碱性成纤维细胞生长因子单克隆抗体ELISA微量检测技术

The development of a quantitative indirect ELISA for basic fibroblast growth factor by monoclonal antibodies against recombiment bovine bFGF

目的:用本室制备的单克隆抗体建立间接ELISA检测微量bFGF的技术。方法:将抗重组牛碱性成纤维细胞生长因子(bFGF)的杂交瘤细胞株,经再克隆化,选择高亲和力的抗bFGF杂交瘤细胞2H2。用ProteinA亲和层析法对2H2腹水型单抗进行纯化。结果:用2H2mAb建立了高灵敏度的间接、竞争ELISA检测bFGF方法,灵敏度分别达到10和1.0pg/孔,并用间接ELISA检测神经胶质瘤细胞SWO38上清中的bFGF含量。结论:为实验研究和临床应用提供了微量bFGF的检测方法。

Objective：To development a quantitative indirect ELISA for basic fibroblast growth factor by using monoclonal antibodies against recombiment bovine bFGF. Methods：Hybridoma cell lines secreting against bFGF monoclonal antibody（mAb） were developed from immunized BALB/c mice with recombinant bovine bFGF. One of these hybridomas cell line designated 2H2 was identified using Western blot analysis and inhibition of biological activity of bFGF. Purified high-affinity monoclonal antibody by Protein A was obtained from acsite of BALB/c mouse treated by anti-bFGF hybridoma cells. Results：An sensitive indirect competitive enzyme immunoassay for bFGF was developed employing this mAb. The detection limit of this assay system were 10 and 1.0 pg/well. Conclusion：A quantitative indirect ELISA for bFGF have been developed.