Rapid isotyping of murine monoclonal antibodies by enzyme-linked immunofiltration assay |
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Authors: | K K Hines C R Clark |
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Institution: | (1) Research and Development, Pierce Chemical Company, PO Box 117, 61105 Rockford, IL, USA |
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Abstract: | Summary As a part of the initial characterization of monoclonal antibodies, the isotype is routinely identified by enzyme-linked immunosorbent assay (ELISA). In the present study, we describe an isotyping methodology that uses the technique referred to as enzyme-linked immunofiltration assay (ELIFA) for greatly accelerating the assay compared with ELISA. In the ELIFA method, solutions are filtered through a nitrocellulose membrane using a controlled flow rate to bind proteins. By using this technology, the time required to isotype a monoclonal antibody was reduced from a minimum of 4 to 8 hr using a standard ELISA assay to 30 min with ELIFA. |
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