Neutralizing monoclonal antibodies to different clades of Influenza A H5N1 viruses

• Published in: Journal of virological methods Vol. 157; no. 2; pp. 161 - 167
• Main Authors: Oh, Sawyin, Selleck, Paul, Temperton, Nigel J., Chan, Paul K.S., Capecchi, Barbara, Manavis, Jim, Higgins, Geoff, Burrell, Christopher J., Kok, Tuckweng
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01-05-2009; Elsevier
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - isolation & purification; Antibodies, Viral - immunology; Antibodies, Viral - isolation & purification; Biological and medical sciences; Chickens; Cross Reactions; Fundamental and applied biological sciences. Psychology; H5N1; Haemagglutinin; Hemagglutinins, Viral - immunology; Immunoglobulin G - immunology; Immunoglobulin G - isolation & purification; Influenza; Influenza A Virus, H5N1 Subtype - immunology; Influenza A Virus, H5N1 Subtype - isolation & purification; Inhibitory Concentration 50; Microbiology; Monoclonal antibodies; Neutralization; Neutralization Tests; Pseudotype; Techniques used in virology; Virology; Pseudotype; Monoclonal antibodies; H5N1; Haemagglutinin; Influenza; Neutralization; Microbiology; Hemagglutinin; Orthomyxoviridae; Monoclonal antibody; Method; Neutralizing antibody; Virology; Infection; Virus; Influenzavirus A; Avian influenzavirus; Influenza A virus; Viral disease
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2008.12.016

Four IgG
1κ monoclonal antibodies (mAbs) against Influenza A/Chicken/Vietnam/8/2004 (H5N1) virus are described. Three of these showed neutralizing activities against H5N1 strains from clades 1, 2 and 3 using a retroviral pseudotype or live virus microneutralization assay. In the pseudotype assay, the IC
90 neutralizing titre range was >1600–51,200, and with the microneutralization was 80
≥
10,240. MAb 1C1 showed strong neutralizing activities in both assays. All four mAbs reacted specifically to the immunogen by immunohistochemical staining and to A/Hong Kong/483/1997 (H5N1) and A/Thailand/1(KAN-1)/2004 (H5N1)-infected MDCK cells by immunofluorescence. ELISA titrations of the mAbs showed specificity for H5N1 haemagglutinin (HA) and no cross-reactivity to 15 other Influenza A subtypes. Only mAbs 1C1 and the non-neutralizing 1F7 reacted with HA
1, the cleaved subunit of HA, by Western blot. These results suggest that the mAbs recognize distinct or overlapping epitopes and will be useful reagents for construction of specific rapid point-of-care assays or for therapeutic use.