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

MAbs
Nileena VelappanAndrew R M Bradbury

Abstract

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.

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Citations

Feb 16, 2021·Protein Engineering, Design & Selection : PEDS·Nileena VelappanAndrew R M Bradbury
Jul 27, 2021·Frontiers in Cellular and Infection Microbiology·Kristian Daniel Ralph RothMichael Hust

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Datasets Mentioned

BETA
AF389121.1

Methods Mentioned

BETA
PCR
fluorescence microscopy
flow cytometry
phage display
SMA
electrophoresis

Software Mentioned

Geneious®

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