WO2025193903A1

WO2025193903A1 - Pneumococcal polysaccharide compositions and uses thereof

Info

Publication number: WO2025193903A1
Application number: PCT/US2025/019697
Authority: WO
Inventors: Francesco Berti; Xiuru Li
Original assignee: Affinivax Inc
Current assignee: Affinivax Inc
Priority date: 2024-03-15
Filing date: 2025-03-13
Publication date: 2025-09-18
Anticipated expiration: 2026-09-15

Abstract

The present disclosure provides compositions comprising a purified capsular polysaccharide having one or more repeating units of formula I: wherein each R is independently selected from H and -C(O)-CH3; and uses thereof.

Description

AFV70463FF PNEUMOCOCCAL POLYSACCHARIDE COMPOSITIONS AND USES THEREOF BACKGROUND [0001] Streptococcus pneumoniae infection can lead to invasive disease in humans, such as meningitis, sepsis, and pneumonia. Redin, et al., Microbiol. Spectr., 2021 Nov-Dec; 9(3): e01150- 21 (Dec.8, 2021). Pneumococcal disease is the cause of death for about 500,000 children under the age of 5 die each year and presents substantial risk to elderly and immunocompromised individuals. Takano, et al., Sci. Rep., 9, 19823 (2019). S. pneumoniae produces structurally diverse capsular polysaccharides, which are a major virulence factor. Redin, et al., Microbiol. Spectr., 2021 Nov-Dec; 9(3): e01150-21 (Dec.8, 2021). Over 100 serotypes of S. pneumoniae have been described to date. Ganaie, et al., mBio., 2020 May-Jun; 11(3): e00937-20 (May 19, 2020). SUMMARY [0002] Pneumococcal polysaccharide vaccines (PPVs) and pneumococcal conjugate vaccines (PCVs) targeting multiple S. pneumoniae serotypes have been developed over the last years and are widely available for the prevention of pneumococcal disease. To date, however, the capsular polysaccharide repeating unit structure of several S. pneumoniae serotypes is unknown, including serotype 38. The present disclosure provides, among other things, insights and technologies associated with the discovery of the structure and characterization of a previously uncharacterized serotype of S. pneumoniae, serotype 38. In some embodiments, insights and technologies provided herein are useful to provide immunogenic compositions (e.g., vaccines) with increased immunogenicity to protect against certain diseases, disorders, and conditions, such as those associated with targeting S. pneumoniae serotype 38. [0003] In some embodiments, the present disclosure provides a composition comprising a purified capsular polysaccharide having repeating units of formula I: AFV70463FF HO HO O wherein each R [0004] In some embodiments, about 60 to about 75% of the repeat units of the purified polysaccharide composition of formula I comprise an O-acetylated group at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃). In some embodiments, a purified capsular polysaccharide described herein comprises between 0% and 100% of repeating units being O-acetylated at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃ in formula I). In some embodiments, a purified capsular polysaccharide described herein comprises greater than or equal to 60% of repeating units being O-acetylated at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃ of formula I). In some embodiments, a purified capsular polysaccharide described herein comprises less than 60% of repeating units being O-acetylated at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃ of formula I). In some embodiments, a purified capsular polysaccharide described herein comprises about 68% of repeating units being O-acetylated at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃ in formula I). [0005] In some embodiments, the present disclosure provides a purified capsular polysaccharide that is activated with a chemical reagent to produce reactive groups for conjugation to a suitable conjugate (e.g., a linker or carrier protein). In some embodiments, activation occurs at the C_2, C_3, C_5, or C_6, of the -Galf residue (i.e., at the position designated as R in formula I, above). In some embodiments, activation occurs in less than 100% of repeating units in a purified capsular AFV70463FF polysaccharide of formula I. In some embodiments, activation occurs in about 1% to about 50% of repeating units in a purified capsular polysaccharide of formula I. [0006] In some embodiments, the present disclosure provides an immunogenic composition comprising (i) a purified capsular polysaccharide; and (ii) a polypeptide, wherein the purified capsular polysaccharide has repeating units of formula I: HO HO O and wherein [0007] In some embodiments, the present disclosure provides a method of inducing an immune response to a Streptococcus pneumoniae capsular polysaccharide comprising administering to a subject an immunogenic composition comprising (i) a purified capsular polysaccharide; and (ii) a polypeptide, wherein the purified capsular polysaccharide has repeating units of formula I:

AFV70463FF HO HO O and wherein [0008] In some embodiments, an immunogenic composition described herein is based on a MAPS technology described in PCT Pat App. Nos. PCT/US/2012037412, PCT/US2019/050907, PCT/US2022/042964, and PCT/US2022/043156, each of which is incorporated by reference in the entirety. [0009] In some embodiments, an immunogenic composition described herein comprises a purified polysaccharide composition of formula I conjugated to a polypeptide. In some embodiments, an immunogenic composition described herein comprises a purified polysaccharide composition of formula I non-covalently complexed with a polypeptide. [0010] In some embodiments, the present disclosure provides a method of characterizing an immunogenic composition comprising a purified capsular polysaccharide having repeating units of formula I

AFV70463FF HO HO O wherein each the method comprises steps of: characterizing a number of units of formula I in the purified capsular polysaccharide where R is –C(O)-CH₃; and comparing the number of units of formula I where R is –C(O)-CH₃ to a reference standard. [0011] In some embodiments, the present disclosure provides a method of characterizing an immunogenic composition comprising a purified capsular polysaccharide having repeating units of formula I HO HO

AFV70463FF wherein each R is independently selected from H and –C(O)-CH₃, wherein the method comprises steps of: quantifying a percentage of units of the purified capsular polysaccharide that are of formula I; and comparing the percentage of units of formula I in the purified capsular polysaccharide to a reference standard. [0012] In some embodiments, the present disclosure provides a method of selecting a level of acetylation of a purified capsular polysaccharide for an immunogenic composition, the method comprising steps of: administering to each of a plurality of animals one of a plurality of compositions comprising a purified capsular polysaccharide having repeating units of formula I, HO HO O wherein each R of the plurality of compositions, R is –C(O)-CH3 in a distinct total instances of units of formula I; determining the level of an immune response to the purified capsular polysaccharide; comparing the level of immune response to a reference level; and selecting a purified capsular polysaccharide with a given acetylation level for inclusion in the immunogenic composition, if the determined immune response is higher than the reference level. AFV70463FF BRIEF DESCRIPTION OF THE DRAWINGS [0013] The present teachings described herein will be more fully understood from the following description of various illustrative embodiments, when read together with the accompanying drawings. It should be understood that the drawings described below are for illustration purposes only and are not intended to limit the scope of the present teachings in any way. [0014] FIG. 1 is an HPAEC-PAD profile of S. pneumoniae serotype 38 capsular polysaccharide following acid hydrolysis in 2M TFA at 120 °C for 2 hours. [0015] FIG.2 is an ESI-Q-TOF MS spectrum of fragments generated by base treatment of S. pneumoniae serotype 38 capsular polysaccharide. [0016] FIG. 3 shows GC-MS spectra of PMMA from native and reduced S. pneumoniae serotype 38 capsular polysaccharide. [0017] FIG. 4 depicts PMMAA derivatives corresponding to the reduced Sugp moiety in Spn38 polysaccharide. [0018] FIG.5 depicts MS fragments of PMMA of reduced Sugp at 23.8 and 24.6 min [0019] FIG. 6 is a ¹H NMR spectra of native (trace a) and de-O-acetylated (trace b) S. pneumoniae serotype 38 capsular polysaccharide recorded at 600 MHz and 323 K. [0020] FIG. 7 is a ¹H-¹³C HSQC spectrum of de-O-acetylated S. pneumoniae serotype 38 capsular polysaccharide recorded at 600 MHz and 323 K. Key repeating unit proton/carbon cross- peaks have been labeled. [0021] FIG. 8 depicts results of partial HSQC NMR of de-O-acetylated and reduced de-O- acetylated S. pneumoniae serotype 38 capsular polysaccharide. [0022] FIG. 9 is a ³¹P NMR spectrum of native S. pneumoniae serotype 38 capsular polysaccharide collected at 242 MHz and 323 K. [0023] FIG. 10 is a ¹H 1D TOCSY (120 ms mixing time) and 1D NOESY (200 ms mixing time) NMR spectra collected by selectively irradiating the anomeric signals of de-O-acetylated S. pneumoniae serotype 38 capsular polysaccharide recorded at 600 MHz and 323 K. Trace a) ¹H reference spectrum; trace b) Selective TOCSY H₁ A; trace c) Selective NOESY H₁ A; trace d) Selective TOCSY H₁ B; trace e) Selective NOESY H₁ B; trace f) Selective TOCSY H₁ C; trace g) Selective NOESY H₁ C; trace h) Selective TOCSY H₁ D and E; trace i) Selective NOESY H₁ D and E. AFV70463FF [0024] FIG. 11 is a 2D NOESY spectrum of de-O-acetylated S. pneumoniae serotype 38 capsular polysaccharide recorded at 600 MHz and 323 K. Key repeating unit proton/carbon cross- peak confirming the glycosidic linkages have been labeled. [0025] FIG. 12 is a ¹H-¹³C HMBC spectrum of de-O-acetylated S. pneumoniae serotype 38 capsular polysaccharide recorded at 600 MHz and 323 K. Key repeating unit proton/carbon cross- peak confirming the glycosidic linkages have been labeled. [0026] FIG.13 is a partial ¹H-¹³C HMBC spectrum of de-O-acetylated S. pneumoniae serotype 38 capsular polysaccharide. [0027] [0028] FIGs. 14A-14D are partial ¹H-¹H TOCSY (FIG. 14A), COSY (FIG. 14B), NOESY (FIG. 14C), and HMBC (FIG. 14D) spectra of native S. pneumoniae serotype 38 capsular polysaccharide recorded at 600 MHz and 323 K. Key repeating unit proton/carbon cross-peak confirming the O-acetylation site at the position C₂ of -Galp residue has been labeled. [0029] FIG. 15 is an exemplary structure of S. pneumoniae serotypes 38 capsular polysaccharide repeating unit. [0030] FIG.16 is a schematic of an exemplary CP1 fusion protein. Such an exemplary CP1 fusion protein comprises a biotin-binding protein such as, e.g., a truncated rhizavidin protein (e.g., amino acids 45-179 of a wild-type rhizavidin protein), a first linker (e.g., a GGGGSSS (SEQ ID NO: 30) linker), a SP1500 polypeptide (e.g., amino acids 27-278 of a full-length S. pneumoniae SP1500 polypeptide), a second linker (e.g., the amino acid sequence AAA), and a SP0785 polypeptide (e.g., amino acids 33-399 of a full length S. pneumoniae SP0785 polypeptide). In some embodiments, a CP1 fusion protein may further comprise a detectable or purification tag (e.g., His tag). The amino acid sequence AAA can be from the Not I site on a pET21/24 plasmid, or synthesized. For a GGGGSSS (SEQ ID NO: 30) linker, the SSS amino acid sequence can be from the Sac I site on a pET21/24 plasmid, with the GGGG (SEQ ID NO: 59) amino acid sequence added to create a flexible linker with minimal steric hindrance. Alternatively, the GGGGSSS (SEQ ID NO: 30) linker can be synthesized. [0031] FIG.17 is a schematic of an exemplary SPP2 fusion protein. Such an exemplary SPP2 fusion protein comprises a biotin-binding protein, such as e.g., a truncated rhizavidin protein (e.g., amino acids 45-179 of a wild-type rhizavidin protein, denoted as Rhavi), a first linker (e.g., a GGGGSSS (SEQ ID NO: 30) linker), a pneumolysin (Ply) polypeptide (e.g., amino acids 2-470 AFV70463FF of a full-length S. pneumoniae Ply polypeptide comprising mutations G294P, D385N, C428G, and W433F, denoted as PdT(G294P)), a second linker (e.g., a GGGGSSS (SEQ ID NO: 30) linker), and a SP0435 polypeptide (e.g., amino acids 62-185 of a full length S. pneumoniae SP0435 polypeptide). In some embodiments, a SPP2 fusion protein may further comprise a detection or purification tag (e.g., a His tag). For a GGGGSSS (SEQ ID NO: 30) linker, the SSS amino acid sequence can be from the Sac I site on a pET21/24 plasmid, with the GGGG (SEQ ID NO: 59) amino acid sequence added to create a flexible linker with minimal steric hindrance. Alternatively, the GGGGSSS (SEQ ID NO: 30) linker can be synthesized. [0032] FIG. 18 is a schematic of a plasmid construct composed of the pET-24a(+) vector containing a sequence encoding an exemplary SPP2 fusion protein. [0033] FIGs. 19 and 20 are flow-charts depicting an exemplary upstream (FIG. 19) and downstream (FIG. 20) manufacturing process for fusion proteins as described herein (e.g., CP1 and SPP2). An exemplary SPP2 polypeptide is a fusion protein comprising a truncated rhizavidin [amino acids 45-179 of a full-length rhizavidin protein], a first linker (e.g., a GGGGSSS linker), a pneumolysin (Ply) polypeptide, a second linker (e.g., a GGGGSSS linker), and a SP0435 polypeptide. An exemplary CP1 polypeptide is a fusion protein comprising a truncated rhizavidin [amino acids 45-179 of a full-length rhizavidin protein], a first linker (e.g., a GGGGSSS linker), a SP1500 polypeptide, a second linker (e.g., the amino acid sequence AAA), and a SP0785 polypeptide. SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis; TFF: tangential flow filtration; DS: drug substance. [0034] FIG. 21 is a table showing exemplary structures of S. pneumoniae antigenic polysaccharides of serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, and 35B. [0035] FIG. 22 is a schematic representation of an exemplary Multiple Antigen Presenting System (MAPS). In the exemplary embodiment shown, MAPS immunogenic complexes comprise one or more polypeptide antigens fused to the biotin-binding protein rhizavidin, or a biotin-binding domain or biotin-binding fragment thereof, and a biotinylated antigenic polysaccharide. In this figure, each MAPS complex is formed between one or more fusion proteins and a biotinylated polysaccharide by non-covalent binding of a truncated rhizavidin to biotin. [0036] FIG. 23 is a flow-chart depicting an exemplary manufacturing process for MAPS immunogenic complexes, comprising biotinylation of antigenic polysaccharides and assembly AFV70463FF with fusion protein(s) as described herein (e.g., CP1 or SPP2). MAPS: Multiple Antigen Presenting System. [0037] FIG. 24 is a flow-chart depicting exemplary manufacturing processes for a MAPS vaccine. For example, such exemplary processes can be used to manufacture a MAPS34 vaccine, comprising MAPS immunogenic complexes that comprise capsular polysaccharides from 34 different S. pneumoniae serotypes. DMAP: 4-Dimethylaminopyridine; MAPS: Multiple Antigen Presenting System; PS: polysaccharide. † In-process tests [0038] FIG.25 depicts the results of nephrology tests using pooled sera from 10 rabbits inoculated with PS38-CRM197. [0039] FIG.26 depicts ELISA results for rabbit sera raised against PS38-CRM197. [0040] FIG.27 depicts ELISA results for rabbit sera following inoculation with a multivalent MAPS vaccine including PS-38 complexed with CP1 or SPP2. Sera were collected from male NZW rabbits (n=20) before the first dose (P0), second dose (P1), and two weeks after the second dose (P2). Anti-ST-38 specific IgG concentrations were determined by comparing samples to a reference standard of known concentration. Horizontal bars are the geometric mean concentrations of the groups and error bars are 95% Cis. DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS [0041] The present disclosure provides, among other things, a purified capsular polysaccharide of S. pneumoniae serotype 38, and immunogenic compositions comprising said purified capsular polysaccharide. Definitions [0042] Compounds of this disclosure include those described generally above and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75^th Ed. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5^th Ed., Ed.: Smith, M.B. and AFV70463FF March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference. [0043] Unless otherwise indicated, structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including replacement of hydrogen by deuterium or tritium, or replacement of a carbon by ¹³C- or ¹⁴C-enriched carbon are within the scope of this disclosure. [0044] About or approximately: As used herein, the term "approximately" or "about," as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In general, those skilled in the art, familiar within the context, will appreciate the relevant degree of variance encompassed by "about" or "approximately" in that context. For example, in some embodiments, the term "approximately" or "about" may encompass a range of values that are within (i.e., ±) 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value. [0045] Administering: As used herein, the term "administering" or "administration" typically refers to the administration of a composition to a subject to achieve delivery of an agent that is, or is included in, a composition to a target site or a site to be treated. Those of ordinary skill in the art will be aware of a variety of routes that may, in appropriate circumstances, be utilized for administration to a subject, for example a human. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, etc. In some particular embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, dermal (which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreal, etc. In some embodiments, administration may be parenteral. In some embodiments, administration may be oral. In some particular embodiments, administration may be intravenous. In some particular embodiments, administration may be subcutaneous. In some embodiments, administration may involve only a single dose. In some embodiments, administration may involve application of a fixed number of doses. In some embodiments, administration may involve dosing that is intermittent (e.g., a plurality of doses AFV70463FF separated in time) and/or periodic (e.g., individual doses separated by a common period of time) dosing. In some embodiments, administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time. In some embodiments, administration may comprise a prime- and-boost protocol. A prime-and-boost protocol can include administration of a first dose of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) followed by, after an interval of time, administration of a second or subsequent dose of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine). In the case of an immunogenic composition, a prime-and-boost protocol can result in an increased immune response in a patient. [0046] Agent: The term “agent”, as used herein, may be used to refer to a compound or entity of any chemical class including, for example, a polypeptide, nucleic acid, saccharide, lipid, small molecule, metal, or combination or complex thereof. In appropriate circumstances, as will be clear from context to those skilled in the art, the term may be utilized to refer to an entity that is or comprises a cell or organism, or a fraction, extract, or component thereof. Alternatively, or additionally, as context will make clear, the term may be used to refer to a natural product in that it is found in and/or is obtained from nature. In some instances, again as will be clear from context, the term may be used to refer to one or more entities that is man-made in that it is designed, engineered, and/or produced through action of the hand of man and/or is not found in nature. In some embodiments, an agent may be utilized in isolated or pure form; in some embodiments, an agent may be utilized in crude form. In some embodiments, potential agents may be provided as collections or libraries, for example that may be screened to identify or characterize active agents within them. In some cases, the term “agent” may refer to a compound or entity that is or comprises a polymer; in some cases, the term may refer to a compound or entity that comprises one or more polymeric moieties. In some embodiments, the term “agent” may refer to a compound or entity that is not a polymer and/or is substantially free of any polymer and/or of one or more particular polymeric moieties. In some embodiments, the term may refer to a compound or entity that lacks or is substantially free of any polymeric moiety. [0047] Amino Acid: The term “amino acid”, as used herein, refers to any compound and/or substance that can be incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H₂N–C(H)(R)– COOH. In some embodiments, an amino acid is a naturally-occurring embodiments, an amino acid is a non-natural amino acid; in some embodiments, an amino acid is AFV70463FF a D-amino acid; in some embodiments, an amino acid is an L-amino acid. “Standard amino acid” refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. “Non-standard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. In some embodiments, an amino acid, including a carboxy- and/or amino-terminal amino acid in a polypeptide, can contain a structural modification as compared with the general structure above. For example, in some embodiments, an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and/or the hydroxyl group) as compared with the general structure. In some embodiments, such modification may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid. In some embodiments, such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid. As will be clear from context, in some embodiments, the term “amino acid” may be used to refer to a free amino acid; in some embodiments, the term “amino acid” may be used to refer to an amino acid residue of a polypeptide. [0048] Antigen: The term “antigen”, as used herein, refers to (i) an agent that induces an immune response; and/or (ii) an agent that binds to a T cell receptor (e.g., when presented by an MHC molecule) or to an antibody. In some embodiments, an antigen induces a humoral response (e.g., including production of antigen-specific antibodies); in some embodiments, an antigen induces a cellular response (e.g., involving T cells whose receptors specifically interact with the antigen). In some embodiments, an antigen induces a humoral response and a cellular response. In some embodiments, an antigen binds to an antibody and may or may not induce a particular physiological response in an organism. In general, an antigen may be or include any chemical entity such as, for example, a small molecule, a nucleic acid, a polypeptide, a carbohydrate, a lipid, a polymer (in some embodiments, other than a biologic polymer (e.g., other than a nucleic acid or amino acid polymer)), etc. In some embodiments, an antigen is or comprises a polypeptide. In some embodiments, an antigen is or comprises a polysaccharide. Those of ordinary skill in the art will appreciate that, in general, an antigen may be provided in isolated or pure form, or alternatively may be provided in crude form (e.g., together with other materials, for example in an AFV70463FF extract such as a cellular extract or other relatively crude preparation of an antigen-containing source). In some embodiments, antigens utilized in accordance with the present disclosure are provided in a crude form. In some embodiments, an antigen is a recombinant antigen. In some embodiments, an antigen is a polypeptide or a polysaccharide that, upon administration to a subject, induces a specific and/or clinically relevant immune response to such polypeptide or polysaccharide. In some embodiments, an antigen is selected to induce a specific and/or clinically relevant immune response to such polypeptide or polysaccharide. As will be appreciated by a skilled artisan, an antigen can be a full-length polypeptide antigen, or a portion thereof comprising one or more B-cell epitopes and/or one or more T-cell epitopes. [0049] Associated With: As used herein, two entities are “associated” with one another if the presence, level and/or form of one is correlated with that of the other. In some embodiments, two or more entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and/or remain in physical proximity with one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another. In some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non-covalently associated, for example by means of affinity interactions, electrostatic interactions, hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof. [0050] Biotin-Binding Moiety: The term “biotin-binding moiety”, as used herein, refers to a biotin-binding protein, a biotin-binding fragment thereof, or a biotin-binding domain thereof. [0051] Binding: It will be understood that the term “binding”, as used herein, typically refers to a non-covalent association between or among two or more entities. “Direct” binding involves physical contact between entities or moieties; indirect binding involves physical interaction by way of physical contact with one or more intermediate entities. Binding between two or more entities can typically be assessed in any of a variety of contexts – including where interacting entities or moieties are studied in isolation or in the context of more complex systems (e.g., while covalently or otherwise associated with a carrier entity and/or in a biological system cell). [0052] Biological sample: As used herein, the term “biological sample” typically refers to a sample obtained or derived from a biological source (e.g., a tissue or organism or cell culture) of interest, as described herein. In some embodiments, a source of interest comprises an organism, such as an animal or human. In some embodiments, a biological sample is or comprises biological AFV70463FF tissue or fluid. In some embodiments, a biological sample may be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; cell-containing body fluids; free floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; washings or lavages such as a ductal lavages or bronchoalveolar lavages; aspirates; scrapings; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions, and/or excretions; and/or cells therefrom, etc. In some embodiments, a biological sample is or comprises cells obtained from an individual. In some embodiments, obtained cells are or include cells from an individual from whom the sample is obtained. In some embodiments, a sample is a “primary sample” obtained directly from a source of interest by any appropriate means. For example, in some embodiments, a primary biological sample is obtained by methods selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluid (e.g., blood, lymph, feces etc.), etc. In some embodiments, as will be clear from context, the term “sample” refers to a preparation that is obtained by processing (e.g., by removing one or more components of and/or by adding one or more agents to) a primary sample. For example, filtering using a semi-permeable membrane. Such a “processed sample” may comprise, for example, nucleic acids or proteins extracted from a sample or obtained by subjecting a primary sample to techniques such as amplification or reverse transcription of mRNA, isolation and/or purification of certain components, etc. [0053] Carrier: As used herein, the term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which a composition is administered. In some exemplary embodiments, carriers can include sterile liquids, such as, for example, water and oils, including oils of petroleum, animal, vegetable or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil and the like. In some embodiments, carriers are or include one or more solid components. [0054] Carrier protein: As used herein, the term “carrier protein” refers to a protein or peptide that is coupled, or complexed, or otherwise associated with a hapten (e.g., a small peptide or lipid) or less immunogenic antigen (e.g., a polysaccharide) and that induces or improves an immune response to such a coupled, or complexed, or otherwise associated hapten (e.g., a small peptide or lipid) or less immunogenic antigen (e.g., a polysaccharide). In some embodiments, such an immune response is or comprises a response to a hapten or less immunogenic antigen that is coupled, or complexed, or otherwise associated with such a carrier protein. In some embodiments, AFV70463FF such an immune response is or comprises a response to both a carrier protein and a hapten or less immunogenic antigen that is coupled, or complexed, or otherwise associated with such a carrier protein. In some embodiments, no significant immune response to a carrier protein itself occurs. In some embodiments, immune response to a carrier protein may be detected; in some embodiments, immune response to such a carrier protein is strong. In some embodiments, a carrier protein is coupled, or complexed, or otherwise associated with one or more other molecules. [0055] Colonization: As used herein, the term “colonization” generally refers to the ability of a microbe to grow at a target site or surface. For example, the terms “colonization” refers to the ability of a microbe (e.g., a bacterium) to grow at an anatomical site (e.g., a mucosal membrane, gastrointestinal tract, injury site, organ, etc.) of a host. [0056] Combination therapy: As used herein, the term “combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents or modality(ies)). In some embodiments, the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, “administration” of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination. For clarity, combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some embodiments, two or more agents, or active moieties thereof, may be administered together in a combination composition, or even in a combination compound (e.g., as part of a single chemical complex or covalent entity). [0057] Comparable: As used herein, the term “comparable” refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison therebetween so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features. Those of ordinary skill in the art will understand, in context, what degree of identity is required in any given circumstance for two or more such agents, entities, situations, sets of AFV70463FF conditions, etc. to be considered comparable. For example, those of ordinary skill in the art will appreciate that sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied. [0058] Composition: Those skilled in the art will appreciate that the term “composition” may be used to refer to a discrete physical entity that comprises one or more specified components. In general, unless otherwise specified, a composition may be of any form – e.g., gas, gel, liquid, solid, etc. [0059] Domain: The term “domain” as used herein refers to a section or portion of an entity. In some embodiments, a “domain” is associated with a particular structural and/or functional feature of the entity so that, when the domain is physically separated from the rest of its parent entity, it substantially or entirely retains the particular structural and/or functional feature. Alternatively or additionally, a domain may be or include a portion of an entity that, when separated from that (parent) entity and linked with a different (recipient) entity, substantially retains and/or imparts on the recipient entity one or more structural and/or functional features that characterized it in the parent entity. In some embodiments, a domain is a section or portion of a molecule (e.g., a small molecule, carbohydrate, lipid, nucleic acid, or polypeptide). In some embodiments, a domain is a section of a polypeptide; in some such embodiments, a domain is characterized by a particular structural element (e.g., a particular amino acid sequence or sequence -_{-sheet character, coiled-coil character, random coil character, etc.),} and/or by a particular functional feature (e.g., binding activity, enzymatic activity, folding activity, signaling activity, etc.). [0060] Dosage form or unit dosage form: Those skilled in the art will appreciate that the term “dosage form” may be used to refer to a physically discrete unit of an active agent (e.g., a therapeutic or diagnostic agent) for administration to a subject. Typically, each such unit contains a predetermined quantity of active agent. In some embodiments, such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen). AFV70463FF [0061] Dosing regimen or therapeutic regimen: Those skilled in the art will appreciate that the terms “dosing regimen” and “therapeutic regimen” may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen comprises a plurality of doses each of which is separated in time from other doses. In some embodiments, individual doses are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen). [0062] Excipient: As used herein, the term “excipient” refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. [0063] Fragment: A “fragment” of a material or entity as described herein has a structure that includes a discrete portion of the whole, but lacks one or more moieties found in the whole. In some embodiments, a fragment consists of such a discrete portion. In some embodiments, a fragment includes a discrete portion of the whole which discrete portion shares one or more functional characteristics found in the whole. In some embodiments, a fragment consists of such a discrete portion. In some embodiments, a fragment consists of or comprises a characteristic structural element or moiety found in the whole. In some embodiments, a fragment of a polymer, e.g., a polypeptide or a polysaccharide, comprises or consists of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, AFV70463FF 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 or more monomeric units (e.g., residues) as found in the whole polymer. In some embodiments, a polymer fragment comprises or consists of at least about 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more of the monomeric units (e.g., residues) found in the whole polymer. The whole material or entity may, in some embodiments, be referred to as the “parent” of the whole. [0064] Homology: As used herein, the term “homology” refers to the overall relatedness between polymeric molecules, e.g., between nucleic acid molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embodiments, polymeric molecules are considered to be “homologous” to one another if their sequences are at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical. In some embodiments, polymeric molecules are considered to be “homologous” to one another if their sequences are at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% similar (e.g., containing residues with related chemical properties at corresponding positions). For example, as is well known by those of ordinary skill in the art, certain amino acids are typically classified as similar to one another as “hydrophobic” or “hydrophilic” amino acids, and/or as having “polar” or “non-polar” side chains. Substitution of one amino acid for another of the same type may often be considered a “homologous” substitution. [0065] Identity: As used herein, the term “identity” refers to the overall relatedness between polymeric molecules, e.g., between nucleic acid molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embodiments, polymeric molecules are considered to be “substantially identical” to one another if their sequences are at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical. Calculation of the percent identity of two nucleic acid or polypeptide sequences, for example, can be performed by aligning the two AFV70463FF sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence. The nucleotides at corresponding positions are then compared. When a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix. [0066] Improve, increase, inhibit or reduce: As used herein, the terms “improve”, “increase”, “inhibit’, “reduce”, or grammatical equivalents thereof, indicate values that are relative to a baseline or other reference measurement. In some embodiments, an appropriate reference measurement may be or comprise a measurement in a particular system (e.g., in a single subject) under otherwise comparable conditions absent presence of (e.g., prior to and/or after) a particular agent or treatment, or in presence of an appropriate comparable reference agent. In some embodiments, an appropriate reference measurement may be or comprise a measurement in comparable system known or expected to respond in a particular way, in presence of the relevant agent or treatment. [0067] Immunologically effective amount or immunologically effective dose: As used herein, “immunologically effective amount” or “immunologically effective dose” refers to an amount of an antigenic or immunogenic substance, e.g., an antigen, immunogen, immunogenic AFV70463FF agent, immunogenic composition, vaccine composition, or pharmaceutical composition, which when administered to a subject, either in a single dose or as part of a series of doses, that is sufficient to enhance a subject’s own immune response against a subsequent exposure to a pathogen. In some embodiments, the pathogen is S. pneumoniae. In some embodiments, the immune response is against one or more different serotypes of S. pneumoniae. In some embodiments, the immune response is against two or more different serotypes of S. pneumoniae. In some embodiments, the immune response is against nine or more different serotypes of S. pneumoniae. In some embodiments, the immune response is against thirteen or more different serotypes of S. pneumoniae. In some embodiments, the immune response is against fifteen or more different serotypes of S. pneumoniae. In some embodiments, the immune response is against twenty-three or more different serotypes of S. pneumoniae. In some embodiments, the immune response is against twenty-four or more different serotypes of S. pneumoniae. An immunologically effective amount may vary based on the subject to be treated, the species of the subject, the degree of immune response desired to induce, etc. In some embodiments, an immunologically effective amount is sufficient for treatment or protection of a subject having or at risk of having disease. In some embodiments, an immunologically effective amount refers to a non-toxic but sufficient amount that can be an amount to treat, attenuate, or prevent infection and/or disease (e.g., bacterial infection, pneumococcal infection, bacterial colonization, pneumococcal colonization, complications associated with bacterial infection, complications associated with pneumococcal infection, etc.) in any subject. In some embodiments, an immunologically effective amount is sufficient to induce an immunoprotective response upon administration to a subject. [0068] Immunoprotective response or protective response: As used herein, “immunoprotective response” or “protective response” refers to an immune response that mediates antigen or immunogen-induced immunological memory. In some embodiments, an immunoprotective response is induced by the administration of a substance, e.g., an antigen, immunogen, immunogenic agent, immunogenic composition, vaccine composition, or pharmaceutical composition to a subject. In some embodiments, immunoprotection involves one or more of active immune surveillance, a more rapid and effective response upon immune activation as compared to a response observed in a naïve subject, efficient clearance of the activating agent or pathogen, followed by rapid resolution of inflammation. In some embodiments, AFV70463FF an immunoprotective response is an adaptive immune response. In some embodiments, an immunoprotective response is sufficient to protect an immunized subject from productive infection by a particular pathogen or pathogens to which a vaccine is directed (e.g., S. pneumoniae infection). [0069] Immunization: As used herein, “immunization”, or grammatical equivalents thereof, refers to a process of inducing an immune response to an infectious organism or agent in a subject (“active immunization”), or alternatively, providing immune system components against an infectious organism or agent to a subject (“passive immunization”). In some embodiments, immunization involves the administration of one or more antigens, immunogens, immunogenic agents, immunogenic compositions, vaccine compositions, immune molecules such as antibodies, immune sera, immune cells such as T cells or B cells, or pharmaceutical compositions to a subject. In some embodiments, immunization is performed by administering an immunologically effective amount of a substance, e.g., an antigen, immunogen, immunogenic agent, immunogenic composition, vaccine composition, immune molecule such as an antibody, immune serum, immune cell such as a T cell or B cell, or pharmaceutical composition to a subject. In some embodiments, immunization results in an immunoprotective response in the subject. In some embodiments, active immunization is performed by administering to a subject an antigenic or immunogenic substance, e.g., an antigen, immunogen, immunogenic agent, immunogenic composition, vaccine composition, or pharmaceutical composition. In some embodiments, passive immunization is performed by administering to a subject an immune system component, e.g., an immune molecule such as an antibody, immune serum, or immune cell such as a T cell or B cell. [0070] Isolated: As used herein, the term “isolated”, or grammatical equivalents thereof, refers to a substance and/or entity that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature and/or in an experimental setting), and/or (2) designed, produced, prepared, and/or manufactured by the hand of man. Isolated substances and/or entities may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which they were initially associated. In some embodiments, isolated agents are about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about AFV70463FF 99% pure. As used herein, a substance is "pure" if it is substantially free of other components. In some embodiments, as will be understood by those skilled in the art, a substance may still be considered "isolated" or even "pure", after having been combined with certain other components such as, for example, one or more carriers or excipients (e.g., buffer, solvent, water, etc.); in such embodiments, percent isolation or purity of the substance is calculated without including such carriers or excipients. To give but one example, in some embodiments, a biological polymer such as a polypeptide or polysaccharide that occurs in nature is considered to be "isolated" when, a) by virtue of its origin or source of derivation is not associated with some or all of the components that accompany it in its native state in nature; b) it is substantially free of other polypeptides or nucleic acids of the same species from the species that produces it in nature; c) is expressed by or is otherwise in association with components from a cell or other expression system that is not of the species that produces it in nature. Thus, for instance, in some embodiments, a polypeptide or polysaccharide that is chemically synthesized or is synthesized in a cellular system different from that which produces it in nature is considered to be an "isolated" polypeptide or polysaccharide. Alternatively or additionally, in some embodiments, a polypeptide or polysaccharide that has been subjected to one or more purification techniques may be considered to be an "isolated" polypeptide or polysaccharide to the extent that it has been separated from other components a) with which it is associated in nature; and/or b) with which it was associated when initially produced. [0071] Linker: As used herein, the term “linker” is used to refer to an entity that connects two or more elements to form a multi-element agent. For example, those of ordinary skill in the art appreciate that a polypeptide whose structure includes two or more functional or organizational domains often includes a stretch of amino acids between such domains that links them to one another. In some embodiments, a polypeptide comprising a linker element has an overall structure of the general form S1-L-S2, wherein S1 and S2 may be the same or different and represent two domains associated with one another by the linker (L). In some embodiments, a polypeptide linker is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more amino acids in length. In some embodiments, a linker is characterized in that it tends not to adopt a rigid three-dimensional structure, but rather provides flexibility to the polypeptide. A variety of different linker elements that can appropriately be used when engineering polypeptides (e.g., fusion polypeptides) are known in the art (Holliger et al, 1993; Poljak, 1994). AFV70463FF [0072] Mutation: As used herein, the term “mutation” is used to refer to a difference in the sequence of a nucleic acid or polypeptide as compared to a reference sequence. Exemplary types of mutations include, but are not limited to, insertions, deletions, and substitutions. For the avoidance of doubt, the term “mutation” is understood by those of skill in the art to include a difference as compared to a reference, and does not necessarily refer to or imply a change having occurred within any particular sequence of interest. [0073] Non-inferior: As used herein, the term “non-inferior” in the context of evaluating a test pharmaceutical composition refers to a test pharmaceutical composition that is (e.g., in terms of immunogenicity and/or functional antibody titer generated by the test composition) at least as effective as a reference composition. In some embodiments, non-inferiority is demonstrated when the lower bound of the 95% confidence interval (CI) for the geometric mean titer (GMT) ratio of a test pharmaceutical composition over a reference composition is at least greater than or equal to 0.4 or above, including, e.g., at least 0.5, at least 0.6, at least 0.7, at least 0.8, at least 0.9, at least 0.95, at least 0.98, at least 1.0, or higher. For example, in some embodiments, an immunogenic composition (e.g., vaccine) described herein is non-inferior to a reference vaccine (e.g., PCV13, PCV20, or PPSV23) when the lower bound of the 95% confidence interval (CI) for the geometric mean titer (GMT) ratio of the immunogenic composition (e.g., vaccine) over the reference vaccine is at least greater than or equal to 0.4 or above, including, e.g., at least 0.5, at least 0.6, at least 0.7, at least 0.8, at least 0.9, at least 0.95, at least 0.98, at least 1.0, or higher. In some embodiments, an immunogenic composition (e.g., vaccine) described herein is non-inferior to a reference vaccine (e.g., PCV13, PCV20, or PPSV23) when the lower bound of the 95% confidence interval (CI) for the geometric mean titer (GMT) ratio of the immunogenic composition (e.g., vaccine) over the reference vaccine is at least greater than or equal to 0.95 or above, including, e.g., at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 1.0, at least 1.1, at least 1.3, at least 1.5, or higher. In some embodiments, an immunogenic composition (e.g., vaccine) described herein is non-inferior to a reference vaccine (e.g., PCV13, PCV20, or PPSV23) when the seroconversion rates, or percentages of vaccine recipients with immune responses, are above a pre-defined threshold, e.g., the lower bound of the 95% confidence interval for the difference between the percentage of subjects who seroconvert, following immunization with an immunogenic composition (e.g., vaccine) described herein or immunization with the reference vaccine (e.g., PCV13, PCV20, or PPSV23), is greater than -0.10. AFV70463FF [0074] Parenteral: The phrases “parenteral administration” and “administered parenterally” as used herein have their art-understood meaning referring to modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion. [0075] Patient or subject: As used herein, the term “patient” or “subject” refers to any organism to which a provided composition is or may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients or subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, a patient is a human. In some embodiments, a patient or a subject is suffering from or susceptible to one or more disorders or conditions. In some embodiments, a patient or subject displays one or more symptoms of a disorder or condition. In some embodiments, a patient or subject has been diagnosed with one or more disorders or conditions. In some embodiments, a patient or a subject is receiving or has received certain therapy to diagnose and/or to treat a disease, disorder, or condition. [0076] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in unit dose amount appropriate for administration in a therapeutic or dosing regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonary, and to other mucosal surfaces. AFV70463FF [0077] Pharmaceutically acceptable: As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. [0078] Prevent or prevention: As used herein, the terms “prevent” or “prevention”, when used in connection with the occurrence of a disease, disorder, and/or condition, refer to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time. [0079] Polysaccharide: The term “polysaccharide” as used herein refers to a polymeric carbohydrate molecule composed of long chains of monosaccharide units bound together by glycosidic, phosphodiester, or other linkages and on hydrolysis give the constituent monosaccharides or oligosaccharides. Polysaccharides range in structure from linear to highly branched. Examples include storage polysaccharides such as starch and glycogen, structural polysaccharides such as cellulose and chitin and microbial polysaccharides, and antigenic polysaccharides found in microorganisms including, but not limited to, capsular polysaccharides (CPS), O polysaccharides (OPS), core O polysaccharides (COPS), and lipopolysaccharides (LPS). [0080] Polypeptide: The term “polypeptide”, as used herein, generally has its art-recognized meaning of a polymer of at least three amino acids, e.g., linked to each other by peptide bonds. Those of ordinary skill in the art will appreciate that the term “polypeptide” is intended to be sufficiently general as to encompass not only polypeptides having a complete sequence recited herein, but also to encompass polypeptides that represent functional fragments (i.e., fragments retaining at least one activity) of such complete polypeptides. Moreover, those of ordinary skill in the art understand that protein sequences generally tolerate some substitution without destroying activity. Thus, any polypeptide that retains activity and shares at least about 30-40% overall sequence identity, often greater than about 50%, 60%, 70%, or 80%, and further usually including at least one region of much higher identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99% in one or more highly conserved regions, usually encompassing at least 3-4 and often up to 20 or more amino acids, with another polypeptide of the same class, is encompassed within the AFV70463FF relevant term “polypeptide” as used herein. Polypeptides may contain L-amino acids, D-amino acids, or both and may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, e.g., terminal acetylation, amidation, methylation, etc. In some embodiments, proteins may comprise natural amino acids, non-natural amino acids, synthetic amino acids, and combinations thereof. [0081] Prevention: The term “prevent” or “prevention”, as used herein in connection with a disease, disorder, and/or medical condition, refers to reducing the risk of developing the disease, disorder and/or condition, and/or a delay of onset, and/or reduction in frequency and/or severity of one or more characteristics or symptoms of a particular disease, disorder or condition. In some embodiments, prevention is assessed on a population basis such that an agent is considered to “prevent” a particular disease, disorder or condition if a statistically significant decrease in the development, frequency, and/or intensity of one or more symptoms of the disease, disorder or condition is observed in a population susceptible to the disease, disorder, or condition. In some embodiments, prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a pre-defined period of time. [0082] Protein: As used herein, the term “protein” encompasses a polypeptide. Proteins may include moieties other than amino acids (e.g., may be glycoproteins, proteoglycans, etc.) and/or may be otherwise processed or modified. Those of ordinary skill in the art will appreciate that a “protein” can be a complete polypeptide chain as produced by a cell (with or without a signal sequence), or can be a characteristic portion thereof. Those of ordinary skill will appreciate that a protein can sometimes include more than one polypeptide chain, for example linked by one or more disulfide bonds or associated by other means. Polypeptides may contain l-amino acids, d- amino acids, or both and may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, e.g., terminal acetylation, amidation, methylation, etc. In some embodiments, proteins may comprise natural amino acids, non-natural amino acids, synthetic amino acids, and combinations thereof. The term “peptide” is generally used to refer to a polypeptide having a length of less than about 100 amino acids, less than about 50 amino acids, less than 20 amino acids, or less than 10 amino acids. In some embodiments, proteins are antibodies, antibody fragments, biologically active portions thereof, and/or characteristic portions thereof. AFV70463FF [0083] Recombinant: As used herein, the term “recombinant” is intended to refer to polypeptides that are designed, engineered, prepared, expressed, created, manufactured, and/or isolated by recombinant means, such as polypeptides expressed using a recombinant expression vector transfected into a host cell; polypeptides isolated from a recombinant, combinatorial human polypeptide library; polypeptides isolated from an animal (e.g., a mouse, rabbit, sheep, fish, etc.) that is transgenic for or otherwise has been manipulated to express a gene or genes, or gene components that encode and/or direct expression of the polypeptide or one or more component(s), portion(s), element(s), or domain(s) thereof; and/or polypeptides prepared, expressed, created or isolated by any other means that involves splicing or ligating selected nucleic acid sequence elements to one another, chemically synthesizing selected sequence elements, and/or otherwise generating a nucleic acid that encodes and/or directs expression of the polypeptide or one or more component(s), portion(s), element(s), or domain(s) thereof. In some embodiments, one or more of such selected sequence elements is found in nature. In some embodiments, one or more of such selected sequence elements is designed in silico. In some embodiments, one or more such selected sequence elements results from mutagenesis (e.g., in vivo or in vitro) of a known sequence element, e.g., from a natural or synthetic source such as, for example, in the germline of a source organism of interest (e.g., of a human, a mouse, etc.). [0084] Reference: As used herein, the term “reference” describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, subject, population, sample, sequence or value of interest is compared with a reference or control agent, animal, subject, population, sample, sequence or value. In some embodiments, a reference or control is tested and/or determined substantially simultaneously with the testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and/or comparison to a particular possible reference or control. [0085] Response: As used herein, a “response” to treatment may refer to any beneficial alteration in a subject’s condition that occurs as a result of or correlates with treatment. Such alteration may include stabilization of the condition (e.g., prevention of deterioration that would AFV70463FF have taken place in the absence of the treatment), amelioration of symptoms of the condition, and/or improvement in the prospects for cure of the condition, etc. Subject response may be measured according to a wide variety of criteria, including clinical criteria and objective criteria. Techniques for assessing response include, but are not limited to, clinical examination, positron emission tomography, chest X-ray CT scan, MRI, ultrasound, endoscopy, laparoscopy, presence or level of biomarkers in a sample obtained from a subject, cytology, and/or histology. The exact response criteria can be selected in any appropriate manner, provided that when comparing groups of subjects, the groups to be compared are assessed based on the same or comparable criteria for determining response rate. One of ordinary skill in the art will be able to select appropriate criteria. [0086] Risk: As will be understood from context, “risk” of a disease, disorder, and/or condition refers to a likelihood that a particular subject will develop the disease, disorder, and/or condition. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 up to 100%. In some embodiments, risk is expressed as a risk relative to a risk associated with a reference sample or group of reference samples. In some embodiments, a reference sample or group of reference samples have a known risk of a disease, disorder, condition and/or event. In some embodiments, a reference sample or group of reference samples are from subjects comparable to a particular subject. In some embodiments, relative risk is 0,1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. [0087] Serotype: As used herein, the term “serotype”, also referred to as a serovar, refers to a distinct variation within a species of bacteria or virus or among immune cells of different subjects. These microorganisms, viruses, or cells are classified together based on their cell surface antigens, allowing the epidemiologic classification of organisms to the sub-species level. A group of serovars with common antigens may be referred to as a serogroup or sometimes serocomplex. [0088] Species: As used herein, the term “species” refers to a distinct immunogenic composition comprising (i) a biotinylated polysaccharide antigen and (ii) a fusion protein comprising a biotin-binding moiety and one or more Streptococcus pneumoniae polypeptide antigens. In some embodiments, a distinct species can differ in one or more of (i) a biotinylated polysaccharide antigen and (ii) a fusion protein comprising a biotin-binding moiety and one or more Streptococcus pneumoniae polypeptide antigens. [0089] Treat: As used herein, the terms “treat,” “treatment,” or “treating” refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce AFV70463FF severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition, for example, for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition. [0090] Vaccination: As used herein, the term “vaccination” refers to the administration of a composition intended to generate an immune response, for example to a disease-causing agent. For the purposes of the present disclosure, vaccination can be administered before, during, and/or after exposure to a disease-causing agent, and in certain embodiments, before, during, and/or shortly after exposure to the agent. In some embodiments, vaccination includes multiple administrations, appropriately spaced in time, of a vaccinating composition. In some embodiments, vaccination initiates immunization. [0091] Variant: As used herein in the context of molecules, e.g., nucleic acids, proteins, or small molecules, the term “variant” refers to a molecule that shows significant structural identity with a reference molecule but differs structurally from the reference molecule, e.g., in the presence or absence or in the level of one or more chemical moieties as compared to the reference entity. In some embodiments, a variant also differs functionally from its reference molecule. In general, whether a particular molecule is properly considered to be a “variant” of a reference molecule is based on its degree of structural identity with the reference molecule. As will be appreciated by those skilled in the art, any biological or chemical reference molecule has certain characteristic structural elements. A variant, by definition, is a distinct molecule that shares one or more such characteristic structural elements but differs in at least one aspect from the reference molecule. In some embodiments, a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and/or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid (e.g., that are attached to the polypeptide or nucleic acid backbone). In some embodiments, a variant polypeptide or nucleic acid shows an overall sequence identity with a reference polypeptide or nucleic acid that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99%. In some embodiments, a variant polypeptide or nucleic acid does not share at least one characteristic AFV70463FF sequence element with a reference polypeptide or nucleic acid. In some embodiments, a reference polypeptide or nucleic acid has one or more biological activities. In some embodiments, a variant polypeptide or nucleic acid shares one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid lacks one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid shows a reduced level of one or more biological activities as compared to the reference polypeptide or nucleic acid. In some embodiments, a polypeptide or nucleic acid of interest is considered to be a “variant” of a reference polypeptide or nucleic acid if it has an amino acid or nucleotide sequence that is identical to that of the reference but for a small number of sequence alterations at particular positions. Typically, fewer than about 20%, about 15%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, or about 2% of the residues in a variant are substituted, inserted, or deleted, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 substituted residues as compared to a reference. Often, a variant polypeptide or nucleic acid comprises a very small number (e.g., fewer than about 5, about 4, about 3, about 2, or about 1) number of substituted, inserted, or deleted, functional residues (i.e., residues that participate in a particular biological activity) relative to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises not more than about 5, about 4, about 3, about 2, or about 1 addition or deletion, and, in some embodiments, comprises no additions or deletions, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 additions or deletions as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 modifications (e.g., substitutions, additions or deletions) at the N-terminus portion, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 modifications (e.g., substitutions, additions or deletions) at the C-terminus AFV70463FF portion, as compared to the reference. In some embodiments, a reference polypeptide or nucleic acid is one found in nature. _{[0092] “ ” refers to a point of attachment between two atoms.} Immunogenic Agents Purified Capsular Polysaccharide of Serotype 38 [0093] The present disclosure provides, among other things, insights and technologies associated with the discovery of the structure and characterization of a previously uncharacterized serotype of S. pneumoniae, serotype 38. In some embodiments, insights and technologies provided herein are useful to provide immunogenic compositions (e.g., vaccines) with increased immunogenicity to protect against certain diseases, disorders, and conditions, such as those associated with targeting S. pneumoniae serotype 38. [0094] In some embodiments, the present disclosure provides a composition comprising purified capsular polysaccharide having repeating units of formula I: HO HO wherein each R R is H. In some embodiments, R is –C(O)-CH₃. In some embodiments, a purified capsular polysaccharide comprises between 1 and about 2000 units of formula I. In some embodiments, a purified capsular AFV70463FF polysaccharide comprises between about 1 and about 1000 units of formula I. In some embodiments, a purified capsular polysaccharide comprises between about 1 and about 750 units of formula I. In some embodiments, a purified capsular polysaccharide comprises between about 1 and about 500 units of formula I. In some embodiments, a purified capsular polysaccharide comprises between about 1 and about 250 units of formula I. In some embodiments, a purified capsular polysaccharide comprises between about 1 and about 100 units of formula I. In some embodiments, a purified capsular polysaccharide comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, or about 200 units of formula I. . In some embodiments, a purified capsular polysaccharide comprises between about 4 and about 500 units of formula I.. In some embodiments, a purified capsular polysaccharide comprises between about 4 and about 400 units of formula I. . In some embodiments, a purified capsular polysaccharide comprises between about 1 and about 10 units of formula I. In some embodiments, a purified capsular polysaccharide comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, or about 10 units of formula I. [0095] In some embodiments, about 60% of the repeat units in the purified capsular polysaccharide of formula I are O-acetylated at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃ in formula I). In some embodiments, a purified capsular polysaccharide described herein comprises greater than or equal to 60% of repeating units being O-acetylated at the C₂ of the - Gal residue (i.e., R is –C(O)-CH₃ in formula I). In some embodiments, a purified capsular polysaccharide described herein comprises 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% of repeating units being O-acetylated at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃ in formula I). In some embodiments, a purified capsular polysaccharide described herein comprises about 60 to about 70% of repeating units wherein R is –C(O)-CH₃ of formula I. In some embodiments, a purified capsular polysaccharide described herein comprises 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, or 70% of repeating units wherein R is –C(O)-CH₃ of formula I. [0096] In some embodiments, a purified capsular polysaccharide described herein comprises between 0% and 100% of repeating units being O-acetylated at the C₂ of the -Galf residue (i.e., R is –C(O)-CH₃ in formula I). In some embodiments, a purified capsular polysaccharide described herein comprises about 10 to about 75% of repeating units being O-acetylated at the C₂ of the - Gal residue (i.e., R is –C(O)-CH₃ in formula I). AFV70463FF [0097] In some embodiments, a purified capsular polysaccharide described herein comprises less than 60% of repeating units being O-acetylated at the C₂ of the -Gal residue (i.e., R is –C(O)- CH₃). In some embodiments, a purified capsular polysaccharide described herein comprises 59, 58, 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0% of repeating units being O-acetylated at the C₂ of the -Gal residue (i.e., R is – C(O)-CH₃ in formula I). [0098] In some embodiments, a purified capsular polysaccharide of formula I is biotinylated (i.e., is a “biotinylated purified capsular polysaccharide”). [0099] In some embodiments, a purified capsular polysaccharide of formula I has a weight average molecular weight (Mw) of about 300 kDa or greater. In some embodiments, a purified capsular polysaccharide of formula I has a weight average molecular weight (Mw) of about 5000 Daltons or less. In some embodiments, a purified capsular polysaccharide of formula I has a weight average molecular weight (Mw) of about 600kDa to about 4000kDa. In some embodiments, a purified capsular polysaccharide of formula I has a weight average molecular weight (Mw) of about 3675 Daltons to about 4000 kiloDaltons. In an embodiment, a purified capsular polysaccharide of formula I has a weight average molecular weight (Mw) of about 600 kDa. In another embodiment, a purified capsular polysaccharide of formula I has a weight average molecular weight (Mw) of about 3,550 kDa. [0100] In some embodiments, the present disclosure provides a purified capsular polysaccharide of formula I that is or has been activated with a chemical reagent, e.g., an oxidizing agent or a reducing agent. A capsular polysaccharide is “activated”, as described herein, when said polysaccharide has been chemically modified to enable conjugation to a linker or a carrier protein. For example, in some embodiments, a purified capsular polysaccharide of formula I is activated with a chemical reagent (e.g., an oxidizing agent or a reducing agent) to produce reactive groups for conjugation to a suitable conjugate (e.g., a linker or carrier protein). In some embodiments, a reactive group is an aldehyde group (-C(O)H). In some embodiments, a reactive group is a cyanoester group (-OCN). In some embodiments, activation occurs at the C_2, C_3, C_5, or C_6, of the -Gal residue (i.e., at the position designated as R in formula I, above). In some embodiments, activation is achieved by oxidative cleavage of a monosaccharide ring, thereby yielding a moiety having one or more aldehyde, cyanoester, or amide groups. AFV70463FF [0101] In some embodiments, the present disclosure provides an activated capsular polysaccharide comprising one or more units of formula I. In some embodiments, the present disclosure provides an activated serotype 38 capsular polysaccharide. In some embodiments, an activated serotype 38 capsular polysaccharide comprises a reactive cyanoester group or an aldehyde group. [0102] In some embodiments, the present disclosure provides a purified capsular polysaccharide comprising one or more units of formula I, and wherein the purified capsular polysaccharide is conjugated to a carrier protein or biotin. [0103] In some embodiments, the present disclosure provides a purified capsular polysaccharide comprising one or more units of formula I, and wherein the purified capsular polysaccharide is conjugated to a carrier protein. In some embodiments, a carrier protein is or comprises an antigenic polypeptide (e.g., ones described herein including, e.g., SP0785 as described herein, SP1500 as described herein, SP0435 as described herein, Ply as described herein or in WO 05/76696, WO 05/108580, or WO 10/71986, etc.) and/or a fusion protein that includes one or more antigenic polypeptides (e.g., ones described herein including, e.g., CP1 as described herein, SPP2 as described herein, etc.). In some embodiments, a carrier protein is a toxoid or a glycoprotein from a pathogen. In some embodiments, a carrier protein is or comprises PspA as described in WO 19/167008; CRM197 as described in WO 14/92378; variants of CRM197 such as eCRM197 as disclosed in WO 22/178015; Diphtheria toxoid; variants of Diphtheria toxoid as described in WO18/156465; Tetanus toxoid as described in WO 19/152921; ExoProtein A, and Protein D as described in US5858677, and combinations thereof. [0104] In some embodiments, the present disclosure provides a purified capsular polysaccharide comprising repeating units of formula I, and wherein the purified capsular polysaccharide is conjugated to biotin. [0105] In some embodiments, activation of a purified capsular polysaccharide of formula I yields a polysaccharide having one or more monomers of formula II:

