree of activation of the activated saccharide following step a) is about 3%. In an embodiment the degree of activation of the activated saccharide following step a) is about 4%. In an embodiment the degree of activation of the activated saccharide following step a) is about 5%. In an embodiment the degree of activation of the activated saccharide following step a) is about 6%. In an embodiment the degree of activation of the activated saccharide following step a) is about 7%. In an embodiment the degree of activation of the activated saccharide following step a) is about 8%. In an embodiment the degree of activation of the activated saccharide following step a) is about 9%. In an embodiment the degree of activation of the activated saccharide following step a) is about 10%. In an embodiment the degree of activation of the activated saccharide following step a) is about 11%. In an embodiment the degree of activation of the activated saccharide following step a) is about 12%. In an embodiment the degree of activation of the activated saccharide following step a) is about 13%. In an embodiment the degree of activation of the activated saccharide following step a) is about 14%. In an embodiment the degree of activation of the activated saccharide following step a) is about 15%. In an embodiment the degree of activation of the activated saccharide following step a) is about 16%. In an embodiment the degree of activation of the activated saccharide following step a) is about 17%. In an embodiment the degree of activation of the activated saccharide following step a) is about 18%. In an embodiment the degree of activation of the activated saccharide following step a) is about 19%. In an embodiment the degree of activation of the activated saccharide following step a) is about 20%. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 3-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 7.5-10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 3-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 5-7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 3-5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2.5-3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 2-2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1.5-2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-1.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-1.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 1-1.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-1 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.5-1 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-0.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 10 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 7.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 3 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 2.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 2 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 1.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 1 molar equivalent to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 0.5 molar equivalents to the lysines on the carrier protein. In one embodiment step b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is about 0.1 molar equivalents to the lysines on the carrier protein. In one embodiment the degree of activation of the activated carrier following step b) is between 1 and 50. The degree of activation of the activated carrier being defined as the number of lysine residues in the carrier protein that become linked to the agent bearing an N- Hydroxysuccinimide (NHS) moiety and an azido group. In an embodiment, the carrier protein is CRM197, which contains 39 lysine residues. In said embodiment the degree of activation of the activated carrier following step b) may be between 1 to 30. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is between 5 to 20. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is between 9 to 18. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is between 8 to 11. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is between 15 to 20. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 5. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 6. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 7. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 8. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 9. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 10. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 11. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 12. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 13. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 14. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 15. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 16. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 17. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 18. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 19. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 20. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 21. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 22. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 23. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 24. In another embodiment the degree of activation of the activated carrier (CRM197) following step b) is about 25. In an embodiment, the carrier protein is SCP or a fragment thereof. In said embodiment the degree of activation of the activated carrier following step b) may be between 1 to 50. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 5 to 50. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 7 to 45. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 5 to 15. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 20 to 30. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 30 to 50. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 30 to 40. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is between 10 to 40. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 5. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 7. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 10. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 13. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 15. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 20. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 26. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 30. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 35. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 37. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 40. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 45. In another embodiment the degree of activation of the activated carrier (SCP) following step b) is about 50. In an embodiment, the carrier protein is TT or a fragment thereof. In said embodiment the degree of activation of the activated carrier following step b) may be between 1 to 30. In another embodiment the degree of activation of the activated carrier (TT) following step b) is between 5 to 25. In another embodiment the degree of activation of the activated carrier (TT) following step b) is between 7 to 25. In another embodiment the degree of activation of the activated carrier (TT) following step b) is between 10 to 20. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 5. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 7. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 10. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 12. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 15. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 20. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 25. In another embodiment the degree of activation of the activated carrier (TT) following step b) is about 30. In an embodiment, the conjugation reaction c) is carried out in aqueous buffer. In an embodiment, the conjugation reaction c) is carried out in aqueous buffer in the presence of copper (I) as catalyst. In an embodiment, the conjugation reaction c) is carried out in aqueous buffer in the presence an oxidant and of copper (I) as catalyst. In a preferred embodiment, the conjugation reaction c) is carried out in aqueous buffer in the presence of copper (I) as catalyst and ascorbate as oxidant. In an embodiment, THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) and aminoguanidine may be further added to protect the protein from side reactions. Therefore, in a preferred embodiment, the conjugation reaction c) is carried out in aqueous buffer in the presence of copper (I) as catalyst and ascorbate as oxidant, wherein the reaction mixture further comprises THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) and aminoguanidine. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is between 0.1 and 3. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is between 0.5 and 2. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is between 0.6 and 1.5. In a preferred embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is between 0.8 and 1. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 0.5. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 0.6. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 0.7. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 0.8. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 0.9. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.1. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.2. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.3. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.4. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.5. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.6. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.7. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.8. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 1.9. In an embodiment the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step c) is about 2. Following the click conjugation reaction, unreacted alkyne groups may remain present in the conjugates, these may be capped using a suitable alkyne group capping agent. In one embodiment this alkyne group capping agent is an agent bearing an azido group. In an embodiment, said alkyne group capping agent is a compound of formula (VI), (VI) wherein X is (CH2)n wherein n is selected from 1 to 15. In one embodiment this alkyne group capping agent is 3-azido-1-propanol. Therefore, in an embodiment, following step (c) the process further comprises a step of capping the unreacted alkyne groups remained in the conjugates with an alkyne group capping agent. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.05 to 20 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.1 to 15 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.5 to 10 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.5 to 5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.5 to 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.5 to 1 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 1 to 5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 1 to 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 1.5 to 2.5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that about 0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that about 1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that about 1.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that about 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that about 2.5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted alkyne groups is performed with an amount of capping agent that about 5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. Following the click conjugation reaction, there may remain unreacted azido groups in the conjugates, these may be capped using a suitable azido group capping agent. Therefore, in an embodiment, following step c), unreacted azido groups in the conjugates, are capped using a suitable azido group capping agent. In one embodiment this azido group capping agent is an agent bearing an alkyne group. In one embodiment this azido group capping agent is an agent bearing a terminal alkyne. In one embodiment this azido group capping agent is an agent bearing a cycloalkyne. In an embodiment, said azido group capping agent is a compound of formula (V), (V) wherein X is (CH2)n wherein n is selected from 1 to 15. In one embodiment this azido group capping agent is propargyl alcohol. Therefore, in an embodiment, following step (c) the process further comprises a step of capping the unreacted azido groups remained in the conjugates with an azido group capping agent. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.05 to 20 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.1 to 15 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.5 to 10 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.5 to 5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.5 to 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.5 to 1 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 1 to 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.75 to 1.5 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is about 1 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is about 1.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is about 0.5 molar equivalent to the amount of polysaccharide repeat unit of the activated saccharide. In an embodiment the capping of the unreacted azido groups is performed with an amount of capping agent that is about 2 molar equivalents to the amount of polysaccharide repeat unit of the activated saccharide. Following conjugation to the carrier protein, the glycoconjugate can be purified (enriched with respect to the amount of saccharide-protein conjugate) by a variety of techniques known to the skilled person. These techniques include dialysis, concentration/diafiltration operations, tangential flow filtration precipitation/elution, column chromatography (DEAE or hydrophobic interaction chromatography), and depth filtration. Therefore, in one embodiment the process for producing the glycoconjugate of the present invention comprises the step of purifying the glycoconjugate after it is produced. In an aspect, the invention provides a glycoconjugate produced according to any of the methods disclosed herein. In an aspect, the invention provides a capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII): (VII), wherein X is selected from the group consisting of CH2(CH2)n’, (CH2CH2O)mCH2CH2, NHCO(CH2)n’, NHCO(CH2CH2O)mCH2CH2, OCH2(CH2)n’ and O(CH2CH2O)mCH2CH2; where n’ is selected from 0 to 10 and m is selected from 1 to 4, and wherein X' is selected from the group consisting of CH2(CH2)n”, CH2O(CH2)n’’CH2, CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. Where the structure in square brackets represents a repeat unit of the capsular saccharide. Formula (VII) is a schematic representation of glycoconjugates of the invention. It should not be understood that a linkage is present at every repeating unit of the saccharide (the structure in square brackets). Rather, a majority of the saccharide repeating unit remains unmodified and covalent linkages between the carrier protein and the saccharide is for a minority of the saccharide repeat units. Additionally, an individual carrier protein (CP) molecule may be linked to more than one saccharide molecule and an individual saccharide molecule can be linked to more than one individual carrier protein (CP) molecule. The structure in square brackets represents a repeat unit of the capsular saccharide. In a preferred embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH2(CH2)n’, where n’ is 0 and wherein X' is CH2(CH2)n” where n’’ is 0. Therefore, in a preferred embodiment, the invention provides a capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VIII), (VIII). Where the structure in square brackets represents a repeat unit of the capsular saccharide. Formula (VIII) is a schematic representation of the preferred glycoconjugates of the invention. It should not be understood that a linkage is present at every repeat unit of the saccharide (the structure in square brackets). Rather, a majority of the saccharide repeating unit remains unmodified and covalent linkages between the carrier protein and the saccharide is for a minority of the saccharide repeat units. Additionally, an individual carrier protein (CP) molecule may be linked to more than one saccharide molecule and an individual saccharide molecule can be linked to more than one individual carrier protein (CP) molecule. The structure in square brackets is a schematic representation of a repeat unit of the saccharide. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH2(CH2)n’, where n’ is selected from 0 to 10 and wherein X' is CH2(CH2)n” where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 0 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 0 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 0 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 0 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 0 and n’’ is 0. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is à and n’’ is 1. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 0 and n’’ is 2. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 0 and n’’ is 4. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 0 and n’’ is 6. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH2(CH2)n’, where n’ is selected from 0 to 10 and wherein X' is CH2O(CH2)n’’CH2 where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 0 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 0 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 0 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 0 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 0 and n’’ is 0. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is à and n’’ is 1. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 0 and n’’ is 2. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 0 and n’’ is 4. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 0 and n’’ is 6. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH2(CH2)n’, where n’ is selected from 0 to 10 and wherein X' is CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. In an embodiment, n’ is selected from 0 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 0 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 0 to 3, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 0 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In an embodiment, n’ is selected from 0 to 1, m’ is selected from 0 to 1 and n’’ is selected from 0 to 1. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 0. In another embodiment, n’ is 0, m’ is 1 and n’’ is 0. In another embodiment, n’ is 0, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 0. In another embodiment, n’ is 1, m’ is 1 and n’’ is 0. In another embodiment, n’ is 1, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In another embodiment, n’ is 2, m’ is 0 and n’’ is 0. In another embodiment, n’ is 2, m’ is 1 and n’’ is 0. In another embodiment, n’ is 2, m’ is 2 and n’’ is 0. In another embodiment, n’ is 2, m’ is 3 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 0.In yet a further embodiment, n’ is 5, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 0. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 1. In a particular embodiment, n’ is 0, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 1. In another embodiment, n’ is 2, m’ is 0 and n’’ is 1. In another embodiment, n’ is 2, m’ is 1 and n’’ is 1. In another embodiment, n’ is 2, m’ is 2 and n’’ is 1. In another embodiment, n’ is 2, m’ is 3 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 1. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 2. