Characterization of Novel Alpha-1-Antitrypsin Coding Variants in a Mammalian Cellular Model

Andrea Denardo; Emna Ben Khlifa; Mattia Bignotti; Annamaria Fra

doi:10.1007/978-1-0716-3605-3_8

Characterization of Novel Alpha-1-Antitrypsin Coding Variants in a Mammalian Cellular Model

Methods Mol Biol. 2024:2750:79-93. doi: 10.1007/978-1-0716-3605-3_8.

Authors

Andrea Denardo¹, Emna Ben Khlifa¹, Mattia Bignotti¹, Annamaria Fra²

Affiliations

¹ Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy.
² Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy. annamaria.fra@unibs.it.

PMID: 38108969
DOI: 10.1007/978-1-0716-3605-3_8

Abstract

Advances in genetic screening technologies have considerably accelerated the discovery of rare alpha-1-antitrypsin (AAT) variants. Expression in cellular models is an effective approach to evaluate the pathogenic potential of these new AAT variants, whose clinical significance would otherwise remain uncertain. Here we provide a detailed description of established methods for in vitro characterization of AAT coding variants expressed in HEK293T/17 cells. The protocols include determination of secretion efficiency, the tendency to form polymeric chains and the anti-elastase inhibitory activity.

Keywords: Alpha-1-antitrypsin polymers; Alpha-1-antitrypsin rare variants; Anti-elastase inhibitory activity; Pathogenicity prediction; SERPINA1.

MeSH terms

Animals
Cloning, Molecular
HEK293 Cells
Humans
Mammals
Nucleic Acid Amplification Techniques*
Polymers*
Technology

Substances

Polymers