|Terrence, M - CDC-ATLANTA, GA|
Submitted to: Avian Immunology Research Group Abstract
Publication Type: Abstract Only
Publication Acceptance Date: July 14, 2004
Publication Date: September 1, 2004
Citation: Kapczynski, D.R., Terrence, M.T. 2004. Chicken Interferon-Gamma Monoclonal Antibodies and Their Application in a Flow Cytometric Microsphere Assay. Avian Immunology Research Group Abstract. Technical Abstract: Microsphere-based immunoassays read by flow cytometry is an emerging technology with tremendous potential in the clinical and epidemiological management of infectious diseases. Unlike most standard immunoassays such as ELISA, these assays present multiplexing capabilities, availability of multiple assay formats, high throughput capacity, reduced time and labor, and reduced sample volumes. In these studies, three monoclonal antibodies (Mabs) against recombinant chicken interferon-gamma (IFN-g) were produced and characterized by ELISA, Western blot and microsphere assay. All Mab's detected IFN-g in direct ELISA binding assays and as capture antibodies in sandwich ELISA. By Western blot analysis, all antibodies identified a 17-kDa IFN-g protein. The IFN-g ELISA was also used to measure cytokine levels in the sera of chickens experimentally challenged with exotic Newcastle disease (END) virus. The Mabs were covalently-coupled to microspheres (4-8 micron) and tested in a flow cytometric assay. Results indicate the microspheres could detect serum IFN-g from END-infected birds and correlated with challenge. These results indicate that the microsphere-based assay is suitable to measure chicken IFN-g.