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 2. In a particular embodiment, n’ is 0, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 2. In another embodiment, n’ is 2, m’ is 0 and n’’ is 2. In another embodiment, n’ is 2, m’ is 1 and n’’ is 2. In another embodiment, n’ is 2, m’ is 2 and n’’ is 2. In another embodiment, n’ is 2, m’ is 3 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 2. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 3. In a particular embodiment, n’ is 0, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 3. In another embodiment, n’ is 2, m’ is 0 and n’’ is 3. In another embodiment, n’ is 2, m’ is 1 and n’’ is 3. In another embodiment, n’ is 2, m’ is 2 and n’’ is 3. In another embodiment, n’ is 2, m’ is 3 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 3. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 4. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 4. In a particular embodiment, n’ is 0, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 4. In another embodiment, n’ is 2, m’ is 0 and n’’ is 4. In another embodiment, n’ is 2, m’ is 1 and n’’ is 4. In another embodiment, n’ is 2, m’ is 2 and n’’ is 4. In another embodiment, n’ is 2, m’ is 3 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 4. In a particular embodiment, n’ is 0, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 0, m’ is 1 and n’’ is 5. In a particular embodiment, n’ is 0, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 0, m’ is 3 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 5. In another embodiment, n’ is 2, m’ is 0 and n’’ is 5. In another embodiment, n’ is 2, m’ is 1 and n’’ is 5. In another embodiment, n’ is 2, m’ is 2 and n’’ is 5. In another embodiment, n’ is 2, m’ is 3 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 5. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is (CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2(CH2)n”, where n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2. In yet another embodiment, m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5. In another embodiment, m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is (CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2O(CH2)n’’CH2, where n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2. In yet another embodiment, m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5. In another embodiment, m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is (CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. In an embodiment, m is selected from 1 to 3, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 0. In another embodiment, m is 1, m’ is 1 and n’’ is 0. In another embodiment, m is 1, m’ is 2 and n’’ is 0. In another embodiment, m is 1, m’ is 3 and n’’ is 0. In another embodiment, m is 2, m’ is 0 and n’’ is 0. In another embodiment, m is 2, m’ is 1 and n’’ is 0. In another embodiment, m is 2, m’ is 2 and n’’ is 0. In another embodiment, m is 2, m’ is 3 and n’’ is 0. In yet another embodiment, m is 3, m’ is 0 and n’’ is 0. In yet another embodiment, m is 3, m’ is 1 and n’’ is 0. In yet another embodiment, m is 3, m’ is 2 and n’’ is 0. In yet another embodiment, m is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, m is 4, m’ is 0 and n” is 0. In yet a further embodiment, m is 4, m’ is 1 and n” is 0. In yet a further embodiment, m is 4, m’ is 2 and n” is 0. In yet a further embodiment, m is 4, m’ is 3 and n” is 0. In a particular embodiment, m is 1, m’ is 0 and n’’ is 1. In a particular embodiment, m is 1, m’ is 1 and n’’ is 1. In a particular embodiment, m is 1, m’ is 2 and n’’ is 1. In a particular embodiment, m is 1, m’ is 3 and n’’ is 1. In another embodiment, m is 2, m’ is 0 and n’’ is 1. In another embodiment, m is 2, m’ is 1 and n’’ is 1. In another embodiment, m is 2, m’ is 2 and n’’ is 1. In another embodiment, m is 2, m’ is 3 and n’’ is 1. In yet another embodiment, m is 3, m’ is 0 and n’’ is 1. In yet another embodiment, m is 3, m’ is 1 and n’’ is 1. In yet another embodiment, m is 3, m’ is 2 and n’’ is 1. In yet another embodiment, m is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, m is 4, m’ is 0 and n” is 1. In yet a further embodiment, m is 4, m’ is 1 and n” is 1. In yet a further embodiment, m is 4, m’ is 2 and n” is 1. In yet a further embodiment, m is 4, m’ is 3 and n” is 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 2. In a particular embodiment, m is 1, m’ is 1 and n’’ is 2. In a particular embodiment, m is 1, m’ is 2 and n’’ is 2. In a particular embodiment, m is 1, m’ is 3 and n’’ is 2. In another embodiment, m is 2, m’ is 0 and n’’ is 2. In another embodiment, m is 2, m’ is 1 and n’’ is 2. In another embodiment, m is 2, m’ is 2 and n’’ is 2. In another embodiment, m is 2, m’ is 3 and n’’ is 2. In yet another embodiment, m is 3, m’ is 0 and n’’ is 2. In yet another embodiment, m is 3, m’ is 1 and n’’ is 2. In yet another embodiment, m is 3, m’ is 2 and n’’ is 2. In yet another embodiment, m is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, m is 4, m’ is 0 and n” is 2. In yet a further embodiment, m is 4, m’ is 1 and n” is 2. In yet a further embodiment, m is 4, m’ is 2 and n” is 2. In yet a further embodiment, m is 4, m’ is 3 and n” is 2. In a particular embodiment, m is 1, m’ is 0 and n’’ is 3. In a particular embodiment, m is 1, m’ is 1 and n’’ is 3. In a particular embodiment, m is 1, m’ is 2 and n’’ is 3. In a particular embodiment, m is 1, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 0 and n’’ is 3. In another embodiment, m is 2, m’ is 1 and n’’ is 3. In another embodiment, m is 2, m’ is 2 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3. In yet another embodiment, m is 3, m’ is 0 and n’’ is 3. In yet another embodiment, m is 3, m’ is 1 and n’’ is 3. In yet another embodiment, m is 3, m’ is 2 and n’’ is 3. In yet another embodiment, m is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, m is 4, m’ is 0 and n” is 3. In yet a further embodiment, m is 4, m’ is 1 and n” is 3. In yet a further embodiment, m is 4, m’ is 2 and n” is 3. In yet a further embodiment, m is 4, m’ is 3 and n” is 3. In a particular embodiment, m is 1, m’ is 0 and n’’ is 4. In a particular embodiment, m is 1, m’ is 1 and n’’ is 4. In a particular embodiment, m is 1, m’ is 2 and n’’ is 4. In a particular embodiment, m is 1, m’ is 3 and n’’ is 4. In another embodiment, m is 2, m’ is 0 and n’’ is 4. In another embodiment, m is 2, m’ is 1 and n’’ is 4. In another embodiment, m is 2, m’ is 2 and n’’ is 4. In another embodiment, m is 2, m’ is 3 and n’’ is 4. In yet another embodiment, m is 3, m’ is 0 and n’’ is 4. In yet another embodiment, m is 3, m’ is 1 and n’’ is 4. In yet another embodiment, m is 3, m’ is 2 and n’’ is 4. In yet another embodiment, m is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, m is 4, m’ is 0 and n” is 4. In yet a further embodiment, m is 4, m’ is 1 and n” is 4. In yet a further embodiment, m is 4, m’ is 2 and n” is 4. In yet a further embodiment, m is 4, m’ is 3 and n” is 4. In a particular embodiment, m is 1, m’ is 0 and n’’ is 5. In a particular embodiment, m is 1, m’ is 1 and n’’ is 5. In a particular embodiment, m is 1, m’ is 2 and n’’ is 5. In a particular embodiment, m is 1, m’ is 3 and n’’ is 5. In another embodiment, m is 2, m’ is 0 and n’’ is 5. In another embodiment, m is 2, m’ is 1 and n’’ is 5. In another embodiment, m is 2, m’ is 2 and n’’ is 5. In another embodiment, m is 2, m’ is 3 and n’’ is 5. In yet another embodiment, m is 3, m’ is 0 and n’’ is 5. In yet another embodiment, m is 3, m’ is 1 and n’’ is 5. In yet another embodiment, m is 3, m’ is 2 and n’’ is 5. In yet another embodiment, m is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, m is 4, m’ is 0 and n” is 5. In yet a further embodiment, m is 4, m’ is 1 and n” is 5. In yet a further embodiment, m is 4, m’ is 2 and n” is 5. In yet a further embodiment, m is 4, m’ is 3 and n” is 5. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH2)n’, where n’ is selected from 1 to 10 and wherein X' is CH2(CH2)n”, where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH2)n’, where n’ is selected from 1 to 10 and wherein X' is CH2O(CH2)n’’CH2, where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH2)n’, where n’ is selected from 1 to 10 and wherein X' is CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. In an embodiment, n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 0. In another embodiment, n’ is 1, m’ is 1 and n’’ is 0. In another embodiment, n’ is 1, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In another embodiment, n’ is 2, m’ is 0 and n’’ is 0. In another embodiment, n’ is 2, m’ is 1 and n’’ is 0. In another embodiment, n’ is 2, m’ is 2 and n’’ is 0. In another embodiment, n’ is 2, m’ is 3 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 0.In yet a further embodiment, n’ is 5, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 0. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 1. In another embodiment, n’ is 2, m’ is 0 and n’’ is 1. In another embodiment, n’ is 2, m’ is 1 and n’’ is 1. In another embodiment, n’ is 2, m’ is 2 and n’’ is 1. In another embodiment, n’ is 2, m’ is 3 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 2. In another embodiment, n’ is 2, m’ is 0 and n’’ is 2. In another embodiment, n’ is 2, m’ is 1 and n’’ is 2. In another embodiment, n’ is 2, m’ is 2 and n’’ is 2. In another embodiment, n’ is 2, m’ is 3 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 2. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 3. In another embodiment, n’ is 2, m’ is 0 and n’’ is 3. In another embodiment, n’ is 2, m’ is 1 and n’’ is 3. In another embodiment, n’ is 2, m’ is 2 and n’’ is 3. In another embodiment, n’ is 2, m’ is 3 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 3. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 4. In another embodiment, n’ is 2, m’ is 0 and n’’ is 4. In another embodiment, n’ is 2, m’ is 1 and n’’ is 4. In another embodiment, n’ is 2, m’ is 2 and n’’ is 4. In another embodiment, n’ is 2, m’ is 3 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 4. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 5. In another embodiment, n’ is 2, m’ is 0 and n’’ is 5. In another embodiment, n’ is 2, m’ is 1 and n’’ is 5. In another embodiment, n’ is 2, m’ is 2 and n’’ is 5. In another embodiment, n’ is 2, m’ is 3 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 5. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2(CH2)n”, where n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2. In yet another embodiment, m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5. In another embodiment, m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2O(CH2)n’’CH2, where n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2. In yet another embodiment, m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5. In another embodiment, m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is NHCO(CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. In an embodiment, m is selected from 1 to 3, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 0. In another embodiment, m is 1, m’ is 1 and n’’ is 0. In another embodiment, m is 1, m’ is 2 and n’’ is 0. In another embodiment, m is 1, m’ is 3 and n’’ is 0. In another embodiment, m is 2, m’ is 0 and n’’ is 0. In another embodiment, m is 2, m’ is 1 and n’’ is 0. In another embodiment, m is 2, m’ is 2 and n’’ is 0. In another embodiment, m is 2, m’ is 3 and n’’ is 0. In yet another embodiment, m is 3, m’ is 0 and n’’ is 0. In yet another embodiment, m is 3, m’ is 1 and n’’ is 0. In yet another embodiment, m is 3, m’ is 2 and n’’ is 0. In yet another embodiment, m is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, m is 4, m’ is 0 and n” is 0. In yet a further embodiment, m is 4, m’ is 1 and n” is 0. In yet a further embodiment, m is 4, m’ is 2 and n” is 0. In yet a further embodiment, m is 4, m’ is 3 and n” is 0. In a particular embodiment, m is 1, m’ is 0 and n’’ is 1. In a particular embodiment, m is 1, m’ is 1 and n’’ is 1. In a particular embodiment, m is 1, m’ is 2 and n’’ is 1. In a particular embodiment, m is 1, m’ is 3 and n’’ is 1. In another embodiment, m is 2, m’ is 0 and n’’ is 1. In another embodiment, m is 2, m’ is 1 and n’’ is 1. In another embodiment, m is 2, m’ is 2 and n’’ is 1. In another embodiment, m is 2, m’ is 3 and n’’ is 1. In yet another embodiment, m is 3, m’ is 0 and n’’ is 1. In yet another embodiment, m is 3, m’ is 1 and n’’ is 1. In yet another embodiment, m is 3, m’ is 2 and n’’ is 1. In yet another embodiment, m is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, m is 4, m’ is 0 and n” is 1. In yet a further embodiment, m is 4, m’ is 1 and n” is 1. In yet a further embodiment, m is 4, m’ is 2 and n” is 1. In yet a further embodiment, m is 4, m’ is 3 and n” is 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 2. In a particular embodiment, m is 1, m’ is 1 and n’’ is 2. In a particular embodiment, m is 1, m’ is 2 and n’’ is 2. In a particular embodiment, m is 1, m’ is 3 and n’’ is 2. In another embodiment, m is 2, m’ is 0 and n’’ is 2. In another embodiment, m is 2, m’ is 1 and n’’ is 2. In another embodiment, m is 2, m’ is 2 and n’’ is 2. In another embodiment, m is 2, m’ is 3 and n’’ is 2. In yet another embodiment, m is 3, m’ is 0 and n’’ is 2. In yet another embodiment, m is 3, m’ is 1 and n’’ is 2. In yet another embodiment, m is 3, m’ is 2 and n’’ is 2. In yet another embodiment, m is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, m is 4, m’ is 0 and n” is 2. In yet a further embodiment, m is 4, m’ is 1 and n” is 2. In yet a further embodiment, m is 4, m’ is 2 and n” is 2. In yet a further embodiment, m is 4, m’ is 3 and n” is 2. In a particular embodiment, m is 1, m’ is 0 and n’’ is 3. In a particular embodiment, m is 1, m’ is 1 and n’’ is 3. In a particular embodiment, m is 1, m’ is 2 and n’’ is 3. In a particular embodiment, m is 1, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 0 and n’’ is 3. In another embodiment, m is 2, m’ is 1 and n’’ is 3. In another embodiment, m is 2, m’ is 2 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3. In yet another embodiment, m is 3, m’ is 0 and n’’ is 3. In yet another embodiment, m is 3, m’ is 1 and n’’ is 3. In yet another embodiment, m is 3, m’ is 2 and n’’ is 3. In yet another embodiment, m is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, m is 4, m’ is 0 and n” is 3. In yet a further embodiment, m is 4, m’ is 1 and n” is 3. In yet a further embodiment, m is 4, m’ is 2 and n” is 3. In yet a further embodiment, m is 4, m’ is 3 and n” is 3. In a particular embodiment, m is 1, m’ is 0 and n’’ is 4. In a particular embodiment, m is 1, m’ is 1 and n’’ is 4. In a particular embodiment, m is 1, m’ is 2 and n’’ is 4. In a particular embodiment, m is 1, m’ is 3 and n’’ is 4. In another embodiment, m is 2, m’ is 0 and n’’ is 4. In another embodiment, m is 2, m’ is 1 and n’’ is 4. In another embodiment, m is 2, m’ is 2 and n’’ is 4. In another embodiment, m is 2, m’ is 3 and n’’ is 4. In yet another embodiment, m is 3, m’ is 0 and n’’ is 4. In yet another embodiment, m is 3, m’ is 1 and n’’ is 4. In yet another embodiment, m is 3, m’ is 2 and n’’ is 4. In yet another embodiment, m is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, m is 4, m’ is 0 and n” is 4. In yet a further embodiment, m is 4, m’ is 1 and n” is 4. In yet a further embodiment, m is 4, m’ is 2 and n” is 4. In yet a further embodiment, m is 4, m’ is 3 and n” is 4. In a particular embodiment, m is 1, m’ is 0 and n’’ is 5. In a particular embodiment, m is 1, m’ is 1 and n’’ is 5. In a particular embodiment, m is 1, m’ is 2 and n’’ is 5. In a particular embodiment, m is 1, m’ is 3 and n’’ is 5. In another embodiment, m is 2, m’ is 0 and n’’ is 5. In another embodiment, m is 2, m’ is 1 and n’’ is 5. In another embodiment, m is 2, m’ is 2 and n’’ is 5. In another embodiment, m is 2, m’ is 3 and n’’ is 5. In yet another embodiment, m is 3, m’ is 0 and n’’ is 5. In yet another embodiment, m is 3, m’ is 1 and n’’ is 5. In yet another embodiment, m is 3, m’ is 2 and n’’ is 5. In yet another embodiment, m is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, m is 4, m’ is 0 and n” is 5. In yet a further embodiment, m is 4, m’ is 1 and n” is 5. In yet a further embodiment, m is 4, m’ is 2 and n” is 5. In yet a further embodiment, m is 4, m’ is 3 and n” is 5. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is OCH2(CH2)n’, where n’ is selected from 1 to 10 and wherein X' is CH2(CH2)n”, where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is OCH2(CH2)n’, where n’ is selected from 1 to 10 and wherein X' is CH2O(CH2)n’’CH2, where n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, n’ is 1 and n’’ is 0. In another embodiment, n’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4 and n” is 0. In yet a further embodiment, n’ is 5 and n” is 0. In yet a further embodiment, n’ is 6 and n” is 0. In a particular embodiment, n’ is 1 and n’’ is 1. In another embodiment, n’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4 and n” is 1. In yet a further embodiment, n’ is 5 and n” is 1. In yet a further embodiment, n’ is 6 and n” is 1. In a particular embodiment, n’ is 1 and n’’ is 2. In another embodiment, n’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4 and n” is 2. In yet a further embodiment, n’ is 5 and n” is 2. In yet a further embodiment, n’ is 6 and n” is 2. In a particular embodiment, n’ is 1 and n’’ is 3. In another embodiment, n’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4 and n” is 3. In yet a further embodiment, n’ is 5 and n” is 3. In yet a further embodiment, n’ is 6 and n” is 3. In a particular embodiment, n’ is 1 and n’’ is 4. In another embodiment, n’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4 and n” is 4. In yet a further embodiment, n’ is 5 and n” is 4. In yet a further embodiment, n’ is 6 and n” is 4. In a particular embodiment, n’ is 1 and n’’ is 5. In another embodiment, n’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4 and n” is 5. In yet a further embodiment, n’ is 5 and n” is 5. In yet a further embodiment, n’ is 6 and n” is 5. In a particular embodiment, n’ is 1 and n’’ is 6. In another embodiment, n’ is 2 and n’’ is 6. In yet another embodiment, n’ is 3 and n’’ is 6. In yet a further embodiment, n’ is 4 and n” is 6. In yet a further embodiment, n’ is 5 and n” is 6. In yet a further embodiment, n’ is 6 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is OCH2(CH2)n’, where n’ is selected from 1 to 10 and wherein X' is CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. In an embodiment, n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, n’ is selected from 1 to 5, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, n’ is selected from 1 to 3, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, n’ is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 0. In another embodiment, n’ is 1, m’ is 1 and n’’ is 0. In another embodiment, n’ is 1, m’ is 2 and n’’ is 0. In another embodiment, n’ is 1, m’ is 3 and n’’ is 0. In another embodiment, n’ is 2, m’ is 0 and n’’ is 0. In another embodiment, n’ is 2, m’ is 1 and n’’ is 0. In another embodiment, n’ is 2, m’ is 2 and n’’ is 0. In another embodiment, n’ is 2, m’ is 3 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 0. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 0.In yet a further embodiment, n’ is 5, m’ is 2 and n” is 0. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 0. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 1. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 1. In another embodiment, n’ is 2, m’ is 0 and n’’ is 1. In another embodiment, n’ is 2, m’ is 1 and n’’ is 1. In another embodiment, n’ is 2, m’ is 2 and n’’ is 1. In another embodiment, n’ is 2, m’ is 3 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 1. