Purification and establishment of ELISA for vitellogenin of Japanese sardine (Sardinops melanostictus)

Takahiro Matsubara , Tokio Wada , Akihiko Hara
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引用次数: 27

Abstract

Two female-specific serum proteins (FSSPs) were detected immunologically in estradiol-treated Japanese sardine Sardinops melanostictus. The major FSSP was demonstrated to be a high molecular estradiol-inducible glycolipophosphoprotein with an immunological relation to a major yolk protein, and was suggested to be vitellogenin (VTG). VTG was purified using negative immunoaffinity chromatography. The isolated VTG was used for raising the specific antiserum against VTG. A homologous enzyme-linked immunosorbent assay (ELISA) was developed using the antiserum and the isolated VTG. The sensitivity range of the ELISA was 44 ng/ml to 2670 ng/ml of VTG concentration for the conditions used in our investigation.

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日本沙丁鱼(Sardinops melanotictus)卵黄蛋白原酶联免疫吸附试验的纯化与建立
用免疫方法检测了雌二醇处理过的日本沙丁鱼血清中两种雌性特异性血清蛋白(fssp)。主要的FSSP被证明是一种由雌二醇诱导的高分子糖脂磷脂蛋白,与一种主要的卵黄蛋白有免疫学关系,可能是卵黄原蛋白(vitellogenin, VTG)。用免疫亲和层析法纯化VTG。用分离的VTG提高对VTG的特异性抗血清。利用抗血清和分离的VTG建立了同源酶联免疫吸附试验(ELISA)。ELISA的灵敏度范围为44 ng/ml ~ 2670 ng/ml。
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