Title:
AB5 TOXIN B SUBUNIT MUTANTS WITH ALTERED CHEMICAL CONJUGATION CHARACTERISTICS
Document Type and Number:
WIPO Patent Application WO2001027144
Kind Code:
A3
Abstract:
A recombinant AB5 B subunit protein including at least one mutation, wherein the mutation alters the number of amino acid residues available for chemical modification as compared to a wild type AB5 B subunit protein, and wherein said recombinant protein retains an effective target ligand bind affinity. For example, specifically designed mutations are produced in the cholera Toxin B subunit (CTB) such that it can still bind with high affinity to its receptor, Gm-1, but can be specifically covalently linked at lysines or cysteines to an immunogen or vaccine. The vaccine produced from this coupling is a mucosal vaccine which has high immunogenicity due to the interaction with the CTB. The vaccine can be produced inexpensively and easily. Alternatively, a technique is disclosed for treating CTB such that non-covalent coupling to a vaccine or immunogen can occur.
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Inventors:
HANDLEY HAROLD H (US)
HAAPARANTA TAPIO (US)
EWALT KARLA L (US)
HAAPARANTA TAPIO (US)
EWALT KARLA L (US)
Application Number:
PCT/US2000/027607
Publication Date:
January 17, 2002
Filing Date:
October 05, 2000
Export Citation:
Assignee:
ACTIVE BIOTECH AB (SE)
HANDLEY HAROLD H (US)
HAAPARANTA TAPIO (US)
EWALT KARLA L (US)
HANDLEY HAROLD H (US)
HAAPARANTA TAPIO (US)
EWALT KARLA L (US)
International Classes:
C12N15/09; A61K39/00; A61P3/10; A61P29/00; A61P31/00; A61P37/04; A61P37/06; C07K14/235; C07K14/245; C07K14/25; C07K14/28; C12P21/02; (IPC1-7): C07K14/28; C07K14/245; C07K14/25; C07K14/235; C12N15/31; C12N5/10; A61K39/106; A61K39/112; A61K39/10; A61K39/108
Foreign References:
| US5834246A | 1998-11-10 | |||
| EP0556388A1 | 1993-08-25 | |||
| FR2636842A1 | 1990-03-30 |
Other References:
JOBLING M G ET AL: "ANALYSIS OF STRUCTURE AND FUNCTION OF THE B SUBUNIT OF CHOLERA TOXIN BY THE USE OF SITE-DIRECTED MUTAGENESIS", MOLECULAR MICROBIOLOGY, vol. 55, no. 7, 1991, pages 1755 - 1767, XP000992490, ISSN: 0950-382X
BACKSTROM M ET AL: "Characterization of an internal permissive site in the cholera toxin B-subunit and insertion of epitopes from human immunodeficiency virus-1, hepatitis B virus and enterotoxigenic Escherichia coli", GENE,NL,ELSEVIER BIOMEDICAL PRESS. AMSTERDAM, vol. 165, no. 2, 20 November 1995 (1995-11-20), pages 163 - 171, XP004043137, ISSN: 0378-1119
DEBINSKI W ET AL: "AN IMMUNOTOXIN WITH INCREASED ACTIVITY AND HOMOGENEITY PRODUCED BY REDUCING THE NUMBER OF LYSINE RESIDUES IN RECOMBINANT PSEUDOMONASEXOTOXIN", BIOCONJUGATE CHEMISTRY,US,AMERICAN CHEMICAL SOCIETY, WASHINGTON, vol. 5, no. 1, 1994, pages 40 - 46, XP000430384, ISSN: 1043-1802
DE WOLF MARC J S ET AL: "Regeneration of active receptor recognition domains on the B subunit of cholera toxin by formation of hybrids from chemically inactivated derivatives.", BIOCHIMICA ET BIOPHYSICA ACTA, vol. 1223, no. 2, 1994, pages 285 - 295, XP000992489, ISSN: 0006-3002
MERRITT ETHAN A ET AL: "Crystal structure of cholera toxin B-pentamer bound to receptor G-M1 pentasaccharide.", PROTEIN SCIENCE, vol. 3, no. 2, 1994, pages 166 - 175, XP000992494, ISSN: 0961-8368
BACKSTROM M ET AL: "Characterization of an internal permissive site in the cholera toxin B-subunit and insertion of epitopes from human immunodeficiency virus-1, hepatitis B virus and enterotoxigenic Escherichia coli", GENE,NL,ELSEVIER BIOMEDICAL PRESS. AMSTERDAM, vol. 165, no. 2, 20 November 1995 (1995-11-20), pages 163 - 171, XP004043137, ISSN: 0378-1119
DEBINSKI W ET AL: "AN IMMUNOTOXIN WITH INCREASED ACTIVITY AND HOMOGENEITY PRODUCED BY REDUCING THE NUMBER OF LYSINE RESIDUES IN RECOMBINANT PSEUDOMONASEXOTOXIN", BIOCONJUGATE CHEMISTRY,US,AMERICAN CHEMICAL SOCIETY, WASHINGTON, vol. 5, no. 1, 1994, pages 40 - 46, XP000430384, ISSN: 1043-1802
DE WOLF MARC J S ET AL: "Regeneration of active receptor recognition domains on the B subunit of cholera toxin by formation of hybrids from chemically inactivated derivatives.", BIOCHIMICA ET BIOPHYSICA ACTA, vol. 1223, no. 2, 1994, pages 285 - 295, XP000992489, ISSN: 0006-3002
MERRITT ETHAN A ET AL: "Crystal structure of cholera toxin B-pentamer bound to receptor G-M1 pentasaccharide.", PROTEIN SCIENCE, vol. 3, no. 2, 1994, pages 166 - 175, XP000992494, ISSN: 0961-8368
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