心房肽酶免疫测定法（EIA）的开发、验证及应用

Development, validation and application of an enzyme immunoassay (EIA) of atriopeptin.

作者信息

McLaughlin L L, Wei Y F, Stockmann P T, Leahy K M, Needleman P, Grassi J, Pradelles P

出版信息

Biochem Biophys Res Commun. 1987 Apr 14;144(1):469-76. doi: 10.1016/s0006-291x(87)80533-8.

Abstract

A rapid, convenient, and sensitive enzyme immunoassay (EIA) for atriopeptin (AP) has been developed. The tracer-ligand for the assay is the 24-amino acid peptide, AP24, which has been covalently coupled to the tetrameric form of acetylcholinesterase (AChE) (EC 3.1.1.7). Tracer, unknown, and primary antibody are incubated in a 96-well microtiter plate precoated with secondary antibody. After washing, a colorimetric reaction is used to measure acetylcholinesterase activity. A direct linear correlation was obtained when comparing the conventional radioimmunoassay and the EIA by using the same primary antibody to assay: plasma samples (rat or human), HPLC column fractions, or atrial extracts. Besides being technically much less demanding and not requiring the use of the radioisotopes, the EIA is more sensitive than the radioimmunoassay and thereby lends itself to a "flash" same-day assay of samples.

摘要

已开发出一种用于心房肽（AP）的快速、便捷且灵敏的酶免疫测定法（EIA）。该测定法的示踪配体是24个氨基酸的肽AP24，它已共价偶联到乙酰胆碱酯酶（AChE）（EC 3.1.1.7）的四聚体形式上。将示踪剂、未知物和一抗在预包被有二抗的96孔微量滴定板中孵育。洗涤后，使用比色反应来测量乙酰胆碱酯酶活性。当使用相同的一抗检测血浆样本（大鼠或人）、HPLC柱馏分或心房提取物时，通过比较传统放射免疫测定法和EIA获得了直接线性相关性。除了在技术上要求低得多且无需使用放射性同位素外，EIA比放射免疫测定法更灵敏，因此适合对样本进行“快速”当日测定。