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 1. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 1. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 2. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 2. In another embodiment, n’ is 2, m’ is 0 and n’’ is 2. In another embodiment, n’ is 2, m’ is 1 and n’’ is 2. In another embodiment, n’ is 2, m’ is 2 and n’’ is 2. In another embodiment, n’ is 2, m’ is 3 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 2. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 2. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 2. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 3. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 3. In another embodiment, n’ is 2, m’ is 0 and n’’ is 3. In another embodiment, n’ is 2, m’ is 1 and n’’ is 3. In another embodiment, n’ is 2, m’ is 2 and n’’ is 3. In another embodiment, n’ is 2, m’ is 3 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 3. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 3. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 3. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 4. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 4. In another embodiment, n’ is 2, m’ is 0 and n’’ is 4. In another embodiment, n’ is 2, m’ is 1 and n’’ is 4. In another embodiment, n’ is 2, m’ is 2 and n’’ is 4. In another embodiment, n’ is 2, m’ is 3 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 4. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 4. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 4. In a particular embodiment, n’ is 1, m’ is 0 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 1 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 2 and n’’ is 5. In a particular embodiment, n’ is 1, m’ is 3 and n’’ is 5. In another embodiment, n’ is 2, m’ is 0 and n’’ is 5. In another embodiment, n’ is 2, m’ is 1 and n’’ is 5. In another embodiment, n’ is 2, m’ is 2 and n’’ is 5. In another embodiment, n’ is 2, m’ is 3 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 0 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 1 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 2 and n’’ is 5. In yet another embodiment, n’ is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, n’ is 4, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 4, m’ is 3 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 0 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 1 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 2 and n” is 5. In yet a further embodiment, n’ is 5, m’ is 3 and n” is 5. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is O(CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2(CH2)n”, where n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2. In yet another embodiment, m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5. In another embodiment, m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is O(CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2O(CH2)n’’CH2, where n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 3 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2 and n’’ is selected from 0 to 2. In a particular embodiment, m is 1 and n’’ is 0. In another embodiment, m is 2 and n’’ is 0. In yet another embodiment, m is 3 and n’’ is 0. In yet a further embodiment, m is 4 and n” is 0. In a particular embodiment, m is 1 and n’’ is 1. In another embodiment, m is 2 and n’’ is 1. In yet another embodiment, m is 3 and n’’ is 1. In yet a further embodiment, m is 4 and n” is 1. In a particular embodiment, m is 1 and n’’ is 2. In another embodiment, m is 2 and n’’ is 2. In yet another embodiment, m is 3 and n’’ is 2. In yet a further embodiment, m is 4 and n” is 2. In a particular embodiment, m is 1 and n’’ is 3. In another embodiment, m is 2 and n’’ is 3. In yet another embodiment, m is 3 and n’’ is 3. In yet a further embodiment, m is 4 and n” is 3. In a particular embodiment, m is 1 and n’’ is 4. In another embodiment, m is 2 and n’’ is 4. In yet another embodiment, m is 3 and n’’ is 4. In yet a further embodiment, m is 4 and n” is 4. In a particular embodiment, m is 1 and n’’ is 5. In another embodiment, m is 2 and n’’ is 5. In yet another embodiment, m is 3 and n’’ is 5. In yet a further embodiment, m is 4 and n” is 5. In a particular embodiment, m is 1 and n’’ is 6. In another embodiment, m is 2 and n’’ is 6. In yet another embodiment, m is 3 and n’’ is 6. In yet a further embodiment, m is 4 and n” is 6. In an embodiment, the invention provides a glycoconjugate comprising a saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is O(CH2CH2O)mCH2CH2, where m is selected from 1 to 4 and wherein X' is CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. In an embodiment, m is selected from 1 to 3, m’ is selected from 0 to 4 and n’’ is selected from 0 to 10. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 4 and n’’ is selected from 0 to 5. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 3. In an embodiment, m is selected from 1 to 2, m’ is selected from 0 to 2 and n’’ is selected from 0 to 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 0. In another embodiment, m is 1, m’ is 1 and n’’ is 0. In another embodiment, m is 1, m’ is 2 and n’’ is 0. In another embodiment, m is 1, m’ is 3 and n’’ is 0. In another embodiment, m is 2, m’ is 0 and n’’ is 0. In another embodiment, m is 2, m’ is 1 and n’’ is 0. In another embodiment, m is 2, m’ is 2 and n’’ is 0. In another embodiment, m is 2, m’ is 3 and n’’ is 0. In yet another embodiment, m is 3, m’ is 0 and n’’ is 0. In yet another embodiment, m is 3, m’ is 1 and n’’ is 0. In yet another embodiment, m is 3, m’ is 2 and n’’ is 0. In yet another embodiment, m is 3, m’ is 3 and n’’ is 0. In yet a further embodiment, m is 4, m’ is 0 and n” is 0. In yet a further embodiment, m is 4, m’ is 1 and n” is 0. In yet a further embodiment, m is 4, m’ is 2 and n” is 0. In yet a further embodiment, m is 4, m’ is 3 and n” is 0. In a particular embodiment, m is 1, m’ is 0 and n’’ is 1. In a particular embodiment, m is 1, m’ is 1 and n’’ is 1. In a particular embodiment, m is 1, m’ is 2 and n’’ is 1. In a particular embodiment, m is 1, m’ is 3 and n’’ is 1. In another embodiment, m is 2, m’ is 0 and n’’ is 1. In another embodiment, m is 2, m’ is 1 and n’’ is 1. In another embodiment, m is 2, m’ is 2 and n’’ is 1. In another embodiment, m is 2, m’ is 3 and n’’ is 1. In yet another embodiment, m is 3, m’ is 0 and n’’ is 1. In yet another embodiment, m is 3, m’ is 1 and n’’ is 1. In yet another embodiment, m is 3, m’ is 2 and n’’ is 1. In yet another embodiment, m is 3, m’ is 3 and n’’ is 1. In yet a further embodiment, m is 4, m’ is 0 and n” is 1. In yet a further embodiment, m is 4, m’ is 1 and n” is 1. In yet a further embodiment, m is 4, m’ is 2 and n” is 1. In yet a further embodiment, m is 4, m’ is 3 and n” is 1. In a particular embodiment, m is 1, m’ is 0 and n’’ is 2. In a particular embodiment, m is 1, m’ is 1 and n’’ is 2. In a particular embodiment, m is 1, m’ is 2 and n’’ is 2. In a particular embodiment, m is 1, m’ is 3 and n’’ is 2. In another embodiment, m is 2, m’ is 0 and n’’ is 2. In another embodiment, m is 2, m’ is 1 and n’’ is 2. In another embodiment, m is 2, m’ is 2 and n’’ is 2. In another embodiment, m is 2, m’ is 3 and n’’ is 2. In yet another embodiment, m is 3, m’ is 0 and n’’ is 2. In yet another embodiment, m is 3, m’ is 1 and n’’ is 2. In yet another embodiment, m is 3, m’ is 2 and n’’ is 2. In yet another embodiment, m is 3, m’ is 3 and n’’ is 2. In yet a further embodiment, m is 4, m’ is 0 and n” is 2. In yet a further embodiment, m is 4, m’ is 1 and n” is 2. In yet a further embodiment, m is 4, m’ is 2 and n” is 2. In yet a further embodiment, m is 4, m’ is 3 and n” is 2. In a particular embodiment, m is 1, m’ is 0 and n’’ is 3. In a particular embodiment, m is 1, m’ is 1 and n’’ is 3. In a particular embodiment, m is 1, m’ is 2 and n’’ is 3. In a particular embodiment, m is 1, m’ is 3 and n’’ is 3. In another embodiment, m is 2, m’ is 0 and n’’ is 3. In another embodiment, m is 2, m’ is 1 and n’’ is 3. In another embodiment, m is 2, m’ is 2 and n’’ is 3. In another embodiment, m is 2, m’ is 3 and n’’ is 3. In yet another embodiment, m is 3, m’ is 0 and n’’ is 3. In yet another embodiment, m is 3, m’ is 1 and n’’ is 3. In yet another embodiment, m is 3, m’ is 2 and n’’ is 3. In yet another embodiment, m is 3, m’ is 3 and n’’ is 3. In yet a further embodiment, m is 4, m’ is 0 and n” is 3. In yet a further embodiment, m is 4, m’ is 1 and n” is 3. In yet a further embodiment, m is 4, m’ is 2 and n” is 3. In yet a further embodiment, m is 4, m’ is 3 and n” is 3. In a particular embodiment, m is 1, m’ is 0 and n’’ is 4. In a particular embodiment, m is 1, m’ is 1 and n’’ is 4. In a particular embodiment, m is 1, m’ is 2 and n’’ is 4. In a particular embodiment, m is 1, m’ is 3 and n’’ is 4. In another embodiment, m is 2, m’ is 0 and n’’ is 4. In another embodiment, m is 2, m’ is 1 and n’’ is 4. In another embodiment, m is 2, m’ is 2 and n’’ is 4. In another embodiment, m is 2, m’ is 3 and n’’ is 4. In yet another embodiment, m is 3, m’ is 0 and n’’ is 4. In yet another embodiment, m is 3, m’ is 1 and n’’ is 4. In yet another embodiment, m is 3, m’ is 2 and n’’ is 4. In yet another embodiment, m is 3, m’ is 3 and n’’ is 4. In yet a further embodiment, m is 4, m’ is 0 and n” is 4. In yet a further embodiment, m is 4, m’ is 1 and n” is 4. In yet a further embodiment, m is 4, m’ is 2 and n” is 4. In yet a further embodiment, m is 4, m’ is 3 and n” is 4. In a particular embodiment, m is 1, m’ is 0 and n’’ is 5. In a particular embodiment, m is 1, m’ is 1 and n’’ is 5. In a particular embodiment, m is 1, m’ is 2 and n’’ is 5. In a particular embodiment, m is 1, m’ is 3 and n’’ is 5. In another embodiment, m is 2, m’ is 0 and n’’ is 5. In another embodiment, m is 2, m’ is 1 and n’’ is 5. In another embodiment, m is 2, m’ is 2 and n’’ is 5. In another embodiment, m is 2, m’ is 3 and n’’ is 5. In yet another embodiment, m is 3, m’ is 0 and n’’ is 5. In yet another embodiment, m is 3, m’ is 1 and n’’ is 5. In yet another embodiment, m is 3, m’ is 2 and n’’ is 5. In yet another embodiment, m is 3, m’ is 3 and n’’ is 5. In yet a further embodiment, m is 4, m’ is 0 and n” is 5. In yet a further embodiment, m is 4, m’ is 1 and n” is 5. In yet a further embodiment, m is 4, m’ is 2 and n” is 5. In yet a further embodiment, m is 4, m’ is 3 and n” is 5. 1.5 Carrier protein of the glycoconjugates of the invention A component of the glycoconjugate is a carrier protein to which the saccharide is conjugated. The terms "protein carrier" or "carrier protein" or “carrier” may be used interchangeably herein. Carrier proteins should be amenable to standard conjugation procedures. In a preferred embodiment, the carrier protein of the glycoconjugate of the invention is selected in the group consisting of: DT (Diphtheria toxoid), TT (tetanus toxoid) or fragment C of TT, CRM197 (a nontoxic but antigenically identical variant of diphtheria toxin), other DT mutants (such as CRM176, CRM228, CRM45 (Uchida et al. (1973) J. Biol. Chem.218:3838-3844), CRM9, CRM102, CRM103 or CRM107; and other mutations described by Nicholls and Youle in Genetically Engineered Toxins, Ed: Frankel, Maecel Dekker Inc. (1992); deletion or mutation of Glu-148 to Asp, Gln or Ser and/or Ala 158 to GIy and other mutations disclosed in U.S. Patent Nos.4,709,017 and 4,950,740; mutation of at least one or more residues Lys 516, Lys 526, Phe 530 and/or Lys 534 and other mutations disclosed in U.S. Patent Nos. 5,917,017 and 6,455,673; or fragment disclosed in U.S. Patent No.5,843,711, pneumococcal pneumolysin (ply) (Kuo et al. (1995) Infect lmmun 63:2706-2713) including ply detoxified in some fashion, for example dPLY-GMBS (WO 2004/081515, WO 2006/032499) or dPLY-formol, PhtX, including PhtA, PhtB, PhtD, PhtE (sequences of PhtA, PhtB, PhtD or PhtE are disclosed in WO 00/37105 and WO 00/39299) and fusions of Pht proteins, for example PhtDE fusions, PhtBE fusions, Pht A-E (WO 01/98334, WO 03/054007, WO 2009/000826), OMPC (meningococcal outer membrane protein), which is usually extracted from Neisseria meningitidis serogroup B (EP0372501), PorB (from N. meningitidis), PD (Haemophilus influenzae protein D; see, e.g., EP0594610 B), or immunologically functional equivalents thereof, synthetic peptides (EP0378881, EP0427347), heat shock proteins (WO 93/17712, WO 94/03208), pertussis proteins (WO 98/58668, EP0471177), cytokines, lymphokines, growth factors or hormones (WO 91/01146), artificial proteins comprising multiple human CD4+ T cell epitopes from various pathogen derived antigens (Falugi et al. (2001) Eur J Immunol 31:3816-3824) such as N19 protein (Baraldoi et al. (2004) Infect lmmun 72:4884-4887) pneumococcal surface protein PspA (WO 02/091998), iron uptake proteins (WO 01/72337), toxin A or B of Clostridioides difficile (formally Clostridium difficile) (WO 00/61761), transferrin binding proteins, pneumococcal adhesion protein (PsaA), recombinant Pseudomonas aeruginosa exotoxin A (in particular non-toxic mutants thereof (such as exotoxin A bearing a substution at glutamic acid 553 (Douglas et al. (1987) J. Bacteriol.169(11):4967-4971)). Other proteins, such as ovalbumin, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA) or purified protein derivative of tuberculin (PPD) also can be used as carrier proteins. Other suitable carrier proteins include inactivated bacterial toxins such as cholera toxoid (e.g., as described in WO 2004/083251), Escherichia coli LT, E. coli ST, and exotoxin A from P. aeruginosa. Another suitable carrier protein is a C5a peptidase from Streptococcus (SCP). In a preferred embodiment, the carrier protein of the glycoconjugate of the invention is TT, DT, DT mutants (such as CRM197) or a C5a peptidase from Streptococcus (SCP). In an embodiment, the carrier protein of the glycoconjugate of the invention is DT (Diphtheria toxoid). In another embodiment, the carrier protein of the glycoconjugate of the invention is TT (tetanus toxoid). In another embodiment, the carrier protein of the glycoconjugate of the invention is PD (H. influenzae protein D; see, e.g., EP0594610 B). In a preferred embodiment, the carrier protein of the glycoconjugate of the invention is CRM197 or a C5a peptidase from Streptococcus (SCP). In a preferred embodiment, the carrier protein of the glycoconjugate of the invention is CRM197. The CRM197 protein is a nontoxic form of diphtheria toxin but is immunologically indistinguishable from the diphtheria toxin. CRM197 is produced by Corynebacterium diphtheriae infected by the nontoxigenic phage β197tox- created by nitrosoguanidine mutagenesis of the toxigenic corynephage beta (Uchida et al. (1971) Nature New Biology 233:8-11). The CRM197 protein has the same molecular weight as the diphtheria toxin but differs therefrom by a single base change (guanine to adenine) in the structural gene. This single base change causes an amino acid substitution (glutamic acid for glycine) in the mature protein and eliminates the toxic properties of diphtheria toxin. The CRM197 protein is a safe and effective T-cell dependent carrier for saccharides. Further details about CRM197 and production thereof can be found, e.g., in U.S. Patent No.5,614,382. In an embodiment, the carrier protein of the glycoconjugate of the invention is the A chain of CRM197 (see CN103495161). In an embodiment, the carrier protein of the glycoconjugate of the invention is the A chain of CRM197 obtained via expression by genetically recombinant E. coli (see CN103495161). In other preferred embodiments, the carrier protein of the glycoconjugate of the invention is SCP (Streptococcal C5a Peptidase). Two important species of β-hemolytic streptococci, Streptococcus pyogenes (group A Streptococcus, GAS) and Streptococcus agalactiae (group B Streptococcus, GBS), which cause a variety of serious human infections that range from mild cases of pharyngitis and impetigo to serious invasive diseases such as necrotizing fasciitis (GAS) and neonatal sepsis (GBS) have developed a way to defeat this immune response. All human isolates of β-hemolytic streptococci, including GAS and GBS, produce a highly conserved cell- wall protein SCP (Streptococcal C5a Peptidase) that specifically inactivates C5a. The scp genes from GAS and GBS encode a polypeptide containing between 1,134 and 1,181 amino acids (Brown et al., PNAS, 2005, vol.102, no.51 pages 18391–18396). The first 31 residues are the export signal presequence and are removed upon passing through the cytoplasmic membrane. The next 68 residues serve as a pro-sequence and must be removed to produce active SCP. The next 10 residues can be removed without loss of protease activity. At the other end, starting with Lys-1034, are four consecutive 17-residue motifs followed by a cell sorting and cell-wall attachment signal. This combined signal is composed of a 20-residue hydrophilic sequence containing an LPTTND sequence, a 17-residue hydrophobic sequence, and a short basic carboxyl terminus. SCP can be divided in domains (see figure 1B of Brown et al., PNAS, 2005, vol.102, no. 51 pages 18391–18396). These domains are the Pre/Pro domain (which comprises the export signal presequence (commonly the first 31 residues) and the pro-sequence (commonly the next 68 residues)), the protease domain (which is splitted in two part (protease part 1 commonly residues 89–333/334 and protease domain part 2 and commonly residues 467/468–583/584), the protease-associated domain (PA domain) (commonly residues 333/334–467/468), three fibronectin type III (Fn) domains (Fn1, commonly residues 583/584–712/713; Fn2, commonly residues 712/713–928/929/930; commonly Fn3, residues 929/930-1029/1030/1031) and a cell wall anchor domain (commonly residues 1029/1030/1031 to the C-terminus). In an embodiment, the carrier protein of the glycoconjugate of the invention is an SCP from GBS (SCPB). An example of SCPB is provided at SEQ ID.NO: 3 of WO97/26008. See also SEQ ID NO: 3 of WO00/34487. In another preferred embodiments, the carrier protein of the glycoconjugate of the invention is an SCP from GAS (SCPA). Examples of SCPA can be found at SEQ ID.No.1 and SEQ ID.No.2 of WO97/26008. See also SEQ ID NO: 1, 2 and 23 of WO00/34487. In a preferred embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP. In other preferred embodiments, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP from GBS (SCPB). In another preferred embodiments, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP from GAS (SCPA). In an embodiment, the carrier protein of the glycoconjugate of the invention is a fragment of an SCP. In an embodiment, the carrier protein of the glycoconjugate of the invention is a fragment of an SCPA. Preferably, the carrier protein of the glycoconjugate of the invention is a fragment of an SCPB. In an embodiment, the carrier protein of the glycoconjugate of the invention is a fragment of an SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. In an embodiment, the carrier protein of the glycoconjugate of the invention is a fragment of an SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP which comprises the protease domain, the protease- associated domain (PA domain) and two of the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP. In an embodiment, said enzymatically inactive fragment of SCP comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCPA. In an embodiment, said enzymatically inactive fragment of an SCPA comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. In a preferred embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB. Preferably, said enzymatically inactive fragment of SCPB comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. In an embodiment, the enzymatic activity of SCP is inactivated by replacing at least one amino acid of the wild type sequence. