夹心酶联免疫吸附测定法定量检测人α1抗胰蛋白酶总量

Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA.

作者信息

Tang Qiushi, Gruntman Alisha M, Flotte Terence R

机构信息

Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, 01605, USA.

Department of Pediatrics, University of Massachusetts Medical School, Worcester, MA, 01605, USA.

出版信息

Methods Mol Biol. 2017;1639:211-216. doi: 10.1007/978-1-4939-7163-3_20.

DOI:10.1007/978-1-4939-7163-3_20

PMID:28752460

Abstract

In this chapter we describe an enzyme-linked immunosorbent assay (ELISA) to quantitatively measure human alpha-1 antitrypsin (AAT) protein levels in serum, other body fluids or liquid media. This assay can be used to measure the expression of the human AAT (hAAT) gene in a variety of gene transfer or gene downregulation experiments.A hAAT-specific capture antibody and a HRP-conjugated anti-AAT detection antibody are used in this assay. The conjugated anti-AAT used in this protocol, instead of the typical sandwich which employs an unconjugated antibody followed by a specifically conjugated IgG, makes the assay simpler and decreases variability. This provides a useful tool to evaluate the AAT levels in clinical and research samples and can allow fairly rapid testing of a large number of samples.

摘要

在本章中，我们描述了一种酶联免疫吸附测定法（ELISA），用于定量测量血清、其他体液或液体培养基中的人α-1抗胰蛋白酶（AAT）蛋白水平。该测定法可用于在各种基因转移或基因下调实验中测量人AAT（hAAT）基因的表达。本测定法使用hAAT特异性捕获抗体和辣根过氧化物酶（HRP）偶联的抗AAT检测抗体。本方案中使用的偶联抗AAT，而非采用未偶联抗体后接特异性偶联IgG的典型夹心方法，使测定更简单并降低了变异性。这为评估临床和研究样本中的AAT水平提供了一个有用的工具，并且能够对大量样本进行相当快速的检测。