Purification of polyoma virus medium-size tumor antigen by immunoaffinity chromatography.
Purification of polyoma virus medium-size tumor antigen by immunoaffinity chromatography.
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通过免疫亲和层析纯化多瘤病毒中等大小肿瘤抗原。
DOI：
10.1073/pnas.79.13.4025
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发表时间：
1982
影响因子：
11.1
通讯作者：
Eckhart,W
中科院分区：
文献类型：
--
作者：
Walter,G;Hutchinson,MA;Hunter,T;Eckhart,W
我们使用针对合成肽Lys-Arg-Ser-Arg-His-Phe（对应于多瘤病毒中型肿瘤（T）抗原的六个COOH末端氨基酸）的抗体，通过亲和层析纯化中型T抗原。通过在含有混合去污剂的中性pH等渗缓冲液中使用大量过量的肽，在温和条件下实现了中T抗原从抗肽抗体的释放。该程序一步即可将中型 T 抗原纯化 2,500 倍。与中型 T 抗原相关的蛋白激酶活性也被释放，并在溶液中的这种活性状态下进行了研究。沉降分析表明，大部分纯化的培养基T抗原呈单体形式（Mr约42,000），与蛋白激酶活性无关。一小部分中型 T 抗原以快速沉淀形式被发现（Mr 约 200,000），具有蛋白激酶活性。
We have used antibodies against the synthetic peptide Lys-Arg-Ser-Arg-His-Phe, corresponding to the six COOH-terminal amino acids of the polyoma virus medium tumor (T) antigen, to purify the medium T antigen by affinity chromatography. Release of the medium T antigen from the anti-peptide antibody was achieved under mild conditions by using a large excess of the peptide in an isotonic buffer at neutral pH containing mixed detergents. This procedure yielded a 2,500-fold purification of the medium T antigen in a single step. The protein kinase activity associated with the medium T antigen was also released and was studied in this active state in solution. Sedimentation analysis showed that the bulk of the purified medium T antigen was in a monomeric form (Mr about 42,000) not associated with protein kinase activity. A small fraction of the medium T antigen was found in a rapidly sedimenting form (Mr about 200,000) that possessed protein kinase activity.