{"title":"Rapid purification of detergent-solubilized Ia antigens by immunoabsorbent chromatography","authors":"Loran T. Clement, Anne M. Kask, Ethan M. Shevach","doi":"10.1016/0161-5890(78)90137-2","DOIUrl":null,"url":null,"abstract":"<div><p>The Ia antigens from strain 2 and strain 13 guinea pig lymphocytes were solubilized with the non-ionic detergent Nonidet P-40* (NP-40) and purified by two simple affinity chromatography steps. The first step consisted of lentil lectin chromatography from which a glycoprotein-enriched fraction was obtained. The final step consisted of chromatography using la-specific immunoabsorbents, from which the antigens were eluted with acid glycine buffer. Antigen yields were determined by inhibition of cytotoxicity, and mean yields were 21% for the strain 13 Ia antigens and 19% for the strain 2 Ia antigens. Electrophoretic analysis demonstrated the high degree of purification attained. Immunoabsorbent chromatography offers a rapid and effective method for the isolation of Ia antigens.</p></div>","PeriodicalId":13265,"journal":{"name":"Immunochemistry","volume":"15 6","pages":"Pages 393-399"},"PeriodicalIF":0.0000,"publicationDate":"1978-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0161-5890(78)90137-2","citationCount":"8","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Immunochemistry","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0161589078901372","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 8
Abstract
The Ia antigens from strain 2 and strain 13 guinea pig lymphocytes were solubilized with the non-ionic detergent Nonidet P-40* (NP-40) and purified by two simple affinity chromatography steps. The first step consisted of lentil lectin chromatography from which a glycoprotein-enriched fraction was obtained. The final step consisted of chromatography using la-specific immunoabsorbents, from which the antigens were eluted with acid glycine buffer. Antigen yields were determined by inhibition of cytotoxicity, and mean yields were 21% for the strain 13 Ia antigens and 19% for the strain 2 Ia antigens. Electrophoretic analysis demonstrated the high degree of purification attained. Immunoabsorbent chromatography offers a rapid and effective method for the isolation of Ia antigens.
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