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人白细胞干扰素的连续抗体亲和层析法

Sequential antibody affinity chromatography of human leukocyte interferon.

作者信息

Berg K

出版信息

Scand J Immunol. 1977;6(1-2):77-86. doi: 10.1111/j.1365-3083.1977.tb00323.x.

Abstract

Antibodies to crude leukocyte interferon (CIF) were bound to Sepharose 4B, and CIF was chromatographed. A relationship between recovered interferon and percentage binding of the antibodies to Sepharose 4B was observed. Highest recovery was obtained when only 50%-85% of th- antibodies were bound. When CIF was subjected to normal affinity chromatography, specific activity of 1-5 X 10(6) interferon units (IFU) (69/19B-units)/mg protein was obtained. When this purified preparation was further chromatographed on a column mainly directed against normal cellular proteins and then concentrated by normal affinity chromatography, a tenfold increase in 'direct' specific activity was obtained. It is suggested that the 'indirect' specific activity is about 2 X 10(8) IFU/mg protein.

摘要

将抗粗制白细胞干扰素(CIF)抗体与琼脂糖4B结合,对CIF进行层析。观察到回收的干扰素与抗体对琼脂糖4B的结合百分比之间的关系。当仅50%-85%的抗体被结合时,回收率最高。当CIF进行常规亲和层析时,获得的比活性为1-5×10⁶干扰素单位(IFU)(69/19B单位)/mg蛋白质。当这种纯化制剂在主要针对正常细胞蛋白质的柱上进一步层析,然后通过常规亲和层析浓缩时,“直接”比活性提高了10倍。据推测,“间接”比活性约为2×10⁸IFU/mg蛋白质。

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