鸟氨酸脱羧酶的夹心酶免疫测定法。

Sandwich enzyme immunoassay for ornithine decarboxylase.

作者信息

Nishiyama M, Matsufuji S, Kanamoto R, Murakami Y, Hayashi S

机构信息

Department of Nutrition, Jikel University School of Medicine, Tokyo, Japan.

出版信息

J Immunoassay. 1989;10(1):19-35. doi: 10.1080/01971528908053225.

DOI:10.1080/01971528908053225

PMID:2497140

Abstract

A sensitive enzyme immunoassay (EIA) was developed for the determination of ornithine decarboxylase (ODC, EC 4.1.1.17), in the range of 0.02-10 ng, using an affinity-purified anti-ODC-Fab'-peroxidase conjugate. The amount of ODC protein was determined in crude extracts from the kidney of testosterone-treated mice, regenerating rat liver and human thyroid carcinoma, with purified mouse kidney or rat liver enzyme as standard. In all these tissues, similar activity/protein ratios were found for ODC: 1.2 x 10(6)-1.9 x 10(6) nmol CO2/h/mg of ODC protein, which were roughly equivalent to the final specific activity of purified enzymes. ODC inactivated by alpha- difluoro-methylornithine (DFMO) could also be assayed with this method similarly to active ODC protein. However, ODC-antizyme complex gave a somewhat lower value than free ODC protein.

摘要

利用亲和纯化的抗鸟氨酸脱羧酶（ODC）-Fab'-过氧化物酶偶联物，开发了一种灵敏的酶免疫测定法（EIA），用于测定0.02 - 10 ng范围内的鸟氨酸脱羧酶（ODC，EC 4.1.1.17）。以纯化的小鼠肾脏或大鼠肝脏酶为标准品，测定了经睾酮处理的小鼠肾脏、再生大鼠肝脏和人甲状腺癌粗提物中的ODC蛋白量。在所有这些组织中，ODC的活性/蛋白比值相似：1.2×10⁶ - 1.9×10⁶ nmol CO₂/小时/毫克ODC蛋白，这大致相当于纯化酶的最终比活性。用α-二氟甲基鸟氨酸（DFMO）灭活的ODC也可用此方法进行测定，其结果与活性ODC蛋白相似。然而，ODC-抗酶复合物的值略低于游离ODC蛋白。