AFV70463FF HO HO O In some one or more monomers of formula II: HO HO [0106] In formula II further comprises monomers of formula I: AFV70463FF HO HO O (O)-CH3 . [0107] In some embodiments, the present disclosure provides a polysaccharide comprising one or more monomers of formula III: R^aO R^aO where one a carrier protein) or biotin, and the remaining instances of R^a are each H. [0108] In some embodiments, the present disclosure provides a polysaccharide comprising one or more monomers of formula IV-1: AFV70463FF HO R^aO O where one and the remaining instances of R^a are each H. [0109] In some embodiments, the present disclosure provides a polysaccharide comprising one or more monomers of formula IV-2: HO R^aO where one instance of R^c is a site for reaction with an oxidizing agent (e.g., periodate), and the remaining instances of R^c are each -OH. [0110] In some embodiments, the present disclosure provides a polysaccharide comprising one or more monomers of formula IV-3: AFV70463FF HO R^aO O where one instance of R^c is a site for reaction with an oxidizing agent (e.g., periodate), and the remaining instance of R^c is -OH. [0111] In some embodiments, a weight average molecular weight of the polysaccharide is from about 300 Da to about 4000 kDa. In some embodiments, a weight average molecular weight of the polysaccharide is from about 600 kDa to about 4000 kDa. In an embodiment, a weight average molecular weight of the polysaccharide is about 600 kDa. In another embodiment, a weight average molecular weight of the polysaccharide is about 3,550 kDa. [0112] In some embodiments, a purified capsular polysaccharide of formula I is activated by an oxidizing agent. In some embodiments, an oxidizing agent is 1-cyano-4- (dimethylamino)pyridinium tetrafluoroborate (CDAP), periodate (e.g., sodium periodate), 1-ethyl- 3-(3-dimethylaminopropyl)carbodiimide (EDAC). In some embodiments, a purified capsular polysaccharide of formula I is activated by reductive amination. In some embodiments, a purified capsular polysaccharide is activated by contacting the purified capsular polysaccharide with an oxidizing agent. In some embodiments, an oxidizing agent is sodium periodate. In some embodiments, a purified capsular polysaccharide is activated by reductive amination. In some embodiments, a purified capsular polysaccharide is activated by contacting the purified capsular polysaccharide with a reducing agent. In some embodiments, a reducing agent is sodium cyanoborohydride. AFV70463FF [0113] In some embodiments, activation occurs at the terminal hydroxyl group of the purified capsular polysaccharide. [0114] In some embodiments, a purified capsular polysaccharide of formula I is represented by Additional Antigenic Polysaccharides [0115] In some embodiments, an immunogenic agent described herein is or comprises one or more antigenic polysaccharides (other than or in addition to a purified capsular polysaccharide of S. pneumoniae serotype 38 described herein (e.g., a purified capsular polysaccharide of formula I)). [0116] In some embodiments, an antigenic polysaccharide is, or is derived from S. pneumoniae. In some embodiments, one or more antigenic polysaccharides are, or are derived from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 serotypes or strains (variants) of S. pneumoniae. In some embodiments, one or more antigenic polysaccharides are, or are derived from more than 25 serotypes or strains (variants) of S. pneumoniae, e.g., 26, 27, 28, 29, 30, 35, 40, 45, or 50 serotypes or strains. In some embodiments, one or more antigenic polysaccharides are, or are derived from more than 60, 70, 80, 90, or 100 serotypes or strains (variants) of S. pneumoniae. [0117] In some embodiments, an antigenic polysaccharide is a branched polysaccharide, or alternatively, can be a straight chain polysaccharide. [0118] In some embodiments, an antigenic polysaccharide is from, or derived from, a S. pneumoniae polysaccharide. In some embodiments, an antigenic polysaccharide present in an immunogenic agent described herein includes one S. pneumoniae polysaccharide. Capsular AFV70463FF polysaccharides are used to distinguish serotypes of S. pneumoniae. There are at least 100 distinct serotypes of S. pneumoniae polysaccharides, each having a different chemical structure. [0119] In some embodiments, an antigenic polysaccharide present in an immunogenic agent includes one or more S. pneumoniae capsular polysaccharides from, or derived from, one or more S. pneumoniae serotypes selected from 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 6E, 6F, 6G, 6H, 7A, 7B, 7C, 7F, 8, 9A, 9L, 9N, 9V, 10A, 10B, 10C, 10F, 11A, 11B, 11C, 11D, 11E, 11F, 12A, 12B, 12F, 13, 14, 15A, 15B, 15C, 15F, 16A, 16F, 17A, 17F, 18A, 18B, 18C, 18F, 19A, 19B, 19C, 19F, 20A, 20B, 21, 22A, 22F, 23A, 23B, 23F, 24A, 24B, 24F, 25A, 25F, 27, 28A, 28F, 29, 31, 32A, 32F, 33A, 33B, 33C, 33D, 33E, 33F, 34, 35A, 35B, 35C, 35F, 36, 37, 39, 40, 41A, 41F, 42, 43, 44, 45, 46, 47A, 47F, and 48. [0120] In some embodiments, an antigenic polysaccharide present in an immunogenic agent includes one or more S. pneumoniae capsular polysaccharides from, or derived from, one or more S. pneumoniae serotypes selected from 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F, and 35B. Antigenic Polypeptides [0121] In some embodiments, an immunogenic agent described herein is or comprises one or more polypeptide antigens. [0122] In some embodiments, a polypeptide antigen is or comprises a bacterial polypeptide antigen. In some embodiments, a bacterial polypeptide antigen is a polypeptide antigen of, or derived from, a Gram-negative or a Gram-positive bacterium. In some embodiments, a bacterial polypeptide antigen is a polypeptide antigen of, or derived from, S. pneumoniae. [0123] In some embodiments, a polypeptide antigen is a polypeptide antigen of, or derived from S. pneumoniae. In some embodiments, an immunogenic agent includes one or more of the following S. pneumoniae antigenic polypeptides, or portions thereof. Exemplary Pneumococcal Polypeptide Antigens: Pneumolysin Polypeptides [0124] In some embodiments, an antigenic polypeptide described herein is or comprises a pneumococcal polypeptide antigen. In some embodiments, a pneumococcal polypeptide antigen is or comprises pneumolysin polypeptide antigen. Pneumolysin (Ply) is a S. pneumoniae protein toxin. Ply is a cholesterol-dependent toxin of the thiol-activated cytolysin family. In some AFV70463FF embodiments, a Ply polypeptide antigen is or comprises a full-length Ply polypeptide. For example, in some embodiments, a full-length Ply polypeptide has 470 amino acids (53 kDa) and is represented by the amino acid sequence as set forth in SEQ ID NO: 4. In some embodiments, a Ply polypeptide antigen includes a portion of a Ply polypeptide (e.g., a portion of a Ply polypeptide of SEQ ID NO: 4, which portion includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450 or more contiguous amino acids of SEQ ID NO: 4). In some embodiments, a portion of a Ply polypeptide corresponds to a protein having amino acids 2-470 of the amino acid sequence set forth in SEQ ID NO: 4. In some embodiments, a Ply polypeptide antigen contains one or more amino acid alterations (e.g., deletion, substitution, and/or insertion) from a naturally-occurring wild-type Ply polypeptide sequence. For example, a Ply polypeptide antigen may contain an amino acid sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 4 or a portion thereof (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450 or more consecutive amino acids of the sequence shown in SEQ ID NO: 4). Alternatively, a Ply polypeptide antigen may contain a portion (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450 or more consecutive amino acids) of a sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 4. In some embodiments, a nucleotide sequence encoding a Ply polypeptide is provided herein as SEQ ID NO: 14. [0125] Pneumolysins are exotoxins produced by bacteria that can cause hemolytic activity and complement activation. While highly immunogenic, their use in vaccines can be limited because they cause lysis of red blood cells. Accordingly, in some embodiments, a Ply polypeptide antigen is or comprises a Ply variant (also designated herein as a mutant Ply or “mPly”) that is substantially non-hemolytic. As used herein, the phrase “substantially non-hemolytic” means the ability of lysing red blood cells being reduced by at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or up to 100%, as compared to that of an equivalent concentration of a reference Ply (e.g., a wild-type Ply). In some embodiments, hemolytic activity of substantially non-hemolytic Ply is at least 5%, at least 10%, at least 15%, at least 20%, at least AFV70463FF 20%, at least 30%, at least 30%, at least 35%, least 40 %, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% lower than an equivalent concentration of a reference Ply (e.g., a wild-type Ply). In some embodiments, the substantially non-hemolytic Ply has no detectable hemolytic activity. The term “wild-type Ply” is accorded the usual definition associated with such phrase, e.g., in some embodiments, a naturally occurring Ply (e.g., a Ply that is naturally secreted by a capable bacterial source). In some embodiments, a wild-type Ply protein is represented by the amino acid sequence as set forth in SEQ ID NO: 4. [0126] In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) comprises a wild-type Ply amino acid sequence (e.g., an amino acid sequence as set forth in SEQ ID NO: 4) or an antigenic fragment thereof, with one or more amino acid substitutions. In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) comprises a wild-type Ply amino acid sequence (e.g., an amino acid sequence as set forth in SEQ ID NO: 4) or an antigenic fragment thereof, with one or more of the following amino acid substitutions: residue D385 substituted with N; residue C428 substituted with G, and residue W433 substituted with F. See, for example, Berry et al., “Effect of defined point mutations in the pneumolysin gene on the virulence of Streptococcus pneumoniae”. Infect Immune 199563(5):1969-1974). In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) carrying the amino acid substitutions D385N, C428G, and W433F is referred to as PdT. In some embodiments, a PdT is or comprises the amino acid sequence as set forth in SEQ ID NO: 5. [0127] In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) comprises a wild-type Ply amino acid sequence (e.g., an amino acid sequence as set forth in SEQ ID NO: 4) or an antigenic fragment thereof, with G294 substituted with P (See, e.g., Oloo et al., “Structure-guided antigen engineering yields pneumolysin mutants suitable for vaccination against pneumococcal disease”. J Biol Chem. 2011286(14):12133-12140). In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) comprises a wild-type Ply amino acid sequence (e.g., an amino acid sequence as set forth in SEQ ID NO: 4) or an antigenic fragment thereof, with all of the following amino acid substitutions: residue D385 substituted with N; residue C428 substituted with G; residue W433 substituted with F; and G294 substituted with P. In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) carrying the amino acid substitutions G294P, D385N, C428G, and W433F is referred to as PdT(G294P). In some embodiments, a PdT(G294P) is or comprises the amino acid sequence as set forth in SEQ ID NO: 6. AFV70463FF [0128] In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) is a portion of a PdT(G294P) polypeptide (e.g., a portion of the PdT(G294P) polypeptide of SEQ ID NO: 6, which portion includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450 or more contiguous amino acids of SEQ ID NO: 6. In some embodiments, such a portion of PdT(G294P) polypeptide include the four amino acid substitutions described herein. In some embodiments, a portion of a PdT(G294P) polypeptide corresponds to a protein having amino acids 2-470 of the amino acid sequence set forth in SEQ ID NO: 6. In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) contains one or more amino acid alterations (e.g., deletion, substitution, and/or insertion) from the PdT(G294P) polypeptide sequence of SEQ ID NO: 6. For example, a mutant Ply (e.g., non- hemolytic Ply) may contain an amino acid sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 6 or a portion thereof (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450 or more consecutive amino acids of the sequence shown in SEQ ID NO: 6). Alternatively, a mutant Ply (e.g., non-hemolytic Ply) may contain a portion (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450 or more consecutive amino acids) of a sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 6. In some embodiments, a mutant Ply (e.g., non-hemolytic Ply) may comprises no more than 25 (including, e.g., no more than 20, no more than 15, no more than 10, no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2) amino acid modifications (e.g., deletion, substitution, and/or insertion) within the sequence of SEQ ID NO: 6 or a portion thereof as described herein. In some embodiments, such amino acid modifications may be present in the N-terminal portion and/or C- terminal portion. Exemplary Pneumococcal Polypeptide Antigens: SP0435 Polypeptides [0129] In some embodiments, an antigenic polypeptide described herein is or comprises a pneumococcal polypeptide antigen. In some embodiments, a pneumococcal polypeptide antigen is or comprises a SP0435 polypeptide. SP0435 is a S. pneumoniae protein. In some embodiments, an SP0435 polypeptide is an elongation factor P. In some embodiments, an SP0435 polypeptide is or AFV70463FF comprises a full-length SP0435 polypeptide. For example, in some embodiments, a full-length SP0435 polypeptide has 186 amino acids (20 kDa) and is represented by the amino acid sequence as set forth in SEQ ID NO: 8. Without wishing to be bound by a particular theory, amino acids 1- 61 of SEQ ID NO: 8 are predicted to be a dimerization domain of an SP0435 polypeptide (amino acids 1-61 of the full-length protein). Accordingly, in some embodiments, a SP0435 polypeptide may exclude such a dimerization domain, for example in some embodiments, to minimize or avoid the possibility of crosslinking or interference with a rhizavidin moiety. In some embodiments, an SP0435 polypeptide includes a portion of an SP0435 polypeptide (e.g., a portion of the SP0435 polypeptide of SEQ ID NO: 8, which portion includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, or more contiguous amino acids of SEQ ID NO: 8). In some embodiments, a portion of an SP0435 polypeptide corresponds to a protein having amino acids 62-185 of the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, an SP0435 polypeptide contains one or more amino acid alterations (e.g., deletion, substitution, and/or insertion) from a naturally-occurring wild-type SP0435 polypeptide sequence. For example, an SP0435 polypeptide may contain an amino acid sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 8 or a portion thereof (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, or more consecutive amino acids of the sequence shown in SEQ ID NO: 8). Alternatively, an SP0435 polypeptide may contain a portion (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, or more consecutive amino acids) of a sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 8. In some embodiments, a nucleotide sequence encoding an SP0435 polypeptide is provided herein as SEQ ID NO: 17. In some embodiments, an SP0434 polypeptide may comprises no more than 25 (including, e.g., no more than 20, no more than 15, no more than 10, no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2) amino acid modifications (e.g., deletion, substitution, and/or insertion) within the sequence of SEQ ID NO: 8 or a portion thereof as described herein. In some embodiments, such amino acid modifications may be present in the N-terminal portion and/or C- terminal portion. In some embodiments, an SP0435 polypeptide is one described in WO AFV70463FF 2011/112906, the entire content of which is incorporated herein by reference for the purposes described herein. Exemplary Pneumococcal Polypeptide Antigens: SP0785 Polypeptides [0130] In some embodiments, an antigenic polypeptide described herein is or comprises a pneumococcal polypeptide antigen. In some embodiments, a pneumococcal polypeptide antigen is or comprises a SP0785 polypeptide. SP0785 is a conserved hypothetical S. pneumoniae protein, for example, in some embodiments as described in WO 2014/124228, the entire content of which is incorporated herein by reference for the purposes described herein. In some embodiments, an SP0785 polypeptide is an efflux transporter protein conserved across S. pneumoniae strains. In some embodiments, an SP0785 polypeptide is or comprises a full-length SP0785 polypeptide. For example, in some embodiments, a full-length SP0785 polypeptide has 399 amino acids (38 kDa) and is represented by the amino acid sequence as set forth in SEQ ID NO: 10. Without wishing to be bound by a particular theory, amino acids 1-32 of SEQ ID NO: 10 are predicted to be a signal sequence and transmembrane domain of an SP0785 polypeptide (amino acids 1-32 of the full- length protein). Accordingly, in some embodiments, an SP0785 polypeptide may exclude such a signal sequence and transmembrane domain. In some embodiments, an SP0785 polypeptide includes a portion of an SP0785 polypeptide (e.g., a portion of the SP0785 polypeptide of SEQ ID NO: 10, which portion includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or more contiguous amino acids of SEQ ID NO: 10). In some embodiments, a portion of an SP0785 polypeptide corresponds to a protein having amino acids 33-399 of the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, an SP0785 polypeptide contains one or more amino acid alterations (e.g., deletion, substitution, and/or insertion) from a naturally-occurring wild-type SP0785 polypeptide sequence. For example, an SP0785 polypeptide may contain an amino acid sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 10 or a portion thereof (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or more consecutive amino acids of the sequence shown in SEQ ID NO: 10). Alternatively, an SP0785 polypeptide may contain a portion (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 AFV70463FF consecutive amino acids) of a sequence that is at least 60% or more (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: 10. In some embodiments, a nucleotide sequence encoding an SP0785 polypeptide is provided herein as SEQ ID NO: 18. In some embodiments, an SP0785 polypeptide may comprises no more than 25 (including, e.g., no more than 20, no more than 15, no more than 10, no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2) amino acid modifications (e.g., deletion, substitution, and/or insertion) within the sequence of SEQ ID NO: 10 or a portion thereof as described herein. In some embodiments, such amino acid modifications may be present in the N-terminal portion and/or C-terminal portion. Exemplary Pneumococcal Polypeptide Antigens: SP1500 Polypeptides [0131] In some embodiments, an antigenic polypeptide described herein is or comprises a pneumococcal polypeptide antigen. In some embodiments, a pneumococcal polypeptide antigen is or comprises a SP1500 polypeptide. SP1500 is a S. pneumoniae protein, for example, in some embodiments as described in WO 2014/124228, the entire content of which is incorporated herein by reference for the purposes described herein. In some embodiments, an SP1500 polypeptide is an Amino Acid ABC Transporter, amino acid-binding polypeptide conserved across S. pneumoniae strains. In some embodiments, an SP1500 polypeptide is or comprises a full-length SP1500 polypeptide. For example, in some embodiments, a full-length SP1500 polypeptide has 278 amino acids (28 kDa) and is represented by the amino acid sequence as set forth in SEQ ID NO: 12. Without wishing to be bound by a particular theory, amino acids 1-26 of SEQ ID NO: 12 are predicted to be a signal sequence of an SP1500 polypeptide (amino acids 1-26 of the full-length protein). Accordingly, in some embodiments, a SP1500 polypeptide may exclude such a signal sequence. In some embodiments, an SP1500 polypeptide includes a portion of an SP1500 polypeptide (e.g., a portion of the SP1500 polypeptide of SEQ ID NO: 12, which portion includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, or more contiguous amino acids of SEQ ID NO: 12). In some embodiments, a portion of an SP1500 polypeptide corresponds to a protein having amino acids 27-278 of the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, an SP1500 polypeptide contains one or more amino acid alterations (e.g., deletion, substitution, and/or AFV70463FF insertion) from a naturally-occurring wild-type SP1500 polypeptide sequence. For example, an SP1500 polypeptide may contain an amino acid sequence that is at least 60% or more (e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 98%) identical to SEQ ID NO: 12 or a portion thereof (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, or more consecutive amino acids of the sequence shown in SEQ ID NO: 12). Alternatively, an SP1500 polypeptide may contain a portion (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, or more consecutive amino acids) of a sequence that is at least 60% or more (e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 98%) identical to SEQ ID NO: 12. In some embodiments, a nucleotide sequence encoding an SP1500 polypeptide is provided herein as SEQ ID NO: 19. In some embodiments, an SP1500 polypeptide may comprises no more than 25 (including, e.g., no more than 20, no more than 15, no more than 10, no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2) amino acid modifications (e.g., deletion, substitution, and/or insertion) within the sequence of SEQ ID NO: 12 or a portion thereof as described herein. In some embodiments, such amino acid modifications may be present in the N-terminal portion and/or C-terminal portion. [0132] In some embodiments, nucleic acid sequences encoding a pneumolysin polypeptide (SEQ ID NO: 4), an SP0435 polypeptide (SEQ ID NO: 8), SP0785 polypeptide (SEQ ID NO: 10), and an SP1500 polypeptide (SEQ ID NO: 12) are provided as SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, respectively. Due to degeneracy in the genetic code, those of ordinary skill in the art would understand that other DNA sequences (including codon-optimized sequences) could encode these polypeptides, as well as the others disclosed herein. [0133] In some embodiments, the present disclosure includes nucleic acid sequences encoding any of the amino acids described herein. Due to degeneracy in the genetic code, those of ordinary skill in the art would understand that other DNA sequences (including codon-optimized sequences) could encode these polypeptides, as well as the others disclosed herein. Fusion Proteins that Include Antigenic Polypeptides [0134] In some embodiments, an immunogenic agent described herein is or comprises a fusion protein. In some embodiments, antigenic polypeptides described herein can be part of a fusion AFV70463FF protein. For example, in some embodiments, an immunogenic agent described herein comprises a fusion protein that is or comprises a complementary affinity molecule and one or more antigenic polypeptides described herein. In some embodiments, a fusion protein comprises two antigenic polypeptides described herein. In some embodiments, a fusion protein comprises three antigenic polypeptides described herein. In some embodiments, a fusion protein comprises four antigenic polypeptides described herein. In some embodiments, a fusion protein comprises 6-10 antigenic polypeptides described herein. In some embodiments, a fusion protein comprises 11-15 antigenic polypeptides described herein. In some embodiments, a fusion protein comprises 15-20 antigenic polypeptides described herein. [0135] In some embodiments, a fusion protein has carrier properties. In some embodiments, a fusion protein has antigenic properties. In some embodiments, a fusion protein has carrier properties and antigenic properties. [0136] In some embodiments, a fusion protein comprises an antigenic polypeptide that is or comprises a S. pneumoniae polypeptide antigen (e.g., in some embodiments, a pneumolysin polypeptide antigen, a SP0435 polypeptide antigen, a SP0785 polypeptide antigen, or a SP1500 polypeptide antigen). [0137] In some embodiments, a fusion protein comprises one or more antigenic polypeptides of S. pneumoniae having an amino acid sequence that is least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to any of SEQ ID NOs: 4-13, or antigenic fragments thereof. In some embodiments, a fusion protein comprises one or more antigenic polypeptides of S. pneumoniae having an amino acid sequence comprising any of SEQ ID NOs: 4-13, or antigenic fragments thereof. In some embodiments, a fusion protein comprises two antigenic polypeptides having an amino acid sequence that is least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to any of SEQ ID NOs: 4-13, or antigenic fragments thereof. In some embodiments, a fusion protein comprises two antigenic polypeptides having an amino acid sequence comprising any of SEQ ID NOs: 4-13, or antigenic fragments thereof. In some embodiments, a fusion protein as described herein further comprises a biotin-binding moiety, for example, in some embodiments, a biotin- binding moiety that is or comprises a rhizavidin polypeptide having an amino acid sequence that is least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to any of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or biotin-binding AFV70463FF fragments thereof. In some embodiments, a fusion protein as described herein further comprises a biotin-binding moiety, for example, in some embodiments, a biotin-binding moiety that is or comprises a rhizavidin polypeptide having the amino acid sequence of any of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or biotin-binding fragments thereof. [0138] In some embodiments, a fusion protein comprises a pneumolysin polypeptide as described herein. For example, in some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P) polypeptide), or an antigenic fragment thereof. In some embodiments, a pneumolysin polypeptide described herein may be truncated from its N-terminal portion and/or C-terminal portion. In some embodiments, a fusion protein as described herein further comprises a rhizavidin polypeptide, for example, in some embodiments, a rhizavidin polypeptide having the amino acid sequence of any of SEQ ID NOs: 1-3, or biotin-binding fragments thereof. [0139] In some embodiments, a fusion protein comprises an SP0435 polypeptide. For example, in some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435 polypeptide), or an antigenic fragment thereof. In some embodiments, an SP0435 polypeptide described herein may be truncated from its N- terminal portion and/or C-terminal portion. In some embodiments, a fusion protein as described herein further comprises a rhizavidin polypeptide, for example, in some embodiments, a rhizavidin polypeptide having the amino acid sequence of any of SEQ ID NOs: 1-3, or biotin-binding fragments thereof. [0140] In some embodiments, a fusion protein comprises an SP0785 polypeptide. For example, in some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785 polypeptide), or an antigenic fragment thereof. In some embodiments, an SP0785 polypeptide described herein may be truncated from its N- terminal portion and/or C-terminal portion. In some embodiments, a fusion protein as described AFV70463FF herein further comprises a rhizavidin polypeptide, for example, in some embodiments, a rhizavidin polypeptide having the amino acid sequence of any of SEQ ID NOs: 1-3, or biotin-binding fragments thereof. [0141] In some embodiments, a fusion protein comprises an SP1500 polypeptide. For example, in some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500 polypeptide), or an antigenic fragment thereof. In some embodiments, an SP1500 polypeptide described herein may be truncated from its N- terminal portion and/or C-terminal portion. In some embodiments, a fusion protein as described herein further comprises a rhizavidin polypeptide, for example, in some embodiments, a rhizavidin polypeptide having the amino acid sequence of any of SEQ ID NOs: 1-3, or biotin-binding fragments thereof. [0142] In some embodiments, a fusion protein comprises one or more polypeptides homologous to a polypeptide described herein (e.g., an antigenic polypeptide isolated from different serotypes, strains, or species). Individual serotypes, strains, or species can contain numerous mutations relative to each other, and some of these can result in different protein sequences between the different serotypes, strains, or species. In some embodiments, a fusion protein comprises one or more polypeptides homologous to a S. pneumoniae polypeptide described herein, e.g., a Ply polypeptide, an SP0435 polypeptide, an SP0785 polypeptide or an SP1500 polypeptide isolated from different serotypes of S. pneumoniae. In some embodiments, antigenic polypeptides have at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the polypeptides of any of SEQ ID NOs: 4-13, or antigenic fragments thereof. Serotype, strain, or species variation may be used to design such variants of the antigenic polypeptides described herein. [0143] In some embodiments, fusion proteins described herein comprise one or more fragments of one or more polypeptides described herein, e.g., biotin-binding fragments of rhizavidin or antigenic fragments of an antigenic polypeptide (e.g., a Ply polypeptide or antigenic fragments thereof, an SP0435 polypeptide or antigenic fragments thereof, a SP0785 polypeptide or antigenic fragments thereof (e.g., with or without a signal sequence), or a SP1500 polypeptide or antigenic fragments thereof (e.g., with or without a signal sequence)). In some embodiments, a AFV70463FF fusion protein described herein comprises a truncated version of a polypeptide described herein, which truncated version is close in size to a polypeptide having the amino acid sequences of any of SEQ ID NOs: 1-13. For example, in some embodiments, such a truncated polypeptide may lack at most one, two, three, four, five, ten, or twenty amino acids from one or both termini from its parent polypeptide. In some embodiments, the same number of residues is removed from the N- terminus and the C-terminus, while in other embodiments, a different number of residues is removed from the N-terminus compared to the C-terminus. In some embodiments, a truncated polypeptide has an amino acid sequence that is least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to any of SEQ ID NOs: 1-13 lacking 1-5, 1-10, or 1-20 amino acid residues from the N-terminus, C-terminus, or both. In some embodiments, a truncated polypeptide has the amino acid sequence of any of SEQ ID NOs: 1-13 lacking 1-5, 1-10, or 1-20 amino acid residues from the N-terminus, C-terminus, or both. In some embodiments, a truncated polypeptide has an amino acid sequence that is least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to any of SEQ ID NOs: 1-13 lacking 1-10 acid residues from the N-terminus, C-terminus, or both. In some embodiments, a truncated polypeptide has the amino acid sequence of any of SEQ ID NOs: 1-13 lacking 1-10 amino acid residues from the N-terminus, C-terminus, or both. For instance, a truncated polypeptide may lack 10 amino acid residues at both the N-terminus and C-terminus of any one of SEQ ID NOs: 1-13, resulting in a protein lacking 20 amino acid residues. Internal deletions, e.g., of 1-10, 11-20, 21-30, or 31-40 amino acids, are also contemplated. [0144] In some embodiments, a fusion protein comprises an N-terminal polypeptide and a C- terminal polypeptide. In some embodiments, one or both of the N-terminal polypeptide and the C- terminal polypeptide is an antigenic polypeptide described herein (e.g., a polypeptide having an amino acid sequence comprising one or more of SEQ ID NOs: 4-13), or an antigenic fragment or variant thereof. In some embodiments, one or both of the N-terminal polypeptide and the C- terminal polypeptide is a biotin-binding moiety, for example a polypeptide having an amino acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a biotin-binding fragment thereof. In some embodiments, one of the N-terminal polypeptide or the C-terminal polypeptide is a biotin-binding moiety, e.g., a polypeptide having an amino acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a biotin-binding fragment thereof, and the other terminal polypeptide is an antigenic polypeptide described herein (e.g., a polypeptide AFV70463FF having an amino acid sequence comprising one or more of SEQ ID NOs: 4-13), or an antigenic fragment or variant thereof. [0145] In some embodiments, an N-terminal polypeptide and a C-terminal polypeptide present in a fusion protein are directly bound to each other. In some embodiments, an N-terminal polypeptide and a C-terminal polypeptide present in a fusion protein are linked via a linker peptide (e.g., a linker disclosed herein). The length and/or amino acids of a linker, when present, can be adjusted to obtain a more flexible, semi-rigid, or rigid linker. In some embodiments, a linker can be a GS-enriched linker. In some embodiments, a linker can be an A-enriched linker. A linker can generally be from 1-40, such as 3-10 or 10-30 and specifically 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids in length. Exemplary flexible peptide linkers are shown as SEQ ID NOs: 30-37. [0146] In some embodiments, a fusion protein comprises one or more linkers described herein. In some embodiments, a fusion protein comprises at least one linker. In some embodiments, a fusion protein comprises at least two linkers. In some embodiments, a linker comprises a polypeptide comprising an amino acid sequence of GGGGSSS (SEQ ID NO: 30). In some embodiments, a linker comprises a polypeptide comprising an amino acid sequence of AAA. In some embodiments, a linker comprises a polypeptide comprising an amino acid sequence of AAAA (SEQ ID NO: 58). In some embodiments, a linker comprises a polypeptide comprising an amino acid sequence of GGGGSGGGGSGGGGS (SEQ ID NO: 33). In some embodiments, a fusion protein comprises a first linker having the amino acid sequence of GGGGSSS (SEQ ID NO: 30) and a second linker having the amino acid sequence of AAA. In some embodiments, a linker sequence may be a residual amino acid sequence from a restriction site. For example, in some embodiments, a fusion protein comprises an amino acid sequence AAA residual from a Not I restriction site. In some embodiments, a fusion protein comprises a linker of GGGGSSS (SEQ ID NO: 30) and an amino acid sequence AAA residual from a Not I restriction site. In some embodiments, the one or more linkers are selected from GGGGSSS (SEQ ID NO: 30) and AAAA (SEQ ID NO: 58). In some embodiments, a fusion protein comprises a linker of GGGGSSS (SEQ ID NO: 30) and AAAA (SEQ ID NO: 58). [0147] In some embodiments, a fusion protein described herein comprises a biotin-binding moiety. In some embodiments, the fusion protein comprises a biotin-binding moiety, and one or more polypeptide antigens. In some embodiments, the fusion protein comprises a biotin-binding AFV70463FF moiety and two or more polypeptide antigens. In some embodiments, the biotin-binding moiety of a fusion protein comprises rhizavidin or a biotin-binding fragment thereof, which in some embodiments as further described in WO 2012/155053, the contents of which are herein incorporated by reference in their entirety for the purposes described herein. [0148] In some embodiments, a fusion protein described herein comprises a biotin-binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin), or biotin-binding fragment thereof. In some embodiments, a fusion protein comprises a biotin-binding moiety that is or comprises a polypeptide having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin, denoted Rhavi) or SEQ ID NO: 3, or biotin-binding fragment thereof. [0149] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin), or biotin-binding fragment thereof; (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P) polypeptide), or an antigenic fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435 polypeptide) or an antigenic fragment thereof. In some embodiments, the fusion protein further comprises one or more linkers. In some embodiments, such one or more linkers are independently selected from the group consisting of: the amino acid sequence of any one of SEQ ID NOs: 30-59 and AAA. In some embodiments, such one or more linkers are each GGGGSSS (SEQ ID NO: 30). [0150] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin), or biotin-binding fragment thereof; (ii) a AFV70463FF polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785 polypeptide), or an antigenic fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27- 278 of S. pneumoniae SP1500 polypeptide) or an antigenic fragment thereof. In some embodiments, the fusion protein further comprises one or more linkers. In some embodiments, such one or more linkers are independently selected from the group consisting of: the amino acid sequence of any one of SEQ ID NOs: 30-59 and AAA. In some embodiments, such one or more linkers are selected from GGGGSSS (SEQ ID NO: 30) and AAA. In some embodiments, the fusion protein comprises an amino acid sequence AAA residual from a Not I restriction site. In some embodiments, the fusion protein comprises a linker of GGGGSSS (SEQ ID NO: 30) and an amino acid sequence AAA residual from a Not I restriction site. [0151] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin, denoted Rhavi) or SEQ ID NO: 3, or biotin-binding fragment thereof; (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P) polypeptide) or an antigenic fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435 polypeptide) or an antigenic fragment thereof. In some embodiments, the fusion protein further comprises one or more linkers. In some embodiments, such one or more linkers are independently selected from the group consisting of: the amino acid sequence of any one of SEQ ID NOs: 30-59 and AAA. In some embodiments, such one or more linkers are each GGGGSSS (SEQ ID NO: 30). In some embodiments, a fusion protein described herein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, AFV70463FF at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence SEQ ID NO: 20 or SEQ ID NO: 21. In some embodiments, the fusion protein comprises the amino acid sequence SEQ ID NO: 20 or SEQ ID NO: 21. In some embodiments, the fusion protein consists of the amino acid sequence SEQ ID NO: 20 (SPP2) or SEQ ID NO: 21. [0152] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin, denoted Rhavi) or SEQ ID NO: 3, or biotin-binding fragment thereof; (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785 polypeptide) or an antigenic fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500 polypeptide) or an antigenic fragment thereof. In some embodiments, the fusion protein further comprises one or more linkers. In some embodiments, such one or more linkers are independently selected from the group consisting of: the amino acid sequence of any one of SEQ ID NOs: 30-59 and AAA. In some embodiments, such one or more linkers are selected from GGGGSSS (SEQ ID NO: 30) and AAA. In some embodiments, the fusion protein comprises an amino acid sequence AAA residual from a Not I restriction site. In some embodiments, the fusion protein comprises a linker of GGGGSSS (SEQ ID NO: 30) and an amino acid sequence AAA residual from a Not I restriction site. In some embodiments, a fusion protein described herein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence SEQ ID NO: 24 or SEQ ID NO: 25. In some embodiments, the fusion protein comprises the amino acid sequence SEQ ID NO: 24 or SEQ ID NO: 25. In some embodiments, the fusion protein consists of the amino acid sequence SEQ ID NO: 24 (CP1) or SEQ ID NO: 25. [0153] In some embodiments, the present disclosure provides fusion proteins with at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% sequence identity to a fusion protein having an amino acid sequence of AFV70463FF any one of SEQ ID NOs: 20-25. In some embodiments, a fusion protein is or includes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to SEQ ID NO: 20 or SEQ ID NO: 21. In some embodiments, a fusion protein is or includes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to SPP2. In some embodiments, a fusion protein is or includes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to SEQ ID NO: 24 or SEQ ID NO: 25. In some embodiments, a fusion protein is or includes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to CP1. [0154] In some embodiments, a fusion protein described herein comprises an antigenic fragment of a fusion protein having an amino acid sequence of any one of SEQ ID NOs: 20-25. For example, a fusion protein may lack at most one, two three, four, five, ten, or twenty amino acids from the N-terminus, C-terminus, or both, of any one of SEQ ID NOs: 20-25. In some embodiments, the same number of residues is removed from the N-terminus and the C-terminus, while in other embodiments, a different number of residues is removed from the N-terminus compared to the C-terminus. In some embodiments, a fusion protein is or includes an antigenic fragment of SEQ ID NO: 20 or SEQ ID NO: 21. In some embodiments, a fusion protein is or includes an antigenic fragment of SPP2. In some embodiments, a fusion protein is or includes an antigenic fragment of SEQ ID NO: 24 or SEQ ID NO: 25. In some embodiments, a fusion protein is or includes an antigenic fragment of CP1. [0155] In some embodiments, a fusion protein described herein includes a variant or fragment of a polypeptide having an amino acid sequence of SEQ ID NOs: 1-13. In some embodiments, a fusion protein described herein includes a polypeptide encoded by a variant or fragment of a gene having a nucleic acid sequence of SEQ ID NOs: 1-13. In some embodiments, a fragment included in a fusion protein described herein is close in size to a full-length polypeptide or a polypeptide having an amino acid sequence of SEQ ID NOs: 1-13. For example, they may lack at most one, two, three, four, five, ten, twenty, or thirty amino acids from one or both termini. In some embodiments, the fragment is 25-50 amino acids in length, or 50-100, or 100-150, or 150-200, or 200-250, or 250-300, or 300-350 amino acids in length. In some embodiments, the fragments AFV70463FF result from processing, or partial processing, of signal sequences by an expression host, e.g. E. coli, an insect cell line (e.g., the baculovirus expression system), a yeast (e.g., S. cerevisiae or S. pombe) cell line, or a mammalian (e.g., human or Chinese Hamster Ovary) cell line. The fragments described above or sub-fragments thereof (e.g., fragments of 8-50, 8-30, or 8-20 amino acid residues) preferably have one of the biological activities described below, such as increasing the amount of IL-17 released by at least 1.5-fold or 2-fold or more (e.g., either as an absolute measure or relative to a control protein). [0156] In some embodiments, fusion proteins described herein can be useful in immunogenic agents described herein. Linkers or Spacers [0157] In some embodiments, a polypeptide described herein is coupled to one or more entities of interest (e.g., small molecules, polypeptides, polynucleotides, lipids, polysaccharides, etc.) with one or more linkers. In some embodiments, a fusion protein described herein comprises one or more linkers. In some embodiments, a linker is or comprises one or more amino acids. In some embodiments, a fusion protein comprises an antigenic polypeptide joined to a biotin-binding moiety by a linker. In some embodiments, a fusion protein comprises a first antigenic polypeptide, a second antigenic polypeptide, a biotin-binding moiety, and at least one linker. In some embodiments, the first antigenic polypeptide and the second antigenic polypeptide are joined by a linker. In some embodiments, the first antigenic polypeptide or the second antigenic polypeptide are joined to the biotin-binding moiety by a linker. In some embodiments, the first antigenic polypeptide and the second antigenic polypeptide are joined by a first linker; and the first antigenic polypeptide or the second antigenic polypeptide are joined to the biotin-binding moiety by a second linker. [0158] In some embodiments, a fusion protein described herein comprises a first linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30), and a second linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30). In some embodiments, a fusion protein described herein comprises a first linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30), and a second linker comprising the amino acid sequence of AAA. [0159] In some embodiments, a linker interposes a structure between two protein moieties. In -helix. In some embodiments, the structure AFV70463FF -strand. In some embodiments, the structure is or comprises a coil/bend. In some embodiments, the structure is or comprises a turn. In some embodiments, a linker decreases steric hindrance between two protein moieties joined by the linker. In some embodiments, a linker decreases unfavorable interactions between two protein moieties joined by the linker. In some embodiments, a linker comprises a mixture of glycine and serine residues. In some embodiments, the linker may additionally comprise threonine, proline, and/or alanine residues. In some embodiment, a linker is hydrophilic. In some embodiments, a linker is hydrophobic. In some embodiments, a linker increases the stability of a fusion protein containing the linker. [0160] In some embodiments, a linker does not interfere with the folding or activity of a polypeptide of interest (e.g., in some embodiments, an antigenic polypeptide) to which it is joined. In some embodiments, a linker does not interfere with the antigenicity of an antigenic polypeptide to which it is joined. In some embodiments, a linker does not reduce the antigenicity of an antigenic polypeptide to which it is joined. In some embodiments, a linker does not eliminate the antigenicity of an antigenic polypeptide to which it is joined. In some embodiments, the effect of the linker is determined by comparing the polypeptide with the polypeptide joined to the linker. [0161] In some embodiments, a linker does not interfere with the folding of a biotin-binding moiety to which it is joined. In some embodiments, a linker does not interfere with the biotin- binding ability of a biotin-binding moiety to which it is joined. In some embodiments, a linker does not reduce the biotin-binding ability of a biotin-binding moiety to which it is joined. In some embodiments, a linker does not eliminate the biotin-binding ability of a biotin-binding moiety to which it is joined. In some embodiments, the effect of the linker is determined by comparing the biotin-binding moiety with the biotin-binding moiety joined to the linker. [0162] In some embodiments, a linker is not antigenic. In some embodiments, a linker does not elicit a T cell response. In some embodiments, a linker does not elicit a B cell response. In some embodiments, a linker does not induce a T cell or a B cell response. [0163] In some embodiments, a linker comprises two or more amino acids. In some embodiments, a linker may be 3-100, 5-100, 10-100, 20-10030-100, 40-100, 50-100, 60-100, 70- 100, 80-100, 90-100, 5-55, 10-50, 10-45, 10-40, 10-35, 10-30, 10-25, 10-20, 10-15, 3-10, 3-9, 3- 8, 3-7, 3-6, 3-5, 3-4, or 2-3 amino acids in length. In some embodiments, a linker comprises between 10-100, 10-90, 10-80, 10-70, 10-60, 10-50, 10-40, 10-30, 10-20, or 10-15 amino acids. In some embodiments, the linker comprises at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, AFV70463FF 70, 75, 80, 85, 90, or 95 amino acids. In some embodiments, a linker is or comprises a peptidyl linker comprising 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acids. [0164] In some embodiments, a linker is a flexible linker. Flexible linkers may be useful for joining domains that require a certain degree of movement or interaction and may include small, non-polar (e.g., Gly) or polar (e.g., Ser or Thr) amino acids. Incorporation of Ser or Thr can also maintain the stability of the linker in aqueous solutions by forming hydrogen bonds with water molecules, and therefore reduce unfavorable interactions between the linker and the protein moieties. In some embodiments, a linker comprises small non-polar (e.g. Gly) or polar (e.g. Ser or Thr) amino acids. In some embodiments, a linker is a Gly-Ser linker. [0165] In some embodiments, a linker is or comprises an amino acid sequence of GGGGSSS (SEQ ID NO: 30). In some embodiments, a linker is or comprises a sequence of (GGGGS)_n (SEQ ID NO: 31), where n represents the number of repeating GGGGS (SEQ ID NO: 32) units and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a polypeptide linker may have an amino acid sequence that is or comprises GGGGSGGGGSGGGGS (SEQ ID NO: 33) (i.e., (GGGGS)₃) or GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 34) (i.e., (GGGGS)₆). In some embodiments, a linker is or comprises (Gly)_n (SEQ ID NO: 35) where n represents the number of repeating Gly residues and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a linker is or comprises GGG (i.e., (Gly)₃). In some embodiments, a linker is or comprises (Gly)₆ (SEQ ID NO: 36). In some embodiments, a linker is or comprises (Gly)₈ (SEQ ID NO: 37). In some embodiments, a linker comprises one or more of Gly, Ser, Thr, Ala, Lys, and Glu. In some embodiments, a linker is or comprises KESGSVSSEQLAQFRSLD (SEQ ID NO: 38). In some embodiments, a linker is or comprises EGKSSGSGSESKST (SEQ ID NO: 39). In some embodiments, a linker is or comprises GSAGSAAGSGEF (SEQ ID NO: 40). In some embodiments, a linker is or comprises an amino acid sequence AAA. In some embodiments, such a linker may be synthesized, or derived from amino acid residues from a restriction site (e.g., a Not I restriction site). [0166] In some embodiments, a linker is a rigid linker. Rigid linkers are useful to keep a fixed distance between domains and to maintain their independent functions. Rigid linkers may also be useful when a spatial separation of the domains is critical to preserve the stability or bioactivity of AFV70463FF one or more components in the fusion. In some embodiments, a linker is or comprises (EAAAK)_n (SEQ ID NO: 41) where n represents the number of repeating EAAAK (SEQ ID NO: 42) units and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a linker is or comprises A(EAAAK)_nA, (SEQ ID NO: 43) where n represents the number of repeating EAAAK (SEQ ID NO: 42) units and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a linker is or comprises A(EAAAK)_nA (SEQ ID NO: 44), where n represents the number of repeating EAAAK (SEQ ID NO: 42) units and is 2, 3, 4, or 5. In some embodiments, a linker is or comprises A(EAAAK)₄ALEA(EAAAK)₄A (SEQ ID NO: 45). In some embodiments, a linker is or comprises [A(EAAAK)_nA]_m, (SEQ ID NO: 46) wherein n is 2, 3, or 4 and m is 1 or 2. In some embodiments, a linker is or comprises AEAAAKEAAAKA (SEQ ID NO: 47). [0167] In some embodiments, a linker is or comprises (X-Pro)_n (SEQ ID NO: 48), with X designating any amino acid, where n represents the number of repeating X-Pro units and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a linker is or comprises (Ala-Pro)_n (SEQ ID NO: 49), where n represents the number of repeating Ala-Pro units and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a linker is or comprises (Ala-Pro)_n (SEQ ID NO: 50), where n represents the number of repeating Ala-Pro units and is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17. [0168] In some embodiments, a linker is or comprises (Lys-Pro)_n (SEQ ID NO: 51), where n represents the number of repeating Lys-Pro units and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a linker is or comprises (Glu-Pro)_n (SEQ ID NO: 52), where n represents the number of repeating Glu-Pro units and is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more. In some embodiments, a linker is or comprises (Ala-Pro)₇ (SEQ ID NO: 53). [0169] In some embodiments, a linker is or comprises GAPGGGGGAAAAAGGGGGGAP (GAG linker, SEQ ID NO: 54). In some embodiments, a linker is or comprises GAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGGGAP (GAG2 linker, SEQ ID NO: 55). In some embodiments, a linker is or comprises GAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGG GAP (GAG3 linker, SEQ ID NO: 56). [0170] In some embodiments, a linker is or comprises VSDP (SEQ ID NO: 57). AFV70463FF [0171] In some embodiments, a linker is or comprises AAAA (SEQ ID NO: 58). [0172] In some embodiments, a linker is or comprises GGGG (SEQ ID NO: 59). [0173] Suitable linkers or spacers also include those having an amino acid sequence at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more homologous or identical to the above exemplary linkers. In some embodiments, the linker comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of GGGGSSS (SEQ ID NO: 30). [0174] Additional linkers suitable for use with some embodiments may be found in U.S. Patent Publication No.2012/0232021, filed on March 2, 2012, and [Chen, 2013] the disclosures of which is hereby incorporated by reference in their entireties. Tags [0175] In some embodiments, a polypeptide (e.g., a fusion protein) described herein may comprise one or more tags. In some embodiments, a fusion protein described herein comprises one or more tags. A tag may be present at the N-terminal or C-terminal of a polypeptide. For instance, tags may be added to a polypeptide (e.g., via additions or modifications on the encoding DNA sequence) to facilitate purification, detection, solubility, or confer other desirable characteristics on the polypeptide. In some embodiments, a tag may be a peptide, oligopeptide, or polypeptide that may be used in affinity purification. In some embodiments, a tag is, comprises, or is derived from one or more of poly-histidine (His), Glutathione S-transferase (GST), tandem affinity purification (TAP), FLAG, myc, human influenza hemagglutinin (HA), maltose binding protein (MBP), vesicular Stomatitis viral glycoprotein (VSV-G), thioredoxin, V5, avidin, streptavidin, biotin carboxyl carrier protein (BCCP), calmodulin, Nus, S tags, lipoprotein D, and galactosidase. In some embodiments, a His tag is or comprises an amino acid sequence of H_n, wherein n is an integer between 2 and 10 (SEQ ID NO: 60). Exemplary His tags include HHHHHH (SEQ ID NO: 61) and MSYYHHHHHH (SEQ ID NO: 62). In other embodiments, a polypeptide described herein is free of tags such as protein purification tags, and is purified by a method not relying on affinity for a purification tag. In some embodiments, a fusion protein described herein comprises AFV70463FF (i) a polypeptide of amino acid sequence of any of SEQ ID NOs: 4-13 or fusion protein of amino acid sequence of any of SEQ ID NOs: 20-25, and (ii) a tag having no more than 1, 2, 3, 4, 5, 10, or 20 amino acids on one or both termini of the polypeptide or fusion protein of (i). [0176] In some embodiments, a polypeptide described herein may contain a membrane translocating sequence (MTS), for example, in some embodiments, to facilitate introduction of the fusion protein into a mammalian cell and subsequent stimulation of the cell-mediated immune response. Exemplary membrane translocating sequences include, but are not limited to the hydrophobic region in the signal sequence of Kaposi fibroblast growth factor, the MTS of a synuclein, the third helix of the Antennapedia homeodomain, SN50, integrin 3 h-region, HIV Tat, pAntp, PR-39, abaecin, apidaecin, Bac5, Bac7, P. berghei CS protein, and those MTSs described in U.S. Patent Nos.6,248,558; 6,432,680; and 6,248,558. [0177] In some embodiments, a polypeptide (e.g., a fusion protein) described herein may comprise one or more tags. In some embodiments, a fusion protein comprises one or more tags. A tag may be N-terminal or C-terminal. For instance, tags may be added to a polypeptide (via additions or modifications on the encoding DNA sequence) to facilitate purification, detection, solubility, or confer other desirable characteristics on the protein. In some embodiments, a tag may be a peptide, oligopeptide, or polypeptide that may be used in affinity purification. In some embodiments, a tag is, comprises, or is derived from one or more of polyhistidine (His), Glutathione S-transferase (GST), tandem affinity purification (TAP), FLAG, myc, human influenza hemagglutinin (HA), maltose binding protein (MBP), vesicular Stomatitis viral glycoprotein (VSV-G), thioredoxin, V5, avidin, streptavidin, biotin carboxyl carrier protein (BCCP), Calmodulin, Nus, S tags, lipoprotein D, and galactosidase. In some embodiments, a His tag is or comprises an amino acid sequence of H_n, wherein n is an integer between 2 and 10 (SEQ ID NO: 60). Exemplary His tags include HHHHHH (SEQ ID NO: 61) and MSYYHHHHHH (SEQ ID NO: 62). In other embodiments, a fusion protein described herein is free of tags such as protein purification tags, and is purified by a method not relying on affinity for a purification tag. In some embodiments, a fusion protein described herein comprises (i) a polypeptide of amino acid sequence of any of SEQ ID NOs: 1-13 or fusion protein of amino acid sequence of any of SEQ ID NOs: 20-25, and (ii) a tag having no more than 1, 2, 3, 4, 5, 10, or 20 amino acids on one or both termini of the polypeptide or fusion protein of (i). AFV70463FF [0178] In some embodiments, a polypeptide described herein may contain a membrane translocating sequence (MTS), for example, in some embodiments, to facilitate introduction of the fusion protein into a mammalian cell and subsequent stimulation of the cell-mediated immune response. Exemplary membrane translocating sequences include, but are not limited to the hydrophobic region in the signal sequence of Kaposi fibroblast growth factor, the MTS of a synuclein, the third helix of the Antennapedia homeodomain, SN50, integrin 3 h-region, HIV Tat, pAntp, PR-39, abaecin, apidaecin, Bac5, Bac7, P. berghei CS protein, and those MTSs described in U.S. Patent Nos.6,248,558; 6,432,680; and 6,248,558. Signal Peptides [0179] In some embodiments, a polypeptide described herein comprises a signal peptide. In some embodiments, a fusion protein comprises a signal peptide. Signal peptides, which may also be called leader peptides, function in translocation of polypeptides through a cellular membrane (e.g., the plasma membrane, endoplasmic reticulum, etc.). In some embodiments, a polypeptide comprises a signal peptide at or near its N-terminus. In some embodiments, the signal sequence may or may not be cleaved off (e.g., by a signal peptidase) after translocation through a membrane. [0180] A signal sequence can be directly linked (e.g., by a peptide bond) or indirectly linked (e.g., by a linker) to a polypeptide. In some embodiments, the signal sequence can be linked to a polypeptide by a peptide linker disclosed herein. [0181] In some embodiments, a signal peptide may be a bacterial signal peptide, a viral signal peptide, or eukaryotic signal peptide. In some embodiments, a eukaryotic signal peptide is a mammalian signal peptide (e.g., a signal peptide from, or derived from, a human protein). [0182] In some embodiments, a signal peptide may be a bacterial signal peptide. Bacterial signal peptides may be used to promote translocation of a polypeptide into periplasmic space of bacteria. In some embodiments, the signal peptide is cleaved off from the polypeptide after translocation into the periplasmic space of a bacterium (e.g., E. coli). Signal peptides that direct secretion of proteins from bacterial cells are well known in the art, for example as disclosed in PCT Pat. App. Pub. No. WO 2005/071088. AFV70463FF [0183] Any signal peptide known to one of ordinary skill in the art can be used. In some embodiments, a signal peptide is or comprises the amino acid sequence: MKKIWLALAGLVLAFSASA (SEQ ID NO: 63), MAPFEPLASGILLLLWLIAPSRA (SEQ ID NO: 64), MKKVAAFVALSLLMAGC (SEQ ID NO: 65), MKKIMLVITLILVSPIAQQTEAKD (SEQ ID NO: 66), MKKKIISAILMSTVILSAAAPLSGVYADT (SEQ ID NO: 67), MKKRKVLIPLMALSTILVSSTGNLEVIQAEV (SEQ ID NO: 68), MNMKKATIAATAGIAVTAFAAPTIASAST (SEQ ID NO: 69), MQKTRKERILEALQEEKKNKKSKKFKTGATIAGVTAIATSITVPGIEVIVSADE (SEQ ID NO: 70), MKKLKMASCALVAGLMFSGLTPNAFAED (SEQ ID NO: 71), MAKKFNYKLPSMVALTLVGSAVTAHQVQAAE (SEQ ID NO: 72), MTDKKSENQTEKTETKENKGMTRREMLKLSAVAGTGIAVGATGLGTILNVVDQVDKA LT (SEQ ID NO: 73), MAYDSRFDEWVQKLKEESFQNNTFDRRKFIQGAGKIAGLGLGLTIAQSVGAFG (SEQ ID NO: 74) or a derivative or functional portion thereof. In some embodiments, a signal peptide is or comprises the amino acid sequence of a human Ig heavy chain V-III region VH26 signal peptide or a derivative or functional portion thereof. In some embodiments, a signal peptide is or comprises the amino acid sequence of MEFGLSWLFLVAILKGVQC (SEQ ID NO: 75) or a derivative or functional portion thereof. In some embodiments, a signal peptide is or comprises a human IgG2 heavy chain signal peptide, or a derivative or functional portion thereof. In some embodiments, a signal peptide is or comprises the amino acid sequence of MGWSCIILFLVATATGVHS (SEQ ID NO: 76) or a derivative or functional portion thereof. [0184] Other examples of signal peptides can be found at Signal Peptide Website, a signal peptide database, which is found at www.signalpeptide.de. [0185] In some embodiments, a polypeptide described herein lacks a signal peptide. Affinity Molecule Pairs [0186] As described herein, in some embodiments, immunogenic agents of the present disclosure include complementary affinity molecule pairs comprising an affinity molecule (i.e., a first affinity molecule) and a complementary affinity molecule (i.e., a second affinity molecule that is complementary to the first affinity molecule). In some embodiments, an immunogenic AFV70463FF agents includes a first affinity molecule. In some embodiments, an immunogenic agent includes a second affinity molecule. In some embodiments, the affinity molecule/complementary affinity molecule pair is selected from one or more of biotin/biotin-binding moiety, antibody/antigen, enzyme/substrate, receptor/ligand, metal/metal-binding protein, carbohydrate/carbohydrate binding protein, lipid/lipid-binding protein, and His tag/His tag-binding molecule. In some embodiments, a first affinity molecule is associated with a polymer (e.g., but not limited to a polysaccharide). In some embodiments a second affinity molecule is associated with a polypeptide of interest (e.g., but not limited to a polypeptide antigen). [0187] In some embodiments, the first affinity molecule is biotin (or a derivative or fragment thereof), and the complementary affinity molecule is a biotin-binding moiety, e.g., a biotin-binding protein or polypeptide, or a biotin-binding domain or biotin-binding fragment thereof. In some embodiments, a fusion protein of an immunogenic agent comprises a biotin-binding moiety, and one or more polypeptide antigens. In some embodiments, a fusion protein comprises a biotin- binding moiety and two or more polypeptide antigens. In some embodiments, the biotin-binding moiety is a biotin-binding moiety that can form a dimer. In some embodiments, the biotin-binding moiety is a biotin-binding moiety that can form a tetramer. In some embodiments, the biotin- binding moiety is or comprises rhizavidin, avidin, streptavidin, bradavidin, tamavidin, lentiavidin, zebavidin, NeutrAvidin, CaptAvidin™, or a biotin-binding domain or biotin-binding fragment thereof, or a combination thereof. In some embodiments, the biotin-binding moiety is or comprises rhizavidin, or a biotin-binding domain or biotin-binding fragment thereof. [0188] In some embodiments, immunogenic agents described herein utilize the high affinity -¹⁵ M) non-covalent binding between biotin and rhizavidin, a biotin-binding protein that has no significant predicted homology with human proteins. Rhizavidin, a naturally occurring dimeric protein in the avidin protein family, was first discovered in Rhizobium etli, a symbiotic bacterium of the common bean. Rhizavidin has only a 22% amino acid identity with chicken avidin, a protein commonly found in eggs, but with high conservation of amino acid residues involved in biotin binding. No cross-reactivity to rhizavidin is observed in human serum samples obtained from subjects exposed to avidin [Helppolainen et al, 2007], suggesting that rhizavidin antibodies may not cross-react with chicken avidin. Biotin conjugates have been used in several clinical applications without any reported adverse events [Buller et al, 2014; Paty et al, 2010; Lazzeri et al, 2004]. AFV70463FF [0189] In some embodiments, a biotin-binding moiety that is useful in accordance with the present disclosure comprises rhizavidin or a biotin-binding domain or biotin-binding fragment thereof, for example, in some embodiments as further described in WO 2012/155053, the contents of which are herein incorporated by reference in their entirety for the purposes described herein. In some embodiments, a biotin-binding moiety is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to rhizavidin, or a biotin-binding domain or biotin-binding fragment thereof. In some embodiments, a biotin-binding moiety that is useful in accordance with the present disclosure comprises a polypeptide of SEQ ID NO: 1 or a biotin-binding domain or biotin-binding fragment thereof. In some embodiments, a biotin-binding moiety is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, 100% identity to the sequence of SEQ ID NO: 1, or biotin-binding domain or biotin-binding fragment thereof. In some embodiments, a biotin-binding moiety that is useful in accordance with the present disclosure comprises a polypeptide of SEQ ID NO: 2 or SEQ ID NO: 3, or a biotin- binding domain or biotin-binding fragment thereof. In some embodiments, a biotin-binding moiety is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 2 or SEQ ID NO: 3, or biotin-binding domain or biotin-binding fragment thereof. Fusion Protein SPP2 [0190] In some embodiments, a fusion protein described herein is or comprises a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein), and one or more polypeptides of or derived from S. pneumoniae. In some embodiments, a provided fusion protein is SPP2, further described in International Application No. PCT/US2022/043156 filed on September 9, 2022, the contents of which are incorporated herein by reference in their entirety for the purposes described herein. In some embodiments, a fusion protein comprises (i) a biotin-binding moiety (e.g., a biotin-binding protein), (ii) a non-hemolytic pneumolysin polypeptide comprising mutations at amino acid residues 294, 385, 428, and 433 of wild-type Streptococcus pneumoniae pneumolysin or an antigenic fragment thereof; and (iii) a S. pneumoniae elongation factor P (SP0435) polypeptide or an antigenic fragment thereof. In some AFV70463FF embodiments, a biotin-binding moiety is or comprises a rhizavidin polypeptide or a biotin-binding portion thereof. In some embodiments, a fusion protein comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein) (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P)) or an antigenic variant or fragment thereof, and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435) or an antigenic variant or fragment thereof. In some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 20 or SEQ ID NO: 21. In some embodiments, a fusion protein is or comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein), (ii) a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to pneumolysin or an antigenic variant or fragment thereof, and (iii) a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SP0435 or an antigenic variant or fragment thereof. In some embodiments, a fusion protein is or comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein), (ii) a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a non-hemolytic pneumolysin comprising mutations at amino acid residues 294, 385, 428, and 433 of wild-type S. pneumoniae pneumolysin or an antigenic variant or fragment thereof, and (iii) a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SP0435 or an antigenic variant or fragment thereof. [0191] In some embodiments, a fusion protein described herein comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein) and (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity AFV70463FF to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P)) or an antigenic variant or fragment thereof. In some embodiments, a fusion protein comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein) and (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435) or an antigenic variant or fragment thereof. [0192] In some embodiments, a fusion protein described herein comprises a biotin-binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin), or biotin-binding fragment thereof. In some embodiments, a fusion protein comprises a biotin-binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin) or SEQ ID NO: 3, or biotin-binding fragment thereof. In some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P)) or an antigenic variant or fragment thereof. In some embodiments, the fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435) or an antigenic variant or fragment thereof. [0193] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin), or biotin-binding fragment thereof; (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P)) or an antigenic variant or fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least AFV70463FF 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435) or an antigenic variant or fragment thereof. In some embodiments, such a fusion protein further comprises one or more linkers. [0194] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin) or SEQ ID NO: 3, or biotin-binding fragment thereof; (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P)) or an antigenic variant or fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435) or an antigenic variant or fragment thereof. In some embodiments, such a fusion protein further comprises one or more linkers. [0195] In some embodiments, a fusion protein described herein is or comprises a SPP2 fusion protein. In some embodiments, a SPP2 fusion protein comprises, from N-terminus to C-terminus, (i) a biotin-binding moiety (e.g., a biotin-binding protein), (ii) a first peptide linker, (iii) a pneumolysin (Ply) polypeptide described herein or an antigenic variant or fragment thereof; (iv) a second peptide linker, and (v) an SP0435 polypeptide described herein or an antigenic variant or fragment thereof. In some such embodiments, a biotin-binding moiety is or comprises rhizavidin or a biotin-binding portion thereof. In some embodiments, a SPP2 fusion protein comprises (i) a biotin-binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin) or to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin) or SEQ ID NO: 3, or biotin-binding fragment thereof; (ii) a first polypeptide linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30); (iii) an pneumolysin (Ply) polypeptide described herein; (iv) a second polypeptide linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30); and (v) an SP0435 polypeptide described herein. In some embodiments, a SPP2 fusion protein comprises, from N-terminus to C-terminus, (i) a biotin- AFV70463FF binding moiety that is or comprises the polypeptide of SEQ ID NO: 1 (rhizavidin), SEQ ID NO: 2 (amino acids 45-179 of rhizavidin), or SEQ ID NO: 3, or biotin-binding fragment thereof; (ii) a first polypeptide linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30); (iii) an pneumolysin (Ply) polypeptide described herein; (iv) a second polypeptide linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30); and (v) an SP0435 polypeptide described herein. In some embodiments, such a SPP2 fusion protein may further comprise a detection or purification tag, e.g., a His tag. In some embodiments, a SPP2 fusion protein comprising a His tag is referred to as SPP2-H. In some such embodiments, a SPP2 fusion protein comprises an SP0435 polypeptide (e.g., ones described herein) between a biotin-binding moiety and a Ply polypeptide (e.g., ones described herein). In some embodiments, a SPP2 fusion protein may comprise a Ply polypeptide between a biotin-binding moiety and a SP0435 polypeptide. In some embodiments, a Ply polypeptide included in a fusion protein described herein is or comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 7 (amino acids 2-470 of S. pneumoniae PdT(G294P)) or an antigenic variant or fragment thereof. In some embodiments, an SP0435 polypeptide included in a fusion protein described herein is or comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 9 (amino acids 62-185 of S. pneumoniae SP0435) or an antigenic variant or fragment thereof. In some embodiments, a fusion protein described herein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence SEQ ID NO: 20 or SEQ ID NO: 21. In some embodiments, a fusion protein described herein comprises the amino acid sequence SEQ ID NO: 20 or SEQ ID NO: 21. In some embodiments, a fusion protein described herein consists of the amino acid sequence SEQ ID NO: 20 or SEQ ID NO: 21. Fusion Protein CP1 [0196] In some embodiments, a fusion protein described herein is or comprises a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein), and one or more polypeptides of or derived from S. pneumoniae. In some embodiments, a fusion protein described herein is CP1, further described in the International Patent Publication No. WO AFV70463FF 2020/056127, the contents of which are incorporated herein by reference in their entirety for the purposes described herein. In some embodiments, a fusion protein comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein), (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785) or an antigenic variant or fragment thereof, and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500) or an antigenic variant or fragment thereof. In some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 24 or SEQ ID NO: 25. In some embodiments, a fusion protein is or comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein), (ii) a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SP0785 or an antigenic variant or fragment thereof, and (iii) a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SP1500 or an antigenic variant or fragment thereof. [0197] In some embodiments, a fusion protein described herein comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein) and (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785) or an antigenic variant or fragment thereof. In some embodiments, a fusion protein comprises (i) a complementary affinity molecule described herein (e.g., a biotin-binding moiety described herein) and (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500) or an antigenic variant or fragment thereof. AFV70463FF [0198] In some embodiments, a fusion protein described herein comprises a biotin-binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin), or biotin-binding fragment thereof. In some embodiments, a fusion protein comprises a biotin-binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin) or SEQ ID NO: 3, or biotin-binding fragment thereof. In some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785) or an antigenic variant or fragment thereof. In some embodiments, a fusion protein comprises a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500) or an antigenic variant or fragment thereof. [0199] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin), or biotin-binding fragment thereof; (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785) or an antigenic variant or fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500) or an antigenic variant or fragment thereof. In some embodiments, such a fusion protein further comprises one or more linkers. [0200] In some embodiments, a fusion protein described herein comprises each of: (i) a biotin- binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence AFV70463FF of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin) or SEQ ID NO: 3, or biotin-binding fragment thereof; (ii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785) or an antigenic variant or fragment thereof; and (iii) a polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500) or an antigenic variant or fragment thereof. In some embodiments, such a fusion protein further comprises one or more linkers. [0201] In some embodiments, a fusion protein described herein is or comprises a CP1 fusion protein. In some embodiments, a CP1 fusion protein comprises, from N-terminus to C-terminus, (i) a biotin-binding moiety (e.g., a biotin-binding protein), (ii) a first peptide linker, (iii) an SP1500 polypeptide described herein or an antigenic variant or fragment thereof; (iv) a second peptide linker, and (v) an SP0785 polypeptide described herein or an antigenic variant or fragment thereof. In some such embodiments, a biotin-binding moiety is or comprises rhizavidin or a biotin-binding portion thereof. In some embodiments, a CP1 fusion protein comprises (i) a biotin-binding moiety that is or comprises a polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the sequence of SEQ ID NO: 1 (rhizavidin) or to the sequence of SEQ ID NO: 2 (amino acids 45-179 of rhizavidin) or SEQ ID NO: 3, or biotin-binding fragment thereof; (ii) a first polypeptide linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30); (iii) an SP0785 polypeptide described herein; (iv) a second polypeptide linker comprising the amino acid sequence of AAA; and (v) an SP1500 polypeptide described herein. In some embodiments, a CP1 fusion protein comprises, from N- terminus to C-terminus, (i) a biotin-binding moiety that is or comprises the polypeptide of SEQ ID NO: 1 (rhizavidin), SEQ ID NO: 2 (amino acids 45-179 of rhizavidin), or SEQ ID NO: 3, or biotin- binding fragment thereof; (ii) a first polypeptide linker comprising the amino acid sequence of GGGGSSS (SEQ ID NO: 30); (iii) an SP1500 polypeptide described herein; (iv) a second polypeptide linker comprising the amino acid sequence of AAA; and (v) an SP0785 polypeptide described herein. In some embodiments, such a CP1 fusion protein may further comprise a detection or purification tag, e.g., a His tag. In some such embodiments, a CP1 fusion protein comprises an SP1500 polypeptide between a biotin-binding moiety and an SP0785 polypeptide. AFV70463FF In some embodiments, a CP1 fusion protein may comprise an SP0785 polypeptide between a biotin-binding moiety and a SP1500 polypeptide. In some embodiments, an SP0785 polypeptide included in a fusion protein described herein is or comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 11 (amino acids 33-399 of S. pneumoniae SP0785) or an antigenic variant or fragment thereof. In some embodiments, an SP1500 polypeptide included in a fusion protein described herein is or comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence of SEQ ID NO: 13 (amino acids 27-278 of S. pneumoniae SP1500) or an antigenic variant or fragment thereof. In some embodiments, a fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identity to the sequence SEQ ID NO: 24 or SEQ ID NO: 25. In some embodiments, a fusion protein comprises the amino acid sequence SEQ ID NO: 24 or SEQ ID NO: 25. In some embodiments, a fusion protein consists of the amino acid sequence SEQ ID NO: 24 or SEQ ID NO: 25. Carrier Proteins [0202] In some embodiments, an immunogenic agent described herein is or comprises one or more carrier proteins. In some embodiments, a carrier protein is or comprises an antigenic polypeptide (e.g., ones described herein including, e.g., SP0785 as described herein, SP1500 as described herein, SP0435 as described herein, Ply as described herein or in WO 05/76696, WO 05/108580, or WO 10/71986, etc.) and/or a fusion protein that includes one or more antigenic polypeptides (e.g., ones described herein including, e.g., CP1 as described herein, SPP2 as described herein, etc.). In some embodiments, a carrier protein is a toxoid or a glycoprotein from a pathogen. Additional exemplary carrier proteins include, but are not limited to PspA as described in WO 19/167008; CRM197 as described in WO 14/92378; variants of CRM197 such as eCRM197 as disclosed in WO 22/178015; Diphtheria toxoid; variants of Diphtheria toxoid as described in WO18/156465; Tetanus toxoid as described in WO 19/152921; ExoProtein A, and Protein D as described in US5858677, and combinations thereof. In some embodiments, the carrier protein is CRM197. AFV70463FF [0203] In some embodiments, a fusion protein described herein may comprise a carrier protein. Exemplary Methods of Isolating and Purifying Polysaccharides [0204] In some embodiments, the disclosure provides methods of purifying one or more polysaccharides described herein from one or more cellular components of bacteria. In some embodiments, methods comprise purifying capsular polysaccharides from one or more cellular components of bacteria. [0205] In some embodiments, the bacteria are Gram-negative. In some embodiments, the bacteria are Gram-positive. In some embodiments, the bacteria are S. pneumoniae. [0206] In some embodiments, the cellular components include protein. In some embodiments, the cellular proteins include nucleic acid. In some embodiments, the cellular components include lipids. In some embodiments, the cellular components include polysaccharides. In some embodiments, the cellular components are part of a lysate. [0207] In some embodiments, the polysaccharide purification processes incorporate a series of ethanol precipitations, washes of crude polysaccharide preparations with ethanol, diethyl ether, and/or acetone, and drying under vacuum to furnish purified products. In some embodiments, a phenol extraction step is incorporated for polysaccharide purifications. In some embodiments, the purification process employs a CTAB (cetyltrimethyl ammonium bromide) precipitation step in addition to using ethanol and phenol precipitation steps. Exemplary Methods of Biotinylating Polysaccharides [0208] In some embodiments, the disclosure provides methods of biotinylating one or more polysaccharides described herein. In some embodiments, the method comprises reacting purified polysaccharides with 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) for activation of hydroxyl groups in the polysaccharides followed by the addition of amine PEG biotin under conditions that result in covalent linkage of biotin to the polysaccharides. In some embodiments, the desired level of biotinylation is achieved by varying the ratio of CDAP to polysaccharide. In some embodiments, the biotinylated polysaccharides are purified by filtration to remove process residuals such as unreacted biotin, dimethylaminopyridine, acetonitrile, cyanide AFV70463FF and unreacted glycine. In some embodiments, the level of polysaccharide biotinylation described herein is optimized to reduce the amount of accessible biotin following MAPS complexation. Immunogenic Complexes [0209] In some embodiments, an immunogenic agent described herein is or comprises an immunogenic complex. In some embodiments, an immunogenic complex is or comprises one or more polypeptides described herein non-covalently complexed with a polysaccharide (e.g., a purified capsular polysaccharide of formula I) described herein. In some embodiments, one or more polypeptides are complexed via affinity interaction with a polysaccharide (e.g., a purified capsular polysaccharide of formula I). In some embodiments, immunogenic agents of the present disclosure include one or more polypeptides non-covalently complexed with a polysaccharide (e.g., a purified capsular polysaccharide of formula I) using one or more affinity molecule pairs each comprising a first affinity molecule and a second affinity molecule complementary to the first affinity molecule (“complementary affinity molecule”). Upon association of the first affinity molecule and the complementary affinity molecule, the one or more polypeptides are non- covalently complexed to a purified capsular polysaccharide of formula I. [0210] In some embodiments, a polypeptide of an immunogenic complex is a fusion protein comprising a biotin-binding moiety and at least one polypeptide antigen. In some embodiments, a biotinylated purified capsular polysaccharide is non-covalently associated with a biotin-binding moiety of a fusion protein. [0211] In some embodiments, an immunogenic complex described herein comprises one or more polypeptides (e.g., antigenic polypeptides) that are complexed via affinity interaction with a purified capsular polysaccharide. In some embodiments, immunogenic agents of the disclosure include one or more polypeptides (e.g., antigenic polypeptides) non-covalently complexed with a purified capsular polysaccharide using one affinity molecule/complementary affinity molecule pair. In some embodiments, immunogenic agents of the disclosure include one or more polypeptides (e.g., antigenic polypeptides) non-covalently complexed with a purified capsular polysaccharide using one or more affinity molecule/complementary affinity molecule pairs. In some embodiments, each of the affinity molecule/complementary affinity molecule pairs is the same, e.g., biotin/biotin-binding moiety pairs. In some embodiments, an immunogenic agent includes (i) a purified capsular polysaccharide (e.g., a purified capsular polysaccharide of formula AFV70463FF I) associated with (e.g., in some embodiments, by chemical conjugation) a first affinity molecule described herein, and (ii) a fusion protein that comprises a complementary affinity molecule described herein and one or more polypeptides (e.g., antigenic polypeptides). In some embodiments, an immunogenic agent includes (i) a purified capsular polysaccharide (e.g., a purified capsular polysaccharide of formula I) associated with (e.g., in some embodiments, by chemical conjugation) a plurality of first affinity molecule described herein, and (ii) a fusion protein that comprises a complementary affinity molecule described herein and one or more polypeptides (e.g., antigenic polypeptides). Upon association of the first affinity molecule and the complementary affinity molecule, the one or more polypeptides (e.g., antigenic polypeptides) are non-covalently complexed to a purified capsular polysaccharide (e.g., a purified capsular polysaccharide of formula I). [0212] In some embodiments, the affinity molecule/complementary affinity molecule pair is selected from one or more of biotin/biotin-binding moiety, antibody/antigen, enzyme/substrate, receptor/ligand, metal/metal-binding protein, carbohydrate/carbohydrate binding protein, lipid/lipid-binding protein, and His tag/His tag-binding molecule. [0213] In some embodiments, the first affinity molecule is biotin (or a derivative or fragment thereof), and the complementary affinity molecule is a moiety, e.g., a biotin-binding protein, or a biotin-binding domain or biotin-binding fragment thereof. In some embodiments, the biotin- binding moiety is rhizavidin, avidin, streptavidin, bradavidin, tamavidin, lentiavidin, zebavidin, NeutrAvidin, CaptAvidin™, or a biotin-binding domain or biotin-binding fragment thereof, or a combination thereof. In some embodiments, the biotin-binding moiety is a dimer, e.g., a non- covalent dimer. In some embodiments, the biotin-binding moiety is rhizavidin or a variant thereof, or a biotin-binding domain or biotin-binding fragment thereof. In some embodiments, the biotin- binding moiety is or comprises a polypeptide of SEQ ID NO: 1, or a biotin-binding domain or biotin-binding fragment thereof. In some embodiments, the biotin-binding moiety is or comprises a polypeptide that comprises an amino acid sequence that is at least 80% (including, e.g., at least 90%, at least 95%, at least 98%, at least 99%, and 100%) identical to SEQ ID NO: 1, or a biotin- binding domain or biotin-binding fragment thereof. In some embodiments, the biotin-binding moiety is or comprises a polypeptide of SEQ ID NO: 2 or SEQ ID NO: 3, or a biotin-binding domain or biotin-binding fragment thereof. In some embodiments, the biotin-binding moiety is or comprises a polypeptide that comprises an amino acid sequence that is at least 80% (including, AFV70463FF e.g., at least 90%, at least 95%, at least 98%, at least 99%, 100%) identical to SEQ ID NO: 2 or SEQ ID NO: 3, or a biotin-binding domain or biotin-binding fragment thereof. [0214] In some embodiments, one or more antigenic polypeptides are covalently linked (e.g., fused) to a complementary affinity molecule described herein. In some embodiments, a fusion protein comprises one or more antigenic polypeptides and a complementary affinity molecule disclosed herein. In some embodiments, the complementary affinity molecule is or comprises a biotin-binding moiety. In some embodiments, the biotin-binding moiety is a dimer, e.g., a non- covalent dimer. In some embodiments, the biotin-binding moiety comprises rhizavidin or a biotin- binding portion thereof. [0215] In some embodiments, an immunogenic agent of the present disclosure is an immunogenic complex as described in WO 2018/237221, WO 2020/056202, PCT/US2022/043156, or PCT/US2022/042964. The contents of each of the aforementioned references are incorporated by reference in their entirety for their purposes described herein [0216] In some embodiments, an immunogenic agent described herein is, or is based on, Multiple Antigen Presenting System (MAPS) complexes. Aspects of the MAPS platform have been previously described in WO2012/155007 and WO2020/056202, the contents of which are herein incorporated by reference in their entirety. Immunogenic Conjugates [0217] In some embodiments, an immunogenic agent described herein is or comprises an immunogenic conjugate. In some embodiments, an immunogenic conjugate is or comprises one or more polypeptides described herein covalently conjugated with a polysaccharide (e.g., a purified capsular polysaccharide of formula I) described herein. [0218] In some embodiments, a polypeptide of an immunogenic conjugate is or comprises an antigenic polypeptide described herein. In some embodiments, a polypeptide of an immunogenic conjugate is or comprises a carrier protein described herein. In some embodiments, a polypeptide of an immunogenic conjugate is or comprises a fusion protein described herein. Manufacture of Immunogenic Complexes and Immunogenic Conjugates [0219] The present disclosure includes methods for manufacturing immunogenic complexes described herein. In some embodiments, a method of manufacturing an immunogenic complex AFV70463FF comprises complexing at least one biotinylated polymer (e.g., a biotinylated polysaccharide (e.g., a biotinylated polysaccharide described herein)) with at least one biotin-binding fusion protein described herein. [0220] In some embodiments, the present disclosure includes methods for manufacturing immunogenic conjugates described herein. In some embodiments, a method of manufacturing an immunogenic conjugate comprises conjugating at least one polymer (e.g., a polysaccharide (e.g., an antigenic polysaccharide)) described herein with at least one polypeptide (e.g., an antigenic polypeptide) described herein. [0221] In some embodiments, the average (e.g., the mean) protein (e.g., antigenic protein) to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic compositions or immunogenic conjugates is approximately 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, 6.5:1, 7:1,7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, or 10:1 (weight/weight [w/w]). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 1:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 2:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 3:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 4:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 5:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 6:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 7:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 8:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is approximately 9:1 (w/w). In some embodiments, the average protein to polymer (e.g., polysaccharide) ratio of a plurality of immunogenic complexes or immunogenic conjugates is AFV70463FF approximately 10:1 (w/w). Immunogenic complexes and vaccines of the present disclosure may comprise mixtures of immunogenic complexes or immunogenic conjugates with different average protein to polymer (e.g., polysaccharide) ratios. In some embodiments, the average ratio of protein to polymer in the plurality of immunogenic complexes is chosen to enhance the polymer immunogenicity potential (carrier or presentation function) and/or to elicit protection against, or to inhibit, a pathogen through a protein-specific immune response. Immunogenic compositions (e.g., vaccines) of the present disclosure may comprise mixtures of immunogenic complexes and/or immunogenic conjugates with different average protein to polymer ratios. [0222] In some embodiments, an immunogenic composition (e.g., a vaccine) comprises a plurality of immunogenic complexes comprising a fusion protein described herein and a capsular polysaccharide. In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, 6.5:1, 7:1,7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, or 10:1 (weight/weight [w/w]). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 1:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 2:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 3:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 4:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 5:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 6:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 7:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 8:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 9:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic complexes is approximately 10:1 (w/w). In some embodiments, the average ratio of fusion protein to capsular polysaccharide in the plurality of immunogenic AFV70463FF complexes is chosen to enhance the polysaccharide immunogenicity potential (carrier or presentation function) and/or to elicit protection against, or to inhibit, pneumococcal colonization by any pneumococcus (independent of polysaccharide serotype) through a protein-specific immune response. Immunogenic compositions (e.g., vaccines) of the present disclosure may comprise mixtures of immunogenic complexes with different average fusion protein to capsular polysaccharide ratios. Characterization of Immunogenic Compositions [0223] In some embodiments, the present disclosure provides a method of characterizing an immunogenic composition comprising a purified capsular polysaccharide having repeating units of formula I HO HO O wherein each the method comprises steps of: characterizing a number of units of formula I in the purified capsular polysaccharide where R is –C(O)-CH₃; and comparing the number of units of formula I where R is –C(O)-CH₃ to a reference standard. [0224] Immunogenic compositions described herein can be characterized according to methods known to those of skill in the art. For example, in some embodiments, an immunogenic composition is characterized by determining a structure of a purified capsular polysaccharide (e.g., a purified capsular polysaccharide of formula I). In some embodiments, an immunogenic AFV70463FF composition is characterized by assessment of a weight average molecular weight of a purified capsular polysaccharide in the immunogenic composition. In some embodiment, an immunogenic composition is characterized by assessment of a percentage of O-acetylated groups (i.e., when R is –C(O)-CH₃) in a purified capsular polysaccharide of formula I in the immunogenic composition. In some embodiments, a purified capsular polysaccharide is characterized by nuclear magnetic resonance (NMR). In some embodiments, an immunogenic composition is characterized by comparison to a reference standard, wherein a reference standard is a composition comprising a known form and quantity of a purified capsular polysaccharide (e.g., a purified capsular polysaccharide of formula I). [0225] In some embodiments, the present disclosure provides a method of characterizing an immunogenic composition comprising a purified capsular polysaccharide having repeating units of formula I HO HO wherein each R method comprises steps of: quantifying a percentage of units of the purified capsular polysaccharide that are of formula I; and comparing the percentage of units of formula I in the purified capsular polysaccharide to a reference standard. [0226] In some embodiments, the present disclosure provides a method of selecting a level of acetylation of a purified capsular polysaccharide for an immunogenic composition, the method AFV70463FF comprising steps of: administering to each of a plurality of animals one of a plurality of compositions comprising a purified capsular polysaccharide having repeating units of formula I, HO HO O wherein each R is independently selected from H and –C(O)-CH₃, wherein in each of the plurality of compositions, R is –C(O)-CH₃ in a distinct percentage of the total instances of units of formula I; determining the level of an immune response to the purified capsular polysaccharide; comparing the level of immune response to a reference level; and selecting a purified capsular polysaccharide with a given acetylation level for inclusion in the immunogenic composition, if the determined immune response is higher than the reference level. Immunogenic Compositions (e.g., Vaccines) [0227] Another aspect of the disclosure provides immunogenic compositions that include one or more immunogenic agents described herein. For example, an immunogenic composition, e.g., vaccine composition, can include one or more immunogenic agents described herein. [0228] In some embodiments, the present disclosure provides an immunogenic composition comprising one or more antigenic polysaccharides described herein. In some embodiments, such an immunogenic composition comprises: (i) a purified capsular polysaccharide (e.g., a purified capsular polysaccharide of serotype 38) described herein, and (ii) one or more additional antigenic polysaccharides (e.g., capsular polysaccharides) described herein. In some embodiment, such an immunogenic composition does not comprise a polypeptide (e.g., an antigenic polypeptide and/or AFV70463FF carrier protein described herein). In some embodiment, the one or more antigenic polysaccharide of such an immunogenic composition are not associated with (e.g., covalently or non-covalently) a polypeptide (e.g., an antigenic polypeptide and/or carrier protein described herein). In some embodiments, such an immunogenic composition is a pneumococcal polysaccharide vaccine (PPV). [0229] In some embodiments, the present disclosure provides an immunogenic composition comprising (i) a purified capsular polysaccharide and (ii) a polypeptide, wherein the purified capsular polysaccharide comprises repeating units of formula I: HO HO O wherein each R is independently selected from H and –C(O)-CH₃. In some embodiments, R is H. In some embodiments, R is –C(O)-CH₃. [0230] In some composition comprises: (i) a purified capsular polysaccharide of formula I and (ii) one or more polypeptides described herein (e.g., an antigenic polypeptide and/or carrier protein described herein). In some embodiments, a purified capsular polysaccharide of formula I is associated with (e.g., covalently or non-covalently) a polypeptide (e.g., an antigenic polypeptide and/or carrier protein described herein). In some embodiments, the polypeptide is a carrier protein, for example selected from the group consisting of: CRM197, tetanus toxoid, exoprotein A, PspA, diphtheria toxoid, and protein D. In some embodiments, the carrier protein is CRM197. In some embodiments, the polypeptide is or comprises a pneumococcal AFV70463FF polypeptide antigen, which may be selected from: a pneumolysin polypeptide antigen, an SP0435 polypeptide antigen, an SP0785 polypeptide antigen, and an SP1500 polypeptide antigen. [0231] In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is from about 10:1 to about 1:10. In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is from about 20:1 to about 1:2. In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is from about 5:1 to about 1:5. In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is from about 3:1 to about 1:3. In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is from about 5:1 to about 1:1. In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10. In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is about 3:1, 2:1, 1:1, 1:2, or 1:3. In some embodiments, a weight ratio of polypeptide to purified capsular polysaccharide in an immunogenic composition described herein is about 3:1. [0232] In some embodiments, an immunogenic composition can include a population of one type of immunogenic complex or immunogenic conjugate, where all of the immunogenic complexes or immunogenic conjugates include the same polypeptide (e.g., antigenic polypeptide) and a purified capsular polysaccharide of formula I. In some embodiments, such compositions can include a plurality of one type of immunogenic agent described herein. For example, an immunogenic composition can include a population of one type of immunogenic agent, where all of the immunogenic compositions include the same polypeptide (e.g., antigenic polypeptide) and the same polymer (e.g., antigenic polysaccharide). or alternatively, such compositions can include a plurality of more than one type of immunogenic complex or immunogenic conjugate described herein. For example, an immunogenic composition can include populations of different types of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition can include a population of a first type of immunogenic complex or immunogenic conjugate and a population of a second type of immunogenic complex or immunogenic conjugate, where the first type and the second type of the AFV70463FF immunogenic complex or immunogenic conjugate have different polypeptide (e.g., antigenic polypeptides) and/or different polymers (e.g., antigenic polysaccharides). In some embodiments, an immunogenic composition can include a population of a first type of immunogenic complex or immunogenic conjugate and a population of a second type of immunogenic complex or immunogenic conjugate, where the first type and the second type of the immunogenic complex or immunogenic conjugate include the same polypeptide (e.g., antigenic polypeptide) and different polymers (e.g., antigenic polysaccharides (e.g., polysaccharides of different serotypes)). [0233] In some embodiments, an immunogenic composition is a monovalent immunogenic composition. In some embodiments, an immunogenic composition is a polyvalent or multivalent immunogenic composition. In some embodiments, an immunogenic composition is a monovariant immunogenic composition, comprising one or more antigens from one strain or variant of a pathogen. In some embodiments, an immunogenic composition is a multivariant immunogenic composition, comprising one or more antigens from more than one strain or variant of a pathogen. In some embodiments, the valency of an immunogenic composition refers to the number of species of immunogenic complexes or immunogenic conjugates present in the immunogenic composition. The valency of an immunogenic composition described herein is not limiting with respect to the total antigens present in said immunogenic composition, or to the number of pathogen strains for which administration of said immunogenic composition may induce an immune-protective response. [0234] In some embodiments, an immunogenic composition comprises between 1-60 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-50 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-45 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-40 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-35 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-34 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-33 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-32, 1-31, or 1-30 species of immunogenic complexes or AFV70463FF immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-30 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-30 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-24 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-15 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-9 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises between 1-5 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition is a polyvalent composition. [0235] In some embodiments, an immunogenic composition comprises at least 10, at least 15, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, or at least 34 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, or at least 34 species of immunogenic complexes or immunogenic conjugates. In some embodiments, an immunogenic composition comprises at least 30 (including, e.g., at least 31, at least 32, at least 33, at least 34) species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises 30- 40 species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises 30-35 species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises 25-50 or 25-40 species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises no more than 60 species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises no more than 50 species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises no more than 45 species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises no more than 40 species of immunogenic complexes or immunogenic conjugates AFV70463FF described herein. In some embodiments, an immunogenic composition comprises no more than 35 species of immunogenic complexes or immunogenic conjugates described herein. [0236] In some embodiments, an immunogenic composition comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 species of immunogenic complexes or immunogenic conjugates described herein. In some embodiments, an immunogenic composition comprises an immunogenic complex or immunogenic conjugate comprising a purified capsular polysaccharide of serotype 38 described herein (e.g. of formula I). In some embodiments, an immunogenic composition comprises an immunogenic complex or immunogenic conjugate comprising a purified capsular polysaccharide of serotype 38 described herein (e.g. of formula I) and further comprises immunogenic complexes or immunogenic conjugates comprising one or more polysaccharides of Streptococcus pneumoniae each having a serotype selected from: 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F and 35B. In some embodiments, an immunogenic composition comprises an immunogenic complex or immunogenic conjugate comprising a purified capsular polysaccharide of serotype 38 described herein (e.g. formula I) and further comprises immunogenic complexes or immunogenic conjugates comprising polysaccharides of each of Streptococcus pneumoniae serotypes: 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F, and 35B. [0237] In some embodiments, an immunogenic composition comprises two or more species of immunogenic complexes or immunogenic conjugates in amounts such that the weight of polymers (e.g., polysaccharides) in the composition from each immunogenic complex or immunogenic conjugate is about the same, e.g., present in a w/w ratio of about 1:1. In some embodiments, the weight of polymers (e.g., polysaccharides) in the composition contributed by each immunogenic complex or immunogenic conjugate nts, the weight of polymers (e.g., polysaccharides) in the composition contributed by each immunogenic complex or immunogenic conjugate is more than AFV70463FF e.g. [0238] In some embodiments, an immunogenic composition comprises two or more species of immunogenic complexes or immunogenic conjugates in amounts such that the weight of polymers (e.g., polysaccharides) in the immunogenic composition contributed by each immunogenic complex or immunogenic conjugate is different, e.g., present in a w/w ratio that is not about 1:1. In some embodiments, an immunogenic composition comprises two or more species of immunogenic complexes or immunogenic conjugate in amounts such that the weight of polymers (e.g., polysaccharides) in the immunogenic composition contributed by a first immunogenic complex or immunogenic conjugate and a second immunogenic complex or immunogenic conjugate is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10. In some embodiments, an immunogenic composition comprises a mixture of immunogenic complexes or immunogenic conjugates, such that the weight of polymers (e.g., polysaccharides) in the immunogenic composition contributed by an immunogenic immunogenic conjugate In some embodiments, an immunogenic composition comprises a mixture of immunogenic complexes or immunogenic conjugates, such that the weight of polymers (e.g., polysaccharides) in the immunogenic composition contributed by an immunogenic complex or immunogenic conjugate an immunogenic composition comprises a mixture of immunogenic complexes or immunogenic conjugates, such that the weight of polymers (e.g., polysaccharides) in the immunogenic composition contributed by each immunogenic complex or immunogenic conjugate an immunogenic composition comprises a mixture of immunogenic complexes or immunogenic conjugates, such that the weight of polymers (e.g., polysaccharides) in the immunogenic composition contributed by each immunogenic complex or immunogenic conjugate [0239] In some embodiments, an immunogenic composition comprises two or more species of immunogenic complexes or immunogenic conjugates in amounts such that the combined weight of polymers (e.g., polysaccharides) and polypeptides in the immunogenic composition contributed by each immunogenic complex or immunogenic conjugate is about the same, e.g., present in a w/w protein:polysaccharide ratio of about 1:1. In some embodiments, an immunogenic composition comprises two or more species of immunogenic complexes or immunogenic conjugates in amounts AFV70463FF such that the combined weight of polymers (e.g., polysaccharides) and polypeptides in the immunogenic composition contributed by each immunogenic complex or immunogenic conjugate is present in a w/w protein:polysaccharide ratio of about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, or about 10:1. [0240] In some embodiments, the combined weight of polymers (e.g., polysaccharides) and polypeptides in an immunogenic composition contributed by each immunogenic complex or immunogenic conjugate [0241] In some embodiments, an immunogenic composition comprises two or more species of immunogenic complexes or immunogenic conjugates in amounts such that the combined weight of polymers (e.g., polysaccharides) and polypeptides in the immunogenic composition contributed by each immunogenic complex or immunogenic conjugate is different, e.g., present in a w/w protein:polysaccharide ratio that is not about 1:1, e.g., a protein:polysaccharide ratio that is 2:1, 3:1, 4:1. 