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DK200301155A - Polynucleotide(s) encoding immunoglobulin molecules - used for efficient prodn. of chimeric human or non-human- or class switched-antibodies - Google Patents

Polynucleotide(s) encoding immunoglobulin molecules - used for efficient prodn. of chimeric human or non-human- or class switched-antibodies Download PDF

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Publication number
DK200301155A
DK200301155A DK200301155A DKPA200301155A DK200301155A DK 200301155 A DK200301155 A DK 200301155A DK 200301155 A DK200301155 A DK 200301155A DK PA200301155 A DKPA200301155 A DK PA200301155A DK 200301155 A DK200301155 A DK 200301155A
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DK
Denmark
Prior art keywords
immunoglobulin chain
chimeric
fragment
human
polynucleotide
Prior art date
Application number
DK200301155A
Other languages
Danish (da)
Inventor
Randy R Robinson
Alvin Y Liu
Arnold H Horwitz
Wall Randolph
Better Marc
Original Assignee
Internat Genetic Engineering I
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from DK199000192A external-priority patent/DK174824B1/en
Application filed by Internat Genetic Engineering I filed Critical Internat Genetic Engineering I
Priority to DK200301155A priority Critical patent/DK175654B1/en
Publication of DK200301155A publication Critical patent/DK200301155A/en
Application granted granted Critical
Publication of DK175654B1 publication Critical patent/DK175654B1/en

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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Following are claimed (A) a polynucleotide molecule comprising a promoter region in operable linkage to a dicistronic transcription unit, the unit encoding a chimeric heavy chain immunoglobulin or fragment and a chimeric light chain immunoglobulin, (B) a chimeric heavy chain immunoglobulin molecule or fragment linked to a polypeptide secretion signal; (C) a chimeric light chain immunoglobulin molecule linked to a polypeptide secretion signal; (D) a chimeric Fab immunoglobulin molecule, (E) a chimeric Fd immunoglobulin molecule, (F) a polynucleotide molecule encoding the immunoglobulin molecule of (D) or (E), (G) a polynucleotide molecule encoding the Fd fragment of a chimeric heavy chain immunoglobulin comprising a restriction site at or downstream from a stop codon.