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. The numbers indicate the amino acid residue position in the peptidase according to the numbering of SEQ ID NO: 1 of WO00/34487. Therefore, in an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence. Preferably, said replacement of at least one amino acid is in the protease domain. In an embodiment, said replacement of at least one amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said replacement is selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said replacement is D130A. In another embodiment, said replacement is H193A. In another embodiment, said replacement is N295A. In yet another embodiment, said replacement is S512A. In an embodiment, the enzymatic activity of SCP is inactivated by replacing at least two amino acids of the wild type sequence. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. In an embodiment, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. Therefore, in an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least two amino acid is in part 2 of the protease domain. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least two amino acid is in part 2 of the protease domain. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least two amino acid is in part 2 of the protease domain. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least two amino acid is in part 2 of the protease domain. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least two amino acid is in part 2 of the protease domain. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence. Preferably, said replacement of at least two amino acids is in the protease domain. In an embodiment, said replacement of at least two amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least two amino acids is in part 2 of the protease domain. In an embodiment, said at least two amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least two amino acids replacements are D130A and H193A. In an embodiment, said at least two amino acids replacements are D130A and N295A. Preferably, said at least two amino acids replacements are D130A and S512A. In an embodiment, said at least two amino acids replacements are H193A and N295A. In an embodiment, said at least two amino acids replacements are H193A and S512A. In an embodiment, said at least two amino acids replacements are N295A and S512A. In an embodiment, the enzymatic activity of SCP is inactivated by replacing at least three amino acids of the wild type sequence. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. Therefore, in an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence. Preferably, said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least three amino acid is in part 2 of the protease domain. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence. Preferably, said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least three amino acid is in part 2 of the protease domain. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence. Preferably, said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least three amino acid is in part 2 of the protease domain. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence. Preferably, said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least three amino acid is in part 2 of the protease domain. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence. Preferably, said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least three amino acid is in part 2 of the protease domain. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence. Preferably, said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least three amino acids is in part 2 of the protease domain. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least three amino acids of the wild type sequence. Preferably, said replacement of at least three amino acids is in the protease domain. In an embodiment, said replacement of at least three amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least three amino acids is in part 2 of the protease domain. In an embodiment, said at least three amino acids replacements are selected from the group consisting of D130A, H193A, N295A and S512A. In an embodiment, said at least three amino acids replacements are D130A, H193A and N295A. In an embodiment, said at least three amino acids replacements are D130A, H193A and S512A. In an embodiment, said at least three amino acids replacements are D130A, N295A and S512A. In an embodiment, said at least three amino acids replacements are H193A, N295A and S512A. In an embodiment, the enzymatic activity of SCP is inactivated by replacing at least four amino acids of the wild type sequence. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A. Therefore, in an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCP where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acid is in part 1 of the protease domain. In an embodiment, said replacement of at least four amino acid is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPA where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least four amino acid is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive SCPB where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least four amino acid is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of an SCP where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least four amino acid is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least four amino acid is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPA which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least one amino acid is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A In an embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCPB which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least four amino acids of the wild type sequence. Preferably, said replacement of at least four amino acids is in the protease domain. In an embodiment, said replacement of at least four amino acids is in part 1 of the protease domain. In an embodiment, said replacement of at least four amino acids is in part 2 of the protease domain. In an embodiment, said at least four amino acids replacements are D130A, H193A, N295A and S512A In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which consists of SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP which consists of SEQ ID NO: 42. SEQ ID NO: 41 : MAKTADTPATSKATIRDLNDPSQVKTLQEKAGKGAGTVVAVIAAGFDKNH EAWRLTDKAKARYQSKEDLEKAKKEHGITYGEWVNDKVAYYHDYSKDGKT AVDQEHGTHVSGILSGNAPSETKEPYRLEGAMPEAQLLLMRVEIVNGLAD YARNYAQAIRDAINLGAKVINMSFGNAALAYANLPDETKKAFDYAKSKGV SIVTSAGNDSSFGGKTRLPLADHPDYGVVGTPAAADSTLTVASYSPDKQL TETVTVKTADQQDKEMPVLSTNRFEPNKAYDYAYANRGTKEDDFKDVKGK IALIERGDIDFKDKIAKAKKAGAVGVLIYDNQDKGFPIELPNVDQMPAAF ISRKDGLLLKDNPQKTITFNATPKVLPTASGTKLSRFSSWGLTADGNIKP DIAAPGQDILSSVANNKYAKLSGTAMSAPLVAGIMGLLQEQYETQYPDMT PSERLDLAKKVLMSSATALYDEDEKAYFSPRQQGAGAVDAKKASAATMYV TDKDNTSSKVHLNNVSDKFEVTVTVHNKSDKPQELYYQATVQTDKVDGKH FALAPKALYETSWQKITIPANSSKQVTVPIDASRFSKDLLAQMKNGYFLE GFVRFKQDPKKEELMSIPYIGFRGDFGNLSALEKPIYDSKDGSSYYHEAN SDAKDQLDGDGLQFYALKNNFTALTTESNPWTIIKAVKEGVENIEDIESS EITETIFAGTFAKQDDDSHYYIHRHANGKPYAAISPNGDGNRDYVQFQGT FLRNAKNLVAEVLDKEGNVVWTSEVTEQVVKNYNNDLASTLGSTRFEKTR WDGKDKDGKVVANGTYTYRVRYTPISSGAKEQHTDFDVIVDNTTPEVATS ATFSTEDRRLTLASKPKTSQPVYRERIAYTYMDEDLPTTEYISPNEDGTF TLPEEAETMEGATVPLKMSDFTYVVEDMAGNITYTPVTKLLEGHSNKPEQ SEQ ID NO: 41 is 950 amino acids long. SEQ ID NO: 42 : AKTADTPATSKATIRDLNDPSQVKTLQEKAGKGAGTVVAVIAAGFDKNH EAWRLTDKAKARYQSKEDLEKAKKEHGITYGEWVNDKVAYYHDYSKDGKT AVDQEHGTHVSGILSGNAPSETKEPYRLEGAMPEAQLLLMRVEIVNGLAD YARNYAQAIRDAINLGAKVINMSFGNAALAYANLPDETKKAFDYAKSKGV SIVTSAGNDSSFGGKTRLPLADHPDYGVVGTPAAADSTLTVASYSPDKQL TETVTVKTADQQDKEMPVLSTNRFEPNKAYDYAYANRGTKEDDFKDVKGK IALIERGDIDFKDKIAKAKKAGAVGVLIYDNQDKGFPIELPNVDQMPAAF ISRKDGLLLKDNPQKTITFNATPKVLPTASGTKLSRFSSWGLTADGNIKP DIAAPGQDILSSVANNKYAKLSGTAMSAPLVAGIMGLLQEQYETQYPDMT PSERLDLAKKVLMSSATALYDEDEKAYFSPRQQGAGAVDAKKASAATMYV TDKDNTSSKVHLNNVSDKFEVTVTVHNKSDKPQELYYQATVQTDKVDGKH FALAPKALYETSWQKITIPANSSKQVTVPIDASRFSKDLLAQMKNGYFLE GFVRFKQDPKKEELMSIPYIGFRGDFGNLSALEKPIYDSKDGSSYYHEAN SDAKDQLDGDGLQFYALKNNFTALTTESNPWTIIKAVKEGVENIEDIESS EITETIFAGTFAKQDDDSHYYIHRHANGKPYAAISPNGDGNRDYVQFQGT FLRNAKNLVAEVLDKEGNVVWTSEVTEQVVKNYNNDLASTLGSTRFEKTR WDGKDKDGKVVANGTYTYRVRYTPISSGAKEQHTDFDVIVDNTTPEVATS ATFSTEDRRLTLASKPKTSQPVYRERIAYTYMDEDLPTTEYISPNEDGTF TLPEEAETMEGATVPLKMSDFTYVVEDMAGNITYTPVTKLLEGHSNKPEQ SEQ ID NO: 42 is 949 amino acids long. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 90% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.5% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.8% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.85% identity with SEQ ID NO: 41. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 90% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.5% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.8% identity with SEQ ID NO: 42. In a particular embodiment, the carrier protein of the glycoconjugate of the invention is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 99.85% identity with SEQ ID NO: 42. 2 Immunogenic compositions 2.1 Combinations of glycoconjugates of the invention In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention (as disclosed at section 1 above). In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising from 1 to 25 different glycoconjugates. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising from 26 to 35 different glycoconjugates. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising from 1 to 25 glycoconjugates from different serotypes of S. pneumoniae (1 to 25 pneumococcal conjugates). In one embodiment the invention relates to an immunogenic composition comprising glycoconjugates from 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 different serotypes of S. pneumoniae. In one embodiment the immunogenic composition comprises glycoconjugates from 16 or 20 different serotypes of S. pneumoniae. In an embodiment the immunogenic composition is a 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20-valent pneumococcal conjugate compositions. In an embodiment the immunogenic composition is a 14, 15, 16, 17, 18 or 19-valent pneumococcal conjugate compositions. In an embodiment the immunogenic composition is a 16-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 19- valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 20-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 21, 22, 23, 24 or 25-valent pneumococcal conjugate compositions. In an embodiment the immunogenic composition is a 21- valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 25-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35-valent pneumococcal conjugate compositions. In an embodiment the immunogenic composition is a 26-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 27-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 28-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 29-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 30-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 31-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 32- valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 33-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 34-valent pneumococcal conjugate composition. In an embodiment the immunogenic composition is a 35-valent pneumococcal conjugate composition. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F. In an embodiment the immunogenic composition is an 11-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F. In an embodiment the immunogenic composition is a 13-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and further comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 15-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 20-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and further comprising glycoconjugates from S. pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 2, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 2, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F and 33F. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F and 33F. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F and 33F. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and further comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F and 33F. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F and 33F. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F and 33F. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F and 33F. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 33F and 35B. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F and 35B. In an embodiment the immunogenic composition is a 25-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising twenty one glycoconjugates selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the immunogenic composition is a 22-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising at least one glycoconjugate selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising twenty two glycoconjugates selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising twenty three glycoconjugates selected from the group consisting of glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the immunogenic composition is a 24-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 2, 9N, 15A, 17F, 20, 23A, 23B, 24F and 35B. In an embodiment the immunogenic composition is a 10-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 2, 9N, 15A, 17F, 19A, 19F, 20, 23A, 23B, 24F and 35B. In an embodiment the immunogenic composition is a 12-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the immunogenic composition is a 23-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 2, 7C, 9N, 10B, 15A, 16F, 17F, 19A, 19F, 20, 21, 22A, 23A, 23B, 24B, 24F, 27, 29, 31, 33B, 34, 35B, 35F and 38. In an embodiment the immunogenic composition is a 25-valent pneumococcal conjugate compositions. In a preferred embodiment, the saccharides are each individually conjugated to different molecules of the protein carrier (each molecule of protein carrier only having one type of saccharide conjugated to it). In said embodiment, the capsular saccharides are said to be individually conjugated to the carrier protein. Preferably, all the glycoconjugates of the above immunogenic compositions are individually conjugated to the carrier protein. In an embodiment of any of the above immunogenic compositions, the glycoconjugate of the invention is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate of the invention is conjugated to SCP. In an embodiment of any of the above immunogenic compositions, the glycoconjugate from S. pneumoniae serotype 22F is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate from S. pneumoniae serotype 33F is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate from S. pneumoniae serotype 15B is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate from S. pneumoniae serotype 12F is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate from S. pneumoniae serotype 10A is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate from S. pneumoniae serotype 11A is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate from S. pneumoniae serotype 8 is conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugates from S. pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F and 23F are conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugates from S. pneumoniae serotypes 1, 5 and 7F are conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugates from S. pneumoniae serotypes 6A and 19A are conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugates of any of the above immunogenic compositions are all individually conjugated to CRM197. In an embodiment of any of the above immunogenic compositions, the glycoconjugate of the invention is conjugated to SCP and the other glycoconjugate(s) is/are all individually conjugated to CRM197. In an embodiment, the glycoconjugates from S. pneumoniae serotypes 1, 4, 5, 6B, 7F, 9V, 14 and/or 23F of any of the above immunogenic compositions are individually conjugated to PD. In an embodiment, the glycoconjugate from S. pneumoniae serotype 18C of any of the above immunogenic compositions is conjugated to TT. In an embodiment, the glycoconjugate from S. pneumoniae serotype 19F of any of the above immunogenic compositions is conjugated to DT. In an embodiment, the glycoconjugates from S. pneumoniae serotypes 1, 4, 5, 6B, 7F, 9V, 14 and/or 23F of any of the above immunogenic compositions are individually conjugated to PD, the glycoconjugate from S. pneumoniae serotype 18C is conjugated to TT and the glycoconjugate from S. pneumoniae serotype 19F is conjugated to DT. In an embodiment the above immunogenic compositions comprise from 8 to 25 different serotypes of S. pneumoniae. In an embodiment the above immunogenic compositions comprise from 26 to 35 different serotypes of S. pneumoniae. In an embodiment, the glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 18C, 19A, 19F, 23F and 33F of any of the above immunogenic compositions are individually conjugated to CRM197, the glycoconjugates from S. pneumoniae serotypes 1, 5, 15B and 22F are conjugated to TT. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. In an embodiment the invention relates to an immunogenic composition comprising a glycoconjugate of the invention and comprising glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F and 33F, wherein the glycoconjugates from S. pneumoniae serotypes 1, 3, 4, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 18C, 19A, 19F, 23F and 33F are individually conjugated to CRM197 and the glycoconjugates from S. pneumoniae serotypes 1, 5, 15B and 22F are individually conjugated to TT. In an embodiment the immunogenic composition is a 21-valent pneumococcal conjugate compositions. Compositions of the invention may include a small amount of free carrier. When a given carrier protein is present in both free and conjugated form in a composition of the invention, the unconjugated form is preferably no more than 5% of the total amount of the carrier protein in the composition as a whole, and more preferably present at less than 2% by weight. 2.2 Dosage of the immunogenic compositions of the invention The amount of glycoconjugate(s) in each dose is selected as an amount which induces an immunoprotective response without significant, adverse side effects in typical vaccinees. Such amount will vary depending upon which specific immunogen is employed and how it is presented. The amount of a particular glycoconjugate in an immunogenic composition can be calculated based on total saccharide for that conjugate (conjugated and non-conjugated). For example, a glycoconjugate with 20% free saccharide will have about 80 µg of conjugated saccharide and about 20 µg of nonconjugated saccharide in a 100 µg saccharide dose. The amount of glycoconjugate can vary depending upon the bacteria and bacteria serotype. The saccharide concentration can be determined by the uronic acid assay. The "immunogenic amount" of the different saccharide components in the immunogenic composition, may diverge and each may comprise about 0.5 µg, about 0.75 µg, about 1 µg, about 2 µg, about 3 µg, about 4 µg, about 5 µg, about 6 µg, about 7 µg, about 8 µg, about 9 µg, about 10 µg, about 15 µg, about 20 µg, about 30 µg, about 40 µg, about 50 µg, about 60 µg, about 70 µg, about 80 µg, about 90 µg, or about 100 µg of any particular saccharide antigen. Generally, each dose will comprise 0.1 µg to 100 µg of saccharide. In an embodiment each dose will comprise 0.1 µg to 100 µg of saccharide. In a preferred embodiment each dose will comprise 0.5 µg to 20 µg. In a preferred embodiment each dose will comprise 1.0 µg to 10 µg. In an even preferred embodiment, each dose will comprise 2.0 µg to 5.0 µg of serotype 3 polysaccharide. