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[从妊娠晚期血清中纯化妊娠特异性β1-糖蛋白(SP1)及SP1γ组分的免疫学鉴定]

[The purification of pregnancy specific beta 1-glycoprotein (SP1) from third trimester serum and the immunological identification of gamma-component of SP1].

作者信息

Ito M, Hayakawa S, Imaizumi H, Nishiyama Y, Sugiyama Y

出版信息

Nihon Sanka Fujinka Gakkai Zasshi. 1984 Oct;36(10):1913-20.

PMID:6438255
Abstract

This paper describes the SP1 purification procedures with high recovery from third trimester serum and the finding of a new SP1 antigen with gamma-mobility (tentatively called SP1-gamma). The serum was salted out at 40% saturation with ammonium sulfate and this was followed by Sephadex G-150, DEAE-cellulose, SP1-negative affinity chromatography and the mono-Q column of FPLC system. The anti-serum for the immunoadsorption techniques was made from the fractionated male serum. By these procedures, the final SP1 preparation was obtained in a yield of 16% and with a purity of 99%. The purified SP1 (SP1-beta) had beta-mobility and its molecular weight approximated 72,000 in SDS polyacryl-amide gel electrophoresis. The isoelectric points (pI) ranged between 3.80 to 4.55 using isoelectric focusing and the site at pI 4.15 was strongly stained. Another SP1, which passed through DEAE-cellulose with a low salt concentration, was detected in the gamma-region by means of immunoelectrophoresis using anti-SP1 serum. It was ascertained that the SP1-gamma was different from the SP1-beta treated with neuraminidase. The SP1-gamma precipitation fusing with SP1-beta could be observed even in pregnant serum by usual immunoelectrophoresis. The SP1-gamma preparation showed at least four pIs of 5.3, 5.5, 5.65 and 5.8.

摘要

本文描述了从孕晚期血清中高回收率纯化SP1的方法,以及发现一种具有γ迁移率的新SP1抗原(暂称为SP1-γ)。血清用40%饱和度的硫酸铵盐析,然后依次通过Sephadex G-150、DEAE-纤维素、SP1阴性亲和层析和FPLC系统的Mono-Q柱。用于免疫吸附技术的抗血清由分级分离的男性血清制备。通过这些方法,最终获得的SP1制剂产率为16%,纯度为99%。纯化的SP1(SP1-β)具有β迁移率,在SDS聚丙烯酰胺凝胶电泳中其分子量约为72,000。使用等电聚焦法,其等电点(pI)在3.80至4.55之间,pI 4.15处的条带染色强烈。另一种在低盐浓度下通过DEAE-纤维素的SP1,利用抗SP1血清通过免疫电泳在γ区检测到。已确定SP1-γ与经神经氨酸酶处理的SP1-β不同。即使在孕妇血清中,通过常规免疫电泳也可观察到SP1-γ与SP1-β的沉淀融合。SP1-γ制剂显示至少有四个等电点,分别为5.3、5.5、5.65和5.8。

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