Hybridoma cell lines secreting monoclonal antibodies to foot-and-mouth disease virus type Asia-1

Abstract

Various immunizing regimens, cell culture requirements and cell fusion conditions were examined for efficient production of hybridomas secreting anti-foot-and-mouth disease virus (FMDV) antibodies. A highly sensitive streptavidin-biotin-based enzyme-linked immunosorbent assay (ELISA) was used for screening of hybridomas for specific antibody production as well as for determining the serotype specificity of the antibodies. Six hybridoma cell lines generating antibodies to FMDV type Asia-1 (vaccine strain 63/72) were produced by fusion of SP2/0 mouse myeloma cells and spleen cells from mice immunized with the inactivated viral antigen. The monoclonal antibodies (MoAb) from four producer clones reacted in ELISA specifically with the intact virus antigen of type Asia-1 without any cross-reactivity with strains of other virus types 0, A and C. Two other clones positive for anti-Asia-1 virus antibodies in ELISA cross-reacted with type C virus strain. The MoAb from three of the four Asia-1 virus type specific clones neutralized the infectivity of the immunizing viral strain. One neutralizing MoAb reacted with the separated VP1 from the immunizing viral strain in immunoblotting.