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure. In an embodiment, each dose will comprise about 0.5 µg of saccharide. In an embodiment, each dose will comprise about 0.55 µg of saccharide. In an embodiment, each dose will comprise about 0.75 µg of saccharide. In an embodiment, each dose will comprise about 1.0 µg of saccharide. In an embodiment, each dose will comprise about 1.1 µg of saccharide. In an embodiment, each dose will comprise about 1.5 µg of saccharide. In an embodiment, each dose will comprise about 2.0 µg of saccharide. In an embodiment, each dose will comprise about 2.2 µg of saccharide. In an embodiment, each dose will comprise about 2.5 µg of saccharide. In an embodiment, each dose will comprise about 3.0 µg of saccharide. In an embodiment, each dose will comprise about 3.5 µg of saccharide. In an embodiment, each dose will comprise about 4.0 µg of saccharide. In an embodiment, each dose will comprise about 4.4 µg of saccharide. In an embodiment, each dose will comprise about 5.0 µg of saccharide. In an embodiment, each dose will comprise about 5.5 µg of saccharide. In an embodiment, each dose will comprise about 6.0 µg of saccharide. Generally, each dose will comprise 0.1 µg to 100 µg of saccharide for a given bacteria or serotype. In an embodiment each dose will comprise 0.1 µg to 100 µg of saccharide for a given bacteria or serotype. In a preferred embodiment each dose will comprise 0.5 µg to 20 µg. In a preferred embodiment each dose will comprise 1.0 µg to 10 µg. In an even preferred embodiment, each dose will comprise 2.0 µg to 5.0 µg of saccharide for a given bacteria or serotype. Any whole number integer within any of the above ranges is contemplated as an embodiment of the disclosure. In an embodiment, each dose will comprise about 0.5 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 0.55 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 0.75 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 1.0 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 1.1 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 1.5 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 2.0 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 2.2 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 2.5 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 3.0 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 3.5 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 4.0 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 4.4 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 5.0 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 5.5 µg of saccharide for each particular glycoconjugate. In an embodiment, each dose will comprise about 6.0 µg of saccharide for each particular glycoconjugate. 2.3 Carrier amount Generally, each dose will comprise 10 µg to 150 µg of carrier protein, particularly 15 µg to 100 µg of carrier protein, more particularly 25 µg to 75 µg of carrier protein, and even more particularly 40 µg to 60 µg of carrier protein. In an embodiment, said carrier protein is CRM197. In an embodiment, said carrier protein is SCP. In an embodiment, each dose will comprise about 10 µg, about 15 µg, about 20 µg, about 25 µg, about 26 µg, about 27 µg, about 28 µg, about 29 µg, about 30 µg, about 31 µg, about 32 µg, about 33 µg, about 34 µg, about 35 µg, about 36 µg, about 37 µg, about 38 µg, about 39 µg, about 40 µg, about 41 µg, about 42 µg, about 43 µg, about 44 µg, about 45 µg, about 46 µg, about 47 µg, about 48 µg, about 49 µg, about 50 µg, about 51 µg, about 52 µg, about 53 µg, about 54 µg, about 55 µg, about 56 µg, about 57 µg, about 58 µg, about 59 µg, about 60 µg, about 61 µg, about 62 µg, about 63 µg, about 64 µg, about 65 µg, about 66 µg, about 67 µg, about 68 µg, about 69 µg, about 70 µg, about 71 µg, about 72 µg, about 73 µg, about 74 µg or about 75 µg of carrier protein. In an embodiment, each dose will comprise about 25 µg, about 26 µg, about 27 µg, about 28 µg, about 29 µg, about 30 µg, about 31 µg, about 32 µg, about 33 µg, about 34 µg, about 35 µg, about 36 µg, about 37 µg, about 38 µg, about 39 µg, about 40 µg, about 41 µg, about 42 µg, about 43 µg, about 44 µg, about 45 µg, about 46 µg, about 47 µg, about 48 µg, about 49 µg, about 50 µg, about 51 µg, about 52 µg, about 53 µg, about 54 µg, about 55 µg, about 56 µg, about 57 µg, about 58 µg, about 59 µg, about 60 µg, about 61 µg, about 62 µg, about 63 µg, about 64 µg, about 65 µg, about 66 µg, about 67 µg, about 68 µg, about 69 µg, about 70 µg, about 71 µg, about 72 µg, about 73 µg, about 74 µg or about 75 µg of carrier protein. In an embodiment, each dose will comprise about 30 µg of carrier protein. In an embodiment, each dose will comprise about 31 µg of carrier protein. In an embodiment, each dose will comprise about 32 µg of carrier protein. In an embodiment, each dose will comprise about 33 µg of carrier protein. In an embodiment, each dose will comprise about 34 µg of carrier protein. In an embodiment, each dose will comprise about 45 µg of carrier protein. In an embodiment, each dose will comprise about 40 µg of carrier protein. In an embodiment, each dose will comprise about 41 µg of carrier protein. In an embodiment, each dose will comprise about 42 µg of carrier protein. In an embodiment, each dose will comprise about 43 µg of carrier protein. In an embodiment, each dose will comprise about 44 µg of carrier protein. In an embodiment, each dose will comprise about 45 µg of carrier protein. In an embodiment, each dose will comprise about 48 µg of carrier protein. In an embodiment, each dose will comprise about 49 µg of carrier protein. In an embodiment, each dose will comprise about 50 µg of carrier protein. In an embodiment, each dose will comprise about 51 µg of carrier protein. In an embodiment, each dose will comprise about 52 µg of carrier protein. In an embodiment, each dose will comprise about 53 µg of carrier protein. In an embodiment, said carrier protein is CRM197. In an embodiment, said carrier protein is SCP. 2.4 Further antigens Immunogenic compositions of the invention comprise conjugated saccharide antigen(s) (glycoconjugate(s)). They may also further include antigen(s) from other pathogen(s), particularly from bacteria and/or viruses. Preferred further antigens are selected from: a diphtheria toxoid (D), a tetanus toxoid (T), a pertussis antigen (P), which is typically acellular (Pa), a hepatitis B virus (HBV) surface antigen (HBsAg), a hepatitis A virus (HAV) antigen, a conjugated Haemophilus influenzae type b capsular saccharide (Hib), inactivated poliovirus vaccine (IPV). In an embodiment, the immunogenic compositions of the invention comprise D-T-Pa. In an embodiment, the immunogenic compositions of the invention comprise D-T-Pa-Hib, D-T-Pa- IPV or D-T-Pa-HBsAg. In an embodiment, the immunogenic compositions of the invention comprise D-T-Pa-HBsAg-IPV or D-T-Pa-HBsAg-Hib. In an embodiment, the immunogenic compositions of the invention comprise D-T-Pa-HBsAg-IPV-Hib. Pertussis antigens: Bordetella pertussis causes whooping cough. Pertussis antigens in vaccines are either cellular (whole cell, in the form of inactivated B. pertussis cells) or acellular. Preparation of cellular pertussis antigens is well documented (e.g., it may be obtained by heat inactivation of phase I culture of B. pertussis). Preferably, however, the invention uses acellular antigens. Where acellular antigens are used, it is preferred to use one, two or (preferably) three of the following antigens: (1) detoxified pertussis toxin (pertussis toxoid, or PT); (2) filamentous hemagglutinin (FHA); (3) pertactin (also known as the 69 kilodalton outer membrane protein). FHA and pertactin may be treated with formaldehyde prior to use according to the invention. PT is preferably detoxified by treatment with formaldehyde and/or glutaraldehyde. Acellular pertussis antigens are preferably adsorbed onto one or more aluminum salt adjuvants. As an alternative, they may be added in an unadsorbed state. Where pertactin is added then it is preferably already adsorbed onto an aluminum hydroxide adjuvant. PT and FHA may be adsorbed onto an aluminum hydroxide adjuvant or an aluminum phosphate. Adsorption of all of PT, FHA and pertactin to aluminum hydroxide is most preferred. Inactivated poliovirus vaccine: Poliovirus causes poliomyelitis. Rather than use oral poliovirus vaccine, preferred embodiments of the invention use IPV. Prior to administration to patients, polioviruses must be inactivated, and this can be achieved by treatment with formaldehyde. Poliomyelitis can be caused by one of three types of poliovirus. The three types are similar and cause identical symptoms, but they are antigenically different and infection by one type does not protect against infection by others. It is therefore preferred to use three poliovirus antigens in the invention: poliovirus Type 1 (e.g., Mahoney strain), poliovirus Type 2 (e.g., MEF- 1 strain), and poliovirus Type 3 (e.g., Saukett strain). The viruses are preferably grown, purified and inactivated individually, and are then combined to give a bulk trivalent mixture for use with the invention. Diphtheria toxoid: Corynebacterium diphtheriae causes diphtheria. Diphtheria toxin can be treated (e.g., using formalin or formaldehyde) to remove toxicity while retaining the ability to induce specific anti-toxin antibodies after injection. These diphtheria toxoids are used in diphtheria vaccines. Preferred diphtheria toxoids are those prepared by formaldehyde treatment. The diphtheria toxoid can be obtained by growing C. diphtheriae in growth medium, followed by formaldehyde treatment, ultrafiltration and precipitation. The toxoided material may then be treated by a process comprising sterile filtration and/or dialysis. The diphtheria toxoid is preferably adsorbed onto an aluminum hydroxide adjuvant. Tetanus toxoid: Clostridium tetani causes tetanus. Tetanus toxin can be treated to give a protective toxoid. The toxoids are used in tetanus vaccines. Preferred tetanus toxoids are those prepared by formaldehyde treatment. The tetanus toxoid can be obtained by growing C. tetani in growth medium, followed by formaldehyde treatment, ultrafiltration and precipitation. The material may then be treated by a process comprising sterile filtration and/or dialysis. Hepatitis A virus antigens: Hepatitis A virus (HAV) is one of the known agents which causes viral hepatitis. A preferred HAV component is based on inactivated virus, and inactivation can be achieved by formalin treatment. Hepatitis B virus (HBV) is one of the known agents which causes viral hepatitis. The major component of the capsid is a protein known as HBV surface antigen or, more commonly, HBsAg, which is typically a 226-amino acid polypeptide with a molecular weight of ~24 kDa. All existing hepatitis B vaccines contain HBsAg, and when this antigen is administered to a normal vaccinee it stimulates the production of anti-HBsAg antibodies which protect against HBV infection. For vaccine manufacture, HBsAg has been made in two ways: purification of the antigen in particulate form from the plasma of chronic hepatitis B carriers or expression of the protein by recombinant DNA methods (e.g., recombinant expression in yeast cells). Unlike native HBsAg (i.e., as in the plasma-purified product), yeast-expressed HBsAg is generally non-glycosylated, and this is the most preferred form of HBsAg for use with the invention. Conjugated Haemophilus influenzae type b antigens: Haemophilus influenzae type b (Hib) causes bacterial meningitis. Hib vaccines are typically based on the capsular saccharide antigen, the preparation of which is well documented. The Hib saccharide can be conjugated to a carrier protein in order to enhance its immunogenicity, especially in children. Typical carrier proteins are tetanus toxoid, diphtheria toxoid, CRM197, H.influenzae protein D, and an outer membrane protein complex from serogroup B meningococcus. The saccharide moiety of the conjugate may comprise full-length polyribosylribitol phosphate (PRP) as prepared from Hib bacteria, and/or fragments of full-length PRP. Hib conjugates may or may not be adsorbed to an aluminum salt adjuvant. 2.5 Adjuvant(s) In some embodiments, the immunogenic compositions disclosed herein may further comprise at least one, two or three adjuvants. In some embodiments, the immunogenic compositions disclosed herein may further comprise at least one adjuvant. In some embodiments, the immunogenic compositions disclosed herein may further comprise one adjuvant. In some embodiments, the immunogenic compositions disclosed herein may further comprise two adjuvants. The term "adjuvant" refers to a compound or mixture that enhances the immune response to an antigen. Antigens may act primarily as a delivery system, primarily as an immune modulator or have strong features of both. Suitable adjuvants include those suitable for use in mammals, including humans. Examples of known suitable delivery-system type adjuvants that can be used in humans include, but are not limited to, alum (e.g., aluminum phosphate, aluminum sulfate or aluminum hydroxide), calcium phosphate, liposomes, oil-in-water emulsions such as MF59 (4.3% w/v squalene, 0.5% w/v polysorbate 80 (Tween 80), 0.5% w/v sorbitan trioleate (Span 85)), water-in-oil emulsions such as Montanide, and poly(D,L-lactide-co-glycolide) (PLG) microparticles or nanoparticles. In an embodiment, the immunogenic compositions disclosed herein comprise aluminum salts (alum) as adjuvant (e.g., aluminum phosphate, aluminum sulfate or aluminum hydroxide). In a preferred embodiment, the immunogenic compositions disclosed herein comprise aluminum phosphate or aluminum hydroxide as adjuvant. In a preferred embodiment, the immunogenic compositions disclosed herein comprise aluminum phosphate as adjuvant. Further exemplary adjuvants to enhance effectiveness of the immunogenic compositions as disclosed herein include, but are not limited to: (1) oil-in-water emulsion formulations (with or without other specific immunostimulating agents such as muramyl peptides (see below) or bacterial cell wall components), such as for example (a) SAF, containing 10% Squalene, 0.4% Tween 80, 5% pluronic-blocked polymer L121, and thr-MDP either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion, and (b) RIBI™ adjuvant system (RAS), (Ribi Immunochem, Hamilton, MT) containing 2% Squalene, 0.2% Tween 80, and one or more bacterial cell wall components such as monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL + CWS (DETOX™); (2) saponin adjuvants, such as QS21, STIMULON™ (Cambridge Bioscience, Worcester, MA), ABISCO® (Isconova, Sweden), or ISCOMATRIX® (Commonwealth Serum Laboratories, Australia), may be used or particles generated therefrom such as ISCOMs (immunostimulating complexes), which ISCOMS may be devoid of additional detergent (e.g., WO 00/07621); (3) Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA); (4) cytokines, such as interleukins (e.g., IL-1, IL- 2, IL-4, IL-5, IL-6, IL-7, IL-12 (e.g., WO 99/44636)), interferons (e.g., gamma interferon), macrophage colony stimulating factor (M-CSF), tumor necrosis factor (TNF), etc.; (5) monophosphoryl lipid A (MPL) or 3-O-deacylated MPL (3dMPL) (see, e.g., GB-2220221, EP0689454), optionally in the substantial absence of alum when used with pneumococcal saccharides (see, e.g., WO 00/56358); (6) combinations of 3dMPL with, for example, QS21 and/or oil-in-water emulsions (see, e.g., EP0835318, EP0735898, EP0761231); (7) a polyoxyethylene ether or a polyoxyethylene ester (see, e.g., WO 99/52549); (8) a polyoxyethylene sorbitan ester surfactant in combination with an octoxynol (e.g., WO 01/21207) or a polyoxyethylene alkyl ether or ester surfactant in combination with at least one additional non-ionic surfactant such as an octoxynol (e.g., WO 01/21152); (9) a saponin and an immunostimulatory oligonucleotide (e.g., a CpG oligonucleotide) (e.g., WO 00/62800); (10) an immunostimulant and a particle of metal salt (see, e.g., WO 00/23105); (11) a saponin and an oil-in-water emulsion (e.g., WO 99/11241); (12) a saponin (e.g., QS21) + 3dMPL + IM2 (optionally + a sterol) (e.g., WO 98/57659); (13) other substances that act as immunostimulating agents to enhance the efficacy of the composition. Muramyl peptides include N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-25 acetyl- normuramyl-L-alanyl-D-isoglutamine (nor-MDP), N-acetylmuramyl-L-alanyl-D-isoglutarninyl-L- alanine-2-(1'-2'-dipalmitoyl-sn-gIycero-3-hydroxyphosphoryloxy)-ethylamine MTP-PE), etc. In an embodiment of the present invention, the immunogenic compositions as disclosed herein comprise a CpG Oligonucleotide as adjuvant. A CpG oligonucleotide as used herein refers to an immunostimulatory CpG oligodeoxynucleotide (CpG ODN), and accordingly these terms are used interchangeably unless otherwise indicated. Immunostimulatory CpG oligodeoxynucleotides contain one or more immunostimulatory CpG motifs that are unmethylated cytosine-guanine dinucleotides, optionally within certain preferred base contexts. The methylation status of the CpG immunostimulatory motif generally refers to the cytosine residue in the dinucleotide. An immunostimulatory oligonucleotide containing at least one unmethylated CpG dinucleotide is an oligonucleotide which contains a 5' unmethylated cytosine linked by a phosphate bond to a 3' guanine, and which activates the immune system through binding to Toll- like receptor 9 (TLR-9). In another embodiment the immunostimulatory oligonucleotide may contain one or more methylated CpG dinucleotides, which will activate the immune system through TLR9 but not as strongly as if the CpG motif(s) was/were unmethylated. CpG immunostimulatory oligonucleotides may comprise one or more palindromes that in turn may encompass the CpG dinucleotide. CpG oligonucleotides have been described in a number of issued patents, published patent applications, and other publications, including U.S. Patent Nos. 6,194,388; 6,207,646; 6,214,806; 6,218,371; 6,239,116; and 6,339,068. In an embodiment of the present invention, the immunogenic compositions as disclosed herein comprise any of the CpG Oligonucleotide described at page 3, line 22, to page 12, line 36 of WO 2010/125480. Different classes of CpG immunostimulatory oligonucleotides have been identified. These are referred to as A, B, C and P class, and are described in greater detail at page 3, line 22, to page 12, line 36 of WO 2010/125480. Methods of the invention embrace the use of these different classes of CpG immunostimulatory oligonucleotides. In an embodiment of the present invention, the immunogenic compositions as disclosed herein comprise an A class CpG oligonucleotide. Preferably, the "A class" CpG oligonucleotide of the invention has the following nucleic acid sequence: 5’ GGGGACGACGTCGTGGGGGGG 3’ (SEQ ID NO: 1). Some non-limiting examples of A-Class oligonucleotides include: 5’ G*G*G_G_A_C_G_A_C_G_T_C_G_T_G_G*G*G*G*G*G 3’ (SEQ ID NO: 2); wherein “*” refers to a phosphorothioate bond and “_” refers to a phosphodiester bond. In an embodiment of the present invention, the immunogenic compositions as disclosed herein comprise a B class CpG Oligonucleotide. In one embodiment, the CpG oligonucleotide for use in the present invention is a B class CpG oligonucleotide represented by at least the formula: 5' X1X2CGX3X43’, wherein X1, X2, X3, and X4 are nucleotides. In one