Generation of murine monoclonal antibodies which cross-neutralize human enterovirus genogroup B isolates

Hsuen Wen Chang, Chia Chyi Liu, Min Han Lin, Hui Min Ho, Ya Ting Yang, Yen Hung Chow, Pele Chong, Charles Sia

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

A live enterovirus 71 (EV71) isolate designated, EV71/E59, with genotype B4 produced in Vero cells and purified over a sucrose gradient was used as the immunogen to generate EV71-specific murine monoclonal antibodies. Four hybridoma clones derived from the fusion of splenocytes of EV71/E59-preimmunized BALB/c (H-2d) mice and the NS-1 myeloma cells that exhibit stable growth were selected for detailed characterization. The proof that the hybridomas produced are indeed true independent clones was based on the obervations that they expressed different complementarity-determining regions (CDRs) in their κ light chain genes. Purified ascitic fluids produced by the individual clones reacted against the viral capsid protein, VP1, in Western blot; and recognized distinct sites of a common epitope localized at the C-terminal half of VP1. Each of the monoclonal antibodies exhibited potent neutralizing activities against the immunizing virus strain, as well as two other isolates namely, N0781-TW-01, and N2838, of subgenogroups B4 and B5, respectively, that were found commonly in recent outbreaks in Taiwan. It was also observed the monoclonal antibodies acted cooperatively in neutralizing the EV71/E59 virus.

Original languageEnglish
Pages (from-to)189-195
Number of pages7
JournalJournal of Virological Methods
Volume173
Issue number2
DOIs
Publication statusPublished - May 1 2011
Externally publishedYes

Fingerprint

Human Enterovirus B
Enterovirus
Genotype
Monoclonal Antibodies
Clone Cells
Hybridomas
Complementarity Determining Regions
Viruses
Vero Cells
Ascitic Fluid
Capsid Proteins
Viral Proteins
Taiwan
Disease Outbreaks
Sucrose
Epitopes
Western Blotting
Light
Growth
Genes

Keywords

  • EV71 monoclonal antibodies

ASJC Scopus subject areas

  • Virology

Cite this

Generation of murine monoclonal antibodies which cross-neutralize human enterovirus genogroup B isolates. / Chang, Hsuen Wen; Liu, Chia Chyi; Lin, Min Han; Ho, Hui Min; Yang, Ya Ting; Chow, Yen Hung; Chong, Pele; Sia, Charles.

In: Journal of Virological Methods, Vol. 173, No. 2, 01.05.2011, p. 189-195.

Research output: Contribution to journalArticle

Chang, Hsuen Wen ; Liu, Chia Chyi ; Lin, Min Han ; Ho, Hui Min ; Yang, Ya Ting ; Chow, Yen Hung ; Chong, Pele ; Sia, Charles. / Generation of murine monoclonal antibodies which cross-neutralize human enterovirus genogroup B isolates. In: Journal of Virological Methods. 2011 ; Vol. 173, No. 2. pp. 189-195.
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