Isolation of antigen/antibody complexes through organic solvent (ICOS) method
| Autor: | Gene Kurosawa, Mototaka Sugiura, Yasushi Akahori, Chiho Muramatsu, Keiko Eguchi, Kazuhiro Suzuki, Yoshitaka Iba, Miho Tanaka, Atsushi Sugioka, Yoshikazu Kurosawa, Miwa Morita, Mariko Sumitomo |
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| Rok vydání: | 2009 |
| Předmět: |
Antibody-dependent cell-mediated cytotoxicity
Antigen-Antibody Complex biology medicine.drug_class Chemistry Biophysics Antibodies Monoclonal Membrane Proteins Cell Biology Monoclonal antibody Biochemistry Molecular biology Epitope Antigen Peptide Library Methods Solvents medicine biology.protein Humans Centrifugation Antibody Peptide library Molecular Biology |
| Zdroj: | Biochemical and Biophysical Research Communications. 378:832-835 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2008.11.129 |
| Popis: | We developed a method termed ICOS (isolation of antigen-antibody complexes through organic solvent) for comprehensive isolation of monoclonal antibodies (mAbs) bound to molecules on the cell surface. By mixing a large number of phage particles of an antibody (Ab) library with living cells, antigen (Ag)-Ab complexes were formed on the cell surface. The mixture was overlaid on organic solution in a tube and subjected to centrifugation. Phages bound to cells were recovered from the precipitate. The phage fraction isolated turned out to contain mAbs that bind to very heterogeneous epitopes and show strong binding activity to Ags. The ICOS method was applied to isolation of human mAbs that may be therapeutic against cancers. Sixty percent of clones isolated by the screening of a phage Ab library against cancer cells turned out to bind to various kinds of tumor-associated Ags. The precise protocol of ICOS method and the rationale of efficient screening were described. |
| Databáze: | OpenAIRE |
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