5:1. 6:1, 7:1, 8:1, 9:1, or 10:1. In some embodiments, an immunogenic composition comprises a mixture of immunogenic complexes or immunogenic conjugates, such that the combined weight of polymers (e.g., polysaccharides) and polypeptides in the immunogenic composition contributed by each immunogenic complex or immunogenic conjugate ranges from [0242] In some embodiments, an immunogenic composition described herein comprises an adjuvant. [0243] In some embodiments, an immunogenic composition described herein is a vaccine composition. [0244] In some embodiments, an immunogenic composition is an immunogenic composition described in WO 2020/056202, the contents of which are hereby incorporated by reference in their entirety for purposes described herein. In some embodiments, an immunogenic composition is an immunogenic composition described in International application PCT/US2022/043156, the contents of which are hereby incorporated by reference in their entirety for purposes described herein. AFV70463FF Pharmaceutical Compositions [0245] In some embodiments, a composition (e.g., an immunogenic composition) described herein is formulated into a pharmaceutical composition. In some embodiments, a pharmaceutical composition comprises a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition comprises an adjuvant. [0246] Optimal amounts of components for a particular pharmaceutical composition can be ascertained by standard studies involving observation of appropriate immune responses in subjects. Following an initial immunization, subjects can receive one or several booster immunizations adequately spaced in time. [0247] The immunogenic agents described herein, and/or preparations thereof, may be formulated in a unit dosage form for ease of administration and uniformity of dosage. The specific therapeutically effective dose level for any particular subject or organism may depend upon a variety of factors including the severity or degree of risk of infection; the activity of the specific pharmaceutical composition employed; other characteristics of the specific pharmaceutical composition employed; the age, body weight, general health, sex of the subject, diet of the subject, pharmacokinetic condition of the subject, the time of administration (e.g., with regard to other activities of the subject such as eating, sleeping, receiving other medicines including other pharmaceutical composition doses, etc.), route of administration, rate of excretion of the specific pharmaceutical composition employed; pharmaceutical compositions used in combination or coincidental with the pharmaceutical composition employed; and like factors well known in the medical arts. [0248] Immunogenic agents for use in accordance with the present disclosure may be formulated into compositions (e.g., pharmaceutical compositions) according to known techniques. In some embodiments where a pharmaceutical composition comprises a vaccine composition, vaccine preparation is generally described in Vaccine Design (Powell and Newman, 1995). For example, an immunologically effective amount of a vaccine composition can be formulated together with one or more organic or inorganic, liquid or solid, pharmaceutically suitable carrier materials. [0249] In general, pharmaceutically acceptable carrier(s) include solvents, dispersion media, and the like, which are compatible with pharmaceutical administration. For example, materials AFV70463FF that can serve as pharmaceutically acceptable carriers include, but are not limited to sugars such as lactose, glucose, dextrose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; polyols such as glycerol, propylene glycol, and liquid polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as preservatives, and antioxidants can also be present in the composition, according to the judgment of the formulator (Martin, 1975). [0250] Pharmaceutical compositions may be formulated by combining one or more of the immunogenic agents disclosed herein with carriers and/or other optional components by any available means including, for example, conventional mixing, granulating, dissolving, lyophilizing, or similar processes. [0251] Pharmaceutical compositions useful in the provided methods may be lyophilized up until they are about to be used, at which point they are extemporaneously reconstituted with diluent. In some embodiments, pharmaceutical compositions or components thereof are lyophilized in the presence of one or more other components (e.g., adjuvants), and are extemporaneously reconstituted with saline solution. Alternatively, individual components, or sets of components may be separately lyophilized and/or stored (e.g., in a vaccination kit), the components being reconstituted and either mixed prior to use or administered separately to the subject. [0252] Lyophilization can produce a more stable composition (e.g., by preventing or reducing breakdown of polysaccharide antigens). Lyophilizing of pharmaceutical compositions or components thereof is well known in the art. Typically, a liquid pharmaceutical composition or component thereof is freeze dried, often in the presence of an anti-caking agent (such as, for example, sugars such as sucrose or lactose). In some embodiments, the anti-caking agent is present, for example, at an initial concentration of 10-200 mg/ml. Lyophilization typically occurs over a series of steps, for instance a cycle starting at -69° C, gradually adjusting to -24°C over 3 h, then retaining this temperature for 18 h, then gradually adjusting to -16°C over 1 h, then retaining this AFV70463FF temperature for 6 h, then gradually adjusting to +34°C over 3 h, and finally retaining this temperature over 9 h. [0253] In some embodiments, a pharmaceutical composition is a liquid. In some embodiments, the liquid is a reconstituted lyophylate. In some embodiments, a pharmaceutical composition has a pH of 5, about 6, about 7, or about 8. In some embodiments, a pharmaceutical composition has a pH between about 5 and about 7.5. In some embodiments, a pharmaceutical composition has a pH between 5 and 7.5. In some embodiments, a pharmaceutical composition has a pH between about 5.3 and about 6.3. In some embodiments, a pharmaceutical composition has a pH between 5.3 and 6.3. In some embodiments, a pharmaceutical composition has a pH of about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, or about 7.5. [0254] Pharmaceutical compositions or components thereof for use in accordance with the present disclosure may be incorporated into liposomes, cochleates, biodegradable polymers such as poly-lactide, poly-glycolide and poly-lactide-co-glycolides, or immune-stimulating complexes (ISCOMs). [0255] In certain situations, it may be desirable to prolong the effect of a pharmaceutical composition for use in accordance with the present disclosure, for example by slowing the absorption of one or more pharmaceutical composition components. Such delay of absorption may be accomplished, for example, by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the product then depends upon its rate of dissolution, which in turn, may depend upon size and form. Alternatively, or additionally, delayed absorption may be accomplished by dissolving or suspending one or more pharmaceutical composition components in an oil vehicle. Injectable depot forms can also be employed to delay absorption. Such depot forms can be prepared by forming microcapsule matrices of one or more pharmaceutical composition components a biodegradable polymers network. Depending upon the ratio of polymer to pharmaceutical composition component, and the nature of the particular polymer(s) employed, the rate of release can be controlled. [0256] Examples of biodegradable polymers that can be employed in accordance with the present disclosure include, for example, poly(orthoesters) and poly(anhydrides). One particular exemplary polymer is polylactide-polyglycolide. AFV70463FF [0257] Depot injectable formulations may also be prepared by entrapping the product in liposomes or microemulsions, which are compatible with body tissues. [0258] Polymeric delivery systems can also be employed in non-depot formulations including, for example, oral formulations. For example, biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid, etc., can be used in oral formulations. Polysaccharide antigens or conjugates may be formulated with such polymers, for example to prepare particles, microparticles, extrudates, solid dispersions, admixtures, or other combinations in order to facilitate preparation of useful formulations (e.g., oral). [0259] Pharmaceutical compositions for use in accordance with the present disclosure include immunogenic compositions, and may additionally include one or more additional active agents (i.e., agents that exert a biological effect – not inert ingredients). For example, it is common in pharmaceutical composition preparation to include one or more adjuvants. It will be appreciated that such additional agents may be formulated together with one or more other pharmaceutical composition components, or may be maintained separately and combined at or near the time of administration. In some embodiments, such additional components may be administered separately from some or all of the other pharmaceutical composition components, within an appropriate time window for the relevant effect to be achieved. Adjuvants [0260] The immunogenic compositions (e.g., vaccine compositions) and pharmaceutical compositions described herein may include an adjuvant. Adjuvants, generally, are agents that enhance the immune response to an antigen. Adjuvants can be broadly separated into two classes, based on their principal mechanisms of action: vaccine delivery systems and immunostimulatory adjuvants (see, e.g., Singh et al., 2003). In most immunogenic composition or pharmaceutical composition formulations, the adjuvant provides a signal to the immune system so that it generates a response to the antigen, and the antigen is required for driving the specificity of the response to the pathogen. Vaccine delivery systems are often particulate formulations, e.g., emulsions, microparticles, immune-stimulating complexes (ISCOMs), nanoparticles, which may be, for example, particles and/or matrices, and liposomes. In contrast, immunostimulatory adjuvants are sometimes from or derived from pathogens and can represent pathogen associated molecular AFV70463FF patterns (PAMP), e.g., lipopolysaccharides (LPS), monophosphoryl lipid A (MPL), or CpG- containing DNA, which activate cells of the innate immune system. [0261] Alternatively, adjuvants may be classified as organic and inorganic. Inorganic adjuvants include aluminum-based adjuvants (e.g., alum salts) such as aluminum phosphate, amorphous aluminum hydroxyphosphate sulfate, and aluminum hydroxide, which are commonly used in human immunogenic compositions and pharmaceutical compositions. Organic adjuvants comprise organic molecules including macromolecules. Non-limiting examples of organic adjuvants include cholera toxin/toxoids, other enterotoxins/toxoids or labile toxins/toxoids of Gram-negative bacteria, interleukins (e.g., IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12, IL-15, IL-18, etc.), interferons (e.g., gamma interferon), granulocyte macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), and tumor necrosis factor (TNF). [0262] Adjuvants may also be classified by the response they induce. In some embodiments, an adjuvant induces generation, proliferation, or activation of T_H1 cells or T_H2 cells. In other embodiments, an adjuvant induces generation, proliferation, or activation of B cells. In yet other embodiments, an adjuvant induces activation of antigen-presenting cells. These categories are not mutually exclusive; in some cases, an adjuvant activates more than one type of cell. [0263] In certain embodiments, the adjuvant induces the generation, proliferation, or activation of T_H17 cells. The adjuvant may promote the CD4+ or CD8+ T cells to secrete IL-17. In some embodiments, an adjuvant that induces the generation, proliferation, or activation of T_H17 cells is one that produces at least a 2-fold, and in some cases a 10-fold, experimental sample to control ratio in the following assay. In the assay, an experimenter compares the IL-17 levels secreted by two populations of cells: (1) cells from animals immunized with the adjuvant and a polypeptide known to induce T_H17 generation, proliferation, or activation, and (2) cells from animals treated with the adjuvant and an irrelevant (control) polypeptide. An adjuvant that induces the generation, proliferation, or activation of T_H17 cells may cause the cells of population (1) to produce more than 2-fold, or more than 10-fold more IL-17 than the cells of population (2). IL-17 may be measured, for example, by ELISA or ELISPOT. Certain toxins, such as cholera toxin and labile toxin (produced by enterotoxigenic E. coli, or ETEC), activate a T_H17 response. Thus, in some embodiments, the adjuvant is a toxin or toxoid. Mutant derivates of labile toxin (toxoids) that are active as adjuvants but significantly less toxic can be used as well. Exemplary detoxified mutant derivatives of labile toxin include mutants lacking ADP-ribosyltransferase activity. AFV70463FF Particular detoxified mutant derivatives of labile toxin include LTK7 (Douce et al., 1995) and LTK63 (Williams et al., 2004), LT-G192 (Douce et al., 1999), and LTR72 (Giuliani et al., 1998). [0264] In some embodiments, the adjuvant comprises a VLP (virus-like particle). One such adjuvant platform, Alphavirus replicons, induces the activation of T_H17 cells using alphavirus and is produced by Alphavax. In certain embodiments of the Alphavirus replicon system, alphavirus may be engineered to express an antigen of interest, a cytokine of interest (for example, IL-17 or a cytokine that stimulates IL-17 production), or both, and may be produced in a helper cell line. More detailed information may be found in U.S. Patent Nos. 5,643,576 and 6,783,939. In some embodiments, an immunogenic composition (e.g., vaccine) formulation is administered to a subject in combination with a nucleic acid encoding a cytokine. [0265] Certain classes of adjuvants activate toll-like receptors (TLRs) in order to activate a T_H17 response. TLRs are well known proteins that may be found on leukocyte membranes, and recognize foreign antigens (including microbial antigens). Administering a known TLR ligand together with an antigen of interest (for instance, as a fusion protein) can promote the development of an immune response specific to the antigen of interest. One exemplary adjuvant that activates TLRs comprises Monophosphoryl Lipid A (MPL). Traditionally, MPL has been produced as a detoxified lipopolysaccharide (LPS) endotoxin obtained from Gram-negative bacteria, such as S. minnesota. In particular, sequential acid and base hydrolysis of LPS produces an immunoactive lipid A fraction (which is MPL), and lacks the saccharide groups and all but one of the phosphates present in LPS. A number of synthetic TLR agonists (in particular, TLR-4 agonists) are disclosed in Evans et al., 2003. Like MPL adjuvants, these synthetic compounds activate the innate immune system via TLR. Another type of TLR agonist is a synthetic phospholipid dimer, for example E6020 (Ishizaka et al., 2007). Various TLR agonists (including TLR-4 agonists) have been produced and/or sold by, for example, the Infectious Disease Research Institute (IRDI), Corixa, Esai, Avanti Polar Lipids, Inc., and Sigma Aldrich. Another exemplary adjuvant that activates TLRs comprises a mixture of MPL, Trehalose Dicoynomycolate (TDM), and dioctadecyldimethylammonium bromide (DDA). Another TLR-activating adjuvant is R848 (resiquimod). [0266] In some embodiments, the adjuvant is or comprises a saponin. Typically, the saponin is a triterpene glycoside, such as those isolated from the bark of the Quillaja saponaria tree. A saponin extract from a biological source can be further fractionated (e.g., by chromatography) to AFV70463FF isolate the portions of the extract with the best adjuvant activity and with acceptable toxicity. Typical fractions of extract from Quillaja saponaria tree used as adjuvants are known as fractions A and C. [0267] In certain embodiments, combinations of adjuvants are used. Three exemplary combinations of adjuvants are MPL and alum, E6020 and alum, and MPL and an ISCOM. [0268] Adjuvants may be covalently or non-covalently bound to antigens. In some embodiments, the adjuvant may comprise a protein which induces inflammatory responses through activation of antigen-presenting cells (APCs). In some embodiments, one or more of these proteins can be recombinantly fused with an antigen of choice, such that the resultant fusion molecule promotes dendritic cell maturation, activates dendritic cells to produce cytokines and chemokines, and ultimately, enhances presentation of the antigen to T cells and initiation of T cell responses (e.g., see Wu et al., 2012). [0269] In some embodiments, immunogenic agents described herein are formulated and/or administered in combination with an adjuvant. In some embodiments, such an adjuvant is or comprises an aluminum-based adjuvant (e.g., an aluminum salt); see e.g., Danielsson and Eriksson, “Aluminum adjuvants in vaccines – a way to modulate the immune response”, Semin Cell Dev Biol, 115:3-9 (2021) and HogenEsch et al., “Optimizing the utilization of aluminum adjuvants in vaccines: you might just get what you want”, NPJ Vaccines, 3:51 (2018), the contents of which are incorporated herein by reference in their entirety for purposes described herein. Examples of an aluminum-based adjuvant include, but are not limited to, aluminum phosphate, aluminum hydroxide, potassium aluminum sulfate (alum), aluminum hydroxide phosphate, aluminum hydroxyphosphate sulfate, and combinations thereof. In some embodiments, an aluminum-based adjuvant comprises aluminum phosphate. In some embodiments, an aluminum-based adjuvant is aluminum phosphate. [0270] In some embodiments, the same adjuvant or mixture of adjuvants is present in each dose of an immunogenic composition or pharmaceutical composition. Optionally, however, an adjuvant may be administered with the first dose of an immunogenic composition or pharmaceutical composition and not with subsequent doses (i.e., booster shots). Alternatively, a strong adjuvant may be administered with the first dose of an immunogenic composition or pharmaceutical composition and a weaker adjuvant or lower dose of the strong adjuvant may be administered with subsequent doses. The adjuvant can be administered before the administration AFV70463FF of the antigen, concurrent with the administration of the antigen or after the administration of the antigen to a subject (sometimes within 1, 2, 6, or 12 hours, and sometimes within 1, 2, or 5 days). Certain adjuvants are appropriate for human subjects, non-human animals, or both. [0271] Immunogenic compositions and pharmaceutical compositions for use in accordance with the present disclosure may include, or be administered concurrently with, other antimicrobial, or anti-inflammatory therapies. For example, such immunogenic compositions and pharmaceutical compositions may include or be administered with one or more agents that kills or retards growth of a pathogen. Such agents include, for example, remdesivir, lopinavir and/or ritonavir (e.g., Kaletra), oseltamivir (Tamiflu), favipiravir, umifenovir, galidesivir, dexamethasone, colchicine, convalescent plasma, monoclonal antibodies (e.g., one or more of bamlanivimab, LY-CoV016, etesevimab, casirivimab, indevimab, sarilumab, tocilizumab), IL-6 inhibitors, kinase inhibitors, interferons, penicillin, vancomycin, erythromycin, azithromycin, and clarithromycin, cefotaxime, ceftriaxone, levoflaxin, gatifloxacin. [0272] Alternatively or additionally, immunogenic compositions and pharmaceutical compositions for use in accordance with the present disclosure may include, or be administered with, one or more other immunogenic compositions, pharmaceutical compositions, or therapies. Additional Components and Excipients [0273] In addition to the antigens and the adjuvants described above, an immunogenic composition (e.g., a vaccine composition) or pharmaceutical composition may include one or more additional components. [0274] In certain embodiments, an immunogenic composition or pharmaceutical composition may include one or more stabilizers such as sugars (such as sucrose, glucose, or fructose), phosphate (such as sodium phosphate dibasic, potassium phosphate monobasic, dibasic potassium phosphate, or monosodium phosphate), glutamate (such as monosodium L-glutamate), gelatin (such as processed gelatin, hydrolyzed gelatin, or porcine gelatin), amino acids (such as arginine, asparagine, histidine, L-histidine, alanine, valine, leucine, isoleucine, serine, threonine, lysine, phenylalanine, tyrosine, and the alkyl esters thereof), inosine, or sodium borate. [0275] In certain embodiments, an immunogenic composition or pharmaceutical composition includes one or more buffers such as histidine, glycine, succinate, Tris, or a mixture AFV70463FF of sodium bicarbonate and ascorbic acid. In some embodiments, the vaccine formulation may be administered in saline, such as phosphate buffered saline (PBS), or distilled water. [0276] In certain embodiments, an immunogenic composition or pharmaceutical composition includes one or more surfactants, for example, but not limited to, polysorbate 80 (TWEEN 80), polysorbate 20 (TWEEN 20), Polyethylene glycol p-(1,1,3,3-tetramethylbutyl)- phenyl ether (TRITON X-100), and 4-(1,1,3,3-Tetramethylbutyl)phenol polymer with formaldehyde and oxirane (TYLOXAPOL). A surfactant can be ionic or non-ionic. [0277] In certain embodiments, an immunogenic composition or pharmaceutical composition includes one or more salts such as sodium chloride, ammonium chloride, calcium chloride, or potassium chloride. [0278] In certain embodiments, a preservative is included in the immunogenic composition or pharmaceutical composition. In other embodiments, no preservative is used. A preservative is most often used in multi-dose immunogenic composition (e.g., vaccine) or pharmaceutical composition vials, and is less often needed in single-dose immunogenic composition (e.g., vaccine) or pharmaceutical composition vials. In certain embodiments, the preservative is 2-phenoxyethanol, thimerosal, methyl and propyl parabens, benzyl alcohol, and/or sorbic acid. Uses of Immunogenic Compositions and Pharmaceutical Compositions [0279] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein that includes one or more antigenic polysaccharides is characterized in that one or more of the opsonization potential, or immune response to one or more antigenic polysaccharides is increased relative to a predetermined level, as measured by ELISA and or by a functional antibody assay. In some embodiments, one or more of the opsonization potential, immune response to the one or more antigenic polysaccharides is increased at least 1-fold, 2-fold, 3-fold, 4-fold, or 5-fold relative to a predetermined level, as measured by ELISA and or by a functional antibody assay. In some embodiments, the predetermined level is a pre-immune level. In some embodiments, one or more polypeptides (e.g., antigenic polypeptides) are carrier proteins for one or more antigenic polysaccharides. [0280] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response AFV70463FF against one or more pathogens in the subject at a level greater than a composition comprising an antigenic polysaccharide alone. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or pathogens in the subject at a level greater than a composition comprising an antigenic polypeptide alone. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a protective immune response. [0281] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more strains (or variants, or serotypes) of a pathogen. In some embodiments, a pathogen is a bacterial pathogen. In some embodiments, a bacterial pathogen is S. pneumoniae. [0282] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against S. pneumoniae. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more serotypes of S. pneumoniae. In some embodiments, such an immune response may be directed against one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein includes polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). In some embodiments, such an immune response may be directed against one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein does not include polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). In some embodiments, such an immune response may be directed against two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein (i) includes polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s); and (ii) does not include polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a AFV70463FF protective immune response against one or more serotypes of S. pneumoniae. In some embodiments, such a protective response may be directed against one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein includes polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). In some embodiments, such a protective response may be directed against one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein does not include polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). In some embodiments, such a protective response may be directed against two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein (i) includes polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s); and (ii) does not include polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). [0283] In some embodiments, the immune response is an antibody or B cell response. In some embodiments, the antibody or B cell response is a memory B cell response. In some embodiments, the immune response is a T cell response. In some embodiments, the T cell response is a memory T cell response. In some embodiments, the immune response is an innate immune response. In some embodiments, the immune response is a CD4+ T cell response, including T_H1, T_H2, or T_H17 response, or a CD8+ T cell response, or a CD4+ and CD8+ T cell response, or a CD4-/CD8- T cell response. In some embodiments, the immune response is an antibody or B cell response, and a T cell response. In some embodiments, the immune response is an antibody or B cell response, a T cell response, and an innate immune response. In some embodiments, the immune response is a protective immune response. In some embodiments, the immune response comprises neutralizing antibodies. In some embodiments, the immune response is a memory response. [0284] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces antibody production against one or more pathogens in the subject at a level greater than a composition comprising an antigenic polysaccharide alone. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, AFV70463FF induces antibody production against one or more pathogens in the subject at level greater than a composition comprising a polypeptide antigen alone. [0285] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more pathogens in the subject at a level greater than a composition comprising an antigenic polysaccharide alone. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more pathogens in the subject at a level greater than a composition comprising a polypeptide antigen alone. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a protective immune response. [0286] Compositions described herein may be used for prophylactic and/or therapeutic treatment of pathogen infection and/or disease. Accordingly, the present disclosure provides a method for immunizing a subject suffering from or susceptible to pathogen infection, comprising administering an immunologically effective amount of any of the compositions described herein. The subject receiving the immunization may be a male or a female, and may be an infant, child, adolescent, or adult. In some embodiments, the subject being treated is a human. In other embodiments, the subject is a non-human animal. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a protective immune response against pathogen infection and/or disease. [0287] In prophylactic embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein is administered to a subject to induce an immune response that can help protect against the establishment of one or more strains (variants) of a pathogen, for example by protecting against asymptomatic infection. In some aspects, the method inhibits infection by a pathogen in an uninfected subject. In another aspect, the method may reduce transmission, replication, and/or pathogen load of one or more strains (variants) of a pathogen in a subject who is already infected. [0288] In prophylactic embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein is administered to a subject to induce an immune response that can help protect against the establishment of S. pneumoniae, for example by protecting against colonization, the first and necessary step in disease. In some embodiments, such AFV70463FF an immune response may be directed against one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein includes polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). In some embodiments, such an immune response may be directed against one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein does not include polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s) (non-vaccine types, NVTs). In some embodiments, such an immune response may be directed against two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) serotypes of Streptococcus pneumoniae, wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein (i) includes polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s); and (ii) does not include polysaccharide(s) present in at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of such serotype(s). Thus, in some aspects, the method inhibits infection by S. pneumoniae in a non-colonized or uninfected subject. In another aspect, the method may reduce the duration of colonization in a subject who is already colonized. [0289] In therapeutic embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition may be administered to a subject suffering from a pathogen infection, in an amount sufficient to treat the subject. Treating the subject, in this case, refers to reducing pathogen symptoms and/or pathogen load and/or sequelae in the subject. In some embodiments, treating the subject refers to reducing the duration of symptoms or sequelae, or reducing the intensity of symptoms or sequelae. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces transmissibility of one or more strains (variants) of a pathogen from the immunized subject to another subject. In certain embodiments, the reductions described above are at least 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%, e.g., relative to a control, e.g., a control subject. [0290] In therapeutic embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein is administered to a subject post-pathogen infection. Compositions described herein may be administered shortly after pathogen infection, e.g. before symptoms or sequelae manifest, or may be administered during or after manifestation of symptoms or sequelae. AFV70463FF [0291] In some embodiments, compositions described herein confer protective immunity, allowing an immunized subject to exhibit delayed onset of symptoms or sequelae, or reduced severity of symptoms or sequelae, as the result exposure to an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein. In certain embodiments, the reduction in severity of symptoms or sequelae is at least 25%, 40%, 50%, 60%, 70%, 80%, or 90%, e.g., relative to a control. In particular embodiments, immunized subjects may display no symptoms or sequelae upon development of an infection with a pathogen (asymptomatic infection), or do not become infected by a pathogen. Protective immunity is typically achieved by one or more of the following mechanisms: mucosal, humoral, or cellular immunity. Mucosal immunity is primarily the result of secretory IgA (sIGA) antibodies on mucosal surfaces of the respiratory, gastrointestinal, and genitourinary tracts. The sIGA antibodies are generated after a series of events mediated by antigen-processing cells, B and T lymphocytes, that result in sIGA production by B lymphocytes on mucosa-lined tissues of the body. Humoral immunity is typically the result of IgG antibodies and IgM antibodies in serum. Cellular immunity can be achieved through cytotoxic T lymphocytes or through delayed-type hypersensitivity that involves macrophages and T lymphocytes, as well as other mechanisms involving T cells without a requirement for antibodies. In particular, cellular immunity may be mediated by T_H1 or T_H17 cells. [0292] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more strains (variants) of a pathogen. [0293] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against S. pneumoniae. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or more) serotypes of S. pneumoniae. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against all serotypes of S. pneumoniae comprised in such compositions. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a protective immune response against AFV70463FF one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or more) serotypes of S. pneumoniae. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a protective immune response against all serotypes of S. pneumoniae comprised in such immunogenic composition (e.g., vaccine). [0294] In some embodiments, the immune response is an antibody or B cell response. In some embodiments, the antibody or B cell response is a memory B cell response. In some embodiments, the immune response is a T cell response. In some embodiments, the T cell response is a memory T cell response. In some embodiments, the immune response is an innate immune response. In some embodiments, the immune response is a CD4+ T cell response, including T_H1, T_H2, or T_H17 response, or a CD8+ T cell response, or a CD4+ and CD8+ T cell response, or CD4-/CD8- T cell response. In some embodiments, the immune response is an antibody or B cell response, and a T cell response. In some embodiments, the immune response is an antibody or B cell response, a T cell response, and an innate immune response. In some embodiments, the immune response is a protective immune response. In some embodiments, the immune response comprises neutralizing antibodies. In some embodiments, the immune response is a memory response. [0295] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an antibody or B cell response against one or more pathogens in the subject at a level greater than an immunogenic composition (e.g., vaccine) or pharmaceutical composition comprising an antigenic polysaccharide alone. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an antibody or B cell response against one or more pathogens in the subject at level greater than an immunogenic composition (e.g., vaccine) or pharmaceutical composition comprising a polypeptide antigen alone. In some embodiments, the immune response is a protective immune response. In some embodiments, the immune response comprises neutralizing antibodies. [0296] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a T cell response against one or more pathogens in the subject at a level greater than a composition comprising an antigenic polysaccharide alone. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a T cell AFV70463FF response against one or more pathogens in the subject at level greater than a composition comprising a polypeptide antigen alone. In some embodiments, the immune response is a protective immune response. In some embodiments, the immune response comprises neutralizing antibodies. [0297] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein treats or prevents infection by one or more strains (variants) of a pathogen. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein treats or prevents disease due to infection by one or more strains (variants) of a pathogen. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein treats or prevents one or more symptoms due to infection by one or more strains (variants) of a pathogen. In some embodiments, the one or more symptoms include pneumonia, organ damage, upper respiratory symptoms, gastro-intestinal symptoms, neurological symptoms, myocarditis, inflammation, fever, chills, fatigue, headache, nausea, muscle or body ache, shortness of breath or difficulty breathing, loss of sense of smell (hyposmia, anosmia), loss of sense of taste (hypogeusia, ageusia), and/or multi-inflammatory syndrome of children or adults (MIS-C, MIS-A). In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein treats or prevents one or more of Invasive Pneumococcal Disease (IPD), bacteremia, sepsis, organ damage, meningitis, pneumonia, otitis media, and sinusitis due to infection by S. pneumoniae. [0298] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits or reduces the rate of occurrence or recurrence of infection by one or more strains (variants) of a pathogen. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits or reduces the rate of occurrence or recurrence of disease due to infection by one or more strains (variants) of a pathogen. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits or reduces the rate of occurrence or recurrence of one or more symptoms due to infection by one or more strains (variants) of a pathogen. In some embodiments, the one or more symptoms include pneumonia, organ damage, AFV70463FF upper respiratory symptoms, gastro-intestinal symptoms, neurological symptoms, myocarditis, inflammation, fever, chills, fatigue, headache, nausea, muscle or body ache, shortness of breath or difficulty breathing, loss of sense of smell (hyposmia, anosmia), loss of sense of taste (hypogeusia, ageusia), and/or multi-inflammatory syndrome of children or adults (MIS-C, MIS-A). In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits or reduces the rate of occurrence of one or more of Invasive Pneumococcal Disease (IPD), bacteremia, sepsis, organ damage, meningitis, pneumonia, otitis media, and sinusitis due to infection by S. pneumoniae. [0299] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces the severity of infection by one or more strains (variants) of a pathogen. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces the severity of disease due to infection by one or more strains (variants) of a pathogen. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces the severity of one or more symptoms due to infection by one or more strains (variants) of a pathogen. In some embodiments, the one or more symptoms include pneumonia, organ damage, upper respiratory symptoms, gastro- intestinal symptoms, neurological symptoms, myocarditis, inflammation, fever, chills, fatigue, headache, nausea, muscle or body ache, shortness of breath or difficulty breathing, loss of sense of smell (hyposmia, anosmia), loss of sense of taste (hypogeusia, ageusia), and/or multi- inflammatory syndrome of children or adults (MIS-C, MIS-A). In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces the severity of one or more of Invasive Pneumococcal Disease (IPD), bacteremia, sepsis, organ damage, meningitis, pneumonia, otitis media, and sinusitis due to infection by S. pneumoniae. [0300] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits transmission of one or more strains (variants) of a pathogen from the subject to another subject. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits asymptomatic infection by one or more strains (variants) of a pathogen in the subject. In some embodiments, upon administration to a subject, an immunogenic AFV70463FF composition (e.g., vaccine) or pharmaceutical composition described herein inhibits replication and/or reduces pathogen load of one or more strains (variants) of a pathogen in the subject. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits colonization by one or more strains (variants) of a pathogen in the subject. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits colonization by one or more strains (variants) of a pathogen of a mucosal surface of the subject. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits colonization by one or more strains (variants) of a pathogen in the nasopharynx of the subject. [0301] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more pathogens in the subject at a level greater than a control composition. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a protective immune response against one or more pathogens in the subject at a level greater than a control composition. In some embodiments, the level greater is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. In some embodiments, the subject is a human. In some embodiments, the human is between about 2 weeks of age and about 6 weeks of age. In some embodiments, the human is between about 6 weeks of age and about 6 years of age. In some embodiments, the human is between about 6 years of age and about 18 years of age. In some embodiments, the human is between about 18 years of age and about 50 years of age. In some embodiments, the human is about 50 years of age and about 75 years of age. In some embodiments, the human is about 75 years of age or older. [0302] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits immunogenicity non- AFV70463FF inferior to (e.g., immunogenicity at least as effective as) that elicited by administration of a control composition. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits immunogenicity non-inferior to (e.g., immunogenicity at least as effective as) that elicited by administration of a control composition against one or more pathogens. In some embodiments, the immunogenicity is determined by measuring the amount of anti-polysaccharide antibody of one or more polysaccharides present in the immunogenic composition (e.g., vaccine) or pharmaceutical composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. In some embodiments, the subject is a human. In some embodiments, the human is between about 2 weeks of age and about 6 weeks of age. In some embodiments, the human is between about 6 weeks of age and about 6 years of age. In some embodiments, the human is between about 6 years of age and about 18 years of age. In some embodiments, the human is between about 18 years of age and about 50 years of age. In some embodiments, the human is about 50 years of age and about 75 years of age. In some embodiments, the human is about 75 years of age or older. [0303] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits immunogenicity superior to that elicited by administration of a control composition. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits immunogenicity superior to that elicited by administration of a control composition against one or more pathogens. In some embodiments, the immunogenicity is determined by measuring the amount of anti-polysaccharide antibody of one or more polysaccharides present in the immunogenic composition (e.g., vaccine) or pharmaceutical composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. In some embodiments, the subject is a human. In some embodiments, the human is between about 2 weeks of age and about 6 weeks of age. In some embodiments, the human is between about 6 weeks of age and about 6 years of age. In some embodiments, the human is between about 6 years of age and about 18 years of age. In some embodiments, the human is between about 18 years of age and about 50 years of age. In some embodiments, the human is about 50 years of age and about 75 years of age. In some embodiments, the human is about 75 years of age or older. [0304] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits a functional antibody titer AFV70463FF non-inferior to (e.g., a functional antibody titer at least as effective as) that elicited by administration of a control composition. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits a functional antibody titer non-inferior to (e.g., a functional antibody titer at least as effective as) that elicited by administration of a control composition against one or more or pathogens. In some embodiments, the functional antibody titer is measured by an opsonophagocytic assay (e.g., ones as described herein). In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits a functional antibody titer non-inferior to (e.g., a functional antibody titer at least as effective as) that elicited by administration of a control composition against one or more pathogens, wherein such a composition does not include polysaccharide(s) present in at least one of the pathogens (non- vaccine types, NVTs). In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. In some embodiments, the subject is a human. In some embodiments, the human is between about 2 weeks of age and about 6 weeks of age. In some embodiments, the human is between about 6 weeks of age and about 6 years of age. In some embodiments, the human is between about 6 years of age and about 18 years of age. In some embodiments, the human is between about 18 years of age and about 50 years of age. In some embodiments, the human is about 50 years of age and about 75 years of age. In some embodiments, the human is about 75 years of age or older. [0305] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits a functional antibody titer superior to that elicited by administration of a control composition. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits a functional antibody titer superior to that elicited by administration of a control composition against one or more pathogens. In some embodiments, the functional antibody titer is measured by an opsonophagocytic assay (e.g., ones as described herein). In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, elicits a functional antibody titer superior to that elicited by administration of a control composition against one or more pathogens, wherein such a composition does not include polysaccharide(s) present in at least one of the pathogens (non-vaccine types, NVTs). In some embodiments, the control composition may be AFV70463FF PCV13, PCV20, or PPSV23. In some embodiments, the subject is a human. In some embodiments, the human is between about 2 weeks of age and about 6 weeks of age. In some embodiments, the human is between about 6 weeks of age and about 6 years of age. In some embodiments, the human is between about 6 years of age and about 18 years of age. In some embodiments, the human is between about 18 years of age and about 50 years of age. In some embodiments, the human is about 50 years of age and about 75 years of age. In some embodiments, the human is about 75 years of age or older. [0306] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an immune response against one or more strains (variants) of a pathogen at a level greater than a control composition. In some embodiments, the immune response is an antibody or B cell response. In some embodiments, the immune response is a T cell response. In some embodiments, the immune response is an innate immune response. In some embodiments, the immune response is a CD4+ T cell response, including T_H1, T_H2, or T_H17 response, or a CD8+ T cell response, or a CD4+ and CD8+ T cell response, or CD4-/CD8- T cell response. In some embodiments, the immune response is an antibody or B cell response, and a T cell response. In some embodiments, the immune response is an antibody or B cell response, a T cell response, and an innate immune response. In some embodiments, the immune response is a protective immune response. In some embodiments, the level greater is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. [0307] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces an antibody or B cell response against one or more pathogens in the subject at a level greater than a control composition. In some embodiments, the level greater is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about AFV70463FF 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. [0308] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a T cell response against one or more pathogens in the subject at a level greater than a control composition. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein, upon administration to a subject, induces a T cell response against one or more pathogens in the subject at level greater than a control composition. In some embodiments, the level greater is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. [0309] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein treats or prevents infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein treats or prevents disease due to infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein treats or prevents one or more symptoms due to infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, the one or more symptoms include pneumonia, organ damage, upper respiratory symptoms, gastro-intestinal symptoms, neurological symptoms, myocarditis, inflammation, fever, chills, fatigue, headache, nausea, muscle or body ache, shortness of breath or difficulty breathing, loss of sense of smell (hyposmia, anosmia), loss of sense of taste (hypogeusia, ageusia), and/or multi-inflammatory syndrome of children or adults (MIS-C, MIS-A). In some embodiments, the level greater is about 1%, about AFV70463FF 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. [0310] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits or reduces the rate of occurrence or recurrence of infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits or reduces the rate of occurrence or recurrence of disease due to infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits or reduces the rate of occurrence or recurrence of one or more symptoms due to infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, the one or more symptoms include pneumonia, organ damage, upper respiratory symptoms, gastro- intestinal symptoms, neurological symptoms, myocarditis, inflammation, fever, chills, fatigue, headache, nausea, muscle or body ache, shortness of breath or difficulty breathing, loss of sense of smell (hyposmia, anosmia), loss of sense of taste (hypogeusia, ageusia), and/or multi- inflammatory syndrome of children or adults (MIS-C, MIS-A). In some embodiments, the level greater is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. [0311] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces the severity of infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control AFV70463FF composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces the severity of disease due to infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein reduces the severity of one or more symptoms due to infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, the one or more symptoms include pneumonia, organ damage, upper respiratory symptoms, gastro- intestinal symptoms, neurological symptoms, myocarditis, inflammation, fever, chills, fatigue, headache, nausea, muscle or body ache, shortness of breath or difficulty breathing, loss of sense of smell (hyposmia, anosmia), loss of sense of taste (hypogeusia, ageusia), and/or multi- inflammatory syndrome of children or adults (MIS-C, MIS-A). In some embodiments, the level greater is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. [0312] In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits transmission of one or more strains (variants) of a pathogen from the subject to another subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits colonization by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits colonization by one or more strains (variants) of a pathogen in the nasopharynx of the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits asymptomatic infection by one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, upon administration to a subject, an AFV70463FF immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein inhibits replication and/or reduces pathogen load of one or more strains (variants) of a pathogen in the subject at a level greater than a control composition. In some embodiments, the level greater is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the control composition. In some embodiments, the control composition may be PCV13, PCV20, or PPSV23. [0313] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein is administered to a subject between about 6 weeks and about 5 years (e.g., prior to the 6^th birthday) for active immunization for the prevention of disease caused by one or more strains (variants) of a pathogen. In some embodiments, such one or more strains (variants) of a pathogen are selected from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F, 35B, and 38. [0314] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein is administered to a subject between about 6 years and about 17 years (e.g., prior to the 18^th birthday) for active immunization for the prevention of disease caused by one or more strains (variants) of a pathogen. In some embodiments, such one or more strains (variants) of a pathogen are selected from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F, 35B, and 38. [0315] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein is administered to a subject 18 years or older for active immunization for the prevention of disease caused by one or more strains (variants) of a pathogen. In some embodiments, such one or more strains (variants) of a pathogen are selected from Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F, 35B, and 38. AFV70463FF Antibody Compositions [0316] Some embodiments provide for an antibody composition comprising antibodies raised in a mammal immunized with an immunogenic agent or immunogenic composition of the present disclosure. In some embodiments, an antibody comprises at least one antibody selected from the group consisting of mAbs and anti-idiotype antibodies. In some embodiments, an antibody composition comprises neutralizing antibodies. In some embodiments, an antibody composition comprises an isolated gamma globulin fraction. In some embodiments, an antibody composition comprises polyclonal antibodies. In some embodiments, the antibody composition is administered to a subject. In some embodiments, the antibody composition administered to a subject confers passive immunization. Methods of Administration [0317] In some embodiments, immunogenic compositions (e.g., vaccines) or pharmaceutical compositions described herein are administered to a subject (e.g. an infant, a toddler, a juvenile, or an older adult) at risk of developing disease due to one or more pathogens. In some embodiments, the subject is a human. In some embodiments, the human is between about 2 weeks of age and about 6 weeks of age. In some embodiments, the human is between about 6 weeks of age and about 6 years of age. In some embodiments, the human is between about 6 years of age and about 18 years of age. In some embodiments, the human is between about 18 years of age and about 50 years of age. In some embodiments, the human is about 50 years of age or older. In some embodiments, immunogenic compositions (e.g., vaccines) or pharmaceutical compositions described herein are administered to a subject at elevated risk of developing a disease due to one or more pathogens, e.g., immunocompromised subjects, subjects having sickle cell disease or other hemoglobinopathies, congenital or acquired asplenia, splenic dysfunction, chronic renal failure or nephrotic syndrome, diseases associated with treatment with immunosuppressive drugs or radiation therapy (including malignant neoplasm, leukemia, lymphomas, Hodgkin's disease, or solid organ transplantation), congenital or acquired immunodeficiency, HIV infection, cerebrospinal fluid leaks, cochlear implant(s), chronic heart disease, chronic lung disease, diabetes mellitus, alcoholism, chronic liver disease, cigarette smoking, asthma, generalized malignancy, multiple myeloma, or solid organ transplantation. It will be appreciated that a subject can be AFV70463FF considered at risk for developing a disease without having been diagnosed with any symptoms of the disease. For example, if the subject is known to have been, or to be intended to be, in situations with relatively high risk of infection, that subject will be considered at risk for developing the disease. [0318] Any effective route of administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein may be utilized such as, for example, oral, nasal, enteral, parenteral, intramuscular or intravenous, subcutaneous, transdermal, intradermal, rectal, vaginal, topical, ocular, pulmonary, or by contact application. In some embodiments, immunogenic compositions (e.g., vaccines) or pharmaceutical compositions may be injected (e.g., via intramuscular, intraperitoneal, intradermal and/or subcutaneous routes); or delivered via the mucosa (e.g., to the oral/alimentary, respiratory, and/or genitourinary tracts). In some embodiments, it may be desirable to administer different doses of an immunogenic composition (e.g., vaccine) or pharmaceutical composition by different routes; in some embodiments, it may be desirable to administer different components of one dose via different routes. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein is administered intramuscularly. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein is administered subcutaneously. [0319] In some embodiments, immunogenic compositions (e.g., vaccines) or pharmaceutical compositions are administered intradermally. Conventional technique of intradermal injection, the "Mantoux procedure", comprises steps of cleaning the skin, and then stretching with one hand, and with the bevel of a narrow gauge needle (26-31 gauge) facing upwards the needle is inserted at an angle of between 10-15°. Once the bevel of the needle is inserted, the barrel of the needle is lowered and further advanced while providing a slight pressure to elevate it under the skin. The liquid is then injected very slowly thereby forming a bleb or bump on the skin surface, followed by slow withdrawal of the needle. [0320] Devices that are specifically designed to administer liquid agents into or across the skin have been described, for example the devices described in WO 99/34850 and EP 1092444, also the jet injection devices described for example in WO 01/13977; US Patent No. 5,480,381, US Patent No. 5,599,302, US Patent No. 5,334,144, US Patent No. 5,993,412, US Patent No. 5,649,912, US Patent No. 5,569,189, US Patent No. 5,704,911, US Patent No. 5,383,851, US AFV70463FF Patent No. 5,893,397, US Patent No. 5,466,220, US Patent No. 5,339,163, US Patent No. 5,312,335, US Patent No. 5,503,627, US Patent No. 5,064,413, US Patent No. 5,520,639, US Patent No. 4,596,556, US Patent No. 4,790,824, US Patent No. 4,941,880, US Patent No. 4,940,460, WO 97/37705 and WO 97/13537. Other methods of intradermal administration of the vaccine preparations may include conventional syringes and needles, or devices designed for ballistic delivery of solid vaccines (WO 99/27961), or transdermal patches (WO 97/48440; WO 98/28037); or applied to the surface of the skin (transdermal or transcutaneous delivery WO 98/20734; WO 98/28037). [0321] As described above, immunogenic compositions (e.g., vaccines) or pharmaceutical compositions may be administered as a single dose or as multiple doses. It will be appreciated that an administration is a single “dose” so long as all relevant components are administered to a subject within a window of time; it is not necessary that every component be present in a single composition. For example, administration of two different immunogenic compositions (e.g., vaccines) or pharmaceutical compositions, within a period of less than 24 h, is considered a single dose. To give but one example, immunogenic compositions (e.g., vaccines) or pharmaceutical compositions having different antigenic components may be administered in separate compositions, but as part of a single dose. As noted above, such separate compositions may be administered via different routes or via the same route. Alternatively or additionally, in embodiments wherein an immunogenic composition (e.g., vaccine) or pharmaceutical composition comprises a combination of immunogenic agents and additional types of active agents, immunogenic agents may be administered via one route, and an additional active agent may be administered by the same route or by a different route. [0322] Immunogenic compositions (e.g., vaccines) and pharmaceutical compositions are administered in such amounts and for such time as is necessary to achieve a desired result. In certain embodiments of the present invention, an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein comprises an immunologically effective amount of at least immunogenic agent. The exact amount required to achieve an immunologically effective amount may vary, depending on the immunogenic agent, and from subject to subject, depending on the species, age, and general condition of the subject, the stage of the disease, the particular pharmaceutical mixture, its mode of administration, and the like. AFV70463FF [0323] The amount of polypeptide(s) (e.g., antigenic polypeptide(s)), polymer(s) (e.g., antigenic polysaccharide(s)), or conjugate(s) in each immunogenic composition (e.g., vaccine) or pharmaceutical composition dose is selected to allow the composition, when administered as described herein, to induce an appropriate immune-protective response without significant, adverse side effects. [0324] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein induces a T_H1 and/or T_H17 cell response upon administration to a subject. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein induces an opsonic/bactericidal response against S. pneumoniae upon administration to a subject. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein reduces rate of transmission and/or colonization by S. pneumoniae upon administration to a subject. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein reduces rate of transmission and/or colonization of the mucosal surfaces by S. pneumoniae upon administration to a subject. [0325] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein reduces rate of transmission and/or colonization of the nasopharynx or the lungs by S. pneumoniae upon transmission. In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein inhibits or reduces asymptomatic infection by S. pneumoniae upon administration to a subject. [0326] Some embodiments provide for a method of immunizing a subject against S. pneumoniae infection comprising administering to the subject an immunologically effective amount of an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein. Some embodiments provide for a method of immunizing a subject against S. pneumoniae infection comprising administering to the subject an immunologically effective amount of an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein. [0327] Some embodiments provide an immunogenic composition (e.g., vaccine) or pharmaceutical composition as described herein for use in therapy. For example, an immunogenic composition (e.g., vaccine) or pharmaceutical composition as described herein for use in the treatment or prevention of disease due to one or more pathogens in a subject such as a human subject. Some embodiments provide an immunogenic composition (e.g., vaccine) or AFV70463FF pharmaceutical composition as described herein for use in the treatment or prevention of disease due to one or more pathogens in a subject such as a human subject. Further embodiments provide a use of an immunogenic composition (e.g., vaccine) or pharmaceutical composition as described herein in the manufacture of a medicament for the treatment or prevention of disease due to one or more pathogens. In some embodiments, such a disease is due to infection and/or colonization by Streptococcus pneumoniae. The characteristics of methods of treatment or prevention disclosed herein are equally applicable to an immunogenic composition (e.g., vaccine) or pharmaceutical composition for use, or a use of an immunogenic composition (e.g., vaccine) or pharmaceutical composition, in the manufacture of a medicament. Combination Prophylaxis or Combination Therapy [0328] In some embodiments, an immunogenic composition (e.g., vaccine) or pharmaceutical composition disclosed herein may be administered in combination with one or more additional agents. In some embodiments, an additional agent may be or comprise a therapeutic agent. In some embodiments, such a therapeutic agent may be or comprise an antibacterial agent. In some embodiments, an additional agent may be or comprise one or more additional vaccines. In some embodiments, an additional agent may be or comprise PCV13. In some embodiments, an additional agent may be or comprise PCV20. In some embodiments, an additional agent may be or comprise PPSV23. In some embodiments, an additional agent may be or comprise an antibiotic. Dosing [0329] In some embodiments, administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition described herein may involve the delivery of a single dose. In some embodiments, administration may involve an initial dose followed by one or several additional immunization doses, adequately spaced. An immunization schedule or regimen is a program for the administration of one or more specified doses of one or more specified immunogenic compositions (e.g., vaccines) or pharmaceutical compositions, by one or more specified routes of administration, at one or more specified ages of a subject. AFV70463FF [0330] Immunization schedules of the present disclosure are provided to induce an immune response (e.g., an immunoprotective response) in a subject sufficient to reduce at least one measure selected from the group consisting of incidence, prevalence, frequency, and/or severity of at least one infection, disease, or disorder, and/or at least one surrogate marker of the infection, disease, or disorder, in a population and/or subpopulation of the subject(s). A supplemental immunization schedule is one which has this effect relative to the standard schedule which it supplements. A supplemental schedule may call for additional administrations and/or supra-immunogenic doses of the immunogenic compositions (e.g., vaccines) or pharmaceutical compositions disclosed herein, found in the standard schedule, or for the administration of immunogenic compositions (e.g., vaccines) or pharmaceutical compositions not part of the standard schedule. A full immunization schedule of the present invention may comprise both a standard schedule and a supplemental schedule. Exemplary sample immunization schedules are provided for illustrative purposes. Detailed descriptions of methods to assess immunogenic response discussed herein allow one to develop alterations to the sample immunization schedules without undue experimentation. [0331] In some embodiments of the present disclosure, a first administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition occurs when a subject is more than about 2 weeks old, more than about 5 weeks old, more than about 1 year old, more than about 2 years old, more than about 15 years old, or more than about 18 years old. [0332] In some embodiments, a first administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition occurs when a subject is about two months old. In some embodiments, a second administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition occurs when a subject is about four months old. In some embodiments, a third administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition occurs when a subject is about six months old. In some embodiments, a fourth administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition occurs when a subject is between about twelve months old and about fifteen months old. [0333] In some embodiments of the present disclosure, a first administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition occurs when a subject is AFV70463FF more than about 18 years old, more than about 50 years old, more than about 55 years old, more than about 60 years old, more than about 65 years old, or more than about 70 years old. [0334] In some embodiments of the disclosure, a single administration of an immunogenic composition (e.g., vaccine) or pharmaceutical composition is employed. It is possible that the purposes of the present disclosure can be served with a single administration, especially when one or more utilized polypeptide(s) (e.g., antigenic polypeptide(s)), polymers (e.g., polysaccharide(s)), and/or immunogenic complex(es) or combinations thereof is/are strong, and in such a situation a single dose schedule is sufficient to induce a lasting immune-protective response. [0335] In certain embodiments, it is desirable to administer two or more doses of an immunogenic composition (e.g., vaccine) or pharmaceutical composition, for greater immune- protective efficacy and coverage. Thus, in some embodiments, a number of doses is at least two, at least three or more doses. There is no set maximum number of doses; however, it is good clinical practice not to immunize more often than necessary to achieve the desired effect. [0336] Without being bound by theory, a first dose of an immunogenic composition (e.g., vaccine) or pharmaceutical composition administered according to the disclosure may be considered a “priming” dose. In certain embodiments, more than one dose is included in an immunization schedule. In such a scenario, a subsequent dose may be considered a “boosting” dose. [0337] A priming dose may be administered to a naïve subject (a subject who has never previously received an immunogenic composition (e.g., vaccine) or pharmaceutical composition). In some embodiments, a priming dose may be administered to a subject who has previously received an immunogenic composition (e.g., vaccine) or pharmaceutical composition at least five or more years previous to administration of an initial immunogenic composition (e.g., vaccine) or pharmaceutical composition dose according to the invention. In other embodiments, a priming dose may be administered to a subject who has previously received an immunogenic composition (e.g., vaccine) or pharmaceutical composition at least twenty or more years previous to administration of a priming immunogenic composition (e.g., vaccine) or pharmaceutical composition according to the disclosure. [0338] When an immunization schedule calls for two or more separate doses, the interval between doses is considered. The interval between two successive doses may be the same throughout an immunization schedule, or it may change as the subject ages. In immunization AFV70463FF schedules of the present disclosure, once a first immunogenic composition (e.g., vaccine) or pharmaceutical composition dose has been administered, there is a first interval before administration of a subsequent dose. A first interval is generally at least about 2 weeks, 1 month, 6 weeks, 2 months, 3 months, 6 months, 9 months, 12 months, or longer. Where more than one subsequent dose(s) are administered, second (or higher) intervals may be provided between such subsequent doses. In some embodiments, all intervals between subsequent doses are of the same length; in other embodiments, second intervals may vary in length. In some embodiments, the interval between subsequent doses may be at least about 12 months, at least about 15 months, at least about 18 months, at least about 21 months or at least about 2 years. In certain embodiments, the interval between doses may be up to 3 years, up to about 4 years, or up to about 5 years or 10 years or more. In certain embodiments, intervals between subsequent doses may decrease as the subject ages. [0339] It will be appreciated by those skilled in the art that a variety of possible combinations and sub-combinations of the various conditions of timing of the first administration, shortest interval, largest interval and total number of administrations (in absolute terms, or within a stated period) exist, and all of these combinations and sub-combinations should be considered to be within the inventor's contemplation though not explicitly enumerated here. Exemplary Assays for Determining Immune Response [0340] In some embodiments, a method of assessing the immunogenicity of an immunogenic composition described herein comprises evaluating, measuring, and/or comparing an immune response using one or more in vitro bioassays, including B cell and T cell responses such as antibody levels by ELISA, multiplex ELISA, MSD, Luminex, flow cytometry, T_H1/T_H17 cell response, cytokine level measurement and functional antibody levels as measured by opsonophagocytic killing (OPK, OPA), plaque reduction neutralization test (PRNT), serum bactericidal killing (SBA), agglutination, motility, cytotoxicity, or adherence; and in vivo assays in animal models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance from mucosal surfaces or bloodstream, reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction AFV70463FF of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the immunogenic composition.. In some embodiments, the immune response is compared to a control composition. In some embodiments, a control composition may comprise a polymer (e.g., an antigenic polysaccharide) present in the immunogenic composition and not comprise a polypeptide (e.g., an antigenic polypeptide) present in the immunogenic composition. In some embodiments, a control composition may comprise a polypeptide (e.g., an antigenic polypeptide) present in the immunogenic composition and not comprise a polymer (e.g., an antigenic polysaccharide) present in the immunogenic composition. In some embodiments, a control composition may comprise an adjuvant present in the immunogenic composition, and not comprise a polymer (e.g., an antigenic polysaccharide) and/or a polypeptide (e.g., an antigenic polypeptide) present in the immunogenic composition. [0341] In some embodiments, a method of assessing the potency of an immunogenic composition described herein comprises evaluating, measuring, and/or comparing an immune response using one or more in vitro bioassays, including B cell and T cell responses such as antibody levels by ELISA, multiplex ELISA, MSD, Luminex, flow cytometry, T_H1/T_H17 cell response, cytokine level measurement and functional antibody levels as measured by OPK (OPA), plaque reduction neutralization test (PRNT), serum bactericidal killing (SBA), internalization, activity neutralization, agglutination, motility, cytotoxicity, or adherence; and in vivo assays in animal models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance or reduction from mucosal surfaces or bloodstream, reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the immunogenic composition In some embodiments, the immune response is compared to a control composition. In some embodiments, a control composition may comprise a polymer (e.g., an antigenic polysaccharide) present in the immunogenic composition and not comprise a polypeptide (e.g., an antigenic polypeptide) present in the immunogenic composition. In some embodiments, a control composition may comprise a polypeptide (e.g., an antigenic polypeptide) present in the immunogenic composition and not comprise a polymer (e.g., an antigenic polysaccharide) present in the immunogenic composition. In some embodiments, a control composition may comprise an adjuvant present in the AFV70463FF immunogenic composition, and not comprise a polymer (e.g., an antigenic polysaccharide) and/or a polypeptide (e.g., an antigenic polypeptide) present in the immunogenic composition. [0342] In some embodiments, a method of assessing the immunogenicity of an vaccine composition described herein comprises evaluating, measuring, and/or comparing an immune response using one or more in vitro bioassays, including B cell and T cell responses such as antibody levels by ELISA, multiplex ELISA, MSD, Luminex, flow cytometry, T_H1/T_H17 cell response, cytokine level measurement and functional antibody levels as measured by opsonophagocytic killing (OPK, OPA), plaque reduction neutralization test (PRNT), serum bactericidal killing (SBA), agglutination, motility, cytotoxicity, or adherence; and in vivo assays in animal models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance from mucosal surfaces or bloodstream, reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the vaccine composition. In some embodiments, the immune response is compared to a control composition. In some embodiments, a control composition may comprise a polymer (e.g., an antigenic polysaccharide) present in the vaccine composition and not comprise a polypeptide (e.g., an antigenic polypeptide) present in the vaccine composition. In some embodiments, a control composition may comprise a polypeptide (e.g., an antigenic polypeptide) present in the vaccine composition and not comprise a polymer (e.g., an antigenic polysaccharide) present in the vaccine composition. In some embodiments, a control composition may comprise an adjuvant present in the vaccine composition, and not comprise a polymer (e.g., an antigenic polysaccharide) and/or a polypeptide (e.g., an antigenic polypeptide) present in the vaccine composition. [0343] In some embodiments, a method of assessing the potency of an vaccine composition described herein comprises evaluating, measuring, and/or comparing an immune response using one or more in vitro bioassays, including B cell and T cell responses such as antibody levels by ELISA, multiplex ELISA, MSD, Luminex, flow cytometry, T_H1/T_H17 cell response, cytokine level measurement and functional antibody levels as measured by OPK (OPA), plaque reduction neutralization test (PRNT), serum bactericidal killing (SBA), internalization, activity neutralization, agglutination, motility, cytotoxicity, or adherence; and in vivo assays in animal AFV70463FF models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance or reduction from mucosal surfaces or bloodstream, reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the vaccine composition. In some embodiments, the immune response is compared to a control composition. In some embodiments, a control composition may comprise a polymer (e.g., an antigenic polysaccharide) present in the vaccine composition and not comprise a polypeptide (e.g., an antigenic polypeptide) present in the vaccine composition. In some embodiments, a control composition may comprise a polypeptide (e.g., an antigenic polypeptide) present in the vaccine composition and not comprise a polymer (e.g., an antigenic polysaccharide) present in the vaccine composition. In some embodiments, a control composition may comprise an adjuvant present in the vaccine composition, and not comprise a polymer (e.g., an antigenic polysaccharide) and/or a polypeptide (e.g., an antigenic polypeptide) present in the vaccine composition. [0344] In some embodiments, a method of assessing the immunogenicity of an pharmaceutical composition described herein comprises evaluating, measuring, and/or comparing an immune response using one or more in vitro bioassays, including B cell and T cell responses such as antibody levels by ELISA, multiplex ELISA, MSD, Luminex, flow cytometry, T_H1/T_H17 cell response, cytokine level measurement and functional antibody levels as measured by opsonophagocytic killing (OPK, OPA), plaque reduction neutralization test (PRNT), serum bactericidal killing (SBA), agglutination, motility, cytotoxicity, or adherence; and in vivo assays in animal models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance from mucosal surfaces or bloodstream, reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the pharmaceutical composition. In some embodiments, the immune response is compared to a control composition. In some embodiments, a control composition may comprise a polymer (e.g., an antigenic polysaccharide) present in the pharmaceutical composition and not comprise a polypeptide (e.g., an antigenic polypeptide) AFV70463FF present in the pharmaceutical composition. In some embodiments, a control composition may comprise a polypeptide (e.g., an antigenic polypeptide) present in the pharmaceutical composition and not comprise a polymer (e.g., an antigenic polysaccharide) present in the pharmaceutical composition. In some embodiments, a control composition may comprise an adjuvant present in the pharmaceutical composition, and not comprise a polymer (e.g., an antigenic polysaccharide) and/or a polypeptide (e.g., an antigenic polypeptide) present in the pharmaceutical composition. [0345] In some embodiments, a method of assessing the potency of an pharmaceutical composition described herein comprises evaluating, measuring, and/or comparing an immune response using one or more in vitro bioassays, including B cell and T cell responses such as antibody levels by ELISA, multiplex ELISA, MSD, Luminex, flow cytometry, T_H1/T_H17 cell response, cytokine level measurement and functional antibody levels as measured by OPK (OPA), plaque reduction neutralization test (PRNT), serum bactericidal killing (SBA), internalization, activity neutralization, agglutination, motility, cytotoxicity, or adherence; and in vivo assays in animal models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance or reduction from mucosal surfaces or bloodstream, reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the pharmaceutical composition. In some embodiments, the immune response is compared to a control composition. In some embodiments, a control composition may comprise a polymer (e.g., an antigenic polysaccharide) present in the pharmaceutical composition and not comprise a polypeptide (e.g., an antigenic polypeptide) present in the pharmaceutical composition. In some embodiments, a control composition may comprise a polypeptide (e.g., an antigenic polypeptide) present in the pharmaceutical composition and not comprise a polymer (e.g., an antigenic polysaccharide) present in the pharmaceutical composition. In some embodiments, a control composition may comprise an adjuvant present in the pharmaceutical composition, and not comprise a polymer (e.g., an antigenic polysaccharide) and/or a polypeptide (e.g., an antigenic polypeptide) present in the pharmaceutical composition. [0346] In some embodiments, a method of assessing the immunogenicity and/or potency of an immunogenic complex comprises evaluating an immune response to immunogenic or vaccine AFV70463FF compositions comprising one or more immunogenic complexes. In some embodiments, the method of assessing the immunogenicity and/or potency of an immunogenic complex described herein comprises evaluating, measuring, and/or comparing an immune response using one or more in vitro bioassays, including B cell and T cell responses such as antibody levels by ELISA, multiplex ELISA, MSD, Luminex, flow cytometry, T_H1/T_H17 cell response, cytokine level measurement and functional antibody levels as measured by OPK, plaque reduction neutralization test (PRNT), serum bactericidal killing (SBA), agglutination, motility, cytotoxicity, or adherence; and in vivo assays in animal models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance or reduction from mucosal surfaces or bloodstream, reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the immunogenic composition. [0347] Generally speaking, it may be desirable to assess humoral responses, cellular responses, and/or interactions between the two. Where humoral responses are being assessed, antibody titers and/or types (e.g., total IgG, IgG1, IgG2, IgM, IgA, etc.) to specific pathogen polysaccharides or polypeptides (either serotype-specific or conserved across two or more serotypes) may be determined, for example before and/or after administration of an initial or a boosting dose of vaccine (and/or as compared with antibody levels in the absence of antigenic stimulation). Cellular responses may be assessed by monitoring reactions such as delayed type hypersensitivity responses, etc. to the carrier protein. Cellular responses can also be measured directly by evaluating the response of peripheral blood mononuclear cells (PBMCs) monocytes to stimulation with the antigens of interest. Precursor and memory B cell populations may be assessed in enzyme linked immunospot (ELISpot) assays directed against specific pathogen polysaccharides or polypeptides. [0348] Any of a variety of assays may be employed to detect levels and/or activity of antibodies in subject sera. Suitable assays include, for example, ligand binding assays, such as radioimmunoassay (RIAs), ELISAs, and multiplex assays (Luminex, Bioplex, MSD); functional assays, such as opsonophagocytic assays or internalization assays; and in vivo assays in animal models of disease. In some embodiments, where the disease is a bacterial disease, parameters of in vivo assays include bacterial clearance or reduction from mucosal surfaces or bloodstream, AFV70463FF reduction or prevention of bacteremia, meningitis, sepsis, or otitis media, reduction or prevention of colonization of the nasopharynx, inhibition or reduction of asymptomatic infection, reduction of mortality, and passive and active protection following challenge with one or more strains (variants) of bacteria that are the targets of the immunogenic composition. [0349] The RIA method detects specific antibodies through incubation of sera with radio- labeled polysaccharides or polypeptides in suspension (e.g., Schiffiman et al, 1980). The antigen- antibody complexes are then precipitated with ammonium sulfate and the radiolabeled pellets assayed for counts per minute (cpm). [0350] In the ELISA detection method, specific antibodies from the sera of vaccinated subjects are quantitated by incubation with polysaccharides or polypeptides (either serotype-specific or conserved across two or more serotypes) which have been adsorbed to a solid support (e.g., Koskela and Leinonen, 1981; Kojima et al., 1990; Concepcion and Frasch, 2001). The bound antibody is detected using enzyme-conjugated secondary detection antibodies. The ELISA also allows isotyping and subclassing of the immune response (i.e., IgM vs. IgG or IgG1 vs. IgG2) by using isotype- or subclass-specific secondary antibodies and can be adapted to evaluate the avidity of the antibodies (Anttila et al, 1998; Romero-Steiner et al, 2005). Multiplex assays (e.g., Luminex) facilitate simultaneous detection of antibodies to multiple antigens. Capsular polysaccharide(s) or polypeptides are conjugated to spectrally distinct microspheres that are mixed and incubated with serum. The antibodies bound to the polysaccharides or polypeptides on the coated microspheres are detected using a secondary antibody (e.g., R-Phycoerythrin-conjugated goat anti-human IgG). [0351] An approach for assessing functional antibody in serum is an opsonophagocytic assay (OPA) or a concentrated opsonophagocytic assay (COPA), which quantitates only the antibodies that can opsonize the bacteria, leading to ingestion and killing of the bacteria. The standard assay utilizes a human phagocytic effector cell, a source of complement, bacteria, and diluted sera. The assay readout is the serum endpoint titer at which there is >50% killing compared to bacteria incubated with complement and human cells alone (Romero-Steiner et al., 1997). This killing OPA can also be multiplexed by utilizing target strains of pathogen that carry different antibiotic resistance markers (Kim et al., 2003). Another type of multiplex opsonic assay is a nonkilling assay in which the uptake by phagocytic effector cells of fluorescent stained encapsulated pathogen or fluorescent microspheres conjugated with antigenic polysaccharides or polypeptides from a AFV70463FF target pathogen in the presence of diluted sera plus a complement source is evaluated by flow cytometry (Martinez et al., 1999). Opsonic activity of serum antibody plus complement can also be evaluated by measuring the oxidative response of phagocytic human effector cells to ingested pathogen (Munro et al., 1985; Ojo-Amaize et al., 1995). [0352] Certain in vivo model systems can be used to evaluate the protection afforded by serum antibodies induced by vaccines of the present disclosure. In such passive protection systems, mice or rats are challenged with the pathogen plus diluted sera, and the endpoint titer of the sera which provides protection against pneumonia, bacteremia, colonization of organs or tissues, or mortality is determined (Stack et al., 1998; Saeland et al., 2000). [0353] In some embodiments, efficacy of immunization may be determined by assaying one or more cytokine levels by stimulating T cells from a subject after immunization. The one or more cytokine levels may be compared to the one or more cytokine levels in the same subject before immunization. Increased levels of the one or more cytokine, such as a 1.5-fold, 2-fold, 5-fold, 10- fold, 20-fold, 50-fold or 100-fold or more increase over pre-immunization cytokine levels, would indicate an increased response to the immunogenic composition (e.g., vaccine) or pharmaceutical composition. In some embodiments, the one or more cytokines are selected from GM-CSP; IL- _{IL- -2; IL-3; IL-4; IL-5; IL-6; IL-7; IL-8; IL-10; IL-12; IL-17A, IL-17F or other members of} _{the IL-17 family; IL-22; IL-23; IFN- - - - - - -} In a non-limiting example, efficacy of immunization may be determined by assaying IL-17 levels (particularly IL-17A) by stimulating T cells from a subject after immunization. The IL-17 levels may be compared to IL-17 levels in the same subject before immunization. Increased IL-17 (e.g., IL-17A) levels, such as a 1.5-fold, 2-fold, 5-fold, 10-fold, 20-fold, 50-fold or 100-fold or more increase, would indicate an increased response to the immunogenic composition (e.g., vaccine) or pharmaceutical composition. [0354] In some embodiments, one may assay neutrophils in the presence of T cells or antibodies from the patient for pathogen (e.g., bacterial) killing. Increased pathogen (e.g., bacterial) killing, such as a 1.5-fold, 2-fold, 5-fold, 10-fold, 20-fold, 50-fold or 100-fold or more increase, would indicate an increased response to the immunogenic composition (e.g., vaccine) or pharmaceutical composition. For example, one may measure T_H17 cell activation, where increased T_H17 cell activation, such as a 1.5-fold, 2-fold, 5-fold, 10-fold, 20-fold, 50-fold or 100-fold or more increase, correlates with an increased response to the immunogenic composition (e.g., AFV70463FF vaccine) or pharmaceutical composition. In another non-limiting example, one may measure T_H1 cell activation, where increased T_H1 cell activation, such as a 1.5-fold, 2-fold, 5-fold, 10-fold, 20- fold, 50-fold or 100-fold or more increase, correlates with an increased response to the immunogenic composition (e.g., vaccine) or pharmaceutical composition. One may also measure levels of an antibody specific to the vaccine, where increased levels of the specific antibody, such as a 1.5-fold, 2-fold, 5-fold, 10-fold, 20-fold, 50-fold or 100-fold or more increase, are correlated with increased immunogenic composition (e.g., vaccine) or pharmaceutical composition efficacy. In certain embodiments, two or more of these assays are used. For example, one may measure IL- 17 levels and the levels of immunogenic composition (e.g., vaccine) or pharmaceutical composition -specific antibody. Alternatively, one may follow epidemiological markers such as incidence of, severity of, or duration of pathogen (e.g., bacterial) infection in vaccinated individuals compared to unvaccinated individuals. [0355] Immunogenic composition (e.g., vaccine) or pharmaceutical composition efficacy may also be assayed in various model systems such as the mouse challenge model. For instance, BALB/c or C57BL/6 strains of mice may be used. After administering the test immunogenic composition (e.g., vaccine) or pharmaceutical composition to a subject (as a single dose or multiple doses), the experimenter administers a challenge dose of pathogen. In some cases, a challenge dose administered intranasally or intratracheally is sufficient to cause pathogen infection and/or a high rate of lethality in unvaccinated animals. In some cases, a challenge dose administered intranasally is sufficient to cause pathogen colonization (especially nasal colonization) in an unvaccinated animal, and in some cases a challenge dose administered via aspiration is sufficient to cause sepsis and a high rate of lethality in unvaccinated animals. In some cases, a challenge dose administered via intraperitoneal injection is sufficient to cause sepsis and a high rate of lethality in unvaccinated animals. In some cases, a challenge dose administered via intravenous injection is sufficient to cause sepsis and a high rate of lethality in unvaccinated animals. [0356] Certain in vivo model systems can be used to evaluate the protection afforded by serum antibodies induced by immunogenic compositions (e.g., vaccines) or pharmaceutical compositions of the present disclosure. In such passive protection systems, mice or rats are challenged with the pathogen plus diluted sera, and the endpoint titer of the sera which provides protection against bacteremia, colonization of organs or tissues, or mortality is determined (see, e.g., Stack et al. 1998; Saeland et al.2000). AFV70463FF Kits or Delivery Devices [0357] The present disclosure also provides for kits for producing an immunogenic agent as disclosed herein which is useful for an investigator to tailor an immunogenic agent (e.g., an immunogenic complex) with their preferred polymer (e.g., an antigenic polysaccharide) and polypeptide (e.g., an antigenic polypeptide), e.g., for research purposes to assess the effect of an antigen, or a combination of antigens on immune response. Such kits can be prepared from readily available materials and reagents. For example, such kits can comprise any one or more of the following materials: a container comprising a polymer (e.g., an antigenic polysaccharide) cross- linked with a plurality of first affinity molecules; a container comprising a complementary affinity molecule which associates with the first affinity molecule, wherein the complementary affinity molecule associates with a polypeptide (e.g., an antigenic polypeptide or a carrier protein); a container comprising an antigenic polypeptide; a container comprising a carrier protein; a container comprising a fusion protein; a container comprising an antigenic polypeptide associated with a complementary affinity molecule; a container comprising a fusion protein associated with a complementary affinity molecule. [0358] In another embodiment, the kit comprises a container comprising a polymer (e.g., a polysaccharide); a container comprising a plurality of first affinity molecules; and a container comprising a cross-linking reagent for cross-linking the first affinity molecules to the polysaccharide, for example, but not limited to, CDAP (l-cyano-4- dimethylaminopyridinium tetrafluoroborate), and EDC (l-Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride). [0359] In another embodiment, the kit comprises a container comprising a polypeptide (e.g., an antigenic polypeptide or carrier protein), and a container comprising a complementary affinity molecule, which associates with a first affinity molecule. In some embodiments, the kit further comprises a means to attach the complementary affinity molecule to the polypeptide (e.g., the antigenic polypeptide or carrier protein), where the means can be by a cross-linking reagent or by some intermediary protein. [0360] In some embodiments, the kit can comprise at least one co-stimulation factor, which can be added to the polymer (e.g., a polysaccharide) or another polymer. In some embodiments, the kit comprises a cross-linking reagent, for example, but not limited to, CDAP (l-cyano-4- dimethylaminopyridinium tetrafluoroborate); EDC (l-Ethyl-3-[3-dimethylaminopropyl] AFV70463FF carbodiimide hydrochloride); sodium cyanoborohydride; cyanogen bromide; and ammonium bicarbonate/iodoacetic acid, for linking the co-factor to the polymer (e.g., a polysaccharide) or to another polymer. [0361] A variety of kits and components can be prepared for use in the methods described herein, depending upon the intended use of the kit, the particular target polymer (e.g., antigenic polysaccharide) and polypeptide (e.g., antigenic polypeptide) and the needs of the user. [0362] In some embodiments, a kit can further include informational material. The informational material can be descriptive, instructional, marketing, or other material that relates to the methods described herein and/or the use of compositions described herein. The informational material of a kit is not limited in its form. In some instances, the informational material can include information about production of a composition described herein, amino acid sequence of a polypeptide described herein, nucleic acid sequence encoding a polypeptide described herein, molecular weight of composition described herein, concentration, date of expiration, batch or production site information, and so forth. [0363] In some cases, the informational material, e.g., instructions, is provided in printed matter, e.g., a printed text, drawing, and/or photograph, e.g., a label or printed sheet. The informational material can also be provided in other formats, such as Braille, computer readable material, video recording, or audio recording. In other instances, the informational material of the kit is contact information, e.g., a physical address, email address, website, or telephone number, where a user of the kit can obtain substantive information about a composition described herein and/or their use in the methods described herein. The informational material can also be provided in any combination of formats. [0364] In addition to the components described above, a kit can include other ingredients, such as a solvent or buffer, a salt, a surfactant, a stabilizer, or a preservative. A kit can also include other agents, e.g., a second or third agent. The components can be provided in any form, e.g., liquid, dried or lyophilized form. The components can be substantially pure (although they can be combined together or delivered separate from one another) and/or sterile. When the components are provided in a liquid solution, the liquid solution can be an aqueous solution, such as a sterile aqueous solution. When the components are provided as a dried form, reconstitution generally is by the addition of a suitable solvent. The solvent, e.g., sterile water or buffer, can optionally be provided in the kit. AFV70463FF [0365] The present disclosure also provides for containers (e.g., vials and syringes) containing immunogenic compositions (e.g., vaccines) or pharmaceutical compositions described herein. In some embodiments, such immunogenic compositions (e.g., vaccines) or pharmaceutical compositions are formulated with other ingredients such as a solvent or buffer, a salt, a surfactant, a stabilizer, or a preservative. In some embodiments where the container is a vial, the components can be provided in liquid, or dried or lyophilized form. In some embodiments, where the container is a syringe the components can be provided in liquid form. When the components are provided in a liquid solution, the liquid solution can be an aqueous solution, such as a sterile aqueous solution. When the components are provided as a dried form, reconstitution generally is by the addition of a suitable solvent. The solvent, e.g., sterile water or buffer, can optionally be provided in the kit. In some embodiments, a container includes a single dose of the immunogenic composition (e.g., vaccine) or pharmaceutical composition (e.g., a syringe or vial containing a single dose). In some embodiments, a container that includes a single dose may include an overfill sufficient to deliver a single dose of the immunogenic composition (e.g., vaccine) or pharmaceutical composition. In some embodiments, a container includes multiple doses of the immunogenic composition (e.g., vaccine) or pharmaceutical composition (e.g., a vial containing multiple doses). The invention is further described in the following paragraphs: 1. A composition comprising a purified capsular polysaccharide having repeating units of formula I:

