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An enzyme immunoassay for the detection of staphylococcal protein A in affinity-purified products |
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| Authors: | M T Dertzbaugh M C Flickinger W B Lebherz |
| Affiliation: | 1. Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA;2. Institute for Advanced Studies in Biological Process Technology, University of Minnesota, St. Paul, MN 55108, USA;3. National Cancer Institute - Frederick Cancer Research Facility, Fermentation Production Facility, Frederick, MD 21701, USA |
| Abstract: | Rabbit antiserum, specific for protein A from Staphylococcus aureus, was conjugated to alkaline phosphatase and used in a double antibody solid-phase enzyme immunoassay. The assay was developed to monitor eluate from a large-scale protein A-Sepharose affinity column used to purify monoclonal antibodies for human clinical trials. The assay detected soluble protein A in the presence of immunoglobulin at concentrations as low as 4 ng/ml. Analysis of the product purified by affinity chromatography revealed the presence of protein A at ng/ml concentrations. The assay developed here can provide a reliable and convenient method for detecting soluble protein A. |
| Keywords: | protein A enzyme immunoassay affinity chromatography monoclonal antibodies cancer therapy Ig immunoglobulin RIA radioimmunoassay ELISA enzyme-linked immunosorbent assay PBS phosphate-buffered saline BSA bovine serum albumin NMWCO nominal molecular weight cut-off cGMP current Good Manufacturing Practices |
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