Quantification of intact 146S foot-and-mouth disease antigen for vaccine production by a double antibody sandwich ELISA using monoclonal antibodies.
A double antibody sandwich (DAS) enzyme-linked immunosorbent assay (ELISA) was developed to quantify 146S antigen of foot-and-mouth disease virus (FMDV) strain A10 Holland grown in suspension cultures of surviving bovine tongue epithelium. When virus harvests were incubated with trypsin--which affects VP1, the most immunogenic structural protein of FMDV--the concentration of 146S antigen as determined by ELISA was reduced by greater than 90%. Therefore, the test detected essentially only those virus particles with intact VP1. When the test was compared with the sucrose density gradient method, concentrations of 146S antigen correlated well (r = 0.87). The rate of variation in both tests was the same. In contrast to the sucrose density gradient method, the DAS-ELISA can simultaneously quantify 146S antigen in many samples, and also indicates when VP1 of 146S particles has disintegrated by the action of proteases.
Van Maanen
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Central Veterinary Institute, Department of Virology, Lelystad, The Netherlands.
Terpstra
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D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000032:analysis* / Q000737:chemistry; D005537:Aphthovirus / Q000276:immunology*; D046508:Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D014357:Trypsin; D014765:Viral Vaccines / Q000032:analysis*; D014776:Virus Cultivation
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D014765:Viral Vaccines; D014357:Trypsin