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J Gen Virol 69 (1988), 473-480; DOI 10.1099/0022-1317-69-2-473
© 1988 Society for General Microbiology
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Identification of Haemagglutinin-Neuraminidase Antibody Binding Sites by Western Blot Analysis of Antibody-resistant Mutants and Partial Digest Fragments of Newcastle Disease Virus

A. C. R. Samson1, M. Nesbit1, A. M. Lyon1 and G. Meulemans2

1 Department of Genetics, University of Newcastle upon Tyne, Newcastle upon Tyne NE1 7RU, U.K.
and2 National Institute for Veterinary Research, Groselenberg 99, 1180 Brussels, Belgium

A collection of monoclonal antibodies (MAbs) which react with the haemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) has been used to isolate MAb-resistant mutants of the Beaudette C strain of NDV. The patterns of cross-reactivity of the HN proteins of these mutants against the collection of MAbs determined by Western blotting allowed the MAbs to be sorted into different groups. Protease V8 partial digest fragments of purified wild-type virions and subsequent reaction against the collection of MAbs using Western blotting provided an alternative method of grouping MAbs which broadly agreed with the former method. Chemical cleavage of the HN protein at aspartate-proline bonds followed by Western blotting of the fragments allowed the approximate position of certain MAb binding sites to be determined.

Keywords: Newcastle disease virus, haemagglutinin-neuraminidase, monoclonal antibody

Received 26 June 1987; accepted 8 October 1987.




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Copyright © 1988 by the Society for General Microbiology.