Monoclonal antibodies to the haemagglutinin HA1 subunit of the pandemic influenza A/H1N1 2009 virus and potential application to serodiagnosis (pages 559–567)

摘要

In order to provide specific serological reagentsnfor pandemic influenza A/H1N1 2009 virus,nmonoclonal antibodies (Mabs) to recombinantnhaemagglutinin component HA1 (rHA1) werengenerated after fusing spleen cells from a mousenimmunized with rHA1 protein derived fromninfluenza strain A/California/06/09 H1N1 with anmouse myeloma cell line. Five hybridomanclones secreting Mabs specific for the rHA1nprotein derived from pandemic influenza AH1N1 2009 and not for rHA1 from seasonalnH1N1 influenza strains A/Brisbane/59/07 and ASolomon Islands/03/06 were identified by EIA.nMabs 7H4, 9A4, and 9E12 were reactive in Westernnblots with full length rHA and/or rHA1 subunitnderived from A/California/06/09 strain. Only Mabn1F5 inhibited haemagglutination of turkey rednblood cells with recombinant NIBRG-121 virusnderived from A/California/07/09, but did not reactnin Western blots. Immunostaining of MDCK cellsninfected with NIBRG-121 was localized to thenmembrane/cytoplasm for four of the reactivenMabs. The differing reactivity of the Mabs innWestern blots, immunostaining, EIA, and haemagglutinationninhibition assay suggest that atnleast four of the five Mabs recognize differentnepitopes on HA1 of the pandemic influenza AH1N1 2009 virus. Ferret antisera to pandemicninfluenza A/H1N1 2009 (A/England/195/09 and ACalifornia/07/09 strains) and sera from humannsubjects vaccinated with Influenza A (H1N1) 2009nMonovalent Vaccine (CELTURA1, Novartis Vaccines,nGermany), inhibit binding of 1F5-HRP tonbiotinylated rHA1 derived from A/California/0609 in a competitive EIA, suggesting that thenepitope recognized by this Mab also evokesnan antibody response in infected ferrets andnvaccinated humans. J. Med. Virol. 83:559–n567, 2011. u0001 2011 Wiley-Liss, Inc.