化学发光夹心酶免疫分析中非特异性结合的减少

The reduction of nonspecific binding in chemiluminescent sandwich enzyme immunoassays.

作者信息

Aoyagi S, Kusumi M, Matsuyuki A, Maeda M, Tsuji A

机构信息

R&D Center, Meidensha Co., Tokyo, Japan.

出版信息

J Immunol Methods. 1991 Mar 1;137(1):73-8. doi: 10.1016/0022-1759(91)90395-v.

DOI:10.1016/0022-1759(91)90395-v

PMID:1707082

Abstract

The reduction in nonspecific binding obtained in EIA procedures based on glucose oxidase (GO)-labelled antibody is determined by the origin of the antibodies used for both the solid phase and for GO labelling. We have examined the relationship between nonspecific binding and various antibody combinations for the purpose of establishing highly sensitive chemiluminescent sandwich enzyme immunoassays. It was shown that nonspecific binding could be reduced by the following combination of solid phase and GO labelled antibodies: (i) guinea pig IgG-guinea pig IgG, (ii) goat IgG-rabbit IgG, (iii) goat IgG-rabbit Fab'. On the basis of these results, we succeeded in establishing highly sensitive chemiluminescent sandwich enzyme immunoassays for human chorionic gonadotropin (hCG) and alpha-fetoprotein (AFP). The detection limit was 2.5 mIU/ml for hCG using combination (i), 1 ng/ml for AFP using combination (ii), and 0.05 ng/ml (70 amol/assay) for AFP using combination (iii).

摘要

基于葡萄糖氧化酶（GO）标记抗体的酶免疫分析（EIA）程序中，非特异性结合的减少取决于用于固相和GO标记的抗体来源。为了建立高灵敏度的化学发光夹心酶免疫分析方法，我们研究了非特异性结合与各种抗体组合之间的关系。结果表明，通过以下固相和GO标记抗体的组合可以减少非特异性结合：（i）豚鼠IgG - 豚鼠IgG，（ii）山羊IgG - 兔IgG，（iii）山羊IgG - 兔Fab'。基于这些结果，我们成功建立了针对人绒毛膜促性腺激素（hCG）和甲胎蛋白（AFP）的高灵敏度化学发光夹心酶免疫分析方法。使用组合（i）时，hCG的检测限为2.5 mIU/ml；使用组合（ii）时，AFP的检测限为1 ng/ml；使用组合（iii）时，AFP的检测限为0.05 ng/ml（70 amol/测定）。