摘要
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.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2006年第4期337-341,共5页
Chinese Journal of Immunology
基金
国家计委部门重点资助项目([1997]2071号)
广东省自然科学基金资助项目(粤财Z[1998]278)