AFV70463FF HO HO O wherein each R 2. The composition of paragraph 1, wherein the purified capsular polysaccharide comprises from about 1 to about 4000 repeating units of formula I, optionally from about 1 to about 500, about 1 to about 250, from about 1 to about 100, from about 4 to about 500 or from about 4 to about 400 units of formular I. 3. The composition of paragraph 1, wherein the purified capsular polysaccharide comprises from about 4 to about 500 units of formular I. 4. The composition of any one of paragraphs 1-3, wherein the purified capsular polysaccharide comprises between 0% and 100% of repeating units wherein R is –C(O)-CH_3. 5. The composition of any one of paragraphs 1-4, wherein the purified capsular polysaccharide comprises about 10% to 75% of repeating units wherein R is –C(O)-CH_3. 6. The composition of any one of paragraphs 1-5, wherein the purified capsular polysaccharide comprises greater than or equal to 60% of repeating units wherein R is –C(O)-CH₃, optionally about 60% to about 70% of repeating units wherein R is –C(O)-CH₃. AFV70463FF 7. The composition of any one of paragraphs 1-6, wherein the purified capsular polysaccharide comprises between 1 and about 2000 units of formular I wherein R is –C(O)-CH₃, optionally between about 1 and about 250 units of formula I wherein R is –C(O)-CH₃, or between about 1 and about 100 units of formular I wherein R is –C(O)-CH₃. 8. The composition of any one of paragraphs 1-5, wherein the purified capsular polysaccharide comprises less than 60% of repeating units wherein R is –C(O)-CH₃. 9. The composition of paragraph 8, wherein the purified capsular polysaccharide comprises 1% to 55%, 2% to 50%, 5% to 40% or 10% to 30% of repeating units wherein R is –C(O)-CH_3. 10. The composition of paragraph 8, wherein the purified capsular polysaccharide comprises 1% to 50% of repeating units wherein R is –C(O)-CH_3. 11. The composition of any one of paragraphs 1-10, wherein a weight average molecular weight of the purified capsular polysaccharide is from about 900 Daltons to about 4,000 kiloDaltons. 12. The composition of any one of paragraphs 1-11, wherein the weight average molecular weight of the purified capsular polysaccharide is about 300kDa or greater. 13. The composition of any one of paragraphs 1-12, wherein the weight average molecular weight of the purified capsular polysaccharide is about 600kDa to about 4000kDa. 14. The composition of any one of paragraphs 1-13, wherein the purified capsular polysaccharide is activated with a chemical reagent to produce reactive groups for conjugation to a suitable conjugate (e.g., a linker or carrier protein). 15. The composition of paragraph 14, wherein the purified capsular polysaccharide is activated with an oxidizing agent to produce a reactive group for conjugation to a linker or a carrier protein. AFV70463FF 16. The composition of paragraph 14 or 15, wherein the reactive group is an aldehyde group (-C(O)H). 17. The composition of paragraph 14 or 15, wherein the reactive group is a cyanoester group (-OCN). 18. The composition of any one of paragraphs 14-17, wherein activation occurs at the C_2, C_3, C_5, or C₆ of the -Galf residue. 19. The composition of any one of paragraphs 14-18 wherein activation is achieved by oxidative cleavage of a monosaccharide ring, optionally yielding a moiety having one or more aldehyde, cyanoester or amide groups. 20. The composition of any one of paragraphs 14-19, wherein activation of the purified capsular polysaccharide yields a monosaccharide moiety having one or more aldehyde groups. 21. The composition of any one of paragraphs 14-20, wherein activation of the capsular polysaccharide of formula I yields a polysaccharide having one or more monomers of formula II: HO HO

