Generation and Characterization of the Monoclonal Antibody Panel Specific to the NS1 Protein of the Influenza A Virus

Abstract

⎯The paper reports on the production of a panel of seven new monoclonal antibodies (MAbs) capable of specifically binding to the NS1 1-124 fragment of the influenza virus A/Puerto Rico/8/1934 (H1N1) NS1 protein in indirect enzyme-linked immunosorbent assay (ELISA), as well as to the full-size native NS1 protein in cells infected with different subtypes of the influenza A virus (A(H1N1), A(H1N1)pdm09, A(H2N2), A(H3N2), and A(H9N2)). A sensitive variant of the microcultural ELISA based on the use of the generated MAbs is proposed; it may be utilized to detect NS1 protein in infected cells and to study its antigenic variability. The results of the performed microcultural ELISA demonstrate type-and subtype-specific antigenic variability of the influenza virus NS1 protein.

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“…The cell spots were lysed by heating at 100 °C for 7 min in 80 μL of 1X Laemmli buffer (Bio Rad, Hercules, CA, USA). Proteins from 5 μL of samples were separated by SDS-PAGE and stained by Coomassie blue (Serva, EU) or transferred to nitrocellulose membrane (Bio Rad, USA) and probed with anti-NS1 1H7 monoclonal antibody [ 21 ].…”