Detergent solubilization, purification, and separation of specificities of HLA antigens from a cultured human lymphoblastoid line, RPMI 4265.
作者： T A Springer , D L Mann , A L DeFranco , J L Strominger
DOI: 10.1016/S0021-9258(17)40215-8
关键词:
摘要: HLA antigens have been purified to homogeneity after detergent solubilization from RPMI 4265, a human lymphoblastoid line. The inhibition of cytotoxicity assay for antigen was modified, using preincubation with bovine serum albumin samples containing prevent lysis target cells by detergent. Solubilization tested many types detergents. A polyethyleneglycol oleyl ether nonionic mixture, Brij 99:Brij 97 (2:1) selected solubilization, since it selectively solubilized antigens, had low absorbance at 280 nm and uncharded. were then Lens culinaris lectin affinity chromatography Bio-Gel A-5m filtration. specifity HLA-A2 separated specificities HLA-B7,12 isoelectric focusing. Purified contained subunit Mr=44,000 NH2-terminal glycine, Mr=12,000, beta2-microglobulin, isoleucine.
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