Butchaiah G, Rao B U
Southern Regional Station, Indian Veterinary Research Institute Bangalore.
Acta Virol. 1989 Mar;33(2):121-30.
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.
为了高效生产分泌抗口蹄疫病毒(FMDV)抗体的杂交瘤,研究了各种免疫方案、细胞培养要求和细胞融合条件。一种基于链霉亲和素-生物素的高灵敏度酶联免疫吸附测定(ELISA)用于筛选产生特异性抗体的杂交瘤,以及确定抗体的血清型特异性。通过将SP2/0小鼠骨髓瘤细胞与用灭活病毒抗原免疫的小鼠脾脏细胞融合,产生了六种针对亚洲1型口蹄疫病毒(疫苗株63/72)产生抗体的杂交瘤细胞系。来自四个产生克隆的单克隆抗体(MoAb)在ELISA中与亚洲1型完整病毒抗原特异性反应,与其他病毒型0、A和C的毒株无任何交叉反应。另外两个在ELISA中抗亚洲1型病毒抗体呈阳性的克隆与C型病毒株发生交叉反应。来自四个亚洲1型病毒特异性克隆中的三个克隆的MoAb中和了免疫病毒株的感染性。一种中和性MoAb在免疫印迹中与免疫病毒株分离的VP1发生反应。