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1 March 2010 Production of H5-Specific Monoclonal Antibodies and the Development of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of H5 Antibodies in Multiple Species
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Abstract

The hemagglutinin gene of an avian influenza virus (AIV) A/duck/NC/674964/07 (H5N2) was cloned and expressed in a baculovirus system (H5-Bac). In parallel, a recombinant hemagglutinin of A/Vietnam/1203/04 (H5N1) was expressed in mammalian cells, purified, and used for generation of H5-specific monoclonal antibodies (MAb). The purified H5-Bac was used to develop a competitive enzyme-linked immunosorbent assay (cELISA) to detect H5 antibodies in a species-independent approach using one of the established H5-specific MAbs as the competitor antibody. The cELISA performed with influenza antibody-free sera or with sera of animals infected with other than H5-encoding AIV showed no significant inhibition of H5-MAb binding, indicating high test specificity. In contrast, sera of poultry (chickens, turkeys, ducks) experimentally infected with H5-encoding AIV were able to significantly inhibit the binding of the MAb in a species-independent approach. Comparison of the results of the cELISA with results obtained by a hemagglutination inhibition assay showed a gradient of the sensitivity (turkeys > ducks > chicken). The described results show that H5-specific antibodies in sera can be detected in a species-independent approach by using a recombinant protein.

Daniel Dlugolenski, Rüdiger Hauck, Robert J. Hogan, Frank Michel, and Egbert Mundt "Production of H5-Specific Monoclonal Antibodies and the Development of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of H5 Antibodies in Multiple Species," Avian Diseases 54(s1), 644-649, (1 March 2010). https://doi.org/10.1637/8683-030909-ResNote.1
Received: 11 March 2009; Accepted: 1 July 2009; Published: 1 March 2010
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