Description

PATENTKRAV 1. Immunoglobulinkæde eller et fragment af en immunoglobulinkæde omfattende mindst en funktionelt virkende region af en variabel region af et antistof, idet den variable region ved dens N-ende direkte er forbundet til et prokaryot sekretionssignalpeptid for at muliggøre sekretion af immunoglobulinkæden eller fragmentet af en immunoglobulinkæde fra en prokaryot organisme. 2. Immunoglobulinkæde ifølge krav 1, som er en let immunoglobulinkæde. 3. Immunoglobulinkæde ifølge krav 1, som er en tung immunoglobulinkæde. 4. Immunoglobulinkæde ifølge et hvilket som helst af kravene, hvor det prokaryote sekretionspeptid er et pectatlyase-signal-peptid. 5. Immunoglobulinkæde ifølge krav 1 eller 4, hvor fragmentet er et Fd-fragment. 6. Immunoglobulinkæde eller fragment af en immunoglobulinkæde ifølge et hvilket som helst af de foregående krav, som er kimært. 7. Polynucleotidmolekyle, som koder for en immunoglobulinkæde eller et fragment af en immunoglobulinkæde ifølge et hvilket som helst af de foregående krav. 8. Molekyle ifølge krav 7, som er DNA, eventuelt omfattende en promotor. 9. Vektor, såsom en ekspressionsvektor, omfattende et molekyle ifølge kravene 7 eller 8. 10. Fremgangsmåde til fremstilling af en immunoglobulinkæde eller et fragment af en immunoglobulinkæde, idet fremgangsmåden omfatter: a) transformering eller transfektering af en prokaryot vært med en vektor som defineret i krav 9 omfattende en promotor, som er i stand til at dirigere transskription i værten; b) dyrkning af værten under passende forhold, sådan at immunoglobulinkæden eller fragmentet af en immunoglobulinkæde udtrykkes, idet den variable region direkte er forbundet ved dens N-ende til et prokaryot sekretionssignalpeptid; og c) opnåelse af en immunoglobulinkæde eller et fragment af en immunoglobulinkæde, som forud for sekretion fra værten oprindeligt var forbundet til signalpepti-det. 11. Fremgangsmåde til fremstilling af en immunoglobulinkæde eller et fragment af en immunoglobulinkæde, som defineret i et hvilket som helst af kravene 1 til 6, idet fremgangsmåden omfatter: a) dyrkning af en prokaryot vært, som er i stand til at udtrykke kæden eller fragmentet af en immunoglobulinkæde under dyrkningsbetingelser; og b) indvinding af kæden eller fragmentet af en immunoglobulinkæde fra kulturen. 12. Fremgangsmåde til fremstilling af en immunoglobulinkæde eller et fragment af en immunoglobulinkæde, idet fremgangsmåden omfatter: (a) dyrkning af en transformeret eller transfekteret prokaryot vært under passende betingelser, sådan at immunoglobulinkæden eller fragmentet af en immunoglobulinkæde udtrykkes som et resultat af transkription i værten af en polynucleotidsekvens, som koder for en immunoglobulinkæde, omfattende mindst én funktionelt virkende region af en variabel region af et antistof, idet den variable region ved dens N-ende direkte er forbundet til et prokaryot sekretionssignalpeptid for at muliggøre sekretion af immunoglobulinkæden eller fragmentet af en immunoglobulinkæde fra en prokaryot organisme; og (b) opnåelse af en immunoglobulinkæde eller et fragment af en immunoglobulinkæde, som forud for sekretionen fra værten oprindeligt var forbundet til signalpeptidet. 13. Fremgangsmåde ifølge et hvilket som helst af kravene 10 til 12, hvor sekretions-signalpeptidet er et pectatlyase-sekretionssignal. 14. Fremgangsmåde ifølge et hvilket som helst af kravene 10 til 13, hvor immu-noglobulinkæden eller fragmentet af en immunoglobulinkæde isoleres fra det pe-riplasmiske rum. 15. Fremgangsmåde ifølge et hvilket som helst af kravene 10 til 13, hvor immu-noglobulinkæden eller fragmentet af en immunoglobulinkæde isoleres fra dyrknings-vækstmediet. 16. Vært, som er transformeret eller transfekteret med en vektor ifølge krav 9.A immunoglobulin chain or fragment of an immunoglobulin chain comprising at least one functionally active region of a variable region of an antibody, the variable region at its N end being directly linked to a prokaryotic secretion signal peptide to allow secretion of the immunoglobulin chain or fragment of a immunoglobulin chain from a prokaryotic organism. The immunoglobulin chain of claim 1, which is a light immunoglobulin chain. The immunoglobulin chain of claim 1, which is a heavy immunoglobulin chain. An immunoglobulin chain according to any one of the claims, wherein the prokaryotic secretion peptide is a pectalasease signal peptide. An immunoglobulin chain according to claim 1 or 4, wherein the fragment is an Fd fragment. An immunoglobulin chain or fragment of an immunoglobulin chain according to any of the preceding claims which is chimeric. A polynucleotide molecule encoding an immunoglobulin chain or a fragment of an immunoglobulin chain according to any one of the preceding claims. The molecule of claim 7, which is DNA, optionally comprising a promoter. A vector, such as an expression vector, comprising a molecule according to claims 7 or 8. A method for producing an immunoglobulin chain or a fragment of an immunoglobulin chain, the method comprising: a) transforming or transfecting a prokaryotic host with a vector as defined claim 9, comprising a promoter capable of directing transcription in the host; b) culturing the host under appropriate conditions such that the immunoglobulin chain or fragment of an immunoglobulin chain is expressed, the variable region being directly linked at its N end to a prokaryotic secretion signal peptide; and c) obtaining an immunoglobulin chain or a fragment of an immunoglobulin chain originally associated with the signal peptide prior to secretion from the host. A method of producing an immunoglobulin chain or a fragment of an immunoglobulin chain as defined in any one of claims 1 to 6, the method comprising: a) culturing a prokaryotic host capable of expressing the chain or fragment of an immunoglobulin chain under culture conditions; and b) recovering the chain or fragment of an immunoglobulin chain from the culture. A method of producing an immunoglobulin chain or a fragment of an immunoglobulin chain, the method comprising: (a) culturing a transformed or transfected prokaryotic host under appropriate conditions such that the immunoglobulin chain or fragment of an immunoglobulin chain is expressed as a result of transcription in the host a polynucleotide sequence encoding an immunoglobulin chain comprising at least one functionally active region of a variable region of an antibody, the variable region at its N-terminus being directly linked to a prokaryotic secretion signal peptide to enable secretion of the immunoglobulin chain or fragment of a immunoglobulin chain from a prokaryotic organism; and (b) obtaining an immunoglobulin chain or a fragment of an immunoglobulin chain initially associated with the signal peptide prior to secretion from the host. A method according to any one of claims 10 to 12, wherein the secretion signal peptide is a pectalasease secretion signal. The method of any one of claims 10 to 13, wherein the immunoglobulin chain or fragment of an immunoglobulin chain is isolated from the periplasmic compartment. The method of any one of claims 10 to 13, wherein the immunoglobulin chain or fragment of an immunoglobulin chain is isolated from the culture growth medium. A host transformed or transfected with a vector according to claim 9.

DK200301155A 1987-07-24 2003-08-12 Polynucleotide(s) encoding immunoglobulin molecules - used for efficient prodn. of chimeric human or non-human- or class switched-antibodies DK175654B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
DK200301155A DK175654B1 (en) 1987-07-24 2003-08-12 Polynucleotide(s) encoding immunoglobulin molecules - used for efficient prodn. of chimeric human or non-human- or class switched-antibodies

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US7752887A 1987-07-24 1987-07-24
US7752887 1987-07-24
DK199000192A DK174824B1 (en) 1987-07-24 1990-01-24 Polynucleotide molecule, vector comprising polynucleotide molecule and prokaryotic host transfected with this vector
DK19290 1990-01-24
DK200301155A DK175654B1 (en) 1987-07-24 2003-08-12 Polynucleotide(s) encoding immunoglobulin molecules - used for efficient prodn. of chimeric human or non-human- or class switched-antibodies
DK200301155 2003-08-12

Publications (2)

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DK200301155A true DK200301155A (en) 2003-08-12
DK175654B1 DK175654B1 (en) 2005-01-03

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DK200301156A DK175581B1 (en) 1987-07-24 2003-08-12 Polynucleotide(s) encoding immunoglobulin molecules - used for efficient prodn. of chimeric human or non-human- or class switched-antibodies
DK200301155A DK175654B1 (en) 1987-07-24 2003-08-12 Polynucleotide(s) encoding immunoglobulin molecules - used for efficient prodn. of chimeric human or non-human- or class switched-antibodies

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DK200301156A (en) 2003-08-12
DK175581B1 (en) 2004-12-13
DK175654B1 (en) 2005-01-03

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