embodiment, X2 is adenine, guanine, or thymine. In another embodiment, X3 is cytosine, adenine, or thymine. The B class CpG oligonucleotide sequences of the invention are those broadly described above as well as disclosed in WO 96/02555, WO 98/18810 and U.S. Patent Nos. 6,194,388; 6,207,646; 6,214,806; 6,218,371; 6,239,116 and 6,339,068. Exemplary sequences include but are not limited to those disclosed in these latter applications and patents. In an embodiment, the "B class" CpG oligonucleotide of the invention has the following nucleic acid sequence: 5’ TCGTCGTTTTTCGGTGCTTTT 3’ (SEQ ID NO: 3), or 5’ TCGTCGTTTTTCGGTCGTTTT 3’ (SEQ ID NO: 4), or 5’ TCGTCGTTTTGTCGTTTTGTCGTT 3’ (SEQ ID NO: 5), or 5’ TCGTCGTTTCGTCGTTTTGTCGTT 3’ (SEQ ID NO: 6), or 5’ TCGTCGTTTTGTCGTTTTTTTCGA 3’ (SEQ ID NO: 7). In any of these sequences, all of the linkages may be all phosphorothioate bonds. In another embodiment, in any of these sequences, one or more of the linkages may be phosphodiester, preferably between the “C” and the “G” of the CpG motif making a semi-soft CpG oligonucleotide. In any of these sequences, an ethyl-uridine or a halogen may substitute for the 5' T; examples of halogen substitutions include but are not limited to bromo-uridine or iodo-uridine substitutions. Some non-limiting examples of B-Class oligonucleotides include: 5’ T*C*G*T*C*G*T*T*T*T*T*C*G*G*T*G*C*T*T*T*T 3’ (SEQ ID NO: 8), or 5’ T*C*G*T*C*G*T*T*T*T*T*C*G*G*T*C*G*T*T*T*T 3’ (SEQ ID NO: 9), or 5’ T*C*G*T*C*G*T*T*T*T*G*T*C*G*T*T*T*T*G*T*C*G*T*T 3’ (SEQ ID NO: 10), or 5’ T*C*G*T*C*G*T*T*T*C*G*T*C*G*T*T*T*T*G*T*C*G*T*T 3’ (SEQ ID NO: 11), or 5’ T*C*G*T*C*G*T*T*T*T*G*T*C*G*T*T*T*T*T*T*T*C*G*A 3’ (SEQ ID NO: 12). wherein “*” refers to a phosphorothioate bond. In an embodiment of the present invention, the immunogenic compositions as disclosed herein comprise a C class CpG Oligonucleotide. In an embodiment, the "C class" CpG oligonucleotides of the invention have the following nucleic acid sequence: 5’ TCGCGTCGTTCGGCGCGCGCCG 3’ (SEQ ID NO: 13), or 5’ TCGTCGACGTTCGGCGCGCGCCG 3’ (SEQ ID NO: 14), or 5’ TCGGACGTTCGGCGCGCGCCG 3’ (SEQ ID NO: 15), or 5’ TCGGACGTTCGGCGCGCCG 3’ (SEQ ID NO: 16), or 5’ TCGCGTCGTTCGGCGCGCCG 3’ (SEQ ID NO: 17), or 5’ TCGACGTTCGGCGCGCGCCG 3’ (SEQ ID NO: 18), or 5’ TCGACGTTCGGCGCGCCG 3’ (SEQ ID NO: 19), or 5’ TCGCGTCGTTCGGCGCCG 3’ (SEQ ID NO: 20), or 5’ TCGCGACGTTCGGCGCGCGCCG 3’ (SEQ ID NO: 21), or 5’ TCGTCGTTTTCGGCGCGCGCCG 3’ (SEQ ID NO: 22), or 5’ TCGTCGTTTTCGGCGGCCGCCG 3’ (SEQ ID NO: 23), or 5’ TCGTCGTTTTACGGCGCCGTGCCG 3’ (SEQ ID NO: 24), or 5’ TCGTCGTTTTCGGCGCGCGCCGT 3’ (SEQ ID NO: 25). In any of these sequences, all of the linkages may be all phosphorothioate bonds. In another embodiment, in any of these sequences, one or more of the linkages may be phosphodiester, preferably between the “C” and the “G” of the CpG motif making a semi-soft CpG oligonucleotide. Some non-limiting examples of C-Class oligonucleotides include: 5’ T*C_G*C_G*T*C_G*T*T*C_G*G*C*G*C_G*C*G*C*C*G 3’ (SEQ ID NO: 26), or 5’ T*C_G*T*C_G*A*C_G*T*T*C_G*G*C*G*C_G*C*G*C*C*G 3’ (SEQ ID NO: 27), or 5’ T*C_G*G*A*C_G*T*T*C_G*G*C*G*C_G*C*G*C*C*G 3’ (SEQ ID NO: 28), or 5’ T*C_G*G*A*C_G*T*T*C_G*G*C*G*C*G*C*C*G 3’ (SEQ ID NO: 29), or 5’ T*C_G*C_G*T*C_G*T*T*C_G*G*C*G*C*G*C*C*G 3’ (SEQ ID NO: 30), or 5’ T*C_G*A*C_G*T*T*C_G*G*C*G*C_G*C*G*C*C*G 3’ (SEQ ID NO: 31), or 5’ T*C_G*A*C_G*T*T*C_G*G*C*G*C*G*C*C*G 3’ (SEQ ID NO: 32), or 5’ T*C_G*C_G*T*C_G*T*T*C_G*G*C*G*C*C*G 3’ (SEQ ID NO: 33), or 5’ T*C_G*C_G*A*C_G*T*T*C_G*G*C*G*C_G*C*G*C*C*G 3’ (SEQ ID NO: 34), or 5’ T*C*G*T*C*G*T*T*T*T*C*G*G*C*G*C*G*C*G*C*C*G 3’ (SEQ ID NO: 35), or 5’ T*C*G*T*C*G*T*T*T*T*C*G*G*C*G*G*C*C*G*C*C*G 3’ (SEQ ID NO: 36), or 5’ T*C*G*T*C_G*T*T*T*T*A*C_G*G*C*G*C*C_G*T*G*C*C*G 3’ (SEQ ID NO: 37), or 5’ T*C_G*T*C*G*T*T*T*T*C*G*G*C*G*C*G*C*G*C*C*G*T 3’ (SEQ ID NO: 38) wherein “*” refers to a phosphorothioate bond and “_” refers to a phosphodiester bond. In any of these sequences, an ethyl-uridine or a halogen may substitute for the 5' T; examples of halogen substitutions include but are not limited to bromo-uridine or iodo-uridine substitutions. In an embodiment of the present invention, the immunogenic compositions as disclosed herein comprise a P class CpG Oligonucleotide. In an embodiment, the CpG oligonucleotide for use in the present invention is a P class CpG oligonucleotide containing a 5' TLR activation domain and at least two palindromic regions, one palindromic region being a 5' palindromic region of at least 6 nucleotides in length and connected to a 3' palindromic region of at least 8 nucleotides in length either directly or through a spacer, wherein the oligonucleotide includes at least one YpR dinucleotide. In an embodiment, said oligonucleotide is not T*C_G*T*C_G*A*C_G*T*T*C_G*G*C*G*C_G*C*G*C*C*G (SEQ ID NO: 27). In one embodiment the P class CpG oligonucleotide includes at least one unmethylated CpG dinucleotide. In another embodiment the TLR activation domain is TCG, TTCG, TTTCG, TYpR, TTYpR, TTTYpR, UCG, UUCG, UUUCG, TTT, or TTTT. In yet another embodiment the TLR activation domain is within the 5' palindromic region. In another embodiment the TLR activation domain is immediately 5' to the 5' palindromic region. In an embodiment, the "P class" CpG oligonucleotides of the invention have the following nucleic acid sequence: 5’ TCGTCGACGATCGGCGCGCGCCG 3’ (SEQ ID NO: 39). In said sequences, all of the linkages may be all phosphorothioate bonds. In another embodiment, one or more of the linkages may be phosphodiester, preferably between the “C” and the “G” of the CpG motif making a semi-soft CpG oligonucleotide. In any of these sequences, an ethyl-uridine or a halogen may substitute for the 5' T; examples of halogen substitutions include but are not limited to bromo-uridine or iodo-uridine substitutions. A non-limiting example of P-Class oligonucleotides include: 5’ T*C_G*T*C_G*A*C_G*A*T*C_G*G*C*G*C_G*C*G*C*C*G 3’ (SEQ ID NO: 40) wherein “*” refers to a phosphorothioate bond and “_” refers to a phosphodiester bond. In one embodiment the oligonucleotide includes at least one phosphorothioate linkage. In another embodiment all internucleotide linkages of the oligonucleotide are phosphorothioate linkages. In another embodiment the oligonucleotide includes at least one phosphodiester-like linkage. In another embodiment the phosphodiester-like linkage is a phosphodiester linkage. In another embodiment a lipophilic group is conjugated to the oligonucleotide. In one embodiment the lipophilic group is cholesterol. In an embodiment, all the internucleotide linkages of the CpG oligonucleotides disclosed herein are phosphodiester bonds (“soft” oligonucleotides, as described in WO 2007/026190). In another embodiment, CpG oligonucleotides of the invention are rendered resistant to degradation (e.g., are stabilized). A "stabilized oligonucleotide" refers to an oligonucleotide that is relatively resistant to in vivo degradation (e.g., via an exo- or endo-nuclease). Nucleic acid stabilization can be accomplished via backbone modifications. Oligonucleotides having phosphorothioate linkages provide maximal activity and protect the oligonucleotide from degradation by intracellular exo- and endo-nucleases. The immunostimulatory oligonucleotides may have a chimeric backbone, which have combinations of phosphodiester and phosphorothioate linkages. For purposes of the instant invention, a chimeric backbone refers to a partially stabilized backbone, wherein at least one internucleotide linkage is phosphodiester or phosphodiester-like, and wherein at least one other internucleotide linkage is a stabilized internucleotide linkage, wherein the at least one phosphodiester or phosphodiester-like linkage and the at least one stabilized linkage are different. When the phosphodiester linkage is preferentially located within the CpG motif such molecules are called “semi-soft” as described in WO 2007/026190. Other modified oligonucleotides include combinations of phosphodiester, phosphorothioate, methylphosphonate, methylphosphorothioate, phosphorodithioate, and/or p- ethoxy linkages. Mixed backbone modified ODN may be synthesized as described in WO 2007/026190. The size of the CpG oligonucleotide (i.e., the number of nucleotide residues along the length of the oligonucleotide) also may contribute to the stimulatory activity of the oligonucleotide. For facilitating uptake into cells, CpG oligonucleotide of the invention preferably have a minimum length of 6 nucleotide residues. Oligonucleotides of any size greater than 6 nucleotides (even many kb long) are capable of inducing an immune response if sufficient immunostimulatory motifs are present, because larger oligonucleotides are degraded inside cells. In certain embodiments, the CpG oligonucleotides are 6 to 100 nucleotides long, preferentially 8 to 30 nucleotides long. In important embodiments, nucleic acids and oligonucleotides of the invention are not plasmids or expression vectors. In an embodiment, the CpG oligonucleotide disclosed herein comprise substitutions or modifications, such as in the bases and/or sugars as described at paragraphs 134 to 147 of WO 2007/026190. In an embodiment, the CpG oligonucleotide of the present invention is chemically modified. Examples of chemical modifications are known to the skilled person and are described, for example in Uhlmann et al. (1990) Chem. Rev. 90:543; S. Agrawal, Ed., Humana Press, Totowa, USA 1993; Crooke et al. (1996) Annu. Rev. Pharmacol. Toxicol. 36:107-129; and Hunziker et al. (1995) Mod. Synth. Methods 7:331-417. An oligonucleotide according to the invention may have one or more modifications, wherein each modification is located at a particular phosphodiester internucleoside bridge and/or at a particular β-D-ribose unit and/or at a particular natural nucleoside base position in comparison to an oligonucleotide of the same sequence which is composed of natural DNA or RNA. In some embodiments of the invention, CpG-containing nucleic acids might be simply mixed with immunogenic carriers according to methods known to those skilled in the art (see, e.g., WO 03/024480). In a particular embodiment of the present invention, any of the immunogenic compositions disclosed herein comprise from 2 μg to 100 mg of CpG oligonucleotide. In a particular embodiment of the present invention, the immunogenic composition of the invention comprises 0.1 mg to 50 mg of CpG oligonucleotide, preferably from 0.2 mg to 10 mg CpG oligonucleotide, more preferably from 0.3 mg to 5 mg CpG oligonucleotide.. In a particular embodiment of the present invention, the immunogenic composition of the invention comprises from 0.3 mg to 5 mg CpG oligonucleotide. Even preferably, the immunogenic composition of the invention may comprise from 0.5 to 2 mg CpG oligonucleotide. Most preferably, the immunogenic composition of the invention may comprise from 0.75 to 1.5 mg CpG oligonucleotide. In a preferred embodiment, any of the immunogenic composition disclosed herein may comprise about 1 mg CpG oligonucleotide. 3 Formulation The immunogenic compositions of the invention may be formulated in liquid form (i.e., solutions or suspensions) or in a lyophilized form. In an embodiment, the immunogenic composition of the invention is formulated in a liquid form. In an embodiment, the immunogenic composition of the invention is formulated in a lyophilized form. Liquid formulations may advantageously be administered directly from their packaged form and are thus ideal for injection without the need for reconstitution in aqueous medium as otherwise required for lyophilized compositions of the invention. Formulation of the immunogenic composition of the present disclosure can be accomplished using art-recognized methods. For instance, the individual polysaccharides and/or conjugates can be formulated with a physiologically acceptable vehicle to prepare the composition. Examples of such vehicles include, but are not limited to, water, buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol) and dextrose solutions. The present disclosure provides an immunogenic composition comprising any of combination of glycoconjugates disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent. In an embodiment, the immunogenic composition of the disclosure is in liquid form, preferably in aqueous liquid form. Immunogenic compositions of the disclosure may comprise one or more of a buffer, a salt, a divalent cation, a non-ionic detergent, a cryoprotectant such as a sugar, and an anti-oxidant such as a free radical scavenger or chelating agent, or any multiple combinations thereof. In an embodiment, the immunogenic compositions of the disclosure comprise a buffer. In an embodiment, said buffer has a pKa of about 3.5 to about 7.5. In some embodiments, the buffer is phosphate, succinate, histidine or citrate. In some embodiments, the buffer is succinate. In some embodiments, the buffer is histidine. In certain embodiments, the buffer is succinate at a final concentration of 1 mM to 10 mM. In one particular embodiment, the final concentration of the succinate buffer is about 5 mM. In an embodiment, the immunogenic compositions of the disclosure comprise a salt. In some embodiments, the salt is selected from the groups consisting of magnesium chloride, potassium chloride, sodium chloride and a combination thereof. In one particular embodiment, the salt is sodium chloride. In one particular embodiment, the immunogenic compositions of the invention comprise sodium chloride at 150 mM. In an embodiment, the immunogenic compositions of the disclosure comprise a surfactant. In an embodiment, the surfactant is selected from the group consisting of polysorbate 20 (TWEENTM20), polysorbate 40 (TWEENTM40), polysorbate 60 (TWEEN™60), polysorbate 65 (TWEEN™65), polysorbate 80 (TWEEN™80), polysorbate 85 (TWEEN™85), TRITON™ N-101, TRITON™ X-100, oxtoxynol 40, nonoxynol-9, triethanolamine, triethanolamine polypeptide oleate, polyoxyethylene-660 hydroxystearate (PEG-15, Solutol H 15), polyoxyethylene-35- ricinoleate (CREMOPHOR® EL), soy lecithin and a poloxamer. In one particular embodiment, the surfactant is polysorbate 80. In some said embodiment, the final concentration of polysorbate 80 in the formulation is at least 0.0001% to 10% polysorbate 80 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 80 in the formulation is at least 0.001% to 1% polysorbate 80 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 80 in the formulation is at least 0.01% to 1% polysorbate 80 weight to weight (w/w). In other embodiments, the final concentration of polysorbate 80 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.02% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.01% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.03% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.04% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 0.05% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 80 in the formulation is 1% polysorbate 80 (w/w). In one particular embodiment, the surfactant is polysorbate 20. In some said embodiment, the final concentration of polysorbate 20 in the formulation is at least 0.0001% to 10% polysorbate 20 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 20 in the formulation is at least 0.001% to 1% polysorbate 20 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 20 in the formulation is at least 0.01% to 1% polysorbate 20 weight to weight (w/w). In other embodiments, the final concentration of polysorbate 20 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.02% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.01% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.03% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.04% polysorbate 80 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 0.05% polysorbate 20 (w/w). In another embodiment, the final concentration of the polysorbate 20 in the formulation is 1% polysorbate 20 (w/w). In one particular embodiment, the surfactant is polysorbate 40. In some said embodiment, the final concentration of polysorbate 40 in the formulation is at least 0.0001% to 10% polysorbate 40 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 40 in the formulation is at least 0.001% to 1% polysorbate 40 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 40 in the formulation is at least 0.01% to 1% polysorbate 40 weight to weight (w/w). In other embodiments, the final concentration of polysorbate 40 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 40 (w/w). In another embodiment, the final concentration of the polysorbate 40 in the formulation is 1% polysorbate 40 (w/w). In one particular embodiment, the surfactant is polysorbate 60. In some said embodiment, the final concentration of polysorbate 60 in the formulation is at least 0.0001% to 10% polysorbate 60 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 60 in the formulation is at least 0.001% to 1% polysorbate 60 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 60 in the formulation is at least 0.01% to 1% polysorbate 60 weight to weight (w/w). In other embodiments, the final concentration of polysorbate 60 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 60 (w/w). In another embodiment, the final concentration of the polysorbate 60 in the formulation is 1% polysorbate 60 (w/w). In one particular embodiment, the surfactant is polysorbate 65. In some said embodiment, the final concentration of polysorbate 65 in the formulation is at least 0.0001% to 10% polysorbate 65 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 65 in the formulation is at least 0.001% to 1% polysorbate 65 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 65 in the formulation is at least 0.01% to 1% polysorbate 65 weight to weight (w/w). In other embodiments, the final concentration of polysorbate 65 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 65 (w/w). In another embodiment, the final concentration of the polysorbate 65 in the formulation is 1% polysorbate 65 (w/w). In one particular embodiment, the surfactant is polysorbate 85. In some said embodiment, the final concentration of polysorbate 85 in the formulation is at least 0.0001% to 10% polysorbate 85 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 85 in the formulation is at least 0.001% to 1% polysorbate 85 weight to weight (w/w). In some said embodiments, the final concentration of polysorbate 85 in the formulation is at least 0.01% to 1% polysorbate 85 weight to weight (w/w). In other embodiments, the final concentration of polysorbate 85 in the formulation is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1% polysorbate 85 (w/w). In another embodiment, the final concentration of the polysorbate 85 in the formulation is 1% polysorbate 85 (w/w). In certain embodiments, the immunogenic composition of the disclosure has a pH of 5.5 to 7.5, more preferably a pH of 5.6 to 7.0, even more preferably a pH of 5.8 to 6.0. In one embodiment, the present disclosure provides a container filled with any of the immunogenic compositions disclosed herein. In one embodiment, the container is selected from the group consisting of a vial, a syringe, a flask, a fermentor, a bioreactor, a bag, a jar, an ampoule, a cartridge and a disposable pen. In certain embodiments, the container is siliconized. In an embodiment, the container of the present disclosure is made of glass, metals (e.g., steel, stainless steel, aluminum, etc.) and/or polymers (e.g., thermoplastics, elastomers, thermoplastic-elastomers). In an embodiment, the container of the present disclosure is made of glass. In one embodiment, the present disclosure provides a syringe filled with any of the immunogenic compositions disclosed herein. In certain embodiments, the syringe is siliconized and/or is made of glass. A typical dose of the immunogenic composition of the invention for injection has a volume of 0.1 mL to 2 mL. In an embodiment, the immunogenic composition of the invention for injection has a volume of 0.2 mL to 1 mL, even more preferably a volume of about 0.5 mL. 4 Uses of the glycoconjugate and immunogenic compositions of the invention The glycoconjugates disclosed herein may be use as antigens. For example, they may be part of a vaccine. Therefore, in an embodiment, the immunogenic compositions of the invention are for use as a medicament. In an embodiment, the immunogenic compositions of the invention are for use as a vaccine. Therefore, in an embodiment, the immunogenic compositions described herein are for use in generating an immune response in a subject. In one aspect, the subject is a mammal, such as a human, non-human primate, cat, sheep, pig, horse, bovine or dog. In one aspect, the subject is a human. The immunogenic compositions described herein may be used in therapeutic or prophylactic methods for preventing, treating or ameliorating a bacterial infection, disease or condition in a subject. Thus, in one aspect, the disclosure provides a method of preventing, treating or ameliorating an infection, disease or condition associated with a bacterial infection in a subject, comprising administering to the subject an immunologically effective amount of an immunogenic composition of the disclosure. The immunogenic composition of the present disclosure can be used to protect or treat a human susceptible to a bacterial infection, by means of administering the immunogenic composition via a systemic or mucosal route. In an embodiment, the immunogenic composition of the invention is administered by intramuscular, intraperitoneal, intradermal or subcutaneous routes. In an embodiment, the immunogenic composition of the invention is administered by intramuscular, intraperitoneal, intradermal or subcutaneous injection. In an embodiment, the immunogenic composition of the invention is administered by intramuscular or subcutaneous injection. In an embodiment, the immunogenic composition of the invention is administered by intramuscular injection. In an embodiment, the immunogenic composition of the invention is administered by subcutaneous injection. 