用免疫吸收层析法快速纯化洗涤剂溶解型Ia抗原

Loran T. Clement, Anne M. Kask, Ethan M. Shevach
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引用次数: 8

摘要

用非离子清洁剂Nonidet P-40* (NP-40)溶解豚鼠淋巴细胞中的Ia抗原,通过两个简单的亲和层析步骤纯化。第一步由扁豆凝集素层析组成,从中获得富含糖蛋白的部分。最后一步是用la特异性免疫吸附剂进行层析,用酸性甘氨酸缓冲液洗脱抗原。通过抑制细胞毒性测定抗原产率,菌株13 Ia抗原的平均产率为21%,菌株2 Ia抗原的平均产率为19%。电泳分析表明获得了高度的纯化。免疫吸收色谱法是一种快速有效的分离Ia抗原的方法。
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Rapid purification of detergent-solubilized Ia antigens by immunoabsorbent chromatography

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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