AFV70463FF 22. The composition of any one of paragraphs 14-21, wherein activation occurs at one or both of the terminal hydroxyl groups of the purified capsular polysaccharide. 23. A composition comprising a polysaccharide having one or more monomers of formula II: HO HO O 24. The composition of paragraph 23, wherein the polysaccharide further comprises monomers of formula I: HO HO

AFV70463FF I wherein each instance of R is independently selected from H and –C(O)-CH₃, and the polysaccharide has a weight average molecular weight of from about 300 Da to about 4000 kDa. 25. An immunogenic composition comprising (i) a purified capsular polysaccharide and (ii) a polypeptide, wherein the purified capsular polysaccharide comprises repeating units of formula I: HO HO O wherein each R 26. The immunogenic composition of paragraph 25, wherein the purified capsular polysaccharide comprises from about 1 to about 4000 repeating units of formula I, optionally from about 1 to about 500, about 1 to about 250, from about 1 to about 100, from about 4 to about 500 or from about 4 to about 400 units of formular I. 27. The immunogenic composition of paragraph 25 or 26, wherein the purified capsular polysaccharide comprises from about 4 to about 500 units of formular I. 28. The immunogenic composition of any one of paragraphs 25-27, wherein the purified capsular polysaccharide comprises between 0% and 100% of repeating units wherein R is –C(O)- CH_3. AFV70463FF 29. The composition immunogenic of any one of paragraphs 25-28, wherein the purified capsular polysaccharide comprises about 10% to 75% of repeating units wherein R is –C(O)-CH_3. 30. The immunogenic composition of any one of paragraphs 25-29, wherein the purified capsular polysaccharide comprises greater than or equal to 60% of repeating units wherein R is – C(O)-CH₃, optionally about 60% to about 70% of relating units wherein R is –C(O)-CH₃. 31. The immunogenic composition of any one of paragraphs 25-30, wherein the purified capsular polysaccharide comprises between 1 and about 2000 units of formular I wherein R is – C(O)-CH₃ , optionally between about 1 and about 250 units of formula I wherein R is –C(O)-CH₃, or between about 1 and about 100 units of formular I wherein R is –C(O)-CH₃. 32. The immunogenic composition of any one of paragraphs 25-31, wherein the purified capsular polysaccharide comprises less than 60% of repeating units wherein R is –C(O)-CH₃. 33. The immunogenic composition of paragraph 32, wherein the purified capsular polysaccharide comprises 1% to 55%, 2% to 50%, 5% to 40% or 10% to 30% of repeating units wherein R is –C(O)-CH_3. 34. The immunogenic composition of paragraph 33, wherein the purified capsular polysaccharide comprises 1% to 50% of repeating units wherein R is –C(O)-CH_3. 35. The immunogenic composition of any one of paragraphs 25-34, wherein a weight average molecular weight of the purified capsular polysaccharide is from about 900 Daltons to about 4,000 kiloDaltons. 36. The immunogenic composition of any one of paragraphs 25-35, wherein the weight average molecular weight of the purified capsular polysaccharide is about 300kDa or greater. AFV70463FF 37. The immunogenic composition of any one of paragraphs 25-36, wherein the weight average molecular weight of the purified capsular polysaccharide is about 600kDa to about 4000kDa. 38. The immunogenic composition of any one of paragraphs 25-37, wherein the polypeptide is associated with the purified capsular polysaccharide. 39. The immunogenic composition of any one of paragraphs 25-38, wherein the polypeptide is or comprises a polypeptide antigen. 40. The immunogenic composition of any one of paragraphs 25-39, wherein the polypeptide is or comprises a carrier protein. 41. The immunogenic composition of any one of paragraphs 25-40, wherein a weight ratio of the polypeptide to the purified capsular polysaccharide is from about 20:1 to about 1:20. 42. The immunogenic composition of paragraph 41, wherein the weight ratio of the polypeptide to the purified capsular polysaccharide is from about 10:1 to about 1:10. 43. The immunogenic composition of paragraph 41, wherein the weight ratio of the polypeptide to the purified capsular polysaccharide is from about 20:1 to about 1:2. 44. The immunogenic composition of paragraph 41, wherein the weight ratio of the polypeptide to the purified capsular polysaccharide is about 3:1. 45. The immunogenic composition of paragraph 41, wherein the weight ratio of the polypeptide to the purified capsular polysaccharide is about 2:1. 46. The immunogenic composition of paragraph 41, wherein the weight ratio of the polypeptide to the purified capsular polysaccharide is about 1:1. AFV70463FF 47. The immunogenic composition of any one of paragraphs 25-46, wherein the polypeptide is a carrier protein, optionally wherein the carrier protein is a toxoid or a glycoprotein from a pathogen, optionally wherein the carrier protein is selected from the group consisting of: CRM197, tetanus toxoid, exoprotein A, PspA, diphtheria toxoid, and protein D. 48. The immunogenic composition of paragraph 47, wherein the polypeptide is CRM197. 49. The immunogenic composition of any one of paragraphs 25-46, wherein the purified capsular polysaccharide having one or more repeating units of formula I is biotinylated. 50. The immunogenic composition of any one of paragraphs 25-46 or 49, wherein the polypeptide is a fusion protein comprising a biotin-binding moiety. 51. The immunogenic composition of paragraph 50, wherein the fusion protein comprises at least one polypeptide antigen. 52. The immunogenic composition of paragraph 50 or 51, wherein the biotinylated purified capsular polysaccharide is non-covalently associated with the biotin-binding moiety of the fusion protein. 53. The immunogenic composition of paragraph 50, 51 or 52, wherein the biotin-binding protein is or comprises rhizavidin or a binding domain thereof. 54. The immunogenic composition of any one of paragraphs 50-53, wherein the biotin-binding moiety comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID NOs: 1-3. 55. The immunogenic composition of any one of paragraphs 50-54, wherein the at least one polypeptide antigen is or comprises a pneumococcal polypeptide antigen. AFV70463FF 56. The immunogenic composition of paragraph 55, wherein the at least one polypeptide antigen is selected from a pneumolysin polypeptide antigen, an SP0435 polypeptide antigen, an SP1500 polypeptide antigen, and an SP0785 polypeptide antigen. 57. The immunogenic composition of paragraph 56, wherein the at least one of the polypeptide antigens comprise an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID NOs: 4-13 or SEQ ID Nos: 20-25. 58. The immunogenic composition of any one of paragraphs 50-57, wherein the fusion protein further comprises a linker positioned between the biotin-binding moiety and at least one of the polypeptide antigen(s). 59. The immunogenic composition of paragraph 58, wherein the linker is GGGGSSS (SEQ ID NO: 30). 60. The immunogenic composition of any one of paragraphs 50-59, wherein the fusion protein comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 24 or SEQ ID NO: 25. 61. The immunogenic composition of paragraph 50-59, wherein the fusion protein comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 20 or SEQ ID NO: 21. 62. The immunogenic composition of any one of paragraphs 25-61, wherein the purified capsular polysaccharide is derived from Streptococcus pneumoniae serotype 38. 63. The immunogenic composition of any one of paragraphs 25-62, wherein the immunogenic composition further comprises one or more polysaccharides of Streptococcus pneumoniae each having a serotype independently selected from: 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 6E, 6F, 6G, 6H, 7A, 7B, 7C, 7F, 8, 9A, 9L, 9N, 9V, 10A, 10B, 10C, 10F, 11A, 11B, 11C, 11D, 11E, 11F, 12A, 12B, 12F, 13, 14, 15A, 15B, 15C, 15F, 16A, 16F, 17A, 17F, 18A, 18B, 18C, 18F, 19A, 19B, 19C, 19F, AFV70463FF 20A, 20B, 21, 22A, 22F, 23A, 23B, 23F, 24A, 24B, 24F, 25A, 25F, 27, 28A, 28F, 29, 31, 32A, 32F, 33A, 33B, 33C, 33D, 33E, 33F, 34, 35A, 35B, 35C, 35F, 36, 37, 39, 40, 41A, 41F, 42, 43, 44, 45, 46, 47A, 47F, and 48. 64. The immunogenic composition of any one of paragraphs 25-63, wherein the composition further comprises one or more polysaccharides of Streptococcus pneumoniae each having a serotype independently selected from: 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F, and 35B. 65. A pharmaceutical composition comprising the immunogenic composition of any one of paragraphs 25-64, and a pharmaceutically acceptable carrier. 66. A method of inducing an immune response to a Streptococcus pneumoniae capsular polysaccharide comprising administering to a subject the immunogenic composition of any one of paragraphs 25-64 or the pharmaceutical composition of paragraph 65. 67. The method of paragraph 66, wherein the immune response is at a level that is higher than a corresponding reference level, wherein the reference level is a level of an immune response induced in a subject following administration of a reference composition. 68. The method of paragraph 67, wherein the reference composition comprises (i) a purified capsular polysaccharide comprising one or more units of formula I, and (ii) the polypeptide, wherein in the purified capsular polysaccharide of the reference composition: R is –C(O)-CH₃ in about 60% of the total instances of units of formula I. 69. The method of any one of paragraphs 66-68, wherein the immune response is against the capsular polysaccharide and/or the polypeptide. 70. The method of any one of paragraphs 66-69, wherein the immune response is or comprises an antibody and/or B cell response. AFV70463FF 71. The composition of any one of paragraphs 1-24, the immunogenic composition of any one of paragraphs 25-64 or the pharmaceutical composition of paragraph 65 for use in the treatment or prevention of S. pneumoniae infection in a subject. 72. The composition, the immunogenic composition or pharmaceutical composition for use of paragraph 71 wherein the S. pneumoniae infection includes one or more of Invasive Pneumococcal Disease (IPD), bacteremia, sepsis, organ damage, meningitis, pneumonia, otitis media and sinusitis due to infection by S. pneumoniae. 73. The composition of any one of paragraphs 1-24, the immunogenic composition of any one of paragraphs 25-64 or the pharmaceutical composition of paragraph 65 for use in protecting against colonization of a subject. 74. The composition of any one of paragraphs 1-24, the immunogenic composition of any one of paragraphs 25-64 or the pharmaceutical composition of paragraph 65 for use in inducing an immune response against a S. pneumoniae bacterial pathogen, optionally against a S. pneumoniae capsular polysaccharide, in a subject. 75. The composition, immunogenic composition or pharmaceutical composition for use of paragraph 74 wherein the immune response is an antibody or B cell response. 76. The composition, the immunogenic composition or pharmaceutical composition for use of paragraph 74 or 75 wherein the immune response is a T cell response, for example a CD4+ T cell response, including a T_H1, T_H2 or T_H17 response. 77. The composition, immunogenic composition or pharmaceutical composition for use of any one of paragraphs 74-76 wherein the immune response is a protective immune response. 78. The composition, immunogenic composition or pharmaceutical composition for use of any one of paragraphs 74-77 wherein the immune response comprises neutralizing antibodies. AFV70463FF 79. The composition, the immunogenic composition or pharmaceutical composition for use of any one of paragraphs 74-77 wherein the immune response to one or more antigenic polypeptides is increased at least 1-fold, 2-fold, 3-fold, 4-fold or 5-fold relative to a predetermined level as measured by ELISA. 80. The composition, immunogenic composition or pharmaceutical composition for use of any one of paragraphs 74-79 wherein the immune response to one or more antigenic polypeptides is increased at least 1-fold, 2-fold, 3-fold, 4-fold or 5-fold relative to a predetermined level as measured by a functional assay. 81. The composition, immunogenic composition or pharmaceutical composition for use of paragraph 79 or 80 wherein the predetermined level is a pre-immune level. 81. The composition, immunogenic composition or pharmaceutical composition for use of any one of paragraphs 74-81 wherein the subject is a human subject. 82. The composition, immunogenic composition or pharmaceutical composition for use of paragraph 81 wherein the human subject is about 50 years of age or older. 83. The composition, immunogenic composition or pharmaceutical composition for use of paragraph 81 wherein the human subject is between about 6 weeks of age and about 6 years of age. 84. A use of the composition of any one of paragraphs 1-24, the immunogenic composition of any one of claims 25-64, or the pharmaceutical composition of claim 65 in the manufacture of a medicament for the prevention or treatment of S. pneumoniae infection. 85. A use of the composition of any one of paragraphs 1-24, the immunogenic composition of any one of claims 25-64, or the pharmaceutical composition of claim 65 in the manufacture of a medicament for inducing an immune response to a S. pneumoniae capsular polysaccharide in a subject following administration of the composition. AFV70463FF 86. A method of characterizing an immunogenic composition comprising a purified capsular polysaccharide comprising one or more units of formula I: HO HO O wherein each R wherein the method comprises steps of: characterizing a number of units of formula I in the purified capsular polysaccharide where R is –C(O)-CH₃; and comparing the number of units of formula I where R is –C(O)-CH₃ to a reference standard. 87. A method of characterizing an immunogenic composition comprising a purified capsular polysaccharide comprising one or more units of formula I:

