肾上腺素能β1受体亚型特异性抗体的制备及鉴定  

Preparation and Identification of Subtype Specific Antibody Against Beta-1 Adrenergic Receptor

作　　者：任京力[1] 赵树进[2] 杨太成[2] 陈汝筑[1] 孙桂玲 

机构地区：[1]中山医科大学药理教研室,广东广州510089 [2]广州军区广州总医院药学部,广东广州510010 [3]广州市药检所,广东广州510160

出　　处：《中山医科大学学报》2002年第1期37-39,43,共4页Academic Journal of Sun Yat-sen University of Medical Sciences

基　　金：全军"九五"医学科研规划科研基金资助项目 ( 98M0 37)

摘　　要：【目的】制备和鉴定 β1受体亚型特异性抗体。【方法】人工合成 β1受体细胞膜外第二环 197 2 2 2位氨基酸序列作为抗原。连接钥孔冒贝血蓝蛋白 (KLH)增加抗原性后免疫兔获得抗血清。通过凝胶双扩散实验和ELISA法鉴定其效价 ;通过免疫荧光法及ELISA法鉴定其特异性 ;通过离体蛙心灌流实验鉴定其药理活性。【结果】该抗血清效价高 (分别为 1∶6 4和 1∶10 6)、特异性强 ,能和心肌 β1受体发生特异性结合 (1∶10 4 ～ 1∶10 5) ,为异丙肾上腺素的非竞争性拮抗剂。 (pD2 ′ =1 6 2 )。【结论】成功制备的 β1受体亚型特异性抗体可能成为进一步研究 β1受体分布、功能和定量的有力工具。To prepare and identify subtype specific antibody against beta 1 adrenergic receptor. The antigen was artificially synthesized according to the amino acid sequence of the second extra celluar loop of beta 1 adrenergic receptor(197 222) and coupled to KLH to improve its antigenicity. The antibody was obtained after immunizing rabbits by peptide KLH compound. The titre of the antibody was determined by double agarose diffusion and ELISA, the specificity by immunofluorescence and ELISA, and pharmacological activity by isolated toad heart irrigation test. The antibody was proved to be high titre (1∶10 6 for ELISA and 1∶64 for double agarose diffusion), specific(1∶10 4～1∶10 5 for ELISA) and able to non competitively antagonize isoprenalines's interaction with beta adrenergic receptor(pD 2′=1 62). [Conclusion] The subtype specific antibody against beta 1 adrenergic receptor was successfully prepared and be served as a valuable tool for further study on quantity, distribution, and function of beta 1 adrenergic receptor.

关 键 词：肾上腺素能受体 多克隆抗体 分离 提纯 特异性抗体 制备 鉴定 

分 类 号：R392－33[医药卫生—免疫学]