5 Subject to be treated with the immunogenic compositions of the invention As disclosed herein, the immunogenic compositions described herein may be used in various therapeutic or prophylactic methods for preventing, treating or ameliorating a bacterial infection, disease or condition in a subject. In a preferred embodiment, said subject is a human. In a most preferred embodiment, said subject is a new born (i.e., under three months of age), an infant (i.e., from 3 months to one year of age) or a toddler (i.e., from one year to four years of age). In an embodiment, the immunogenic compositions disclosed herein are for use as a vaccine. In such embodiment, the subject to be vaccinated may be less than 1 year of age. For example, the subject to be vaccinated can be about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11 or about 12 months of age. In an embodiment, the subject to be vaccinated is about 2, about 4 or about 6 months of age. In another embodiment, the subject to be vaccinated is less than 2 years of age. For example, the subject to be vaccinated can be about 12 to about 15 months of age. In some cases, as little as one dose of the immunogenic composition according to the invention is needed, but under some circumstances, a second, third or fourth dose may be given (see section 8 below). In an embodiment of the present invention, the subject to be vaccinated is a human adult 50 years of age or older, more preferably a human adult 55 years of age or older. In an embodiment, the subject to be vaccinated is a human adult 65 years of age or older, 70 years of age or older, 75 years of age or older or 80 years of age or older. In an embodiment the subject to be vaccinated is an immunocompromised individual, in particular a human. An immunocompromised individual is generally defined as a person who exhibits an attenuated or reduced ability to mount a normal humoral or cellular defense to challenge by infectious agents. In an embodiment of the present invention, the immunocompromised subject to be vaccinated suffers from a disease or condition that impairs the immune system and results in an antibody response that is insufficient to protect against or treat pneumococcal disease. In an embodiment, said disease is a primary immunodeficiency disorder. Preferably, said primary immunodeficiency disorder is selected from the group consisting of combined T- and B- cell immunodeficiencies, antibody deficiencies, well-defined syndromes, immune dysregulation diseases, phagocyte disorders, innate immunity deficiencies, autoinflammatory disorders, and complement deficiencies. In an embodiment, said primary immunodeficiency disorder is selected from the one disclosed on page 24, line 11, to page 25, line 19 of WO 2010/125480. In a particular embodiment of the present invention, the immunocompromised subject to be vaccinated suffers from a disease selected from the groups consisting of: HIV-infection, acquired immunodeficiency syndrome (AIDS), cancer, chronic heart or lung disorders, congestive heart failure, diabetes mellitus, chronic liver disease, alcoholism, cirrhosis, spinal fluid leaks, cardiomyopathy, chronic bronchitis, emphysema, chronic obstructive pulmonary disease (COPD), spleen dysfunction (such as sickle cell disease), lack of spleen function (asplenia), blood malignancy, leukemia, multiple myeloma, Hodgkin’s disease, lymphoma, kidney failure, nephrotic syndrome and asthma. In an embodiment of the present invention, the immunocompromised subject to be vaccinated suffers from malnutrition. In a particular embodiment of the present invention, the immunocompromised subject to be vaccinated is taking a drug or treatment that lowers the body’s resistance to infection. In an embodiment, said drug is selected from the one disclosed on page 26, line 33, to page 26, line 4 of WO 2010/125480. In a particular embodiment of the present invention, the immunocompromised subject to be vaccinated is a smoker. In a particular embodiment of the present invention, the immunocompromised subject to be vaccinated has a white blood cell count (leukocyte count) below 5 x 109 cells per liter, or below 4 x 109 cells per liter, or below 3 x 109 cells per liter, or below 2 x 109 cells per liter, or below 1 x 109 cells per liter, or below 0.5 x 109 cells per liter, or below 0.3 x 109 cells per liter, or below 0.1 x 109 cells per liter. White blood cell count (leukocyte count): The number of white blood cells (WBC) in the blood. The WBC is usually measured as part of the CBC (complete blood count). White blood cells are the infection-fighting cells in the blood and are distinct from the red (oxygen-carrying) blood cells known as erythrocytes. There are different types of white blood cells, including neutrophils (polymorphonuclear leukocytes; PMN), band cells (slightly immature neutrophils), T-type lymphocytes (T-cells), B-type lymphocytes (B-cells), monocytes, eosinophils, and basophils. All the types of white blood cells are reflected in the white blood cell count. The normal range for the white blood cell count is usually between 4,300 and 10,800 cells per cubic millimeter of blood. This can also be referred to as the leukocyte count and can be expressed in international units as 4.3 - 10.8 x 109 cells per liter. In a particular embodiment of the present invention, the immunocompromised subject to be vaccinated suffers from neutropenia. In a particular embodiment of the present invention, the immunocompromised subject to be vaccinated has a neutrophil count below 2 x 109 cells per liter, or below 1 x 109 cells per liter, or below 0.5 x 109 cells per liter, or below 0.1 x 109 cells per liter, or below 0.05 x 109 cells per liter. A low white blood cell count or “neutropenia” is a condition characterized by abnormally low levels of neutrophils in the circulating blood. Neutrophils are a specific kind of white blood cell that help to prevent and fight infections. The most common reason that cancer patients experience neutropenia is as a side effect of chemotherapy. Chemotherapy-induced neutropenia increases a patient’s risk of infection and disrupts cancer treatment. In a particular embodiment of the present invention, the immunocompromised subject to be vaccinated has a CD4+ cell count below 500/mm3, or CD4+ cell count below 300/mm3, or CD4+ cell count below 200/mm3, CD4+ cell count below 100/mm3, CD4+ cell count below 75/mm3, or CD4+ cell count below 50/mm3. CD4 cell tests are normally reported as the number of cells in mm3. Normal CD4 counts are between 500 and 1,600, and CD8 counts are between 375 and 1,100. CD4 counts drop dramatically in people with HIV. In an embodiment of the invention, any of the immunocompromised subjects disclosed herein is a human male. In an embodiment of the invention, any of the immunocompromised subjects disclosed herein is a human female. 6 Regimen In some cases, as little as one dose of the immunogenic composition according to the invention is needed, but under some circumstances, such as conditions of greater immune deficiency, a second, third or fourth dose may be given. Following an initial vaccination, subjects can receive one or several booster immunizations adequately spaced. In an embodiment, the schedule of vaccination of the immunogenic composition according to the invention is a single dose. In a particular embodiment, said single dose schedule is for healthy persons being at least 2 years of age. In an embodiment, the schedule of vaccination of the immunogenic composition according to the invention is a multiple dose schedule. In a particular embodiment, said multiple dose schedule consists of a series of 2 doses separated by an interval of about 1 month to about 2 months. In a particular embodiment, said multiple dose schedule consists of a series of 2 doses separated by an interval of about 1 month, or a series of 2 doses separated by an interval of about 2 months. In another embodiment, said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month to about 2 months. In another embodiment, said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month, or a series of 3 doses separated by an interval of about 2 months. In another embodiment, said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month to about 2 months followed by a fourth dose about 10 months to about 13 months after the first dose. In another embodiment, said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month followed by a fourth dose about 10 months to about 13 months after the first dose, or a series of 3 doses separated by an interval of about 2 months followed by a fourth dose about 10 months to about 13 months after the first dose. In an embodiment, the multiple dose schedule consists of at least one dose (e.g., 1, 2 or 3 doses) in the first year of age followed by at least one toddler dose. In an embodiment, the multiple dose schedule consists of a series of 2 or 3 doses separated by an interval of about 1 month to about 2 months (for example 28-56 days between doses), starting at 2 months of age, and followed by a toddler dose at 12-18 months of age. In an embodiment, said multiple dose schedule consists of a series of 3 doses separated by an interval of about 1 month to about 2 months (for example 28-56 days between doses), starting at 2 months of age, and followed by a toddler dose at 12-15 months of age. In another embodiment, said multiple dose schedule consists of a series of 2 doses separated by an interval of about 2 months, starting at 2 months of age, and followed by a toddler dose at 12-18 months of age. In an embodiment, the multiple dose schedule consists of a 4-dose series of vaccine at 2, 4, 6, and 12-15 months of age. In an embodiment, a prime dose is given at day 0 and one or more boosts are given at intervals that range from about 2 to about 24 weeks, preferably with a dosing interval of 4-8 weeks. In an embodiment, a prime dose is given at day 0 and a boost is given about 3 months later. 7. The invention also provides the following embodiments as defined in the following numbered paragraphs 1 to 86 1. A method of making a capsular saccharide glycoconjugate, comprising the steps of: (a) reacting an isolated capsular saccharide with a carbonic acid derivative and an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide, (b) reacting a carrier protein with an agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group where the NHS moiety reacts with the amino groups to form an amide linkage thereby obtaining an azido functionalized carrier protein, (c) reacting the activated alkynyl saccharide of step (a) with the azido functionalized carrier protein of step (b) by Cu+1 mediated azide-alkyne cycloaddition reaction to form a glycoconjugate. 2. The method of paragraph 1 wherein, the isolated saccharide is sized before the activation step (a). 3. The method of paragraph 2 wherein, the isolated capsular saccharide is sized to a weight average molecular weight of between 50 kDa and 500 kDa. 4. The method of any one of paragraphs 1-3 wherein, said carbonic acid derivative is selected from the group consisting of 1,1’-carbonyldiimidazole (CDI), 1,1’-carbonyl-di-(1,2,4-triazole) (CDT), disuccinimidyl carbonate (DSC) and N-hydroxysuccinimidyl chloroformate. 5. The method of any one of paragraphs 1-4 wherein, said carbonic acid derivative is 1,1’- carbonyldiimidazole (CDI). 6. The method of any one of paragraphs 1-4 wherein, said carbonic acid derivative is 1,1'- Carbonyl-di-(1,2,4-triazole) (CDT). 7. The method of any one of paragraphs 1-6 wherein said alkyne linker is a compound of formula (I), H2N X (I) wherein X is selected from the group consisting of CH2, CH2(CH2)n, (CH2CH2O)mCH2CH2, NHCO(CH2)n, NHCO(CH2CH2O)mCH2CH2, OCH2(CH2)n and O(CH2CH2O)mCH2CH2; where n is selected from 1 to 10 and m is selected from 1 to 4. 8. The method of any one of paragraphs 1-6 wherein said alkyne linker is a compound of formula (II), 9. The method of any one of paragraphs 1-8 wherein, said agent bearing an N- Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (III), where X is selected from the group consisting of (CH2)nCH2C=O and (CH2CH2O)mCH2CH2=O; where n is selected from 0 to 10 and m is selected from 0 to 4. 10. The method of paragraph 9 wherein said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (III), wherein X is (CH2)nCH2C=O, where n is selected from 0 to 10. 11. The method of paragraph 9 wherein said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (III), wherein X is (CH2CH2O)mCH2CH2=O, where m is selected from 0 to 4. 12. The method of any one of paragraphs 1-8 wherein, said agent bearing an N- Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (IV): 13. The method of any one of paragraphs 1-12 wherein, step (a) comprises reacting the capsular saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative-activated capsular saccharide with an alkyne linker in an aprotic solvent to produce an activated alkynyl capsular saccharide. 14. The method of any one of paragraphs 1-13 wherein, at step (a) the isolated capsular saccharide is reacted with said carbonic acid derivative in an aprotic solvent. 15. The method of any one of paragraphs 1-13 wherein, at step (a) the isolated capsular saccharide is reacted with a carbonic acid derivative in a solution comprising or consisting essentially of dimethylsulphoxide (DMSO). 16. The method of any one of paragraphs 1-14 wherein, at step (a) the isolated capsular saccharide is reacted with CDI in an aprotic solvent comprising 0.1% to 1% (v/v) water. 17. The method of any one of paragraphs 1-14 wherein, at step (a) the isolated capsular saccharide is reacted with CDI in DMSO comprising 0.1% to 1% (v/v) water. 18. The method of any one of paragraphs 1-17 wherein at step (a) carbonic acid derivative activation is followed by the addition of water. 19. The method of paragraph 18 wherein water is added to bring the total water content in the mixture to between about 1% to about 10% (v/v). 20. The method of any one of paragraphs 1-19 wherein step (a) further comprises reacting the carbonic acid derivative-activated saccharide with an amount of alkyne linker that is between 0.01-10 molar equivalent to the amount of saccharide Repeat Unit of the activated saccharide. 21. The method of any one of paragraphs 1-20 wherein the degree of activation of the activated saccharide following step (a) is between 0.5 to 50%. 22. The method of any one of paragraphs 1-21 wherein step (b) comprises reacting the carrier protein with an amount of agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group that is 0.1-10 molar equivalents to the lysines on the carrier. 23. The method of any one of paragraphs 1-22 wherein the degree of activation of the activated carrier following step (b) is between 1 and 50. 24. The method of any one of paragraphs 1-23 wherein the conjugation reaction step (c) is carried out in aqueous buffer in the presence of copper (I) as catalyst. 25. The method of any one of paragraphs 1-23 wherein the conjugation reaction step (c) is carried out in aqueous buffer in the presence of an oxidant and of copper (I) as catalyst. 26. The method of any one of paragraphs 1-23 wherein the conjugation reaction step (c) is carried out in aqueous buffer in the presence of copper (I) as catalyst and ascorbate as oxidant, wherein the reaction mixture further comprises THPTA (tris(3- hydroxypropyltriazolylmethyl)amine) and aminoguanidine. 27. The method of any one of paragraphs 1-26 wherein the initial input ratio (weight by weight) of activated alkynyl saccharide to activated azido-carrier at step (c) is between 0.1 and 3. 28. The method of any one of paragraphs 1-27 wherein following step (c), the method further comprises a step of capping the unreacted azido groups remained in the conjugate with an azido group capping agent. 29. The method of paragraph 28 wherein, said azido group capping agent is a compound of formula (V), (V) wherein X is (CH2)n wherein n is selected from 1 to 15. 30. The method of paragraph 28 wherein, said azido group capping agent is propargyl alcohol. 31. The method of any one of paragraphs 28-30 wherein the capping of the unreacted azido groups is performed with an amount of capping agent that is between 0.05 to 20 molar equivalents to the amount of saccharide repeat unit of the activated saccharide. 32. The method of any one of paragraphs 1-31 wherein following step (c), the method further comprises a step of capping the unreacted alkyne groups remained in the conjugate with an alkyne group capping agent. 33. The method of paragraph 32 wherein said alkyne group capping agent is an agent bearing an azido group. 34. The method of paragraph 33 wherein said alkyne group capping agent is a compound of formula (VI), (VI) wherein X is (CH2)n wherein n is selected from 1 to 15. 35. The method of paragraph 32 wherein said alkyne group capping agent is 3-azido-1-propanol. 36. The method of any one of paragraphs 32-35 wherein the capping of the unreacted alkyne groups is performed with an amount of capping agent that is between 0.05 to 20 molar equivalents to the amount of saccharide repeat unit of the activated saccharide. 37. The method of any one of paragraphs 1-36 wherein the method further comprises the step of purifying the glycoconjugate after it is produced. 38. A capsular saccharide glycoconjugate produced according to any one of the methods of paragraphs 1 to 37. 