豚鼠血清中抗胰岛素IgG的新型酶免疫测定法。

A novel enzyme immunoassay of anti-insulin IgG in guinea pig serum.

作者信息

Kohno T, Ishikawa E

出版信息

Biochem Biophys Res Commun. 1987 Sep 15;147(2):644-9. doi: 10.1016/0006-291x(87)90979-x.

Abstract

A novel enzyme immunoassay of anti-insulin IgG in guinea pig serum is described. Guinea pig anti-insulin serum diluted with nonspecific guinea pig serum was incubated with dinitrophenyl biotinyl nonspecific rabbit IgG-insulin conjugate and a rabbit (anti-dinitrophenyl bovine serum albumin) IgG-coated polystyrene ball. After washing to eliminate nonspecific guinea pig IgG in the diluted serum, the polystyrene ball was incubated with dinitrophenyl-L-lysine to elute the complex of anti-insulin IgG and the conjugate. The eluate was incubated with an avidin-coated polystyrene ball. Finally, the amount of guinea pig anti-insulin IgG in the complex trapped onto the avidin-coated polystyrene ball was measured by incubation with rabbit (anti-guinea pig IgG) Fab'-peroxidase conjugate. This enzyme immunoassay was 10,000-fold more sensitive than the conventional enzyme immunoassay using insulin-coated polystyrene ball and rabbit (anti-guinea pig IgG) Fab'-peroxidase conjugate.

摘要

本文描述了一种检测豚鼠血清中抗胰岛素IgG的新型酶免疫测定法。用非特异性豚鼠血清稀释的豚鼠抗胰岛素血清与二硝基苯基生物素化非特异性兔IgG-胰岛素缀合物以及包被有兔（抗二硝基苯基牛血清白蛋白）IgG的聚苯乙烯球一起孵育。洗涤以去除稀释血清中的非特异性豚鼠IgG后，将聚苯乙烯球与二硝基苯基-L-赖氨酸一起孵育，以洗脱抗胰岛素IgG与缀合物的复合物。将洗脱液与包被有抗生物素蛋白的聚苯乙烯球一起孵育。最后，通过与兔（抗豚鼠IgG）Fab'-过氧化物酶缀合物孵育，测定捕获在包被有抗生物素蛋白的聚苯乙烯球上的复合物中豚鼠抗胰岛素IgG的量。这种酶免疫测定法比使用包被有胰岛素的聚苯乙烯球和兔（抗豚鼠IgG）Fab'-过氧化物酶缀合物的传统酶免疫测定法灵敏10000倍。