Recombinant scFv Antibodies against M2e Protein of Influenza A virus

Introduction and Objectives: The M gene of Influenza A virus encodes two conserved proteins, M1 (a capsid protein) and M2 (an ion-channel protein). The M2 protein is an integral membrane protein consisting of an ectodomain (M2e) at the N-terminus. M2e, which consists of 24 N-terminal residues, is remarkably conserved, and the N-terminal epitope SLLTEVET (residues 2-9) is conserved across almost all studied subtypes of Influenza A viruses (99.3%). Recombinant antibodies are new generation of monoclonal antibodies, which are isolated via phage display technology from immune or non-immune phage libraries against target antigens. These antibodies are used for diagnosis of many different antigens and therapeutics proposes. The object of the present study was to isolate recombinant monoclonal antibodies against this important virus.Materials and Methods: The purified Influenza A virus surface antigen, M2e, was coated to immunotubes and used as a target for selection of antibodies from the Tomlinson I and J phage display libraries of single-chain fragment variable (scFv) antibodies. Clones that were able to recognize antigen were isolated in three rounds of binding, elution and amplification. The specificity of scFv antibodies chosen from the resulting panel, were confirmed using enzyme-linked immunosorbent assay (ELISA), dot blotting and western blotting methods. Results: Recombinant antibodies capable of recognizing M2e antigen were isolated with high affinity; and their specificity was approved.Conclusion: Isolated soluble single chain antibodies are good candidates to apply as monoclonal recombinant antibodies in diagnostic kits for detection of Influenza A virus in contaminated samples.

کلیدواژه ها:

Recombinant antibody ، scFv ، Influenza A virus ، M2e protein.