Detection, isolation and characterization of staphylococcal enterotoxin B in protein A preparations purified by immunoglobulin G affinity chromatography
Journal of Immunological Methods, ISSN: 0022-1759, Vol: 116, Issue: 1, Page: 37-43
1989
- 8Citations
- 4Captures
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Metrics Details
- Citations8
- Citation Indexes8
- CrossRef7
- Captures4
- Readers4
Article Description
Studies were performed to detect and isolate trace contaminants of staphylococcal enterotoxin B (SEB) in various protein A preparations isolated by affinity chromatography employing human IgG covalently bound to Sepharose 4B. Utilizing an ELISA technique, trace amounts (0.018–0.138%) of SEB could be detected in protein A preparations after separation of the SEB employing a molecular sizing column in a high pressure liquid chromatography (HPLC) system. Trace contamination by SEB could be removed from protein A preparations by an additional DEAE ion exchange chromatography step employing a low ionic strength buffer system (0.005 M NaCl in 0.01 phosphate buffer, pH 7.50). The resulting protein A preparations possessed a purity higher than that observed prior to the final purification step. Polyacrylamide gel electrophoresis (PAGE) analyses of the trace contamination removed from protein A preparations by ion exchange chromatography revealed, in addition to SEB, several additional contaminating polypeptides of an unknown nature. These studies indicate that protein A preparations of high purity can be prepared by employing DEAE ion exchange chromatography in addition to affinity chromatography utilizing immobilized human IgG.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/0022175989903104; http://dx.doi.org/10.1016/0022-1759(89)90310-4; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0024532624&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/2915124; https://linkinghub.elsevier.com/retrieve/pii/0022175989903104; http://dx.doi.org/10.1016/0022-1759%2889%2990310-4; https://dx.doi.org/10.1016/0022-1759%2889%2990310-4
Elsevier BV
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