AFV70463FF HO HO O wherein each R wherein the method comprises steps of: quantifying a percentage of units of the purified capsular polysaccharide that are of formula I; and comparing the percentage of units of formula I in the purified capsular polysaccharide to a reference standard. 88. A method of selecting a level of acetylation of a purified capsular polysaccharide for an immunogenic composition, the method comprising steps of: administering to each of a plurality of animals one of a plurality of compositions comprising a purified capsular polysaccharide comprising one or more units of formula I,

AFV70463FF HO HO O wherein each R in each of the plurality of compositions, R is –C(O)-CH₃ in a distinct percentage of the total instances of units of formula I; determining the level of an immune response to the purified capsular polysaccharide; comparing the level of immune response to a reference level; and selecting a purified capsular polysaccharide with a given acetylation level for inclusion in the immunogenic composition, if the determined immune response is higher than the reference level. 89. An activated capsular polysaccharide comprising one or more units of formula I,

AFV70463FF HO HO O wherein each R 90. An activated serotype 38 capsular polysaccharide. 91. An activated capsular polysaccharide comprising one or more units of formula I, HO HO wherein each the purified capsular polysaccharide is conjugated to a carrier AFV70463FF 92. A purified capsular polysaccharide comprising one or more monomers of formula III: R^aO R^aO O where one a carrier protein) and biotin, and the remaining instances of R^a are each H. 93. The purified capsular polysaccharide of paragraph 92, wherein one instance of R^a is CN, or -C(NH)N-R^b, and R^b is independently selected from a protein (e.g., a carrier protein) and biotin, and the remaining instances of R^a are each H. 94. A purified capsular polysaccharide comprising one or more monomers of formula IV- 1:

AFV70463FF HO R^aO O where one and the remaining instances of R^a are each H. 95. A purified capsular polysaccharide comprising one or more monomers of formula IV- 2: HO R^aO where one instance of R^c is a site for reaction with an oxidizing agent (e.g., periodate), and the remaining instances of R^c are each -OH. AFV70463FF 96. A purified capsular polysaccharide comprising one or more monomers of formula IV- 3: HO R^aO O where one instance of R^c is a site for reaction with an oxidizing agent (e.g., periodate), and the remaining instance of R^c is -OH.

AFV70463FF EXAMPLES Example 1: Characterization of a Purified Capsular Polysaccharide [0366] Streptococcus pneumoniae (Spn) is an important encapsulated human pathogen circulating globally, well recognized as the etiological agent of community-acquired pneumonia (CAP), otitis media, septicemia, and meningitis (together referred to as invasive pneumococcal disease [IPD]). See Centers for Disease Control and Prevention. Epidemiology and prevention of vaccine-preventable diseases, 13^th ed.; Public Health Foundation: Washington, DC, USA, 2015. [0367] Pneumococcal polysaccharide vaccines (PPVs) and pneumococcal conjugate vaccines (PCVs) targeting multiple Spn serotypes have been developed and are widely available for the prevention of pneumococcal disease. To date PPV23 and PCV10, PCV13, PCV15, PCV20 vaccines are available on the market. See M. Masomian, Z. Ahmad, L.T. Gew, C.L. Poh, Vaccines (Basel) 8 (2020) 132; K. Cannon, C. Elder, M. Young, D.A. Scott, I.L. Scully, G. Baugher, Y. Peng, K.U. Jansen, W.C. Gruber, W. Watson, W., Vaccine 39 (2021) 7494-7502. [0368] Spn synthesizes >100 types of capsular polysaccharides (CPSs), classified according to the unique glycan components and linkages that constitute each serotype. While the CPS repeating units of the serotypes included in the licensed vaccines have been determined, the CPS repeating unit structure of a large proportion of the other serotypes, including Type 38, has not been determined yet. See F. Ganaie, J.S. Saad, L. McGee, A.J. van Tonder, S.D. Bentley, S.W. Lo, R.A; Gladstone, P. Turner, J.D. Keenan, R.F. Breiman, M.H. Nahm, mBio 11 (2020). [0369] Spn type 38 (Type 38 in Danish nomenclature; Type 71 in US nomenclature) was probably first recognized in the 1940’s. See F. Kauffmann, E. Morch, K. Schmith. On the serology of the pneumococcus group. J Immunol 39 (1940) 397-426; Public Health Reports Vol.59 April 7, 1944 No.14.; Public Health Reports Vol.59 April 14, 1944 No.15. [0370] In some studies, Type 38 pneumococcus strains have been isolated from cerebrospinal fluid as one of the most common pneumococcal serotypes. See A.E. Taunay, R. Austrian, I.M. Landgraf, M.F. Vieira, C.E. Melles. Serotypes of Streptococcus pneumoniae isolated from cerebrospinal fluid in 1977-1988 in São Paulo City. Brazil Rev Inst Med Trop Sao Paulo 32 (1990) 11-15; L. Ter Horst, M.C. Brouwer, A. van der Ende, D. van de Beek. Community-acquired AFV70463FF bacterial meningitis in adults with cerebrospinal fluid leakage. Clin Infect Dis. 70 (2020) 2256- 2261. [0371] In 2017-18, the IPD frequency of Type 38 in USA was 102 and 342 cases in pediatrics (<5 years) and adults (>65 years), respectively. [0372] The present example describes use of NMR Spectroscopy and other analytical techniques to reveal the Type 38 polysaccharide repeating unit structure which is composed of a _{penta-saccharide repeat unit -[ -D-Galf - -D-GalpA6(Ser)- - -D-GlcpNAc-} _{3)- -Sugp- 4)- -D-Galp(2OAc)- . The polysaccharide is O-acetylated at the position} C₂ of the -Galp residue at approximately 68% to 87% of the repeat units. Results and Discussion Composition analysis [0373] Monosaccharide composition of Spn type 38 CPS was determined by HPAEC-PAD analysis following standard procedures using 2 M TFA hydrolysis. Galactose, Glucosamine, Galacturonic acid, and a constituent which was tentatively identified as Serine were detected in the hydrolysate (FIG. 1). Black particulates observed during TFA hydrolysis indicated the presence of an acid sensitive sugar moiety. Incorporation of a sodium borohydride reduction step prior to TFA hydrolysis eliminated the black particulate formation but also led to the formation of several new components. One of these new components was identified as Fucosamine by matching the retention time with a standard. This finding confirmed the presence of an acid labile keto sugar in Spn type 38 CPS, with carbonyl group located at the C₄ position of a 2-acetamido- 2,6 dideoxy- hexose residue. [0374] A novel keto sugar, ‘Sugp’, has been previously identified as a constituent of pneumococcal Type 5 capsular polysaccharide. The structure of Sugp in that study was established as 2-acetamido-2,6 dideoxy-D-xylo-hexos-4-ulose. See P.E. Jansson, B. Lindberg, U. Lindquist. Structural studies of the capsular polysaccharide from Streptococcus pneumoniae type 5. Carbohydr Res 140 (1985) 101-110. Our current results point also to the presence of the Sugp residue in Spn type 38 CPS. [0375] Amino acid analysis confirmed the presence of Serine as a constituent of S. pneumoniae serotype 38 capsular polysaccharide. Further evidence that serine is an integral part of the repeat unit was obtained through mass spectral analysis of oligosaccharide fragments generated by base AFV70463FF treatment. As shown in FIG.2, the presence of several oligosaccharide entities with incremental masses of 87 confirmed that serine is covalently linked to the polysaccharide repeat units. Linkage analysis [0376] Glycosyl linkage analyses of the native and the sodium borodeuteride reduced Spn type 38 CPS were performed following methylation and subsequent conversion of the methylated products into permethylated alditol acetates. Residues identified as 3-linked GlcpNAc, 4-linked Galp and t-Galf were detected in the native and reduced polysaccharide samples. [0377] Two additional constituents with identical mass fragmentation patterns observed in the reduced Spn type 38 CPS sample were identified as isomers of a 2-acetamido-2,6-dideoxy-hexose moiety (Figs.3-5). These additional species are derived from the reduction of the keto sugar, Sugp. Deuterium incorporation at C₄ indicates that the keto function is located at C₄ of Sugp and therefore the Sugp residue must be linked through the 3-position. [0378] Significant quantities of t-Galp were also observed during the linkage 81 analysis of the native polysaccharide. A possible explanation for this observation is that a significant proportion of the Sugp residue in the native Spn type 38 CPS had undergone degradation during methylation (due to the basic conditions) with concomitant release of the aglycone, which in this case presumably is the 4-Galp residue. The liberated Galp is then identified as t-Galp in the linkage analysis. This observation agrees with the previous findings of base catalyzed degradation of the Sugp moiety in Spn type 5 CPS during methylation (see P.E. Jansson, B. Lindberg, U. Lindquist. Structural studies of the capsular polysaccharide from Streptococcus pneumoniae type 5. _{Carbohydr Res 140 (1985) 101-110) and provided a clue to the presence of a Sugp (1 4)-Galp} linkage in Spn type 38 CPS. Linkage analysis of a sodium borohydride reduced polysaccharide sample did not show any t-Galp residues. NMR assignments for Spn type 38 CPS repeating unit [0379] ¹H NMR spectrum of native type 38 CPS (FIG.6; trace a) showed several unresolved anomeric protons, ring signals, O- and N-acetyl methyl signals and methyl signals from a deoxy sugar, together with signals from residual cell wall polysaccharide (CWPS). In the proton anomeric region, the de-O-acetylated CPS shows three well resolved signals at 5.30, 5.21, 5.15 and two signal partially overlapped at 4.77 and 4.79 ppm (FIG.6; trace b). The integral ratio of these signals resulted 1:1:1:2, confirming the presence of five monosaccharides. AFV70463FF [0380] ¹³C 1D spectrum also confirm the presence of five anomeric signals at 107.64, 100.87, 99.55, 98.22, and 94.17 ppm, which were correlated to the relative proton signal by ¹H-¹³C HSQC spectrum (FIG.7). In addition, three signals for nitrogen-bearing carbon atoms (51-56 ppm) and four carbonyl resonances in the 170-175 ppm region were detected in the HSQC and HMBC experiments. A signal observed at 11.1 ppm can be attributable to the C-6 methyl group of Sug residue as had been previously reported for Spn serotype 5 (see P.E. Jansson, B. Lindberg, U. Lindquist. Structural studies of the capsular polysaccharide from Streptococcus pneumoniae type 5. Carbohydr Res 140 (1985) 101-110). Upon reduction of the polysaccharide with sodium borodeuteride, this signal at 12511.1 ppm shifted to 15.7 ppm (FIG.8) as observed previously for the keto sugar of type 5 polysaccharide. [0381] ³¹P NMR spectrum of native type 38 CPS collected at 242 MHz and 323 K revealed signals of residual CWPS di-substituted with phosphocholine (FIG. 9). See C. Karlsson, P.E. Jansson, U.B. Skov Sørensen, Eur J Biochem 265 (1999) 1091-1097. Removal of the CWPS impurity by nitrous acid treatment of Spn type 38 CPS followed by purification abolished all the ³¹P NMR signals. [0382] Glycosyl composition and linkage analyses, as discussed above have shown the presence of t-Galf, 4-Galp, 3-GlcpNAc and 3-Sugp moieties. The linkage position(s) for the GalpA moiety could not be readily established. Assignment of the anomeric signals of all five monosaccharides in the repeat unit was accomplished using various NMR techniques as discussed below. [0383] The signal at 5.16 ppm with cross peak at 107.87 ppm in the HSQC (FIG.7) spectrum of the de-O-acetylated Spn type 38 CPS was attributable to the -configuration of the Galf residue. Upon reduction with sodium borohydride, the signal at 4.79 ppm was observed to convert into two new anomeric signals at 4.80 and 4.85 ppm (data not shown). This signal at 4.79 ppm therefore -Sugp residue, as the reduction of the keto group in Sugp is expected to affect its anomeric proton position. The signal at 5.23 pm was assigned to the -GlcpNAc residue by first identifying the H₂ proton of the GlcpNAc through ¹H-¹³C HSQC followed by correlating the H₂ and the anomeric proton using ¹H-¹H COSY data. In addition. the GlcpNAc signal at 5.23 ppm was observed to shift to 5.01 ppm in the spectrum of the reduced polysaccharide which gave the clue that the GlcpNAc is linked to 3-position of Sugp. AFV70463FF [0384] Smith degradation of a sample of de-O-acetylated Spn type 38 CPS resulted in the loss of all Galactose residues and caused the disappearance of the signal at 5.3 ppm. Since the chemical shift of the Galf residue has been previously established to be at 5.16 ppm, the signal at 5.3 ppm can be assigned to the presence of an -linked Galp molecule. The remaining anomeric signal at 4.77 ppm was then assigned to a GalpA residue with -configuration. [0385] Proton 1D and 2D TOCSY, 2D COSY, HSQC, 1D and 2D NOESY and HMBC experimental data (Figs. 6-8 and 10-12) were used to establish the repeating unit and the assignment of protons and carbons are summarized in Table 1-1. Table 1-1. ¹H and ¹³C NMR chemical shifts (ppm) for the de-O-acetylated type 38 CPS repeating unit. J_C1-H1 are reported in parentheses. Residue C₁ C₂ C₃ C₄ C₅ C₆ C CH₃ ^NAc C CO^NAc H1 H2 H3 H4 H5 H6 H - C 99.54 [174] 53.53 77.71 68.28 76.06 63.77 22.54 174.26 -GlcpNAc 5.23 4.08 3.86 4.08 3.86 3.72/3.86 2.07 NA B 98.45 [174] 51.49 78.24 ND* 69.53 11.10 22.54 174.26 -Sugp 4.79 4.23 3.91 NA 4.38 1.18 2.07 NA A 94.35 [168] 68.72 70.36 77.94 72.51 60.49 NA NA -Galp 5.29 3.96 3.72 4.01 3.78 3.70/3.86 NA NA D 101.04 [162] 73.05 76.33 68.13 74.83 170.16 NA NA -GalpA 4.77 3.86 4.07 3.71 4.24 NA NA NA Ser 174.45 55.85 61.86 NA NA NA NA NA NA 4.48 3.92/3.96 NA NA NA NA NA E 107.86 [174] 82.67 81.26 77.74 70.47 60.36 NA NA -Galf 5.16 4.02 4.01 3.97 3.84 3.77/3.70 NA NA * Keto group at position C₄ remains as hydrated and ¹³C chemical shift has not been assigned NA: not applicable ND: not determined [0386] ¹H and ¹³C NMR chemical shifts (ppm) for native type 38 CPS repeating unit assignments are provided in Table 1-2, illustrating the presence of the O-acetyl group in the native species on residue A: Table 1-2.¹H and ¹³C NMR chemical shifts (ppm) for native type 38 CPS repeating unit. JC1- H1 are reported in parentheses. Residue C1 C2 C3 C4 C5 C6 C CH3^NAc C CO^NAc C CO^OAc AFV70463FF H₁ H₂ H₃ H₄ H₅ H₆ H - - C 99.76 [168] 53.75 77.81 68.63 74.25 63.79 22.77 174.26 NA -GlcpNAc 5.21 4.10 3.71 3.91 3.85 3.73/3.86 2.06 NA NA B 98.53 [174] 51.57 78.46 ND* 69.59 11.16 22.63 174.45 NA -Sugp 4.78 4.22 3.90 NA 4.35 1.18 2.08 NA NA A 94.30 [174] 66.45 73.82 77.91 72.09 60.58 NA NA 172.89 -Galp 5.24 5.88 4.25 4.00 3.88 3.70/3.91 NA NA NA (2OAc) 2.12/20.45 D 101.26 [162] 73.41 77.23 68.22 73.68 168.80 NA NA NA -GalpA 4.84 3.87 4.11 3.70 4.40 NA NA NA NA Ser 175.23 56.39 62.58 NA NA NA NA 4.31 3.88/3.91 NA NA NA E 107.95 [174] 82.83 81.19 77.91 70.47 60.44 NA NA NA -Galf^c 5.14 4.00 4.04 3.99 3.84 3.74/3.86 NA NA NA * Keto group at position C₄ remains as hydrated and ¹³C chemical shift has not been assigned NA: not applicable ND: not determined [0387] The sequence of the repeating unit was deduced from NOESY and HMBC experimental data. Pertinent NMR data along with assignments of inter-residue linkage positions are summarized in Table 2. Inter residue linkages between Sugp to Galp, Galf to GalpA, GalpA to GlcpNAc, and GlcpNAc to Sugp were confirmed with HMBC (and NOESY) data. The linkage between Galp and the GalpA moieties was identified using NOE data (FIG.11). Table 2. Identification of inter-residue linkage positions within the de-O-acetylated S. pneumoniae serotype 38 capsular polysaccharide repeat unit. Anomeric Position HMBC NOE (¹H and ¹³C Chemical shift, ppm) -Galp A (5.30, 94.27) * D2, D3 -Sug B (4.79, 98.49) C_4- A A3, A4 -GlcNAc C (5.23, 99.55) C_3- B B1, B2, B3 -GalpA D (4.77, 101.20) H_3- C C3, C2 -Galf E (5.16, 107.87) C_2- D, H_2- D D2, D3 * Not readily assignable AFV70463FF Site attachment of the Serine residue [0388] Amino acid analysis data showed the presence of serine in a 1:1 stoichiometric ratio with respect to the penta-saccharide repeat unit. The presence of serine as part of the repeat structure was also able to be established through 2D NMR experiments (Table 1). The nature of linkage and the site of attachment of serine was deduced from the HMBC data (FIG. 13). The GalpA carbonyl signal at 170.16 ppm showed a 2-bond correlation to the H5 of GalA and a 3-bond correlation to the serine methine proton thus confirming that the serine moiety is linked to GalpA via an amide linkage. Previous structural studies reported for the O-polysaccharide of Providencia stuartii O43 had identified a similar GalpA-Serine structure (see O.G. Ovchinnikova, N.A. Kocharova, A. Torzewska, A. Blaszczyk, A.S. Shashkov, Y.A. Knirel, A. Rozalski. Structure of the O-polysaccharide from the lipopolysaccharide of Providencia stuartii O43 containing an amide of D-galacturonic acid with L-serine. Carbohydr Res 340 (2005) 1407-1411) but our current study is the first time an amino acid is shown to be an integral component a Spn CPS. [0389] Based on all the above data obtained through composition, linkage and NMR analyses we propose the following structure for the de-O-acetylated Spn type 38 CPS. -_{[ -D-Galf - -D-GalpA6(Ser)- - -D-GlcpNAc- - -Sugp- - -D-} _Galp- 1H and ¹³ -Galp residues to be partially substituted at position C₂ with O-acetyl group (Figs. 14A-14D). The proton at position C₂ -Galp residues of the native Spn type 38 CPS resonate at lower magnetic field in comparison _{to the signal for the de-O- -} Galp H₁ upon de-O-acetylation agrees with previous observations of influence of O-acetyl groups on protons on vicinal carbons. Further 2D-COSY, TOCSY, NOSEY and HMBC experiments for native 38 confirmed the acetyl group to be on the C₂ of Galp residue (Fig 13). [0391] These data suggest that Spn type 38 CPS is composed of a penta-saccharide repeat unit -_{[ -D-Galf - -D-GalpA6(Ser)- - -D-GlcpNAc- - -Sugp- - -D-} _{Galp- -acetylation (68-87%) at position C2 of the -Gal residue (FIG. 15).} Conclusions [0392] NMR and other analytical data revealed that Spn type 38 CPS is composed of a penta- _{saccharide repeat unit -[ -D-Galf - -D-GalpA6(Ser)- - -D-GlcpNAc- 3)- -} AFV70463FF _{Sugp- 4)- -D-Galp(2OAc)- . The polysaccharide is O-acetylated at the position C2 of the} -Galp residue at approximately 68%-87% of the repeat units. [0393] ‘Decorative groups’ such as pyruvate and O-acetyl have been largely identified in several Spn types. The presence of Serine has been revealed for O-polysaccharide of Providencia stuartii O43 (see O.G. Ovchinnikova, N.A. Kocharova, A. Torzewska, A. Blaszczyk, A.S. Shashkov, Y.A. Knirel, A. Rozalski. Structure of the O-polysaccharide from the lipopolysaccharide of Providencia stuartii O43 containing an amide of D-galacturonic acid with L-serine. Carbohydr Res 340 (2005) 1407-1411) but this is the first case that an amino acid is shown to be an integral component a Spn CPS. This evidence opens other questions on the possible glycobiology mechanism which can explain the presence of amino acids in Spn CPS repeating units. [0394] A cross reaction was shown regularly by type 25 antiserums for type 71 (Spn type 38). See Public Health Reports Vol. 59 April 14, 1944 No. 15. Kauffmann noted cross reactions between types 10 and 71 but none of the type 10 antiserums employed in a following study caused capsular swelling. See F. Kauffmann, E. Morch, K. Schmith. On the serology of the pneumococcus group. J Immunol 39 (1940) 397-426; Public Health Reports Vol.59 April 14, 1944 No.15. [0395] Geno et al. reported that Spn type 25F CPS is composed of Glc, Rha, GlcN, Rib, and Rib-ol-P constituents, while no information is reported for Spn type 25A CPS. See K.A. Geno, G.L. Gilbert, J.Y. Song, I.C. Skovsted, K.P. Klugman, C. Jones, H.B. Konradsen, M.H. Nahm. Pneumococcal Capsules and Their Types: Past, Present, and Future. Clin Microbiol Rev 28 (2015) 871-899. However, further investigation to interrogate cross reactivity and genomic similarities might be useful to better correlate Spn type 38 with other Spn types. Experimental [0396] Purified Spn type 38 CPS was purchased from ATCC (www.atcc.org). For most of the NMR studies the CPS was treated with nitrous acid to remove C-polysaccharide contamination using the following procedure: to the solution of Spn type 38 CPS (10 mg/ml) in 5% of acetic acid, equal volume of 5% of sodium nitrite was added, the mixture was stirred for 4 hours at room temperature and further purified by tangential flow filtration. Monosaccharide composition by HPAEC-PAD and ESI-Q-TOF MS [0397] Spn type 38 CPS before and after NaBD4 treatment, and the monosaccharide standards were subjected to hydrolysis in 2 M TFA at 120°C for 2 hours. Following hydrolysis, the AFV70463FF hydrolyzates were evaporated to dryness, reconstituted in 1 mL of water and diluted prior to HPAEC-PAD analyses. Dionex ICS-3000 system equipped with a CarboPac PA1 analytical column (4 x 250 mm) and CarboPac PA1 guard (4 x 50 mm) was used as reported previously (see Z. Zhang, N.M. Khan, K.M. Nunez, E.K. Chess, C.M. Szabo. Complete monosaccharide analysis by high performance anion-exchange chromatography with pulsed amperometric detection. Anal Chem 84 (2012) 4104-4110. Linkage analysis by PMAA followed by GC/MS analysis [0398] Spn type 38 CPS before and after NaBD4 treatment were permethylated with methyl iodide in dimethyl sulfoxide containing an excess of sodium methylsulfinyl-methanide according to the Hakomori procedure (see S. Hakomori. A rapid permethylation of glycolipid, and polysaccharide catalyzed by methylsulfinyl carbanion in dimethyl sulfoxide. Biochem 55 (1964) 205-208). The methylated products were hydrolyzed, reduced with sodium borodeuteride, and O- acetylated using acetic anhydride in presence of TFA as previously described to give alditol acetates (see J. C. Richards, M. B. Perry, P. J. Kniskern. Structural analysis of the specific polysaccharide of Streptococcus pneumoniae type 9L (American type 49) Can J Biochem Cell Biol 62 (1984) 1309-1320). For identification of amino sugars, the reduced product was subjected to N-acetylation using acetic anhydride and triethylamine before O-acetylation. For identification uronic acid residues, the reduced product was subjected to methanolysis prior to O-acetylation. NMR Spectroscopy [0399] Spn type 38 CPS analytical sample was prepared by dissolving ~3 mg of powder in 600 μL of D₂O (Sigma-Aldrich) and introduced into a 5 mm NMR tube (Wilmad) for data acquisition. Spn type 38 CPS (~3 mg of powder in 600 μL of D₂O) was de-O-acetylated by addition of NaOD (200 mM as final concentration) and incubation at room temperature for 10 min. The sample was then neutralized by addition of DCl. [0400] All the 1D and 2D homo- and hetero-nuclear ¹H, ¹³C, and ³¹P NMR spectra were recorded at 323 0.1 K by a Bruker Avance III 600 MHz spectrometer equipped with a high precision temperature controller using a 5-mm broadband probe (Bruker) and processed using Topspin 3.2 software (Bruker). [0401] All the 1D and 2D ¹H, ¹³C, and ³¹P NMR spectra were recorded by standard Bruker pulse programs with a spectral width of 20 ppm, 200 ppm, and 30 ppm, respectively.1D and 2D TOCSY experiments were performed using mixing times of 120 ms and 180 ms, respectively.1D AFV70463FF and 2D NOESY experiments were performed using mixing times of 200 ms and 300 ms, respectively. 2D ¹H-¹³C HSQC experiment was performed considering a J ¼ of 145 Hz and the HMBC experiment optimized for a coupling constant of 6 Hz. 2D HSQC-TOCSY and HSQC- NOESY experiments were recorded using mixing times of 180 ms and 300 ms, respectively. [0402] Polysaccharide spectra were referenced to the residual phosphocholine cell wall polysaccharide signals (¹H signal at 3.23 ppm, ¹³C signal at 54.5, ³¹P signal of -D-GalpNAc residue at 0.22 ppm). Abbreviations [0403] Gal: Galactose; GalA: Galacturonic acid; Sug: 2-acetamido-2,6-dideoxy-xylo-hexos- 4-ulose; GlcNAc: N-acetyl-glucosamine; FucNAc: N-acetyl-fucosamine; NMR: Nuclear Magnetic Resonance; TFA: Trifluoroacetic acid; HPAEC-PAD: High Performance Anionic Exchange Chromatography-Pulsed Amperometric Detection. Example 2: Preparation of Exemplary Fusion Proteins CP1 and SPP2 Vector Constructions [0404] In some embodiments, fusion proteins CP1 and SPP2 each comprise a biotin-binding portion of rhizavidin spanning amino acids 45 to 179 of the full-length protein, wherein the predicted signal sequences (amino acids 1-44) of rhizavidin were not incorporated. To optimize the expression level of CP1 in E. coli, in some embodiments, the gene sequence that encodes rhizavidin polypeptide (e.g., in some embodiments, amino acids 45-179; SEQ ID NO: 2) was redesigned using E. coli-preferred expression codons, synthesized and cloned into plasmid pET24a(+). This synthetic rhizavidin gene was designated Rhavi. [0405] To construct the fusion proteins CP1 and SPP2, in some embodiments, a DNA sequence encoding a flexible linker (e.g, in some embodiments, a flexible linker of GGGGSSS; SEQ ID NO: 30) was directly inserted into the 3’ end of the synthetic Rhavi gene to provide separation from Rhavi and promote proper folding of the subsequent fusion protein. [0406] For CP1, the genes encoding desired portions of SP1500 and SP0785 protein (not including predicted signal sequences; nucleic acid sequences encoding the polypeptides of SEQ ID NO: 13 and SEQ ID NO: 11, respectively) were synthesized and inserted into the Rhavi expression vector just beyond the linker region. Three residual amino acids (AAA) from a Not I AFV70463FF restriction site of pET24a(+) separate SP1500 and SP0785, and may be referred to as the second linker of CP1. Stop codons were included at the 3’-end of the CP1 coding sequences. After cloning was complete, DNA sequencing of the pET-24a(+):CP1 plasmid was performed to confirm the presence, orientation and sequence of the CP1 DNA insert in the pET-24a(+) vector. A schematic of fusion protein CP1 is shown in FIG.16. [0407] For SPP2, a DNA sequence encoding an exemplary fusion protein SPP2 (Rhavi- GGGGSSS-PdT(G294P)-GGGGSSS-SP0435; e.g., SEQ ID NO: 26 or SEQ ID NO: 27) was cloned into the pET-24a(+) vector using PCR cloning. Stop codons were included at the 3’-end of the coding sequences. After cloning was complete, DNA sequencing of the pET-24a(+):SPP2 plasmid was performed to confirm the presence, orientation and sequence of the SPP2 DNA insert in the pET-24a(+) vector. A schematic of fusion protein SPP2 is shown in FIG.17. The map of plasmid pET24a(+):SPP2 is shown in FIG.18. Establishment of Master Cell Banks (MCB) [0408] To generate the Research Cell Banks (RCB), the pET-24a(+):CP1 and pET24a(+):SPP2 plasmids were transformed into an E. coli expression strain. Transformed E. coli cells were plated in culture plates containing Kanamycin. A single colony from the plate was selected and used to inoculate into liquid medium in a shaker flask. The flasks were placed in an incubator shaker for overnight culture and grown to the desired OD. The bacterial culture was then mixed with glycerol solution. The mixed solution was aliquoted into vials to make RCB. [0409] To generate the MCB, RCB was inoculated into liquid medium in shaker flasks and grown to the desired OD. The bacterial culture was then brought to 15% glycerol, mixed and aliquoted into vials. The MCB vials were then placed at -80°C (-70°C to -90°C) and selected vials were taken for quality assurance testing. Manufacturing Process Overview [0410] The CP1 and SPP2 fusion proteins were expressed in E. coli. The expressed CP1 and SPP2 fusion proteins were released from E. coli cells and purified in a series of chromatographic and filtration steps. The following section describes the 300 L culture scale CP1 manufacturing process. [0411] The process was initiated by thawing and inoculating cells from a MCB vial into cell medium. Initial cell expansion was performed in flask and then the bacterial culture was AFV70463FF transferred to a 300 L fermenter. Bacteria were harvested by centrifugation. The recovered cell paste was resuspended in a lysis buffer and fluidized by a microfluidizer. Bulk CP1 fusion protein in the fluidized cell lysate was purified by precipitation and chromatography steps. The final process stream was concentration and buffer exchange into 20 mM Tris, 150 mM NaCl at pH 8.0 ± 0.1 via ultrafiltration membranes. Lastly, the CP1 fusion protein was 0.22 m filtered immediately prior to bottling and stored at -80 °C. No raw materials contained animal-derived or human-derived components. Upstream Process [0412] A flow chart of a representative upstream manufacturing process for fusion proteins CP1 and SPP2 is provided in FIG.19, and details for the individual steps follow. [0413] The cell culture utilized chemically defined media and components depending on the manufacturing step. The media used were cell media, production media, feed 1 media and feed 2 media. These media were used in inoculum expansion, production culture, and feeding, respectively. [0414] Step 1 – 1: Thawing of Cells [0415] Sufficient cell medium was prepared and transferred into an appropriate culture flask and then frozen MCB is thawed. [0416] Step 1 – 2: Flask Expansion [0417] Thawed MCB was inoculated and grown in a 2.5 L flask. Cell growth was monitored by a spectrophotometer at OD600. [0418] Step 1 – 3: Production Culture [0419] The production culture was run in fed-batch mode. To support product generation and to prolong the cell culture production period, feed 1 and feed 2 media were added after appropriate _{cell growth. Once the fermenter temperature reached the set point, isop -D-1-} thiogalactopyranoside was added to induce fusion protein expression. After approximately 18 hours from induction, harvest was begun by centrifugation. [0420] Step 1 – 4: Harvest [0421] Bacterial culture was harvested by continuous flow centrifugation. AFV70463FF [0422] Step 1 – 5: Cell Lysis and Clarification [0423] The recovered cell paste was resuspended in lysis buffer and fluidized by a microfluidizer. The fluidized cell lysate, including bulk fusion protein, was centrifuged and the supernatant was recovered. [0424] Alternative upstream processes may be employed to manufacture the CP1 and SPP2 fusion proteins. Downstream Process [0425] After harvest and cell lysis, fusion proteins CP1 and SPP2 were purified by several chromatography steps. A flow chart of a representative fusion protein purification process is provided in FIG.20. Details for the individual steps follow. [0426] Step 2 – 1: Ammonium Sulfate Precipitation/Centrifugation and Filtration [0427] Ammonium sulfate solution was added to the clarified lysate and adjusted to a final concentration of 0.8 M. After mixing at room temperature, the clarified lysate with ammonium sulfate was centrifuged and the supernatant was recovered. The recovered supernatant was filtered through 0.22 m filter and collected in an appropriate container. [0428] Step 2 – 2: Hydrophobic Interaction Chromatography [0429] The purpose of this step is to remove process-related contaminants. Fusion protein bound to the resin was washed with high concentration ammonium sulfate buffer then eluted with elution buffer (the concentration of NaCl in the buffer was gradually reduced). A sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis was performed with each fraction to confirm the presence of fusion protein. [0430] Step 2 – 3: Tangential Flow Filtration Concentration/Filtration [0431] The elution pool was concentrated and then buffer exchanged with several-fold volumes of Tris-buffered saline using a 30 kDa filtration membrane. The filtrated tangential flow filtration (TFF) retentate was filtered by 0.22 m filter. [0432] Step 2 – 4: Anion Exchange Chromatography [0433] The purpose of this step is to remove process-related contaminants. Fusion protein bound to the resin was washed with NaCl wash buffer then eluted with NaCl elution buffer (the concentration of NaCl in the buffer was gradually increased). SDS-PAGE analysis was performed with each fraction to confirm the presence of fusion protein. AFV70463FF [0434] Step 2 – 5: Type 1 Chromatography [0435] The purpose of this step is to further remove process-related contaminants. Fusion protein bound to the resin was washed with phosphate wash buffer then eluted with phosphate elution buffer (the concentration of phosphate in the buffer was gradually increased). SDS-PAGE analysis was performed with each fraction to confirm the presence of fusion protein. [0436] Step 2 – 6: TFF Formulation [0437] The Type 1 chromatography fraction pool was concentrated and then buffer exchanged with several-fold volumes of formulation buffer (20 mM Tris, 150 mM sodium chloride, pH 8.0 ± 0.1) using a 30 kDa filtration membrane. The formulated fusion protein was filtered through 0.22 m filter. [0438] Alternative downstream processes may be employed to manufacture the CP1 and SPP2 fusion proteins. Container Closure System [0439] CP1 and SPP2 fusion proteins were stored at -80°C (-70°C to -90°C) in sterile polyethylene terephthalate bottles. Example 3: S. pneumoniae Capsular Polysaccharides (PS) and Preparations of the Same Preparation and Purification [0440] Methods of culturing pneumococci are well known in the art (e.g. Chase, 1967, Methods of Immunology and Immunochemistry 1:52). Methods of preparing pneumococcal capsular polysaccharides are also well known in the art (e.g., European Patent No. EP0497524). Isolates of pneumococcal serotypes are available from the ATCC, the National Collection of Type Cultures operated by Public Health England, and other repositories. [0441] S. pneumoniae is identified as non-motile, Gram-positive, lancet-shaped diplococci that are alpha-hemolytic on blood agar. Most but not all strains are encapsulated. Serotypes are differentiated on the basis of Neufeld Test (Quelling reaction) using specific antisera (e.g., U.S. Pat. No.5,847,112), latex agglutination, or multilocus sequence typing. [0442] A frozen vial representing each of the S. pneumoniae serotypes present in MAPS vaccine candidates of the present disclosure was thawed and used to generate a seed culture in AFV70463FF appropriate pre-sterilized growth media. The seed culture was grown with temperature and pH control. The seed culture was transferred to a production fermenter that contained pre-sterilized growth media. The production culture was grown with temperature, pH and agitation rate control. The growth process was terminated with addition of an inactivating agent with a controlled temperature hold. [0443] The purification process was initiated by removal of cell debris using a combination of centrifugation and filtration. The material was filtered followed by solvent-based fractionations to remove impurities and recover PS. Exemplary polysaccharide structures [0444] Representative polysaccharide (PS) structures are shown in FIG. 21. All PS specifications in Table 3 were obtained from European Pharmacopoeia 9.0 Table 0966.-1, except for molecular size and serotype 6A, which are based on the manufacturer’s certificate of analysis. Table 3. Exemplary Polysaccharide (PS) Specifications Molecular size Molecular ^Protein Nucleic Total g_en ^Phosp _(KD) ^Uronic Hexos- Methyl- O-acetyl t_ype† acids _nitro ^horus acids amines pentoses Groups § 1 2 2 3.5-6 0-1.5 0.15 45 1.8 2 2 2 0-1 0-1.0 0.15 15 38 3 5 2 0-1 0-1.0 0.15 40 4 3 2 4-6 0-1.5 0.15 40 5 7.5 2 2.5-6.0 2 0.60 12 20 6A 1.5 1.5 0-2 2.5-5.0 0.50 6B 2 2 0-2 2.5-5.0 0.50 6C 3 2 7_{C 3 2} 7F 5 2 1.5-4.0 0-1.0 0.20 8 2 2 0-1 0-1.0 0.15 25 9N 2 1 2.2-4 0-1.0 0.20 20 28 9V 2 2 0.5-3 0-1.0 0.45 15 13 10A 7 2 0.5-3.5 1.5-3.5 0.65 12 11A 3 2 0-2.5 2.0-5.0 0.40 9 12F 3 2 3-5 0-1.0 0.25 25 AFV70463FF Molecular size Molecular ^Protein Nucleic Total _(KD) ^Uronic Hexos- Methyl- O-acetyl t_ype† acids _nitrogen ^Phosphorus acids amines pentoses Groups § 14 5 2 1.5-4 0-1.0 0.30 20 1_{5A 3 2} 15B 3 2 1-3 2.0-4.5 0.55 15 1_{6F 3 2} 17F 2 2 0-1.5 0-3.5 0.45 20 18C 3 2 0-1 2.4-4.9 0.15 14 19A 2 2 0.6-3.5 3.0-7.0 0.45 12 20 19F 3 2 1.4-3.5 3.0-5.5 0.20 12.5 20 20 2 2 0.5-2.5 1.5-4.0 0.60 12 22F 0-2 0-1.0 2_{3A 3 2} _{23B 3 2} 23F 0-1 3.0-4.5 2_{4F 3 2} 31 33F 0-2 0-1.0 35B 3 2 38 3 2 Values shown are percentage contents of components of monovalent bulk PS. † The different types are indicated using the Danish nomenclature. Cross-linked agarose for chromatography R. § Cross-linked agarose for chromatography R1. Example 4: Preparation of MAPS Immunogenic Complexes Overview [0445] The MAPS platform provides various advantages including, e.g., high affinity -¹⁵M), non-covalent binding between biotin and rhizavidin, a biotin-binding protein that has no significant predicted homology with human proteins. Rhizavidin, a naturally occurring dimeric protein in the avidin protein family, was first discovered in Rhizobium etli, a symbiotic bacterium of the common bean. Rhizavidin has only 22% amino acid identity with chicken avidin, a protein commonly found in eggs, but with high conservation AFV70463FF of amino acid residues involved in biotin binding. No cross-reactivity to rhizavidin was observed in human serum samples obtained from subjects exposed to avidin [Helppolainen et al, 2007], suggesting that rhizavidin antibodies may not cross-react with chicken avidin. Biotin conjugates have been used in several clinical applications without any reported adverse events [Buller et al, 2014; Paty et al, 2010; Lazzeri et al, 2004]. The biotinylation of the PS and MAPS immunogenic complexing process in MAPS24 were optimized to consistently show no free biotin, thus reducing the potential for generating anti-biotin antibodies. [0446] MAPS vaccine candidates comprise genetically constructed fusion proteins of a biotin- binding moiety (e.g., rhizavidin, or a biotin-binding domain or biotin-binding fragment thereof), and a protein antigens of interest, which are then complexed with biotinylated PS of interest, leading to specific assembly into integrated macromolecular immunogenic complexes that, when processed by the immune system, result in the activation of protective B- and T-cell immune responses, as shown schematically in FIG.22. [0447] MAPS34 is a pneumococcal vaccine candidates based on the proprietary MAPS platform. MAPS34 is a 34-valent MAPS vaccine candidate comprising 34 pneumococcal capsular polysaccharides of serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 35B, and 38, individually biotinylated and complexed with either SPP2 or CP1 fusion protein. MAPS vaccine candidates comprising at least 30 (e.g., 30 [MAPS30], 31 [MAPS31], 32 [MAPS32], or 33 [MAPS33]) of the same 34 pneumococcal capsular polysaccharides as in MAPS34 are also contemplated. MAPS30+ refers to any one of MAPS30, MAPS31, MAPS32, MAPS33 or MAPS34 pneumococcal vaccine candidates. [0448] In some embodiments, an exemplary SPP2 comprises a genetic fusion construct of truncated rhizavidin plus pneumococcal proteins pneumolysin (with a combination of four amino acid substitutions as described herein, denoted PdT(G294P)) and SP0435, joined by linkers, as shown in FIG.17. In some embodiments, an exemplary CP1 comprises a genetic fusion construct of truncated rhizavidin plus pneumococcal proteins SP1500 and SP0785 joined by linkers, as shown in FIG.16. No available vaccines contain these unique combinations of PS and polypeptide antigens which are capable of eliciting an immune response against such a broad range of pneumococcal serotypes. AFV70463FF Assembly of MAPS Immunogenic Complexes [0449] Drug substance (MAPS immunogenic complexes) comprises PS and CP1 and SPP2 fusion protein. In some embodiments, since the PS are biotinylated and CP1 and SPP2 fusion protein have a rhizavidin biotin-binding domain, they are linked by the high affinity biotin- rhizavidin interaction. MAPS immunogenic complexation was conducted individually for each PS serotype. [0450] In some embodiments, each MAPS immunogenic complex was made from 2 key intermediates: PS of each of up to 34 S. pneumoniae serotypes, and rhizavidin fusion proteins CP1 or SPP2. PS of each serotype were activated, for example, by creation of a cyanate ester and then biotinylated. The biotinylated PS were mixed with CP1 or SPP2 fusion protein to create MAPS immunogenic complexes of defined PS serotype and fusion protein, linked by the high affinity biotin-rhizavidin interaction. MAPS immunogenic complexes of defined PS serotype and fusion protein are referred to as a species. [0451] MAPS immunogenic complexes of each species were formulated with 150 mM sodium chloride and surfactant buffer, then 0.2 m filtered immediately prior to bottling and storing at 2°C to 8°C. Formulated MAPS immunogenic complexes of each species are referred to as MAPS drug substance. [0452] No raw materials contained animal-derived or human-derived components. [0453] A flow chart of a representative MAPS complexation process is provided in FIG.23, and details for the individual steps follow. [0454] Step 1: Clean-up of Polysaccharide [0455] The purpose of this step is to remove process residuals. Dissolved PS was purified by filtration then exchanged with several-fold volumes of water for injection, followed by ultra- filtration to concentrate the PS. The cleaned-up PS was filtered by a 0.22 m filter membrane. [0456] Step 2: Biotinylation and Purification of Polysaccharide [0457] The hydroxyl group on the PS was activated with a 1-cyano-4-dimetylamino- pyridinium tetrafluoroborate (CDAP) to create a highly active cyanoester. The cyanoesters were reacted with amine-PEG3-biotin and unreacted cyanoesters were capped with glycine. The biotinylated PS was buffer exchanged into 1 mM PBS, 150mM sodium chloride in order to remove unreacted amine-PEG3-biotin, glycine, hydrolyzed CDAP and residue. After buffer exchange, the biotinylated PS was filtered with a 0.22 m filter. AFV70463FF [0458] Step 3: MAPS immunogenic complexation [0459] The biotinylated PS was mixed with CP1 or SPP2 fusion protein in order to create the desired species of MAPS immunogenic complexes, linked by a high affinity biotin-rhizavidin interaction. The MAPS immunogenic complexes were purified to remove uncomplexed PS and protein by using a filtration membrane. After purification, the MAPS immunogenic complexes were filtered with a 0.22 m filter and stored at 2°C to 8°C. Specifications [0460] Exemplary MAPS immunogenic complex/MAPS drug substance specifications are set forth in Table 4. Table 4. Exemplary MAPS Drug Substance Specifications PS Serotype (1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 33F, 35B, and 38) Quality Attribute Acceptance Criteria Appearance Clear, colorless, no particulate matter pH 5 – 7.5 PS Identity Positive PS Concentration > 0.25 mg/mL Protein Concentration FIO Free Protein FIO Free PS < 30% Endotoxin < 1.25x10^-2 EU/μg PS Bioburden FIO: for information only; MAPS: multiple antigen-presenting system; PS: polysaccharide. AFV70463FF Example 5: MAPS34 Vaccine – Exemplary MAPS30+ Vaccine Drug Product [0461] MAPS34 is a 34-valent MAPS vaccine candidate comprising 34 pneumococcal capsular polysaccharides of serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 35B, and 38, individually biotinylated and complexed with either SPP2 or CP1 fusion protein. MAPS vaccine candidates comprising at least 30 (e.g., 30 [MAPS30], 31 [MAPS31], 32 [MAPS32], 33 [MAPS33]) of the same 34 pneumococcal capsular polysaccharides as in MAPS34 are also contemplated. MAPS30+ refers to any one of MAPS30, MAPS31, MAPS32, MAPS33, or MAPS34 pneumococcal vaccine candidates. [0462] In a representative formulation, a MAPS34 vaccine candidate is formulated so that each 0.5-mL dose of MAPS34 drug product comprises 1, 2, 5, or 6 each of S. pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 17F, 18C, 19A, 19F, 20B, 22F, 23A, 23B, 23F, 24F, 31, 35B, and 38, contributed by each species of MAPS immunogenic complex. MAPS34 is formulated for IM administration with aluminum phosphate adjuvant (Adju-Phos, from Croda Denmark, formerly Brenntag Biosector). In some embodiments, the total amount of aluminum per dose is 0.625 mg, which is below the FDA/WHO maximum recommended dose of 0.85 mg to 1.25 mg. In some embodiments, the total amount of aluminum per dose is less than 0.625 mg. In some embodiments, the total amount of aluminum per dose is between 0.325 mg and 0.175 mg. In some embodiments, the total amount of aluminum per dose is 0.25 mg. In some embodiments, the total amount of aluminum per dose is at least 0.125 mg. FIG.24 shows a representative scheme for MAPS34 drug product manufacturing. Example 6 – Conjugation of PS38 to CRM197 [0463] Polysaccharide (PS) 38 was conjugated to CRM-197 (Cross-Reacting Material, Fina Biosolutions) using 1-cyano-4-dimethylamino-pyridinium tetrafluoroborate (CDAP) chemistry to form PS38-CRM-197 conjugates. Under alkaline conditions, CDAP reacts with hydroxyl groups on the cleaned-up polysaccharides to create a highly reactive cyanoester. When the CRM-197 protein is added, its terminal primary amine reacts with the cyanoester and covalently links the AFV70463FF PS38 and CRM. The CRM-conjugated PS38 was internally evaluated to assess protein loading, free protein and PS content prior to immunization. Example 7 – Immunogenicity results achieved against PS38 MAPS complexes in the context of a multivalent MAPS Vaccine [0464] PS38 MAPS complexes using either CP1 or SPP2 as fusion protein where combined with further S. pneumoniae serotype MAPS complexes and tested for immunogenicity in a rabbit _{model. Groups of 20 New Zealand White rabbit were immunized with 0.2 g of each} _{polysaccharide (representing 1/25 of a human dose of 5 g), adjuvanted with 5 g of} Aluminium (AlPO4 – 1/25 of human dose), using a schedule of administration on days 0 and 14 with blood taken on days 0, 14 and 28. Sera were collected from male NZW rabbits (n=20) before the first does (P0), second dose (P1) and two weeks after the second dose (P2). Anti-ST-38 specific IgG concentrations (ng/mL) were determined by comparing samples to a reference standard of known concentration. IgG Antibody Responses to S. Pneumoniae CPS-38 [0465] Total serotype-specific polysaccharide antibodies was measured by electrochemiluminescence. Quantification of total serotype-specific antibodies was performed using a Meso Scale Discovery (MSD) electrochemiluminescence (ECL) immunoassay. Using this system, the concentration of each serotype-specific antibody was interpolated from a reference standard of known concentrations. Quantification of antibody concentrations [0466] U-plex plates were coated with 10 polysaccharides with different linkers overnight and stored at 2-8°C. After coating, the U-plex plates were blocked with a blocking buffer (1% Casein in 1× Phosphate buffered saline [PBS]) and incubated with shaking at room temperature. Following blocking, the U-plex plates were washed with 1×DPBST, and reference standards, blank, Quality Control samples, and experimental test samples were added to the plates, and incubated with shaking at room temperature for 1 hour. Next, the plates were again washed with 1×DPBST and secondary anti-rabbit SULFO-TAG conjugated antibody was added for another hour incubation with shaking at room temperature. The plates AFV70463FF were washed one last time, and 1×Read Buffer was added onto the plates, which were then immediately read on a MESO™ QuickPlex SQ 120 (Model No. 1300). The resulting ECL signal was proportional to the concentrations of each serotype-specific polysaccharide antibodies. Therefore, the antibody concentrations of the quality controls and experimental test sera can be interpolated from the ECL signals using the series of reference standards of known concentrations. Individual reference standard curves were automatically created by the MSD software (Discovery Workbench version 4.0) and used to determine the concentration of each serotype-specific (including ST-38) polysaccharide antibody in QCs and test sera. [0467] The results are shown in Figure 27 which demonstrates that a good immune response was generated against CPS-38 in a multivalent MAPS technology composition. Figure 27 shows the geometric mean concentrations (GMCs) of the groups and the upper interval (UI) and lower interval (LI) of the 95% Cis. Example 8 – Immunogenicity results achieved against a PS38-CRM197 conjugate Rabbit Immunization Strategy [0468] Antibodies against polysaccharides are generated by immunizing 10 Female New Zealand White Rabbits subcutaneously three times with conjugated polysaccharide 38-CRM197 (25μg dose by PS content). The first injection used Freund’s Complete Adjuvant (FCA), and the 2 booster injections used Freund’s Incomplete Adjuvant (FIA). The booster injections occur every 21 days, and the production bleeds were collected 10 days after the second and third injections (days 31 and 52). Production bleeds were screened prior to the next boost to determine titer and if the study should proceed with the next injections. Rate Nephelometry Assessment [0469] Rate nephelometry is a light scattering based method that measures the rate of formation of antibody and antigen complex. Briefly, in this assessment monovalent reference standards using cleaned native polysaccharide of serotype 38 were prepared and mixed with a pool of antisera from rabbits immunized the CRM-conjugated PS38. As the titers of antibody against type 38 increase due to immunizations the instrument response to the PS will increase. Pools of naïve sera, post-production bleed 1 (P1; post two doses), and post-production bleed 2 (P2; post 3 doses) were evaluated against 5 concentrations of native PS. The intensity of the instrument response (IR) and linearity of the reference curves were evaluated to determine whether sufficient AFV70463FF antibody titers had been achieved. As the rabbits were subjected to the second and third doses of PS38-CRM197, there was a robust increase in the instrument response relative to naïve rabbits as shown in FIG.25 and Table 5. Naïve rabbits (P0,n=4) were not part of the original study. P1 and P2 are a pool of n=10 rabbits (306-315). Antibody titers evaluated by top IR and linearity.5-point reference standard used. Table 5. Nephelometry results showing a robust immune response Reference Standard (μg/ml) P0 (IR) P1 (IR) P2 (IR) 0.988 3.64 32.39 53.26 1.48 5.67 46.13 68.36 2.22 3.49 60.76 94.90 3.33 7.90 88.68 143.06 5.00 17.61 134.32 222.24 Weight Based Concentration of Sera by ELISA [0470] In addition to the nephelometry analysis, the anti-PS38 IgG concentrations were determined by ELISA and results are shown in Table 6 and Figure 26. Individual concentration for each of the 10 rabbits (306 to 315) were tested for IgG conc at post 2 (P1) and post 3 (P2) immunizations. Naïve sera were also evaluated (n=4) and results for this group were below the LLOQ of the method. First, the CRM-38 purified IgG was evaluated to assign a weight-based concentration to the IgG. This purified CRM-38 IgG was used as the reference standard to evaluate the naïve sera, P1, and P2 sera. The ELISA plate was coated with 30 μg/mL of PS38 (antigen), and the rabbit sera was the analyte. For the reference standard curve and test article the secondary antibody was Goat anti-rabbit Fc-HRP (IgG anti-Fc domain). Table 6: ELISA results showing robust immune responses against PS38-CRM197 Rabbit P1-38 IgG Conc. P2-38 IgG Conc. Number (μg/mL) (μg/mL) AFV70463FF 306 505.6 1134.3 307 228.5 833.7 308 165.9 331.0 309 122.2 429.2 310 121.8 801.6 311 104.5 358.6 312 67.4 312.7 313 937.3 2508.9 314 1250.5 5186.1 315 724.6 2020.0 Geometric Mean 265.0 733.1

