一种采用酶标记法和新型固相载体的兔免疫球蛋白G夹心酶免疫测定法。

A sandwich enzyme immunoassay of rabbit immunoglobulin G with an enzyme labeling method and a new solid support.

作者信息

Tanimori H, Ishikawa F, Kitagawa T

出版信息

J Immunol Methods. 1983 Aug 12;62(1):123-31. doi: 10.1016/0022-1759(83)90117-5.

Abstract

A method was devised for enzyme labeling goat antibody to rabbit immunoglobulin G with beta-D-galactosidase (EC 3.2.1.23) using a heterobifunctional cross-linker, N-(gamma-maleimidobutyryloxy)-succinimide. Labeling of the purified antibody was by a continuous 2-step process and including a chromatographic purification procedure could be completed within one day. The partially purified anti-rabbit IgG was coated on Amino-Dylark cylinders, a new solid support, using glutaraldehyde as the coupling reagent. With enzyme-labeled antibody and the solid-phase anti-rabbit IgG, a sandwich enzyme immunoassay for rabbit IgG was developed with a lower limit of detection at 3.5 pM (0.1 ng/tube). The specificity of the assay was excellent and all 4 types of IgG tested showed 0.0001% or less cross-reactivity with rabbit IgG.

摘要

设计了一种方法，使用异双功能交联剂N-(γ-马来酰亚胺丁酰氧基)-琥珀酰亚胺，用β-D-半乳糖苷酶（EC 3.2.1.23）对山羊抗兔免疫球蛋白G进行酶标记。纯化抗体的标记通过连续两步法进行，并且包括色谱纯化步骤，可在一天内完成。使用戊二醛作为偶联试剂，将部分纯化的抗兔IgG包被在一种新型固相载体氨基-Dylark柱上。利用酶标记抗体和固相抗兔IgG，开发了一种兔IgG夹心酶免疫测定法，检测下限为3.5 pM（0.1 ng/管）。该测定法的特异性极佳，所测试的所有4种IgG与兔IgG的交叉反应率均显示为0.0001%或更低。