Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA

Qiushi Tang; Alisha M Gruntman; Terence R Flotte

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

Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA

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

Authors

Qiushi Tang^{1

2}, Alisha M Gruntman³, Terence R Flotte^{3

4

5}

Affiliations

¹ Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, 01605, USA. Qiushi.Tang@umassmed.edu.
² Department of Pediatrics, University of Massachusetts Medical School, Worcester, MA, 01605, USA. Qiushi.Tang@umassmed.edu.
³ Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
⁴ Department of Pediatrics, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
⁵ Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA, 01605, USA.

PMID: 28752460
DOI: 10.1007/978-1-4939-7163-3_20

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.

Keywords: AAT; Alpha-1 antitrypsin; Enzyme-linked immunosorbent assay; Sandwich ELISA.

MeSH terms

Enzyme Assays
Enzyme-Linked Immunosorbent Assay / methods*
Humans
Reference Standards
alpha 1-Antitrypsin / analysis*

Substances

alpha 1-Antitrypsin