Selection and verification of antibodies against the cytoplasmic domain of M2 of influenza, a transmembrane protein

mAbs

Sofiya Micheva-Viteva¹  Antonietta M. Lillo¹  Samantha H. Adikari¹  Nileena Velappan¹  Geoffrey S. Waldo¹  Andrew R.M. Bradbury² 

[1] Biosecurity and Public Health, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM, USA;Specifica Inc., Santa Fe, NM, USA;

关键词: Influenza A;    M2 cytoplasmic domain;    antibodies;    transmembrane proteins;    phage display;    yeast display;   

DOI  :  10.1080/19420862.2020.1843754

来源: DOAJ

【 摘 要 】

Interactions between the cytoplasmic domains of viral transmembrane proteins and host machinery often determine the outcome of viral infection. The M2 protein of influenza A has been identified as a key player in autophagy-mediated viral replication. Here, we describe the engineering and validation of an antibody specific for the cytoplasmic domain of the M2 protein. Through phage and yeast display selection techniques, we obtained an antibody that recognizes: 1) the M2 cytoplasmic domain purified from bacterial inclusion bodies and refolded, 2) full-length M2 recombinant protein expressed in mammalian cells, and 3) native M2 protein in influenza A infected cells. This antibody can serve as a molecular tool to enhance our knowledge of protein–protein interactions between influenza A virus and the host cell machinery. We anticipate the methods described herein will further the development of antibodies specific to the cytoplasmic domains of transmembrane proteins.

【 授权许可】

Unknown