Purification of monoclonal antibodies from ascitic fluid using preparative electrophoresis
| Autor: | S Lim, H.P Manusu, Keith L. Williams, D.B Rylatt, Andrew A. Gooley |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
medicine.drug_class
Immunoglobulin light chain Monoclonal antibody Biochemistry Analytical Chemistry Mice medicine Animals Ascitic Fluid Gel electrophoresis Chromatography biology Chemistry Isoelectric focusing Organic Chemistry Antibodies Monoclonal General Medicine Hydrogen-Ion Concentration Gel electrophoresis of proteins Electrophoresis biology.protein Electrophoresis Polyacrylamide Gel Isoelectric Focusing Antibody Protein A |
| Zdroj: | Journal of Chromatography A. 827:329-335 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/s0021-9673(98)00847-4 |
| Popis: | Four monoclonal antibodies (including Ig subclasses, G1, G2a, and G2b) were purified from murine ascitic fluid by a preparative electrophoresis system using a charge- and size-based strategy. Most of the smaller contaminating proteins were removed at pH 8.3 when the ascitic fluid was passed through a cartridge containing a separating membrane with a pore size of M(r) 100,000. After this single step, the immunoglobulin heavy and light chains were the only significant bands present when analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A second step, involving electrophoresis at pH 6.4-7.5 depending on the antibody can be used to remove residual contaminants. For each of the antibodies tested, the recovery of activity at each step was over 80%. As this technology is directly scalable, purification of antibodies by the method described here could be considered a cost effective alternative to protein A chromatography. |
| Databáze: | OpenAIRE |
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