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目的 :制备抗牛碱性成纤维细胞生长因子 (bFGF)单克隆抗体(mAb) ,并鉴定其亚类 ,建立ELISA检测bFGF含量的方法。方法 :应用基因重组的牛bFGF免疫BALB/c小鼠 ,通过细胞融合 ,建立分泌抗bFGFmAb的杂交瘤细胞株。应用免疫沉淀技术鉴定抗bFGFmAb的亚类 ;应用基因重组的牛bFGF免疫青紫蓝兔 ,制备抗bFGF的多抗血清 ;将抗bFGFmAb及兔抗血清用ProteinA亲和层析纯化后 ,建立检测bFGF含量的ELISA方法。结果 :共获得 3株稳定分泌抗bFGFmAb的杂交瘤细胞株 ;它们所分泌的mAb均为IgG1;采用夹心ELISA法检测bFGF的敏感性达ng水平。 结论 :抗bFGFmAb(IgG1)和多克隆抗体制备为临床应用及相关研究提供了必要的试剂
OBJECTIVE: To prepare anti-bovine basic fibroblast growth factor (mAb) monoclonal antibody (mAb) and to identify its subclasses to establish an ELISA assay for bFGF content. Methods: BALB / c mice were immunized with recombinant bovine serum albumin (bFGF). The hybridoma cell lines secreting anti-bFGF mAb were established by cell fusion. The anti-bFGF mAb subclass was identified by immunoprecipitation. The anti-bFGF multi-antiserum was prepared by immunizing the blue bovine serum with bFGF gene recombinant. The anti-bFGF mAb and rabbit antiserum were purified by Protein A affinity chromatography. ELISA method. Results: Three hybridoma cell lines stably secreting anti-bFGF mAb were obtained. The secreted mAbs were both IgG1. The sensitivity of bFGF was up to ng level by sandwich ELISA. Conclusion: The preparation of anti-bFGF mAb (IgG1) and polyclonal antibody provides the necessary reagents for clinical application and related research