title: Neutralization and Binding Profile of Monoclonal Antibodies Generated Against Influenza A H1N1 Viruses.
abstract: Monoclonal antibodies (MAbs) provide scope for the development of better therapeutics and diagnostic tools. Herein, we describe the binding and neutralization profile(s) for a panel of murine MAbs generated against influenza A H1N1 viruses elicited by immunization with pandemic H1 recombinant hemagglutinin (rHA)/whole virus or seasonal H1 rHA. Neutralizing MAbs, MA-2070 and MA-M, were obtained after pandemic A/California/07/2009 (H1N1) virus/rHA immunization(s). Both MAbs reacted specifically with rHA from A/California/07/2009 and A/England/195/2009 in ELISA. MA-2070 bound rHA of A/California/07/2009 with high affinity (KD = 51.36 ± 9.20 nM) and exhibited potent in vitro neutralization (IC50 = 2.50 μg/mL). MA-2070 bound within the stem domain of HA. MA-M exhibited both hemagglutination inhibition (HI, 1.50 μg/mL) and in vitro neutralization (IC50 = 0.66 μg/mL) activity against the pandemic A/California/07/2009 virus and showed higher binding affinity (KD = 9.80 ± 0.67 nM) than MA-2070. MAb, MA-H generated against the seasonal A/Solomon Islands/03/2006 (H1N1) rHA binds within the head domain and bound the seasonal H1N1 (A/Solomon Islands/03/2006 and A/New Caledonia/20/1990) rHAs with high affinity (KD; 0.72-8.23 nM). MA-H showed high HI (2.50 μg/mL) and in vitro neutralization (IC50 = 2.61 μg/mL) activity against the A/Solomon Islands/03/2006 virus. All 3 MAbs failed to react in ELISA with rHA from various strains of H2N2, H3N2, H5N1, H7N9, and influenza virus B, suggesting their specificity for either pandemic or seasonal H1N1 influenza virus. The MAbs reported here may be useful in developing diagnostic assays.
authors.affiliation: 1 Reproductive Cell Biology Laboratory, National Institute of Immunology , New Delhi, India .
authors.forename: Nachiket
authors.initials: N
authors.lastname: Shembekar
authors.affiliation: 2 Molecular Biophysics Unit, Indian Institute of Science , Bangalore, India .
authors.forename: Vamsee V Aditya
authors.initials: VV
authors.lastname: Mallajosyula
authors.affiliation: 1 Reproductive Cell Biology Laboratory, National Institute of Immunology , New Delhi, India .
authors.forename: Ankita
authors.initials: A
authors.lastname: Malik
authors.affiliation: 1 Reproductive Cell Biology Laboratory, National Institute of Immunology , New Delhi, India .
authors.forename: Ashok
authors.initials: A
authors.lastname: Saini
authors.affiliation: 2 Molecular Biophysics Unit, Indian Institute of Science , Bangalore, India .
authors.forename: Raghavan
authors.initials: R
authors.lastname: Varadarajan
authors.affiliation: 1 Reproductive Cell Biology Laboratory, National Institute of Immunology , New Delhi, India .
authors.forename: Satish Kumar
authors.initials: SK
authors.lastname: Gupta
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D006385:Hemagglutination Inhibition Tests; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology* / Q000472:pathogenicity; D007252:Influenza Vaccines / Q000276:immunology; D007251:Influenza, Human / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D051379:Mice
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines