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Title: Generating and characterizing monoclonal and polyclonal antibodies against avian H5N1 hemagglutinin protein
Authors: Sheng Fan Wang
Kuan Hsuan Chen
Arunee Thitithanyanont
Ling Yao
Yuan Ming Lee
Yu Jiun Chan
Shih Jen Liu
Pele Chong
Wu Tse Liu
Jason C. Huang
Yi Ming Arthur Chen
National Yang-Ming University Taiwan
School of Medicine
Mahidol University
Laboratory Medicine
Veterans General Hospital-Taipei
Taipei City Hospital Taiwan
National Health Research Institutes Taiwan
Keywords: Biochemistry, Genetics and Molecular Biology
Issue Date: 15-May-2009
Citation: Biochemical and Biophysical Research Communications. Vol.382, No.4 (2009), 691-696
Abstract: Accurate and timely diagnoses are central to H5N1 infection control. Here we describe the cloning and expression of the HA1 protein of the A/Vietnam/1203/04 strain in a bacterial system to generate mono-/polyclonal antibodies. All of the eight generated monoclonal antibodies recognized the same linear epitope on the top globular region of the HA structure-a highly conserved epitope among all circulating H5N1 clades identified by amino acid alignment. Results from immunofluorescence staining and Western blotting indicate that all monoclonal antibodies interacted with a denatured form of HA proteins, while the resultant polyclonal antibodies recognized both denatured and native HA proteins on H5N1 reverse-genetics (RG) viruses. Results from flow cytometry and microneutralization assays indicate that the polyclonal antibodies blocked viral binding and neutralized H5N1-RG viruses. Our results may prove useful to establishing future H5N1 mono-and polyclonal antibodies, and perhaps contribute to the development of an alternative H5N1 vaccine. © 2009 Elsevier Inc. All rights reserved.
ISSN: 10902104
Appears in Collections:Scopus 2006-2010

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