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机构地区:[1]暨南大学生物工程学系
出 处:《细胞与分子免疫学杂志》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
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.
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