Generating and characterizing monoclonal and polyclonal antibodies against avian H5N1 hemagglutinin protein

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

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17 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)691-696
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume382
Issue number4
DOIs
Publication statusPublished - May 15 2009
Externally publishedYes

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Keywords

  • Clade
  • Epitope
  • H5N1 virus
  • Hemagglutinin
  • Monoclonal antibodies
  • Polyclonal antibodies

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Wang, S. F., Chen, K. H., Thitithanyanont, A., Yao, L., Lee, Y. M., Chan, Y. J., Liu, S. J., Chong, P., Liu, W. T., Huang, J. C., & Chen, Y. M. A. (2009). Generating and characterizing monoclonal and polyclonal antibodies against avian H5N1 hemagglutinin protein. Biochemical and Biophysical Research Communications, 382(4), 691-696. https://doi.org/10.1016/j.bbrc.2009.03.119