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用重组牛ｂＦＧＦ免疫Ｂａｌｂ／ｃ小鼠，通过细胞融合，以反向间接血凝和间接ＥＬＩＳＡ筛选，以及有限稀释法克隆化，建立了９株稳定分泌抗ｂＦＧＦ单克隆抗体（ｍＡｂ）杂交瘤细胞。对其中３株用Ｗｅｓｔｅｒｎｂｌｏｔ和生物活性抗体中和实验进行了鉴定。结果显示，ｍＡｂ可结合重组牛或人ｂＦＧＦ，并能抑制ｂＦＧＦ刺激Ｂａｌｂ／ｃ３Ｔ３细胞的生长；腹水的ＥＬＩＳＡ滴度为１∶３０００；用其中２株ｍＡｂ建立的双抗体夹心ＥＬＩＳＡ法灵敏度可达５０ｐｇ／孔。本文还讨论了抗ｂＦＧＦｍＡｂ的应用价值。
Balb / c mice were immunized with recombinant bovine bFGF, and 9 strains of hybridoma cells stably secreting anti-bFGF monoclonal antibody (mAb) were established by cell fusion, reverse indirect hemagglutination and indirect ELISA screening, and cloning by limiting dilution method . Three of them were identified by western blot and bioactive antibody neutralization assay. The results showed that mAb could bind recombinant bFGF or bFGF and inhibit the growth of Balb / c3T3 cells induced by bFGF. The ELISA titer of ascites was 1: 3000. The double antibody sandwich ELISA established by 2 mAbs showed 50 pg /hole. This article also discusses the value of anti-bFGF mAb.