AFV70463FF SEQUENCE LISTING [0471] SEQ ID NO: 1, Rhizavidin protein, full-length [aa 1-179]: MIITSLYATFGTIADGRRTSGGKTMIRTNAVAALVFAVATSALAFDASNFKDFSSIASASS SWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLTGRVNGTFIAFSVGWNNS TENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQGQDTFQYVPTTENKSLL KD [0472] SEQ ID NO: 2, truncated rhizavidin protein [aa 45-179], denoted Rhavi: FDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLTGR VNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQGQ DTFQYVPTTENKSLLKD [0473] SEQ ID NO: 3, truncated rhizavidin protein [aa 45-179], denoted Rhavi, includes leading methionine: MFDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLT GRVNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQ GQDTFQYVPTTENKSLLKD [0474] SEQ ID NO: 4, Streptococcus pneumoniae Pneumolysin protein: MANKAVNDFILAMNYDKKKLLTHQGESIENRFIKEGNQLPDEFVVIERKKRSLSTNTSDI SVTATNDSRLYPGALLVVDETLLENNPTLLAVDRAPMTYSIDLPGLASSDSFLQVEDPSN SSVRGAVNDLLAKWHQDYGQVNNVPARMQYEKITAHSMEQLKVKFGSDFEKTGNSLD IDFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPGDVFQDTVTVEDLKQRGISAERPLVYI SSVAYGRQVYLKLETTSKSDEVEAAFEALIKGVKVAPQTEWKQILDNTEVKAVILGGDP SSGARVVTGKVDMVEDLIQEGSRFTADHPGLPISYTTSFLRDNVVATFQNSTDYVETKV TAYRNGDLLLDHSGAYVAQYYITWDELSYDHQGKEVLTPKAWDRNGQDLTAHFTTSIP LKGNVRNLSVKIRECTGLAWEWWRTVYEKTDLPLVRKRTISIWGTTLYPQVEDKVEND [0475] SEQ ID NO: 5, Streptococcus pneumoniae Pneumolysin protein with mutations D385N, C428G, and W433F, denoted PdT: MANKAVNDFILAMNYDKKKLLTHQGESIENRFIKEGNQLPDEFVVIERKKRSLSTNTSDI SVTATNDSRLYPGALLVVDETLLENNPTLLAVDRAPMTYSIDLPGLASSDSFLQVEDPSN AFV70463FF SSVRGAVNDLLAKWHQDYGQVNNVPARMQYEKITAHSMEQLKVKFGSDFEKTGNSLD IDFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPGDVFQDTVTVEDLKQRGISAERPLVYI SSVAYGRQVYLKLETTSKSDEVEAAFEALIKGVKVAPQTEWKQILDNTEVKAVILGGDP SSGARVVTGKVDMVEDLIQEGSRFTADHPGLPISYTTSFLRDNVVATFQNSTDYVETKV TAYRNGDLLLDHSGAYVAQYYITWDELSYNHQGKEVLTPKAWDRNGQDLTAHFTTSIP LKGNVRNLSVKIREGTGLAFEWWRTVYEKTDLPLVRKRTISIWGTTLYPQVEDKVEND [0476] SEQ ID NO: 6, Streptococcus pneumoniae Pneumolysin protein with mutations G294P, D385N, C428G, and W433F, denoted PdT(G294P): MANKAVNDFILAMNYDKKKLLTHQGESIENRFIKEGNQLPDEFVVIERKKRSLSTNTSDI SVTATNDSRLYPGALLVVDETLLENNPTLLAVDRAPMTYSIDLPGLASSDSFLQVEDPSN SSVRGAVNDLLAKWHQDYGQVNNVPARMQYEKITAHSMEQLKVKFGSDFEKTGNSLD IDFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPGDVFQDTVTVEDLKQRGISAERPLVYI SSVAYGRQVYLKLETTSKSDEVEAAFEALIKGVKVAPQTEWKQILDNTEVKAVILGPDP SSGARVVTGKVDMVEDLIQEGSRFTADHPGLPISYTTSFLRDNVVATFQNSTDYVETKV TAYRNGDLLLDHSGAYVAQYYITWDELSYNHQGKEVLTPKAWDRNGQDLTAHFTTSIP LKGNVRNLSVKIREGTGLAFEWWRTVYEKTDLPLVRKRTISIWGTTLYPQVEDKVEND [0477] SEQ ID NO: 7, Streptococcus pneumoniae Pneumolysin PdT(G294P) [aa 2-470] protein: ANKAVNDFILAMNYDKKKLLTHQGESIENRFIKEGNQLPDEFVVIERKKRSLSTNTSDIS VTATNDSRLYPGALLVVDETLLENNPTLLAVDRAPMTYSIDLPGLASSDSFLQVEDPSNS SVRGAVNDLLAKWHQDYGQVNNVPARMQYEKITAHSMEQLKVKFGSDFEKTGNSLDI DFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPGDVFQDTVTVEDLKQRGISAERPLVYIS SVAYGRQVYLKLETTSKSDEVEAAFEALIKGVKVAPQTEWKQILDNTEVKAVILGPDPS SGARVVTGKVDMVEDLIQEGSRFTADHPGLPISYTTSFLRDNVVATFQNSTDYVETKVT AYRNGDLLLDHSGAYVAQYYITWDELSYNHQGKEVLTPKAWDRNGQDLTAHFTTSIPL KGNVRNLSVKIREGTGLAFEWWRTVYEKTDLPLVRKRTISIWGTTLYPQVEDKVEND [0478] SEQ ID NO: 8, Streptococcus pneumoniae SP0435 protein, full length, TIGR4 strain (GenBank: ABJ54475.1): MIEASKLKAGMTFETADGKLIRVLEASHHKPGKGNTIMRMKLRDVRTGSTFDTSYRPEE KFEQAIIETVPAQYLYKMDDTAYFMNTETYDQYEIPVVNVENELLYILENSDVKIQFYGT AFV70463FF EVIGVTVPTTVELTVAETQPSIKGATVTGSGKPATMETGLVVNVPDFIEAGQKLVINTAE GTYVSRA [0479] SEQ ID NO: 9, Streptococcus pneumoniae SP0435 [aa 62-185] protein, TIGR4 strain: EQAIIETVPAQYLYKMDDTAYFMNTETYDQYEIPVVNVENELLYILENSDVKIQFYGTEV IGVTVPTTVELTVAETQPSIKGATVTGSGKPATMETGLVVNVPDFIEAGQKLVINTAEGT YVSR [0480] SEQ ID NO: 10, SP0785 protein, full-length [aa 1-399], TIGR4 strain: (Note: One T394A mismatch with SP0785 NCBI Sequences ABJ54007.1 and YP816180) MKKKNGKAKKWQLYAAIGAASVVVLGAGGILLFRQPSQTALKDEPTHLVVAKEGSVA SSVLLSGTVTAKNEQYVYFDASKGDLDEILVSVGDKVSEGQALVKYSSSEAQAAYDSA SRAVARADRHINELNQARNEAASAPAPQLPAPVGGEDATVQSPTPVAGNSVASIDAQLG DARDARADAAAQLSKAQSQLDATTVLSTLEGTVVEVNSNVSKSPTGASQVMVHIVSNE NLQVKGELSEYNLANLSVGQEVSFTSKVYPDKKWTGKLSYISDYPKNNGEAASPAAGN NTGSKYPYTIDVTGEVGDLKQGFSVNIEVKSKTKAILVPVSSLVMDDSKNYVWIVDEQQ KAKKVEVSLGNADAENQEITSGLTNGAKVISNPTSSLEEGKEVKADEATN [0481] SEQ ID NO: 11, SP0785 protein lacking signal sequence [aa 33-399]: (Note: One T394A mismatch with SP0785 NCBI Sequences ABJ54007.1 and YP816180) FRQPSQTALKDEPTHLVVAKEGSVASSVLLSGTVTAKNEQYVYFDASKGDLDEILVSVG DKVSEGQALVKYSSSEAQAAYDSASRAVARADRHINELNQARNEAASAPAPQLPAPVG GEDATVQSPTPVAGNSVASIDAQLGDARDARADAAAQLSKAQSQLDATTVLSTLEGTV VEVNSNVSKSPTGASQVMVHIVSNENLQVKGELSEYNLANLSVGQEVSFTSKVYPDKK WTGKLSYISDYPKNNGEAASPAAGNNTGSKYPYTIDVTGEVGDLKQGFSVNIEVKSKTK AILVPVSSLVMDDSKNYVWIVDEQQKAKKVEVSLGNADAENQEITSGLTNGAKVISNPT SSLEEGKEVKADEATN [0482] SEQ ID NO: 12, SP1500 protein, full-length [aa 1-278], TIGR4 strain: MKKWMLVLVSLMTALFLVACGKNSSETSGDNWSKYQSNKSITIGFDSTFVPMGFAQKD GSYAGFDIDLATAVFEKYGITVNWQPIDWDLKEAELTKGTIDLIWNGYSATDERREKVA FSNSYMKNEQVLVTKKSSGITTAKDMTGKTLGAQAGSSGYADFEANPEILKNIVANKEA AFV70463FF NQYQTFNEALIDLKNDRIDGLLIDRVYANYYLEAEGVLNDYNVFTVGLETEAFAVGAR KEDTNLVKKINEAFSSLYKDGKFQEISQKWFGEDVATKEVKEGQ [0483] SEQ ID NO: 13, SP1500 [aa 27-278]: TSGDNWSKYQSNKSITIGFDSTFVPMGFAQKDGSYAGFDIDLATAVFEKYGITVNWQPI DWDLKEAELTKGTIDLIWNGYSATDERREKVAFSNSYMKNEQVLVTKKSSGITTAKDM TGKTLGAQAGSSGYADFEANPEILKNIVANKEANQYQTFNEALIDLKNDRIDGLLIDRVY ANYYLEAEGVLNDYNVFTVGLETEAFAVGARKEDTNLVKKINEAFSSLYKDGKFQEISQ KWFGEDVATKEVKEGQ [0484] SEQ ID NO: 14, Ply gene encoding Ply protein, full-length [aa 1-470] ATGGCAAATAAAGCAGTAAATGACTTTATACTAGCTATGAATTACGATAAAAAGAA ACTCTTGACCCATCAGGGAGAAAGTATTGAAAATCGTTTCATCAAAGAGGGTAATC AGCTACCCGATGAGTTTGTTGTTATCGAAAGAAAGAAGCGGAGCTTGTCGACAAAT ACAAGTGATATTTCTGTAACAGCTACCAACGACAGTCGCCTCTATCCTGGAGCACTT CTCGTAGTGGATGAGACCTTGTTAGAGAATAATCCCACTCTTCTTGCGGTCGATCGT GCTCCGATGACTTATAGTATTGATTTGCCTGGTTTGGCAAGTAGCGATAGCTTTCTCC AAGTGGAAGACCCCAGCAATTCAAGTGTTCGCGGAGCGGTAAACGATTTGTTGGCT AAGTGGCATCAAGATTATGGTCAGGTCAATAATGTCCCAGCTAGAATGCAGCATGA AAAAATCACGGCTCACAGCATGGAACAACTCAAGGTCAAGTTTGGTTCTGACTTTGA AAAGATAGGGAATTCTCTTGATATTGATTTTAACTCTGTCCATTCAGGCGAAAAGCA GATTCAGATTGTTAATTTTAAGCAGATTTATTATACAGTCAGCGTAGATGCTGTTAA AAATCCAGGAGATGTGTTTCAAGATACTGTAACGGTAGAGGATTTAAGGCAGAGAG GAATTTCTGCAGAGCGTCCTTTGGTCTATATTTCGAGTGTTGCTTATGGGCGCCAAGT CTATCTCAAGTTGGAAACCACGAGTAAGAGTGATGAAGTAGAGGCTGCTTTTGAATC TTTGATAAAAGGAGTAGCTCCTCAGACAGAGTGGAAGCAGATTTTGGACAATACAG AAGTGAAGGCGGTTATTTTAGGGGGCGACCCAAGTTCGGGTGCCCGAGTTGTAACA GGCAAGGTGGATATGGTAGAGGACTTGATTCAAGAAGGCAGTCGCTTTACAGCCGA TCATCCAGGCTTGCCGATTTCCTATACAACTTCTTTTTTACGTGACAATGTAGTTGCG ACCTTTCAAAACAGTACAGACTATGTTGAGACTAAGGTTACAGCTTACAGAAACGG AGATTTACTGCTGGATCATAGTGGTGCCTATGTTGCTCAATATTATATTACTTGGGAT GAATTATCCTATGATCATCAAGGCAAGGAAGTCTTGACTCCTAAGGCTTGGGACAGA AFV70463FF AATGGGCAGGATTTGACGGCTCACTTTACCACTAGTATTCCTTTAAAAGGGAATGTT CGCAATCTCTCTGTCAAAATTAGAGAGTGTACCGGGCTTGCCTGGGAATGGTGGCGT ACGGTTTATGAAAAAACCGATTTGCCACTAGTGCGTAAGCGGACGATTTCTATTTGG GGAACAACTCTCTATCCTCAGGTAGAGGATAAGGTAGAAAATGATTAG [0485] SEQ ID NO: 15, codon-optimized nucleotide sequence encoding PdT(G294P) [aa 1- 470]: ATGGCGAACAAGGCGGTGAACGATTTTATCCTGGCGATGAACTATGACAAGAAGAA ACTGCTGACCCACCAAGGCGAGAGCATTGAGAACCGTTTCATTAAAGAAGGCAACC AGCTGCCGGACGAGTTTGTGGTTATCGAGCGTAAGAAACGTAGCCTGAGCACCAAC ACCAGCGACATTAGCGTGACCGCGACCAACGATAGCCGTCTGTACCCGGGTGCGCT GCTGGTTGTGGATGAAACCCTGCTGGAAAACAACCCGACCCTGCTGGCGGTGGACC GTGCGCCGATGACCTATAGCATCGATCTGCCGGGTCTGGCGAGCAGCGACAGCTTCC TGCAAGTTGAGGATCCGAGCAACAGCAGCGTGCGTGGTGCGGTTAACGACCTGCTG GCGAAGTGGCACCAGGATTACGGCCAAGTGAACAACGTTCCGGCGCGTATGCAGTA TGAAAAAATCACCGCGCACAGCATGGAGCAACTGAAGGTTAAATTCGGTAGCGACT TTGAAAAGACCGGCAACAGCCTGGACATTGATTTCAACAGCGTGCACAGCGGCGAG AAGCAGATCCAAATCGTTAACTTCAAGCAGATCTACTACACCGTGAGCGTTGACGCG GTGAAGAACCCGGGTGACGTTTTCCAGGATACCGTGACCGTTGAAGATCTGAAACA ACGTGGCATTAGCGCGGAGCGTCCGCTGGTGTACATCAGCAGCGTTGCGTACGGTCG TCAAGTGTATCTGAAGCTGGAAACCACCAGCAAAAGCGATGAGGTTGAAGCGGCGT TTGAGGCGCTGATTAAGGGCGTGAAAGTTGCGCCGCAGACCGAATGGAAGCAAATT CTGGACAACACCGAGGTGAAAGCGGTTATTCTGGGCCCGGATCCGAGCAGCGGCGC GCGTGTGGTTACCGGTAAAGTGGACATGGTTGAGGATCTGATTCAGGAAGGTAGCC GTTTTACCGCGGACCACCCGGGCCTGCCGATCAGCTACACCACCAGCTTCCTGCGTG ACAACGTGGTTGCGACCTTTCAAAACAGCACCGATTACGTGGAAACCAAGGTTACC GCGTATCGTAACGGTGACCTGCTGCTGGACCACAGCGGTGCGTACGTGGCGCAGTA CTATATCACCTGGGATGAACTGAGCTATAACCACCAGGGTAAAGAGGTGCTGACCC CGAAAGCGTGGGACCGTAACGGCCAGGATCTGACCGCGCACTTCACCACCAGCATT CCGCTGAAGGGCAACGTGCGTAACCTGAGCGTTAAAATCCGTGAGGGTACCGGCCT GGCGTTTGAATGGTGGCGTACCGTGTACGAGAAGACCGACCTGCCGCTGGTTCGTAA AFV70463FF ACGTACCATCAGCATTTGGGGTACCACCCTGTATCCGCAGGTGGAGGACAAAGTTG AAAATGAT [0486] SEQ ID NO: 16, codon-optimized nucleotide sequence encoding PdT(G294P) [aa 2- 470]: GCGAACAAGGCGGTGAACGATTTTATCCTGGCGATGAACTATGACAAGAAGAAACT GCTGACCCACCAAGGCGAGAGCATTGAGAACCGTTTCATTAAAGAAGGCAACCAGC TGCCGGACGAGTTTGTGGTTATCGAGCGTAAGAAACGTAGCCTGAGCACCAACACC AGCGACATTAGCGTGACCGCGACCAACGATAGCCGTCTGTACCCGGGTGCGCTGCT GGTTGTGGATGAAACCCTGCTGGAAAACAACCCGACCCTGCTGGCGGTGGACCGTG CGCCGATGACCTATAGCATCGATCTGCCGGGTCTGGCGAGCAGCGACAGCTTCCTGC AAGTTGAGGATCCGAGCAACAGCAGCGTGCGTGGTGCGGTTAACGACCTGCTGGCG AAGTGGCACCAGGATTACGGCCAAGTGAACAACGTTCCGGCGCGTATGCAGTATGA AAAAATCACCGCGCACAGCATGGAGCAACTGAAGGTTAAATTCGGTAGCGACTTTG AAAAGACCGGCAACAGCCTGGACATTGATTTCAACAGCGTGCACAGCGGCGAGAAG CAGATCCAAATCGTTAACTTCAAGCAGATCTACTACACCGTGAGCGTTGACGCGGTG AAGAACCCGGGTGACGTTTTCCAGGATACCGTGACCGTTGAAGATCTGAAACAACG TGGCATTAGCGCGGAGCGTCCGCTGGTGTACATCAGCAGCGTTGCGTACGGTCGTCA AGTGTATCTGAAGCTGGAAACCACCAGCAAAAGCGATGAGGTTGAAGCGGCGTTTG AGGCGCTGATTAAGGGCGTGAAAGTTGCGCCGCAGACCGAATGGAAGCAAATTCTG GACAACACCGAGGTGAAAGCGGTTATTCTGGGCCCGGATCCGAGCAGCGGCGCGCG TGTGGTTACCGGTAAAGTGGACATGGTTGAGGATCTGATTCAGGAAGGTAGCCGTTT TACCGCGGACCACCCGGGCCTGCCGATCAGCTACACCACCAGCTTCCTGCGTGACAA CGTGGTTGCGACCTTTCAAAACAGCACCGATTACGTGGAAACCAAGGTTACCGCGTA TCGTAACGGTGACCTGCTGCTGGACCACAGCGGTGCGTACGTGGCGCAGTACTATAT CACCTGGGATGAACTGAGCTATAACCACCAGGGTAAAGAGGTGCTGACCCCGAAAG CGTGGGACCGTAACGGCCAGGATCTGACCGCGCACTTCACCACCAGCATTCCGCTG AAGGGCAACGTGCGTAACCTGAGCGTTAAAATCCGTGAGGGTACCGGCCTGGCGTT TGAATGGTGGCGTACCGTGTACGAGAAGACCGACCTGCCGCTGGTTCGTAAACGTA CCATCAGCATTTGGGGTACCACCCTGTATCCGCAGGTGGAGGACAAAGTTGAAAAT GAT AFV70463FF [0487] SEQ ID NO: 17, SP0435 gene encoding SP0435 protein, full-length [aa 1-186], TIGR4 strain: ATGATTGAAGCAAGTAAATTAAAAGCTGGTATGACCTTTGAAACAGCTGACGGCAA ATTGATTCGCGTTTTGGAAGCTAGTCACCACAAACCAGGTAAAGGAAACACGATCA TGCGTATGAAATTGCGTGATGTCCGTACTGGTTCTACATTTGACACAAGCTACCGTC CAGAGGAAAAATTTGAACAAGCTATTATCGAGACTGTCCCAGCTCAATACTTGTACA AAATGGATGACACAGCATACTTCATGAATACAGAAACTTATGACCAATACGAAATC CCTGTAGTCAATGTTGAAAACGAATTGCTTTACATCCTTGAAAACTCTGATGTGAAA ATCCAATTCTACGGAACTGAAGTGATCGGTGTCACCGTTCCTACTACTGTTGAGTTG ACAGTTGCTGAAACTCAACCATCTATCAAAGGTGCTACTGTTACAGGTTCTGGTAAA CCAGCAACGATGGAAACTGGACTTGTCGTAAACGTTCCAGACTTCATCGAAGCAGG ACAAAAACTCGTTATCAACACTGCAGAAGGAACTTACGTTTCTCGTGCC [0488] SEQ ID NO: 18, SP0785 gene encoding SP0785 protein, full-length [aa 1-399], TIGR4 strain: ATGAAGAAAAAGAATGGTAAAGCTAAAAAGTGGCAACTGTATGCAGCAATCGGTGC TGCGAGTGTAGTTGTATTGGGTGCTGGGGGGATTTTACTCTTTAGACAACCTTCTCA GACTGCTCTAAAAGATGAGCCTACTCATCTTGTTGTTGCCAAGGAAGGAAGCGTGGC CTCCTCTGTTTTATTGTCAGGGACAGTAACAGCAAAAAATGAACAATATGTTTATTT TGATGCTAGTAAGGGTGATTTAGATGAAATCCTTGTTTCTGTGGGCGATAAGGTCAG CGAAGGGCAGGCTTTAGTCAAGTACAGTAGTTCAGAAGCGCAGGCGGCCTATGATT CAGCTAGTCGAGCAGTAGCTAGGGCAGATCGTCATATCAATGAACTCAATCAAGCA CGAAATGAAGCCGCTTCAGCTCCGGCTCCACAGTTACCAGCGCCAGTAGGAGGAGA AGATGCAACGGTGCAAAGCCCAACTCCAGTGGCTGGAAATTCTGTTGCTTCTATTGA CGCTCAATTGGGTGATGCCCGTGATGCGCGTGCAGATGCTGCGGCGCAATTAAGCA AGGCTCAAAGTCAATTGGATGCAACAACTGTTCTCAGTACCCTAGAGGGAACTGTG GTCGAAGTCAATAGCAATGTTTCTAAATCTCCAACAGGGGCGAGTCAAGTTATGGTT CATATTGTCAGCAATGAAAATTTACAAGTCAAGGGAGAATTGTCTGAGTACAATCTA GCCAACCTTTCTGTAGGTCAAGAAGTAAGCTTTACTTCTAAAGTGTATCCTGATAAA AAATGGACTGGGAAATTAAGCTATATTTCTGACTATCCTAAAAACAATGGTGAAGC AGCTAGTCCAGCAGCCGGGAATAATACAGGTTCTAAATACCCTTATACTATTGATGT AFV70463FF GACAGGCGAGGTTGGTGATTTGAAACAAGGTTTTTCTGTCAACATTGAGGTTAAAAG CAAAACTAAGGCTATTCTTGTTCCTGTTAGCAGTCTAGTAATGGATGATAGTAAAAA TTATGTCTGGATTGTGGATGAACAACAAAAGGCTAAAAAAGTTGAGGTTTCATTGGG AAATGCTGACGCAGAAAATCAAGAAATCACTTCTGGTTTAACGAACGGTGCTAAGG TCATCAGTAATCCAACATCTTCCTTGGAAGAAGGAAAAGAGGTGAAGGCTGATGAA GCAACTAAT [0489] SEQ ID NO: 19, SP1500 gene encoding SP1500 protein, full-length [aa 1-278], TIGR4 strain: ATGAAAAAATGGATGCTTGTATTAGTCAGTCTGATGACTGCTTTGTTCTTAGTAGCTT GTGGGAAAAATTCTAGCGAAACTAGTGGAGATAATTGGTCAAAGTACCAGTCTAAC AAGTCTATTACTATTGGATTTGATAGTACTTTTGTTCCAATGGGATTTGCTCAGAAAG ATGGTTCTTATGCAGGATTTGATATTGATTTAGCTACAGCTGTTTTTGAAAAATACGG AATCACGGTAAATTGGCAACCGATTGATTGGGATTTGAAAGAAGCTGAATTGACAA AAGGAACGATTGATCTGATTTGGAATGGCTATTCCGCTACAGACGAACGCCGTGAA AAGGTGGCTTTCAGTAACTCATATATGAAGAATGAGCAGGTATTGGTTACGAAGAA ATCATCTGGTATCACGACTGCAAAGGATATGACTGGAAAGACATTAGGAGCTCAAG CTGGTTCATCTGGTTATGCGGACTTTGAAGCAAATCCAGAAATTTTGAAGAATATTG TCGCTAATAAGGAAGCGAATCAATACCAAACCTTTAATGAAGCCTTGATTGATTTGA AAAACGATCGAATTGATGGTCTATTGATTGACCGTGTCTATGCAAACTATTATTTAG AAGCAGAAGGTGTTTTAAACGATTATAATGTCTTTACAGTTGGACTAGAAACAGAA GCTTTTGCGGTTGGAGCCCGTAAGGAAGATACAAACTTGGTTAAGAAGATAAATGA AGCTTTTTCTAGTCTTTACAAGGACGGCAAGTTCCAAGAAATCAGCCAAAAATGGTT TGGAGAAGATGTAGCAACCAAAGAAGTAAAAGAAGGACAG [0490] SEQ ID NO: 20, Rhavi-linker-PdT(G294P)-linker-SP0435 [aa 62-185] fusion protein, denoted SPP2: FDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLTGR VNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQGQ DTFQYVPTTENKSLLKDGGGGSSSANKAVNDFILAMNYDKKKLLTHQGESIENRFIKEG NQLPDEFVVIERKKRSLSTNTSDISVTATNDSRLYPGALLVVDETLLENNPTLLAVDRAP MTYSIDLPGLASSDSFLQVEDPSNSSVRGAVNDLLAKWHQDYGQVNNVPARMQYEKIT AFV70463FF AHSMEQLKVKFGSDFEKTGNSLDIDFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPGDV FQDTVTVEDLKQRGISAERPLVYISSVAYGRQVYLKLETTSKSDEVEAAFEALIKGVKVA PQTEWKQILDNTEVKAVILGPDPSSGARVVTGKVDMVEDLIQEGSRFTADHPGLPISYTT SFLRDNVVATFQNSTDYVETKVTAYRNGDLLLDHSGAYVAQYYITWDELSYNHQGKE VLTPKAWDRNGQDLTAHFTTSIPLKGNVRNLSVKIREGTGLAFEWWRTVYEKTDLPLV RKRTISIWGTTLYPQVEDKVENDGGGGSSSEQAIIETVPAQYLYKMDDTAYFMNTETYD QYEIPVVNVENELLYILENSDVKIQFYGTEVIGVTVPTTVELTVAETQPSIKGATVTGSGK PATMETGLVVNVPDFIEAGQKLVINTAEGTYVSR [0491] SEQ ID NO: 21, Rhavi-linker-PdT(G294P)-linker-SP0435 [aa 62-185] fusion protein, denoted SPP2, includes leading methionine: MFDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLT GRVNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQ GQDTFQYVPTTENKSLLKDGGGGSSSANKAVNDFILAMNYDKKKLLTHQGESIENRFIK EGNQLPDEFVVIERKKRSLSTNTSDISVTATNDSRLYPGALLVVDETLLENNPTLLAVDR APMTYSIDLPGLASSDSFLQVEDPSNSSVRGAVNDLLAKWHQDYGQVNNVPARMQYE KITAHSMEQLKVKFGSDFEKTGNSLDIDFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPG DVFQDTVTVEDLKQRGISAERPLVYISSVAYGRQVYLKLETTSKSDEVEAAFEALIKGVK VAPQTEWKQILDNTEVKAVILGPDPSSGARVVTGKVDMVEDLIQEGSRFTADHPGLPIS YTTSFLRDNVVATFQNSTDYVETKVTAYRNGDLLLDHSGAYVAQYYITWDELSYNHQ GKEVLTPKAWDRNGQDLTAHFTTSIPLKGNVRNLSVKIREGTGLAFEWWRTVYEKTDL PLVRKRTISIWGTTLYPQVEDKVENDGGGGSSSEQAIIETVPAQYLYKMDDTAYFMNTE TYDQYEIPVVNVENELLYILENSDVKIQFYGTEVIGVTVPTTVELTVAETQPSIKGATVTG SGKPATMETGLVVNVPDFIEAGQKLVINTAEGTYVSR [0492] SEQ ID NO: 22, Rhavi-linker-PdT(G294P)-linker-SP0435 [aa 62-185]-Hisx6 fusion protein, denoted SPP2-H: FDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLTGR VNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQGQ DTFQYVPTTENKSLLKDGGGGSSSANKAVNDFILAMNYDKKKLLTHQGESIENRFIKEG NQLPDEFVVIERKKRSLSTNTSDISVTATNDSRLYPGALLVVDETLLENNPTLLAVDRAP MTYSIDLPGLASSDSFLQVEDPSNSSVRGAVNDLLAKWHQDYGQVNNVPARMQYEKIT AFV70463FF AHSMEQLKVKFGSDFEKTGNSLDIDFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPGDV FQDTVTVEDLKQRGISAERPLVYISSVAYGRQVYLKLETTSKSDEVEAAFEALIKGVKVA PQTEWKQILDNTEVKAVILGPDPSSGARVVTGKVDMVEDLIQEGSRFTADHPGLPISYTT SFLRDNVVATFQNSTDYVETKVTAYRNGDLLLDHSGAYVAQYYITWDELSYNHQGKE VLTPKAWDRNGQDLTAHFTTSIPLKGNVRNLSVKIREGTGLAFEWWRTVYEKTDLPLV RKRTISIWGTTLYPQVEDKVENDGGGGSSSEQAIIETVPAQYLYKMDDTAYFMNTETYD QYEIPVVNVENELLYILENSDVKIQFYGTEVIGVTVPTTVELTVAETQPSIKGATVTGSGK PATMETGLVVNVPDFIEAGQKLVINTAEGTYVSRHHHHHH [0493] SEQ ID NO: 23, Rhavi-linker-PdT(G294P)-linker-SP0435 [aa 62-185]-Hisx6 fusion protein, denoted SPP2-H, includes leading methionine: MFDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLT GRVNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQ GQDTFQYVPTTENKSLLKDGGGGSSSANKAVNDFILAMNYDKKKLLTHQGESIENRFIK EGNQLPDEFVVIERKKRSLSTNTSDISVTATNDSRLYPGALLVVDETLLENNPTLLAVDR APMTYSIDLPGLASSDSFLQVEDPSNSSVRGAVNDLLAKWHQDYGQVNNVPARMQYE KITAHSMEQLKVKFGSDFEKTGNSLDIDFNSVHSGEKQIQIVNFKQIYYTVSVDAVKNPG DVFQDTVTVEDLKQRGISAERPLVYISSVAYGRQVYLKLETTSKSDEVEAAFEALIKGVK VAPQTEWKQILDNTEVKAVILGPDPSSGARVVTGKVDMVEDLIQEGSRFTADHPGLPIS YTTSFLRDNVVATFQNSTDYVETKVTAYRNGDLLLDHSGAYVAQYYITWDELSYNHQ GKEVLTPKAWDRNGQDLTAHFTTSIPLKGNVRNLSVKIREGTGLAFEWWRTVYEKTDL PLVRKRTISIWGTTLYPQVEDKVENDGGGGSSSEQAIIETVPAQYLYKMDDTAYFMNTE TYDQYEIPVVNVENELLYILENSDVKIQFYGTEVIGVTVPTTVELTVAETQPSIKGATVTG SGKPATMETGLVVNVPDFIEAGQKLVINTAEGTYVSRHHHHHH [0494] SEQ ID NO: 24, Rhavi-linker-SP1500 [aa 27-278]-linker-SP0785 [aa 33-399] fusion protein, denoted CP1: FDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLTGR VNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQGQ DTFQYVPTTENKSLLKDGGGGSSSTSGDNWSKYQSNKSITIGFDSTFVPMGFAQKDGSY AGFDIDLATAVFEKYGITVNWQPIDWDLKEAELTKGTIDLIWNGYSATDERREKVAFSN SYMKNEQVLVTKKSSGITTAKDMTGKTLGAQAGSSGYADFEANPEILKNIVANKEANQ AFV70463FF YQTFNEALIDLKNDRIDGLLIDRVYANYYLEAEGVLNDYNVFTVGLETEAFAVGARKED TNLVKKINEAFSSLYKDGKFQEISQKWFGEDVATKEVKEGQAAAFRQPSQTALKDEPTH LVVAKEGSVASSVLLSGTVTAKNEQYVYFDASKGDLDEILVSVGDKVSEGQALVKYSS SEAQAAYDSASRAVARADRHINELNQARNEAASAPAPQLPAPVGGEDATVQSPTPVAG NSVASIDAQLGDARDARADAAAQLSKAQSQLDATTVLSTLEGTVVEVNSNVSKSPTGA SQVMVHIVSNENLQVKGELSEYNLANLSVGQEVSFTSKVYPDKKWTGKLSYISDYPKN NGEAASPAAGNNTGSKYPYTIDVTGEVGDLKQGFSVNIEVKSKTKAILVPVSSLVMDDS KNYVWIVDEQQKAKKVEVSLGNADAENQEITSGLTNGAKVISNPTSSLEEGKEVKADE ATN [0495] SEQ ID NO: 25, Rhavi-linker-SP1500 [aa 27-278]-linker-SP0785 [aa 33-399] fusion protein, denoted CP1, includes leading methionine: MFDASNFKDFSSIASASSSWQNQSGSTMIIQVDSFGNVSGQYVNRAQGTGCQNSPYPLT GRVNGTFIAFSVGWNNSTENCNSATGWTGYAQVNGNNTEIVTSWNLAYEGGSGPAIEQ GQDTFQYVPTTENKSLLKDGGGGSSSTSGDNWSKYQSNKSITIGFDSTFVPMGFAQKDG SYAGFDIDLATAVFEKYGITVNWQPIDWDLKEAELTKGTIDLIWNGYSATDERREKVAF SNSYMKNEQVLVTKKSSGITTAKDMTGKTLGAQAGSSGYADFEANPEILKNIVANKEA NQYQTFNEALIDLKNDRIDGLLIDRVYANYYLEAEGVLNDYNVFTVGLETEAFAVGAR KEDTNLVKKINEAFSSLYKDGKFQEISQKWFGEDVATKEVKEGQAAAFRQPSQTALKD EPTHLVVAKEGSVASSVLLSGTVTAKNEQYVYFDASKGDLDEILVSVGDKVSEGQALV KYSSSEAQAAYDSASRAVARADRHINELNQARNEAASAPAPQLPAPVGGEDATVQSPTP VAGNSVASIDAQLGDARDARADAAAQLSKAQSQLDATTVLSTLEGTVVEVNSNVSKSP TGASQVMVHIVSNENLQVKGELSEYNLANLSVGQEVSFTSKVYPDKKWTGKLSYISDY PKNNGEAASPAAGNNTGSKYPYTIDVTGEVGDLKQGFSVNIEVKSKTKAILVPVSSLVM DDSKNYVWIVDEQQKAKKVEVSLGNADAENQEITSGLTNGAKVISNPTSSLEEGKEVK ADEATN [0496] SEQ ID NO: 26, codon-optimized nucleotide sequence encoding Rhavi-linker- PdT(G294P)-linker-SP0435 [aa 62-185], denoted SPP2: TTCGACGCATCCAACTTTAAAGACTTTAGCAGCATCGCGTCCGCAAGCTCTAGCTGG CAGAATCAATCTGGTAGCACCATGATTATCCAAGTGGACAGCTTTGGTAACGTCAGC GGTCAATATGTTAATCGTGCACAGGGTACGGGTTGTCAGAATTCTCCGTACCCGCTG AFV70463FF ACCGGTCGTGTTAACGGCACGTTCATCGCTTTCAGCGTCGGTTGGAACAATTCTACT GAAAATTGCAACAGCGCGACCGGTTGGACGGGCTATGCACAAGTGAATGGCAATAA CACCGAAATCGTCACGTCCTGGAATCTGGCGTATGAGGGTGGCAGCGGTCCGGCTAT TGAACAGGGCCAGGATACCTTCCAATACGTCCCTACGACCGAGAATAAGTCCCTTCT GAAAGACGGCGGTGGCGGTTCGAGCTCGGCGAACAAGGCGGTGAACGATTTTATCC TGGCGATGAACTATGACAAGAAGAAACTGCTGACCCACCAAGGCGAGAGCATTGAG AACCGTTTCATTAAAGAAGGCAACCAGCTGCCGGACGAGTTTGTGGTTATCGAGCGT AAGAAACGTAGCCTGAGCACCAACACCAGCGACATTAGCGTGACCGCGACCAACGA TAGCCGTCTGTACCCGGGTGCGCTGCTGGTTGTGGATGAAACCCTGCTGGAAAACAA CCCGACCCTGCTGGCGGTGGACCGTGCGCCGATGACCTATAGCATCGATCTGCCGGG TCTGGCGAGCAGCGACAGCTTCCTGCAAGTTGAGGATCCGAGCAACAGCAGCGTGC GTGGTGCGGTTAACGACCTGCTGGCGAAGTGGCACCAGGATTACGGCCAAGTGAAC AACGTTCCGGCGCGTATGCAGTATGAAAAAATCACCGCGCACAGCATGGAGCAACT GAAGGTTAAATTCGGTAGCGACTTTGAAAAGACCGGCAACAGCCTGGACATTGATT TCAACAGCGTGCACAGCGGCGAGAAGCAGATCCAAATCGTTAACTTCAAGCAGATC TACTACACCGTGAGCGTTGACGCGGTGAAGAACCCGGGTGACGTTTTCCAGGATACC GTGACCGTTGAAGATCTGAAACAACGTGGCATTAGCGCGGAGCGTCCGCTGGTGTA CATCAGCAGCGTTGCGTACGGTCGTCAAGTGTATCTGAAGCTGGAAACCACCAGCA AAAGCGATGAGGTTGAAGCGGCGTTTGAGGCGCTGATTAAGGGCGTGAAAGTTGCG CCGCAGACCGAATGGAAGCAAATTCTGGACAACACCGAGGTGAAAGCGGTTATTCT GGGCCCGGATCCGAGCAGCGGCGCGCGTGTGGTTACCGGTAAAGTGGACATGGTTG AGGATCTGATTCAGGAAGGTAGCCGTTTTACCGCGGACCACCCGGGCCTGCCGATC AGCTACACCACCAGCTTCCTGCGTGACAACGTGGTTGCGACCTTTCAAAACAGCACC GATTACGTGGAAACCAAGGTTACCGCGTATCGTAACGGTGACCTGCTGCTGGACCAC AGCGGTGCGTACGTGGCGCAGTACTATATCACCTGGGATGAACTGAGCTATAACCA CCAGGGTAAAGAGGTGCTGACCCCGAAAGCGTGGGACCGTAACGGCCAGGATCTGA CCGCGCACTTCACCACCAGCATTCCGCTGAAGGGCAACGTGCGTAACCTGAGCGTTA AAATCCGTGAGGGTACCGGCCTGGCGTTTGAATGGTGGCGTACCGTGTACGAGAAG ACCGACCTGCCGCTGGTTCGTAAACGTACCATCAGCATTTGGGGTACCACCCTGTAT CCGCAGGTGGAGGACAAAGTTGAAAATGATGGTGGTGGTGGTAGCAGCAGCGAGCA GGCGATCATTGAAACCGTGCCGGCGCAATACCTGTATAAGATGGACGATACCGCGT AFV70463FF ACTTCATGAACACCGAAACCTACGACCAATATGAAATTCCGGTGGTTAACGTTGAGA ACGAACTGCTGTACATCCTGGAAAACAGCGATGTGAAAATTCAGTTTTATGGTACCG AGGTTATCGGCGTGACCGTTCCGACCACCGTGGAGCTGACCGTTGCGGAAACCCAA CCGAGCATCAAGGGTGCGACCGTGACCGGTAGCGGTAAACCGGCGACCATGGAAAC CGGTCTGGTGGTTAACGTGCCGGACTTCATTGAGGCGGGCCAGAAGCTGGTTATCAA TACCGCGGAGGGTACCTATGTTAGCCGTTAATGA [0497] SEQ ID NO: 27, codon-optimized nucleotide sequence encoding Rhavi-linker- PdT(G294P)-linker-SP0435 [aa 62-185], denoted SPP2, includes leading ATG sequence: ATGTTCGACGCATCCAACTTTAAAGACTTTAGCAGCATCGCGTCCGCAAGCTCTAGC TGGCAGAATCAATCTGGTAGCACCATGATTATCCAAGTGGACAGCTTTGGTAACGTC AGCGGTCAATATGTTAATCGTGCACAGGGTACGGGTTGTCAGAATTCTCCGTACCCG CTGACCGGTCGTGTTAACGGCACGTTCATCGCTTTCAGCGTCGGTTGGAACAATTCT ACTGAAAATTGCAACAGCGCGACCGGTTGGACGGGCTATGCACAAGTGAATGGCAA TAACACCGAAATCGTCACGTCCTGGAATCTGGCGTATGAGGGTGGCAGCGGTCCGG CTATTGAACAGGGCCAGGATACCTTCCAATACGTCCCTACGACCGAGAATAAGTCCC TTCTGAAAGACGGCGGTGGCGGTTCGAGCTCGGCGAACAAGGCGGTGAACGATTTT ATCCTGGCGATGAACTATGACAAGAAGAAACTGCTGACCCACCAAGGCGAGAGCAT TGAGAACCGTTTCATTAAAGAAGGCAACCAGCTGCCGGACGAGTTTGTGGTTATCGA GCGTAAGAAACGTAGCCTGAGCACCAACACCAGCGACATTAGCGTGACCGCGACCA ACGATAGCCGTCTGTACCCGGGTGCGCTGCTGGTTGTGGATGAAACCCTGCTGGAAA ACAACCCGACCCTGCTGGCGGTGGACCGTGCGCCGATGACCTATAGCATCGATCTGC CGGGTCTGGCGAGCAGCGACAGCTTCCTGCAAGTTGAGGATCCGAGCAACAGCAGC GTGCGTGGTGCGGTTAACGACCTGCTGGCGAAGTGGCACCAGGATTACGGCCAAGT GAACAACGTTCCGGCGCGTATGCAGTATGAAAAAATCACCGCGCACAGCATGGAGC AACTGAAGGTTAAATTCGGTAGCGACTTTGAAAAGACCGGCAACAGCCTGGACATT GATTTCAACAGCGTGCACAGCGGCGAGAAGCAGATCCAAATCGTTAACTTCAAGCA GATCTACTACACCGTGAGCGTTGACGCGGTGAAGAACCCGGGTGACGTTTTCCAGG ATACCGTGACCGTTGAAGATCTGAAACAACGTGGCATTAGCGCGGAGCGTCCGCTG GTGTACATCAGCAGCGTTGCGTACGGTCGTCAAGTGTATCTGAAGCTGGAAACCACC AGCAAAAGCGATGAGGTTGAAGCGGCGTTTGAGGCGCTGATTAAGGGCGTGAAAGT AFV70463FF TGCGCCGCAGACCGAATGGAAGCAAATTCTGGACAACACCGAGGTGAAAGCGGTTA TTCTGGGCCCGGATCCGAGCAGCGGCGCGCGTGTGGTTACCGGTAAAGTGGACATG GTTGAGGATCTGATTCAGGAAGGTAGCCGTTTTACCGCGGACCACCCGGGCCTGCCG ATCAGCTACACCACCAGCTTCCTGCGTGACAACGTGGTTGCGACCTTTCAAAACAGC ACCGATTACGTGGAAACCAAGGTTACCGCGTATCGTAACGGTGACCTGCTGCTGGAC CACAGCGGTGCGTACGTGGCGCAGTACTATATCACCTGGGATGAACTGAGCTATAA CCACCAGGGTAAAGAGGTGCTGACCCCGAAAGCGTGGGACCGTAACGGCCAGGATC TGACCGCGCACTTCACCACCAGCATTCCGCTGAAGGGCAACGTGCGTAACCTGAGC GTTAAAATCCGTGAGGGTACCGGCCTGGCGTTTGAATGGTGGCGTACCGTGTACGAG AAGACCGACCTGCCGCTGGTTCGTAAACGTACCATCAGCATTTGGGGTACCACCCTG TATCCGCAGGTGGAGGACAAAGTTGAAAATGATGGTGGTGGTGGTAGCAGCAGCGA GCAGGCGATCATTGAAACCGTGCCGGCGCAATACCTGTATAAGATGGACGATACCG CGTACTTCATGAACACCGAAACCTACGACCAATATGAAATTCCGGTGGTTAACGTTG AGAACGAACTGCTGTACATCCTGGAAAACAGCGATGTGAAAATTCAGTTTTATGGTA CCGAGGTTATCGGCGTGACCGTTCCGACCACCGTGGAGCTGACCGTTGCGGAAACC CAACCGAGCATCAAGGGTGCGACCGTGACCGGTAGCGGTAAACCGGCGACCATGGA AACCGGTCTGGTGGTTAACGTGCCGGACTTCATTGAGGCGGGCCAGAAGCTGGTTAT CAATACCGCGGAGGGTACCTATGTTAGCCGTTAATGA [0498] SEQ ID NO: 28, codon-optimized nucleic acid sequence encoding Rhavi-linker- SP1500 [aa 27-278]-linker-SP0785 [aa 33-399] fusion protein, denoted CP1: TTCGACGCATCCAACTTTAAAGACTTTAGCAGCATCGCGTCCGCAAGCTCTAGCTGG CAGAATCAATCTGGTAGCACCATGATTATCCAAGTGGACAGCTTTGGTAACGTCAGC GGTCAATATGTTAATCGTGCACAGGGTACGGGTTGTCAGAATTCTCCGTACCCGCTG ACCGGTCGTGTTAACGGCACGTTCATCGCTTTCAGCGTCGGTTGGAACAATTCTACT GAAAATTGCAACAGCGCGACCGGTTGGACGGGCTATGCACAAGTGAATGGCAATAA CACCGAAATCGTCACGTCCTGGAATCTGGCGTATGAGGGTGGCAGCGGTCCGGCTAT TGAACAGGGCCAGGATACCTTCCAATACGTCCCTACGACCGAGAATAAGTCCCTTCT GAAAGACGGCGGTGGCGGTTCGAGCTCGACCAGCGGCGACAATTGGTCCAAATACC AGAGCAACAAGAGCATCACGATCGGCTTCGACAGCACTTTTGTGCCGATGGGTTTCG CGCAAAAAGACGGTAGCTACGCGGGTTTCGATATTGACCTGGCGACCGCTGTCTTTG AFV70463FF AGAAATACGGCATTACGGTTAATTGGCAGCCGATTGATTGGGACCTGAAAGAGGCC GAACTCACCAAAGGCACCATCGACCTGATCTGGAATGGTTACTCCGCAACCGATGA GCGTCGCGAAAAAGTTGCCTTCAGCAACAGCTATATGAAGAATGAACAAGTGTTGG TAACCAAGAAATCTAGCGGCATTACGACCGCGAAAGACATGACCGGTAAGACGCTG GGTGCGCAGGCCGGTAGCTCTGGCTATGCGGATTTCGAGGCGAATCCTGAGATTCTG AAAAACATCGTTGCGAATAAAGAGGCGAACCAGTACCAGACCTTTAACGAAGCACT GATCGACCTGAAAAACGATCGCATTGACGGTCTGCTGATCGATCGTGTGTACGCGAA CTATTATCTGGAAGCCGAGGGCGTTCTGAACGATTATAATGTTTTTACCGTGGGTCT GGAGACTGAGGCATTCGCGGTTGGTGCGCGCAAGGAAGATACCAACCTGGTTAAAA AGATTAATGAGGCATTTAGCTCACTGTACAAGGACGGCAAGTTCCAAGAAATTAGC CAGAAGTGGTTCGGTGAAGATGTTGCGACGAAAGAGGTTAAAGAGGGCCAAGCGGC CGCATTTCGCCAACCGAGCCAGACTGCGTTGAAAGATGAGCCGACCCATCTGGTTGT TGCGAAAGAGGGCAGCGTGGCATCGAGCGTGCTGCTGAGCGGTACGGTTACTGCCA AAAACGAACAATACGTGTACTTCGATGCTAGCAAGGGTGATCTGGATGAAATTCTG GTGAGCGTGGGTGACAAAGTTAGCGAAGGCCAGGCACTGGTGAAGTATTCATCCTC CGAGGCACAGGCAGCGTACGACAGCGCAAGCCGCGCAGTGGCGCGTGCCGACCGTC ACATTAACGAATTGAACCAAGCGCGTAACGAGGCCGCAAGCGCGCCAGCACCGCAG CTGCCGGCTCCGGTGGGTGGCGAAGATGCGACGGTGCAGAGCCCGACCCCGGTTGC GGGTAATTCGGTCGCCAGCATCGATGCGCAGCTGGGTGACGCGCGTGATGCCCGTG CGGATGCGGCTGCTCAACTGAGCAAGGCTCAGAGCCAACTGGACGCGACGACGGTG CTGAGCACCTTGGAGGGTACCGTTGTCGAAGTCAACAGCAATGTGAGCAAGAGCCC AACGGGTGCGAGCCAGGTTATGGTCCACATTGTGAGCAATGAAAACTTACAGGTCA AGGGTGAGCTGAGCGAGTATAACCTGGCGAATCTGAGCGTTGGTCAAGAGGTCAGC TTTACCAGCAAGGTCTACCCGGATAAGAAATGGACCGGCAAGTTGAGCTACATCAG CGACTACCCGAAGAACAATGGCGAGGCAGCCTCCCCGGCAGCCGGCAACAATACCG GCTCTAAGTATCCGTACACCATCGACGTAACCGGTGAGGTCGGCGACCTGAAACAG GGTTTTAGCGTGAATATCGAAGTGAAGTCCAAGACCAAGGCAATTTTGGTTCCGGTT AGCTCCCTGGTGATGGACGATAGCAAGAATTATGTGTGGATTGTCGACGAGCAACA GAAAGCGAAAAAAGTTGAAGTGAGCCTGGGCAATGCTGATGCCGAGAACCAAGAA ATCACGTCTGGTCTGACCAACGGTGCGAAAGTTATTAGCAACCCGACCAGCAGCCT GGAAGAGGGTAAAGAGGTCAAAGCCGACGAAGCTACGAAC AFV70463FF [0499] SEQ ID NO: 29, codon-optimized nucleic acid sequence encoding Rhavi-linker- SP1500 [aa 27-278]-linker-SP0785 [aa 33-399] fusion protein, denoted CP1, includes leading ATG sequence: ATGTTCGACGCATCCAACTTTAAAGACTTTAGCAGCATCGCGTCCGCAAGCTCTAGC TGGCAGAATCAATCTGGTAGCACCATGATTATCCAAGTGGACAGCTTTGGTAACGTC AGCGGTCAATATGTTAATCGTGCACAGGGTACGGGTTGTCAGAATTCTCCGTACCCG CTGACCGGTCGTGTTAACGGCACGTTCATCGCTTTCAGCGTCGGTTGGAACAATTCT ACTGAAAATTGCAACAGCGCGACCGGTTGGACGGGCTATGCACAAGTGAATGGCAA TAACACCGAAATCGTCACGTCCTGGAATCTGGCGTATGAGGGTGGCAGCGGTCCGG CTATTGAACAGGGCCAGGATACCTTCCAATACGTCCCTACGACCGAGAATAAGTCCC TTCTGAAAGACGGCGGTGGCGGTTCGAGCTCGACCAGCGGCGACAATTGGTCCAAA TACCAGAGCAACAAGAGCATCACGATCGGCTTCGACAGCACTTTTGTGCCGATGGGT TTCGCGCAAAAAGACGGTAGCTACGCGGGTTTCGATATTGACCTGGCGACCGCTGTC TTTGAGAAATACGGCATTACGGTTAATTGGCAGCCGATTGATTGGGACCTGAAAGA GGCCGAACTCACCAAAGGCACCATCGACCTGATCTGGAATGGTTACTCCGCAACCG ATGAGCGTCGCGAAAAAGTTGCCTTCAGCAACAGCTATATGAAGAATGAACAAGTG TTGGTAACCAAGAAATCTAGCGGCATTACGACCGCGAAAGACATGACCGGTAAGAC GCTGGGTGCGCAGGCCGGTAGCTCTGGCTATGCGGATTTCGAGGCGAATCCTGAGAT TCTGAAAAACATCGTTGCGAATAAAGAGGCGAACCAGTACCAGACCTTTAACGAAG CACTGATCGACCTGAAAAACGATCGCATTGACGGTCTGCTGATCGATCGTGTGTACG CGAACTATTATCTGGAAGCCGAGGGCGTTCTGAACGATTATAATGTTTTTACCGTGG GTCTGGAGACTGAGGCATTCGCGGTTGGTGCGCGCAAGGAAGATACCAACCTGGTT AAAAAGATTAATGAGGCATTTAGCTCACTGTACAAGGACGGCAAGTTCCAAGAAAT TAGCCAGAAGTGGTTCGGTGAAGATGTTGCGACGAAAGAGGTTAAAGAGGGCCAAG CGGCCGCATTTCGCCAACCGAGCCAGACTGCGTTGAAAGATGAGCCGACCCATCTG GTTGTTGCGAAAGAGGGCAGCGTGGCATCGAGCGTGCTGCTGAGCGGTACGGTTAC TGCCAAAAACGAACAATACGTGTACTTCGATGCTAGCAAGGGTGATCTGGATGAAA TTCTGGTGAGCGTGGGTGACAAAGTTAGCGAAGGCCAGGCACTGGTGAAGTATTCA TCCTCCGAGGCACAGGCAGCGTACGACAGCGCAAGCCGCGCAGTGGCGCGTGCCGA CCGTCACATTAACGAATTGAACCAAGCGCGTAACGAGGCCGCAAGCGCGCCAGCAC CGCAGCTGCCGGCTCCGGTGGGTGGCGAAGATGCGACGGTGCAGAGCCCGACCCCG AFV70463FF GTTGCGGGTAATTCGGTCGCCAGCATCGATGCGCAGCTGGGTGACGCGCGTGATGCC CGTGCGGATGCGGCTGCTCAACTGAGCAAGGCTCAGAGCCAACTGGACGCGACGAC GCCCAACGGGTGCGAGCCAGGTTATGGTCCACATTGTGAGCAATGAAAACTTACAG GTCAAGGGTGAGCTGAGCGAGTATAACCTGGCGAATCTGAGCGTTGGTCAAGAGGT CAGCTTTACCAGCAAGGTCTACCCGGATAAGAAATGGACCGGCAAGTTGAGCTACA TCAGCGACTACCCGAAGAACAATGGCGAGGCAGCCTCCCCGGCAGCCGGCAACAAT ACCGGCTCTAAGTATCCGTACACCATCGACGTAACCGGTGAGGTCGGCGACCTGAA ACAGGGTTTTAGCGTGAATATCGAAGTGAAGTCCAAGACCAAGGCAATTTTGGTTCC GGTTAGCTCCCTGGTGATGGACGATAGCAAGAATTATGTGTGGATTGTCGACGAGCA ACAGAAAGCGAAAAAAGTTGAAGTGAGCCTGGGCAATGCTGATGCCGAGAACCAA GAAATCACGTCTGGTCTGACCAACGGTGCGAAAGTTATTAGCAACCCGACCAGCAG CCTGGAAGAGGGTAAAGAGGTCAAAGCCGACGAAGCTACGAAC [0500] SEQ ID NO: 30, linker sequence [7 amino acids]: GGGGSSS [0501] SEQ ID NO: 31, linker sequence [5 amino acid repeats]: (GGGGS)_n [0502] SEQ ID NO: 32, linker sequence [5 amino acids]: GGGGS [0503] SEQ ID NO: 33, linker sequence [15 amino acids]: GGGGSGGGGSGGGGS [0504] SEQ ID NO: 34, linker sequence [30 amino acids]: GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS [0505] SEQ ID NO: 35, linker sequence: (Gly)_n [0506] SEQ ID NO: 36, linker sequence [6 amino acids]: GGGGGG [0507] SEQ ID NO: 37, linker sequence [8 amino acids]: GGGGGGGG [0508] SEQ ID NO: 38, linker sequence [18 amino acids]: KESGSVSSEQLAQFRSLD AFV70463FF [0509] SEQ ID NO: 39, linker sequence [14 amino acids]: EGKSSGSGSESKST [0510] SEQ ID NO: 40, linker sequence [12 amino acids]: GSAGSAAGSGEF [0511] SEQ ID NO: 41, linker sequence [5 amino acid repeats]: (EAAAK)_n [0512] SEQ ID NO: 42, linker sequence: EAAAK [0513] SEQ ID NO: 43, linker sequence: A(EAAAK)_nA [0514] SEQ ID NO: 44, linker sequence: A(EAAAK)_nA , n=1-5 [0515] SEQ ID NO: 45, linker sequence: A(EAAAK)₄ALEA(EAAAK)₄A [0516] SEQ ID NO: 46, linker sequence: [A(EAAAK)_nA]_m , n=1-4, m=1-2 [0517] SEQ ID NO: 47, linker sequence [12 amino acids]: AEAAAKEAAAKA [0518] SEQ ID NO: 48, linker sequence [2 amino acid repeats]: (XP)_n [0519] SEQ ID NO: 49, linker sequence [2 amino acid repeats]: (AP)_n [0520] SEQ ID NO: 50, linker sequence [2 amino acid repeats]: (AP)_n , n=1-17 [0521] SEQ ID NO: 51, linker sequence [2 amino acid repeats]: (KP)_n [0522] SEQ ID NO: 52, linker sequence [2 amino acid repeats]: (QP)_n [0523] SEQ ID NO: 53, linker sequence [14 amino acids]: APAPAPAPAPAPAP AFV70463FF [0524] SEQ ID NO: 54, GAG linker sequence [21 amino acids]: GAPGGGGGAAAAAGGGGGGAP [0525] SEQ ID NO: 55, GAG2 linker sequence [39 amino acids]: GAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGGGAP [0526] SEQ ID NO: 56, GAG3 linker sequence [57 amino acids]: GAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGG GAP [0527] SEQ ID NO: 57, linker sequence [4 amino acids]: VSDP [0528] SEQ ID NO: 58, linker sequence [4 amino acids]: AAAA [0529] SEQ ID NO: 59, linker sequence [4 amino acids]: GGGG [0530] SEQ ID NO: 60, His tag 1: (His)_n , n=2-10 [0531] SEQ ID NO: 61, His tag 2: HHHHHH [0532] SEQ ID NO: 62, His tag 3: MSYYHHHHHH [0533] SEQ ID NO: 63, signal peptide sequence: MKKIWLALAGLVLAFSASA [0534] SEQ ID NO: 64, signal peptide sequence: MAPFEPLASGILLLLWLIAPSRA [0535] SEQ ID NO: 65, signal peptide sequence: MKKVAAFVALSLLMAGC [0536] SEQ ID NO: 66, signal peptide sequence: MKKIMLVITLILVSPIAQQTEAKD [0537] SEQ ID NO: 67, signal peptide sequence: MKKKIISAILMSTVILSAAAPLSGVYADT [0538] SEQ ID NO: 68, signal peptide sequence: MKKRKVLIPLMALSTILVSSTGNLEVIQAEV AFV70463FF [0539] SEQ ID NO: 69, signal peptide sequence: MNMKKATIAATAGIAVTAFAAPTIASAST [0540] SEQ ID NO: 70, signal peptide sequence: MQKTRKERILEALQEEKKNKKSKKFKTGATIAGVTAIATSITVPGIEVIVSADE [0541] SEQ ID NO: 71, signal peptide sequence: MKKLKMASCALVAGLMFSGLTPNAFAED [0542] SEQ ID NO: 72, signal peptide sequence: MAKKFNYKLPSMVALTLVGSAVTAHQVQAAE [0543] SEQ ID NO: 73, signal peptide sequence: MTDKKSENQTEKTETKENKGMTRREMLKLSAVAGTGIAVGATGLGTILNVVDQVDKA LT [0544] SEQ ID NO: 74, signal peptide sequence: MAYDSRFDEWVQKLKEESFQNNTFDRRKFIQGAGKIAGLGLGLTIAQSVGAFG [0545] SEQ ID NO: 75, signal peptide sequence: MEFGLSWLFLVAILKGVQC [0546] SEQ ID NO: 76, signal peptide sequence: MGWSCIILFLVATATGVHS

