Title:
ISOLATION OF IMMUNOGLOBULIN MOLECULES THAT LACK INTER-HEAVY CHAIN DISULFIDE BONDS
Document Type and Number:
WIPO Patent Application WO2004026427
Kind Code:
A3
Abstract:
The current invention features methods for reliably and controllably separating immunoglobulin half antibodies from immunoglobulin whole antibodies, as well as purified immunoglobulin half antibody preparations and purified immunoglobulin whole antibody preparations while preserving biological activity. These dissociated half antibodies can be chromatographically separated from whole antibodies. There are four known subclasses of IgG molecules: IgG1; IgG2; IgG3; and IgG4. IgG4 molecules differ from the other IgG isotypes in that the disulfide bonds that link the two heavy chain subunits together do not always form. Due to the non-covalent interactions that hold the heavy chain subunits together, the heterogeneity of IgG4 molecules is not apparent following gel filtration of purified IgG4 protein. However, when purified IgG4 protein is separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE) under non-reducing conditions, two distinct protein species can be identified - whole antibody and "half-antibodies".
Inventors:
BIRCK-WILSON ESZTER (US)
DAY MARIA (US)
DAY MARIA (US)
Application Number:
PCT/US2003/028543
Publication Date:
October 07, 2004
Filing Date:
September 11, 2003
Export Citation:
Assignee:
GTC BIOTHERAPEUTICS INC (US)
BIRCK-WILSON ESZTER (US)
DAY MARIA (US)
BIRCK-WILSON ESZTER (US)
DAY MARIA (US)
International Classes:
A61K39/395; C07K16/02; C07K16/04; C07K16/06; C07K16/18; C12P21/08; G01N33/53; G01N33/531; G01N33/563; B01D; (IPC1-7): A61K39/395; C07K16/02; C07K16/04; C07K16/06; C12P21/08; G01N33/53; G01N33/531; G01N33/563
Foreign References:
| US4479895A | 1984-10-30 | |||
| US5292668A | 1994-03-08 | |||
| US6329507B1 | 2001-12-11 |
Other References:
KRETZSCHMAR ET AL: "High-level expression in insect cells and purification of secreted monomeric single-chain Fv antibodies", JOURNAL OF IMMUNOLOGICAL METHODS, vol. 195, 1996, pages 93 - 101, XP004021258
See also references of EP 1549342A4
See also references of EP 1549342A4
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