人心肌脂肪酸结合蛋白单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibodies of Human H-FABP

作　　者：郑杰[1] 龚成[1] 赵肃清[1] 

机构地区：[1]广东工业大学轻工化工学院制药工程系,广东广州510006

出　　处：《广东化工》2015年第22期32-33,共2页Guangdong Chemical Industry

基　　金：广东省自然科学基金项目(S2013010016021)

摘　　要：目的制备人心肌脂肪酸结合蛋白(H-FAB)单克隆抗体。方法利用重组H-FABP蛋白免疫BALB/c小鼠,取免疫后的小鼠脾细胞与骨髓瘤细胞(SP2/0细胞)在PEG的作用下进行融合,通过杂交瘤细胞的筛选,细胞的亚克隆以及扩大培养建立分泌抗H-FABP特异性抗体的单克隆细胞株。将细胞接种入小鼠体内制备腹水抗体,纯化后得到抗H-FABP单克隆抗体并对其性质进行鉴定分析。得到3株稳定分泌H-FAB抗体的单克隆细胞株,命名为2C06、2F08、6G03,非竞争性ELISA测定亲和力常数得知三株抗体的亲和力常数都达到108以上,三株抗体的亲和力较好。交叉实验表明三株单克隆抗体与牛血清、猪血清、鼠血清均无交叉反应,western blot实验证明对H-FABP有较好的特异性。成功得到3株特异性识别H-FAB单克隆细胞株,可用于H-FAB的检测研究。Objective To produce and identify monoelone antibodies of human heart type fatty acid-binding protein （H-FABP）. Methods The BALB/C mouse was immuned with intraperitoneal by recombinant humen H-FABP. The spleen cells and myeloma cells of the mouse were fused and cultivated, then monoclonal antibody of human H-FABP was selected. The antibody was purificated by Protein G Sepharose 4 Fast Flow affinity chromatograph and it was identified. Results Three monoclonal cell lines antibodies of hydridoma culture supernatants of 2C06, 2F08 and 6G03 could combine with H-FABP. Conclusion： Three strains monoclonal antibody cell lines were prepared successfully, and they can be used to prepare the reagents for analyzing H-FABP.

关 键 词：人心肌脂肪酸结合蛋白 单克隆抗体 制备 鉴定 

分 类 号：Q513.5[生物学—生物化学]