摘要
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。
Nine hybridoma cell lines secreting anti bFGF mAb have been developed from immunized Balb/c mice with recombinant bovine bFGF, by cell fusion, screened with reverse indirect hemo agglutination and indirect ELISA and cloned with limited dilution method. Three of these hybridoma cell lines were identified by using Western blot analysis and inhibition of biological activity of bFGF. The results showed that anti bFGF mAbs could bind to recombinant bovine or human bFGF and inhibite growth of Balb/c 3T3 cells stimulated by bFGF in vitro . The ELISA titre of the mAbs in ascites was 1∶3 000. The mAbs 3B2 and 3F5 have been used in a two site EIA to detect the complete form of bFGF. The sensitivity of the EIA reached 50 pg/well of bFGF. The application and significance of anti bFGF mAbs were discussed in this paper.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
1998年第4期275-277,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金
关键词
BFGF
单克隆抗体
鉴定
肿瘤
basic fibroblast growth factor monoclonal antibody tumor