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Complete monosaccharide analysis by high performance anion-exchange chromatography with pulsed amperometric detection. Anal Chem 84(9):4104-4110 (2012). EQUIVALENTS [0547] The embodiments of the disclosure described above are intended to be merely exemplary, numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention as defined in any appended claims.

Claims

AFV70463FF CLAIMS 1. A composition comprising a purified capsular polysaccharide having repeating units of formula I: HO HO O wherein each R 2. The composition of claim 1, wherein the purified capsular polysaccharide comprises from about 4 to about 4000 repeating units of formula I. 3. The composition of claims 1 or 2, wherein the purified capsular polysaccharide comprises between 0% and 100% of repeating units wherein R is –C(O)-CH₃. 4. The composition of any one of claims 1-3, wherein a weight average molecular weight of the purified capsular polysaccharide is from about 900 Daltons to about 4,000 kiloDaltons. 5. The composition of any one of claims 1-4, wherein the purified capsular polysaccharide is activated with a chemical reagent to produce reactive groups for conjugation to a suitable conjugate (e.g., a linker or carrier protein). AFV70463FF 6. The composition of claim 5, wherein activation occurs at the C_2, C_3, C_5, or C₆ of the -Galf residue. 7. A composition comprising a polysaccharide having one or more monomers of formula II: HO HO O 8. An immunogenic composition comprising (i) a purified capsular polysaccharide and (ii) a polypeptide, wherein the purified capsular polysaccharide comprises repeating units of formula I: HO HO

AFV70463FF I wherein each R is independently selected from H and –C(O)-CH₃. 9. The immunogenic composition of claim 8, wherein the purified capsular polysaccharide comprises from about 4 to about 4000 repeating units of formula I. 10. The immunogenic composition of claims 8 or 9, wherein the purified capsular polysaccharide comprises between 0% and 100% of repeating units wherein R is –C(O)-CH₃. 11. The immunogenic composition of claims 8 or 9 or 10, wherein the purified capsular polysaccharide comprises about 60% to about 70% of repeating units wherein R is –C(O)-CH₃. 12. The immunogenic composition of any one of claims 8-11, wherein the polypeptide is associated with the purified capsular polysaccharide. 13. The immunogenic composition of any one of claims 8-12, wherein the polypeptide is a carrier protein, optionally wherein the carrier protein is a toxoid or a glycoprotein from a pathogen, optionally wherein the carrier protein is selected from the group consisting of: CRM197, tetanus toxoid, exoprotein A, PspA, diphtheria toxoid, and protein D. 14. The immunogenic composition of any one of claims 8-12, wherein a purified capsular polysaccharide having one or more repeating units of formula I is biotinylated. 15. The immunogenic composition of claim 14, wherein the polypeptide is a fusion protein comprising a biotin-binding moiety and at least one polypeptide antigen, and wherein the biotinylated purified capsular polysaccharide is non-covalently associated with the biotin-binding moiety of the fusion protein. 16. The immunogenic composition of any one of claims 8-15, wherein the immunogenic composition further comprises one or more polysaccharides of Streptococcus pneumoniae each having a serotype independently selected from: 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 6E, 6F, 6G, 6H, 7A, AFV70463FF 7B, 7C, 7F, 8, 9A, 9L, 9N, 9V, 10A, 10B, 10C, 10F, 11A, 11B, 11C, 11D, 11E, 11F, 12A, 12B, 12F, 13, 14, 15A, 15B, 15C, 15F, 16A, 16F, 17A, 17F, 18A, 18B, 18C, 18F, 19A, 19B, 19C, 19F, 20A, 20B, 21, 22A, 22F, 23A, 23B, 23F, 24A, 24B, 24F, 25A, 25F, 27, 28A, 28F, 29, 31, 32A, 32F, 33A, 33B, 33C, 33D, 33E, 33F, 34, 35A, 35B, 35C, 35F, 36, 37, 39, 40, 41A, 41F, 42, 43, 44, 45, 46, 47A, 47F, and 48. 17. A pharmaceutical composition comprising the immunogenic composition of any one of claims 8-16, and a pharmaceutically acceptable carrier. 18. A method of inducing an immune response to a Streptococcus pneumoniae capsular polysaccharide comprising administering to a subject the immunogenic composition of any one of claims 8-16 or the pharmaceutical composition of claim 17. 19. An activated capsular polysaccharide comprising one or more units of formula I, HO HO wherein each R 20. An activated capsular polysaccharide comprising one or more units of formula I, AFV70463FF HO HO O wherein each the purified capsular polysaccharide is conjugated to a carrier protein or biotin. 21. A purified capsular polysaccharide comprising one or more monomers of formula III: R^aO R^aO where one a carrier protein) and biotin, and the remaining instances of R^a are each H.