Expression, purification and characterization of a mouse-human chimeric antibody and chimeric Fab' fragment
| Autor: | D C Low, John Robert Adair, Geoffrey Yarranton, Sarojani Angal, K A Proudfoot, J. C. Lloyd, David J. King, Mark Bodmer |
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| Rok vydání: | 1992 |
| Předmět: |
medicine.drug_class
Gene Expression CHO Cells Immunoglobulin light chain Monoclonal antibody Biochemistry Immunoglobulin Fab Fragments Mice Antigen Antigens Neoplasm Cricetinae medicine Animals Humans Molecular Biology Binding selectivity Chromatography High Pressure Liquid biology Chimera Chinese hamster ovary cell Proteolytic enzymes Antibodies Monoclonal Cell Biology Molecular biology biology.protein Electrophoresis Polyacrylamide Gel Antibody Research Article |
| Zdroj: | The Biochemical journal. 281 |
| ISSN: | 0264-6021 |
| Popis: | B72.3 is a mouse monoclonal antibody against a tumour-associated antigen, TAG72, which recognizes breast, ovarian and colorectal tumour tissue. A mouse-human chimeric version of B72.3 has been expressed in Chinese-hamster ovary cells. This molecule has the binding specificity of B72.3 and constant regions from human IgG4. The chimeric B72.3 assembles to intact IgG and recognizes TAG72 as well as B72.3 in competitive binding assays. A proportion of the chimeric B72.3 (approx. 10%) does not form inter-heavy-chain disulphide bonds but still assembles into the IgG tetramer. This appears to be a general property of human IgG4 molecules. Co-expression of the chimeric light chain with a chimeric Fd' gene resulted in the expression of functional Fab'. Very little F(ab')2 is produced, although the Fab' can be oxidized to the dimeric F(ab')2 in vitro. The production of Fab' and F(ab')2 by this method is an attractive alternative to proteolytic digestion of IgG. The ability to produce these molecules in large quantities will allow the production and testing of a range of anti-tumour antibody and antibody fragment conjugates. |
| Databáze: | OpenAIRE |
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