39. A capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII): (VII), wherein X is selected from the group consisting of CH2(CH2)n’, (CH2CH2O)mCH2CH2, NHCO(CH2)n’, NHCO(CH2CH2O)mCH2CH2, OCH2(CH2)n’ and O(CH2CH2O)mCH2CH2; where n’ is selected from 0 to 10 and m is selected from 1 to 4, and wherein X' is selected from the group consisting of CH2(CH2)n”, CH2O(CH2)n’’CH2, CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4. 40. A capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH2(CH2)n’, where n’ is 0 and wherein X' is CH2(CH2)n” where n’’ is 0. 41. A capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VIII), (VIII). 42. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 41 comprising a capsular saccharide wherein the weight average molecular weight (Mw) of said capsular saccharide before conjugation is between 10 kDa and 2,000 kDa. 43. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 41 comprising a capsular saccharide wherein the weight average molecular weight (Mw) of said capsular saccharide before conjugation is between 50 kDa and 1,000 kDa. 44. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 41 comprising a capsular saccharide wherein the weight average molecular weight (Mw) of said capsular saccharide before conjugation is between 200 kDa and 750 kDa. 45. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 44 having a weight average molecular weight (Mw) of between 250 kDa and 20,000 kDa. 46. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 44 having a weight average molecular weight (Mw) of between 500 kDa and 5,000 kDa. 47. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 44 having a weight average molecular weight (Mw) of between 750 kDa and 2,500 kDa. 48. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 47 wherein, the degree of conjugation of the capsular saccharide glycoconjugate is between 2 and 15. 49. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 48 wherein the ratio of capsular saccharide to carrier protein in the glycoconjugate (w/w) is between 0.5 and 3.0. 50. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 49 wherein the said capsular saccharide glycoconjugate comprises at least one covalent linkage between the carrier protein and the saccharide for every 4 saccharide repeat units of the saccharide. 51. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 49 wherein the said capsular saccharide glycoconjugate comprises at least one covalent linkage between the carrier protein and the saccharide for every 25 saccharide repeat units of the saccharide. 52. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 49 wherein the said capsular saccharide glycoconjugate comprises at least one covalent linkage between the carrier protein and the saccharide for every 5 to 10 saccharide repeat units of the saccharide. 53. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is CRM197. 54. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is SCP. 55. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive SCP. 56. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive SCP from GBS (SCPB). 57. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is a fragment of an SCPB. 58. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is a fragment of an SCP which comprises the protease domain, the protease- associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. 59. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive fragment of an SCP, wherein said enzymatically inactive fragment of SCP comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain. 60. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least one amino acid of the wild type sequence and wherein said replacement is selected from the group consisting of D130A, H193A, N295A and S512A where the numbers indicate the amino acid residue position in the peptidase according to the numbering of SEQ ID NO: 1 of WO00/34487. 61. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive fragment of SCP which comprises the protease domain, the protease-associated domain (PA domain) and the three fibronectin type III (Fn) domains but does not comprise the export signal presequence, the pro-sequence and the cell wall anchor domain, where said inactivation is accomplished by replacing at least two amino acids of the wild type sequence wherein said at least two amino acids replacements are D130A and S512A where the numbers indicate the amino acid residue position in the peptidase according to the numbering of SEQ ID NO: 1 of WO00/34487. 62. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity with SEQ ID NO: 41. 63. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive fragment of SCP consisting of a polypeptide having at least 95% identity with SEQ ID NO: 42. 64. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive fragment of SCP which consists of SEQ ID NO: 41. 65. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is an enzymatically inactive fragment of SCP which consists of SEQ ID NO: 42. 66. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is DT (Diphtheria toxoid). 67. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is TT (tetanus toxoid). 68. The capsular saccharide glycoconjugate of any one of paragraphs 38 to 52 wherein said carrier protein is PD (H. influenzae protein D). 69. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from a pathogenic bacteria. 70. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from a pathogenic Streptococcus, a pathogenic Staphylococcus, a pathogenic Enterococcus, a pathogenic Bacillus, a pathogenic Corynebacterium, a pathogenic Listeria, a pathogenic Erysipelothrix, a pathogenic Clostridium, a pathogenic Haemophilus, a pathogenic Neisseria or a pathogenic Escherichia. 71. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Enterococcus faecalis, Escherichia coli, Staphylococcus aureus or Streptococcus. 72. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Haemophilus influenzae, Neisseria meningitidis, S. pneumoniae, S. pyogenes, S. agalactiae, Group C & G Streptococci or Escherichia coli. 73. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Neisseria meningitidis, S. pneumoniae, S. agalactiae or Escherichia coli. 74. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from S. pneumoniae or S. agalactiae. 75. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from S. pneumoniae. 76. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Staphylococcus aureus. 77. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Enterococcus faecalis. 78. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from is Haemophilus influenzae type b. 79. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Neisseria meningitidis. 80. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Escherichia coli. 81. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Enterococcus faecalis. 82. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Streptococcus agalactiae (Group B streptococcus (GBS)). 83. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from GBS type Ia, Ib, II, III, IV, V, VI, VII, VIII or IX. 84. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from a Streptococcus pneumoniae serotype selected from the group consisting of serotypes 1, 2, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9V, 9N, 10A, 10B, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A, 23B, 23F, 24B, 24F, 27, 29, 31, 33B, 33F, 34, 35B, 35F, 38, 72 and 73. 85. The method of making a capsular saccharide glycoconjugate of any one of paragraphs 1 to 37 or the capsular saccharide glycoconjugate of any one of paragraphs 38 to 68, wherein said capsular saccharide is a capsular saccharide from Streptococcus pneumoniae serotype 3. 86. An immunogenic composition comprising a capsular saccharide glycoconjugate of any one of paragraphs 38 to 85. As used herein, the term "about" means within a statistically meaningful range of a value, such as a stated concentration range, time frame, molecular weight, temperature or pH. Such a range can be within an order of magnitude, typically within 20%, more typically within 10%, and even more typically within 5% or within 1% of a given value or range. Sometimes, such a range can be within the experimental error typical of standard methods used for the measurement and/or determination of a given value or range. The allowable variation encompassed by the term "about" will depend upon the particular system under study, and can be readily appreciated by one of ordinary skill in the art. Whenever a range is recited within this application, every number within the range is also contemplated as an embodiment of the disclosure. The terms "comprising", "comprise" and "comprises" herein are intended by the inventors to be optionally substitutable with the terms “consisting essentially of”, “consist essentially of”, “consists essentially of”, "consisting of', "consist of' and "consists of', respectively, in every instance. An "immunogenic amount", an "immunologically effective amount", a “therapeutically effective amount”, a “prophylactically effective amount”, or "dose", each of which is used interchangeably herein, generally refers to the amount of antigen or immunogenic composition sufficient to elicit an immune response, either a cellular (T cell) or humoral (B cell or antibody) response, or both, as measured by standard assays known to one skilled in the art. Any whole number integer within any of the ranges of the present document is contemplated as an embodiment of the disclosure. All references or patent applications cited within this patent specification are incorporated by reference herein. The invention is illustrated in the accompanying examples. The examples below are carried out using standard techniques, which are well known and routine to those of skill in the art, except where otherwise described in detail. The examples are illustrative, but do not limit the invention. EXAMPLE Example 1: Activation of Serotype 3 Capsular Polysaccharide with an alkyne linker Lyophilized S. pneumoniae serotype 3 capsular polysaccharide (300 mg) was reconstituted with anhydrous DMSO (75 mL). The reaction mixture was then warmed to 35°C, and then CDI (100 mg/mL in DMSO), 288 µL (0.2 MEq) was added. The reaction mixture was stirred at 35°C for 3 hrs. After the reaction mixture was cooled to 23°C, WFI 1.5 mL (2% v/v) was added to quench free CDI and stirred for 30 min at 23°C. Then, propargylamine 113 µL (2 MEq) was added and then stirred at 23°C for 20 hrs. After 20 hrs reaction, the reaction mixture was diluted to chilled (at 5°C) with 300 mL 10 mM Sodium Phosphate Buffer (SPB) in saline (pH 7) (5X, v/v). The diluted reaction mixture was then purified by UF/DF using 10K MWCO PES membrane (Millipore Pellicon 2 Mini) against 10 mM SPB in saline (pH 7) (30X, v/v) and filtered through 0.22 µm and analyzed. Example 2: Activation of a carrier protein (SCP) with an agent bearing an N- Hydroxysuccinimide (NHS) moiety and an azido group (azidoacetic acid NHS ester (AAN)) To the SCP solution (500 mg, 86 mL) WFI 14 mL and 0.5 M SPB (pH 8.3) 25 mL were added. After cooled to 8°C, Azidoacetic acid NHS ester (AAN) (20 mg/mL in DMSO) 2.0 mL (0.5 MEq to lysine on SCP) was added to the reaction mixture dropwise maintaining the reaction temperature at 8 ± 3 ºC. After the reaction mixture was stirred for 30 min at 10 ºC, purified by UF/DF using 10K MWCO PES membrane (Millipore Pellicon 2 Mini) against 10 mM sodium phosphate buffer in saline (pH 7.0) (30X diavolume). After UF/DF, retentate was taken and then sucrose was added as solid and then filtered through 0.22 um filter and analyzed. Example 3: Conjugation of S. pneumoniae serotype 3 Capsular Polysaccharide using click chemistry (see FIG.1) The activated alkynyl polysaccharide (as obtained at example 1) and the azido functionalized carrier protein (as obtained at example 2) are conjugated by Cu+1 mediated azide-alkyne cycloaddition reaction, referred as “Click Reaction”. To the mixture of activated alkynyl serotype 3 polysaccharide (see example 1 above) (200 mg) and azido functionalized SCP (200 mg), 1 M Sodium Phosphate Buffer (SPB) in saline (pH 7) and WFI was added to make 100 mM SPB (pH 7) and 2 mg/mL saccharide concentration. And then, the mixture of 5 mM copper sulfate (CuSO4) (5 mL) and 25 mM Tris(3- hydroxypropyltriazolylmethyl)amine (THPTA) (5 mL) was added to the reaction mixture at 23°C and followed by the addition of 100 mM aminoguanidine (10 mL) and 100 mM ascorbate (10 mL). After the reaction mixture was stirred for 1.5 hours at 23°C, just before capping step, additional mixture of 5 mM copper sulfate (2.5 mL) and 25 mM THPTA (2.5 mL) was added to the reaction mixture at 23°C and followed by the additional addition of 100 mM aminoguanidine (5 mL) and 100 mM ascorbate (5 mL). 3-Azido-propyl alcohol (20 MEq per alkyne on polysaccharide) was added to cap the unreacted alkyne group on the polysaccharide and stirred for 1 hour at the same temperature, and then propargyl alcohol (40 MEq per alkyne on poly) was added to cap the unreacted azido group on SCP and stirred for 1 hour at 23°C. Then, the reaction mixture was purified by first UF/DF using 100K MWCO RC membrane (Millipore Pellicon 2 Mini) against 10 mM EDTA + 10 mM SPB in saline (pH 7.0) (30X diavolume) and followed by 5 mM succinate in saline (pH 6.0) (20X diavolume). After UF/DF, filtered through 0.22 um filter (Millipak 40), and analyzed. Table 1. Attributes of Pn3 Conjugates obtained using click chemistry Conjugate # 1 2 Polysaccharide Polysaccharide MW, kDa 338 338 Activated polysaccharide attribute Activated Polysaccharide MW, kDa 401 412 Activated carrier (SCP) attribute Activated carrier MW, kDa 111 111 Conjugate attributes Carrier protein SCP SCP Yield (%) 59 64 Output SPR 0.64 063 Free Saccharide, % <5 <5 Free Protein, % <1 <1 Conjugate MW, kDa 938 1069 MW: molecular weight; SPR: Saccharide to protein ratio The click chemistry allows for generating serotype 3 conjugates with very low free saccharide, very low free protein, good yield and acceptable output SPR and molecular weights. All publications and patent applications mentioned in the specification are indicative of the level of those skilled in the art to which this invention pertains. All publications and patent applications are hereby incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, certain changes and modifications may be practiced within the scope of the appended claims.

Claims

Claims 1. A method of making a capsular saccharide glycoconjugate, comprising the steps of: (a) reacting an isolated capsular saccharide with a carbonic acid derivative and an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide, (b) reacting a carrier protein with an agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group where the NHS moiety reacts with the amino groups to form an amide linkage thereby obtaining an azido functionalized carrier protein, (c) reacting the activated alkynyl saccharide of step (a) with the azido functionalized carrier protein of step (b) by Cu+1 mediated azide-alkyne cycloaddition reaction to form a glycoconjugate.
2. The method of claim 1 wherein, the isolated saccharide is sized to a weight average molecular weight of between 50 kDa and 500 kDa before the activation step (a).
3. The method of any one of claims 1-2 wherein, said carbonic acid derivative is selected from the group consisting of 1,1’-carbonyldiimidazole (CDI), 1,1’-carbonyl-di-(1,2,4-triazole) (CDT), disuccinimidyl carbonate (DSC) and N-hydroxysuccinimidyl chloroformate.
4. The method of any one of claims 1-3 wherein said alkyne linker is a compound of formula (I), H2N X wherein X is selected from the group consisting of CH2, CH2(CH2)n, (CH2CH2O)mCH2CH2, NHCO(CH2)n, NHCO(CH2CH2O)mCH2CH2, OCH2(CH2)n and O(CH2CH2O)mCH2CH2; where n is selected from 1 to 10 and m is selected from 1 to 4.
5. The method of any one of claims 1-4 wherein, said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (III), wherein X is selected from the group consisting of (CH2)nCH2C=O and (CH2CH2O)mCH2CH2=O; where n is selected from 0 to 10 and m is selected from 0 to 4.
6. The method of any one of claims 1-5 wherein, said agent bearing an N-Hydroxysuccinimide (NHS) moiety and an azido group is a compound of formula (IV): (IV)
7. The method of any one of claims 1-6 wherein, step (a) comprises reacting the capsular saccharide with a carbonic acid derivative followed by reacting the carbonic acid derivative- activated capsular saccharide with an alkyne linker in an aprotic solvent to produce an activated alkynyl saccharide.
8. The method of any one of claims 1-7 wherein at step (a) carbonic acid derivative activation is followed by the addition of water.
9. The method of any one of claims 1-8 wherein the conjugation reaction step (c) is carried out in aqueous buffer in the presence of an oxidant and of copper (I) as catalyst.
10. The method of any one of claims 1-8 wherein the conjugation reaction step (c) is carried out in aqueous buffer in the presence of copper (I) as catalyst and ascorbate as oxidant, wherein the reaction mixture further comprises THPTA (tris(3-hydroxypropyltriazolylmethyl)amine) and aminoguanidine.
11. The method of any one of claims 1-10 wherein the method further comprises the step of purifying the glycoconjugate after it is produced.
12. A capsular saccharide glycoconjugate produced according to any one of the methods of claims 1 to 11.
13. A capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII): (VII), wherein X is selected from the group consisting of CH2(CH2)n’, (CH2CH2O)mCH2CH2, NHCO(CH2)n’, NHCO(CH2CH2O)mCH2CH2, OCH2(CH2)n’ and O(CH2CH2O)mCH2CH2; where n’ is selected from 0 to 10 and m is selected from 1 to 4, and wherein X' is selected from the group consisting of CH2(CH2)n”, CH2O(CH2)n’’CH2, CH2O(CH2CH2O)m’(CH2)n’’CH2, where n’’ is selected from 0 to 10 and m’ is selected from 0 to 4.
14. A capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VII), wherein X is CH2(CH2)n’, where n’ is 0 and wherein X' is CH2(CH2)n” where n’’ is 0.
15. A capsular saccharide glycoconjugate comprising a capsular saccharide covalently conjugated to a carrier protein (CP) through a spacer and having the general formula (VIII), (VIII).
16. The capsular saccharide glycoconjugate of any one of claims 12 to 15 wherein said carrier protein is CRM197, SCP, DT (Diphtheria toxoid), TT (tetanus toxoid), or PD (H. influenzae protein D).
17. The method of making a capsular saccharide glycoconjugate of any one of claims 1 to 11 or the capsular saccharide glycoconjugate of any one of claims 12 to 16, wherein said capsular saccharide is a capsular saccharide from a pathogenic bacteria.
18. The method of making a capsular saccharide glycoconjugate of any one of claims 1 to 11 or the capsular saccharide glycoconjugate of any one of claims 12 to 16, wherein said capsular saccharide is a capsular saccharide from Haemophilus influenzae, Neisseria meningitidis, S. pneumoniae, S. pyogenes, S. agalactiae, Group C & G Streptococci or Escherichia coli.
19. The method of making a capsular saccharide glycoconjugate of any one of claims 1 to 11 or the capsular saccharide glycoconjugate of any one of claims 12 to 16, wherein said capsular saccharide is a capsular saccharide from a Streptococcus pneumoniae serotype selected from the group consisting of serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9V, 9N, 10A, 10B, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A, 23B, 23F, 24B, 24F, 27, 29, 31, 33B, 33F, 34, 35B, 35F, 38, 72 and 73.
20. An immunogenic composition comprising a capsular saccharide glycoconjugate of any one of claims 12 to 19.
EP24704907.5A 2023-02-10 2024-02-07 Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof Pending EP4661911A1 (en)

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