猪生长激素抗独特型抗体的制备及生物活性检测  

Preparation of porcine growth hormone anti-idiotypic antibody and detection of its biological activity

作　　者：兰海楠[1] 李维[1] 郑鑫[1] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118

出　　处：《西北农林科技大学学报（自然科学版）》2012年第11期7-12,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(30871842)

摘　　要：【目的】以鼠肝细胞为模型,在细胞水平上研究猪生长激素抗独特型抗体的作用机制。【方法】以猪生长激素单克隆抗体1A5-F(ab′)2部分免疫BALB/c鼠,采用杂交瘤细胞技术制备猪生长激素抗独特型单克隆抗体。分离Wistar大鼠肝脏细胞,以其为模型,采用流式分析、激光共聚焦显微镜观察、Western-blot等技术手段,研究猪生长激素抗独特型单克隆抗体的生物活性。【结果】成功制备了3株猪生长激素抗独特型单克隆抗体,分别命名为CG1、CG2、CG3。其中,CG1阳性信号最强,经鉴定,CG1为Ab2β类抗独特型抗体,属于IgG2a亚类。CG1能够以二聚体的形式激活鼠肝细胞表面的生长激素受体,并可激活信号转导蛋白JAK2。【结论】CG1抗体可与鼠肝脏细胞表面的生长激素受体特异性地结合,并可磷酸化信号转导蛋白JAK2。【Objective】 This study was done to investigate the mechanism of porcine growth hormone anti-idiotypic antibody using rat hepatocytes as the experimental model.【Method】 We produced the monoclonal anti-idiotypic antibody（named as CG1） by immunization of BALB/c mice with purified monoclonal anti-pGH antibody.This antibody was used as biological tools to explore biological activity of CG1 by flow cytometry,laser confocal microscope and Western-blot.【Result】 Three strains of porcine growth hormone anti-idiotypic monoclonal antibody,named CG1,CG2,and CG3,were produced successfully.CG1 showed the strongest positive signal among them.It was identificated that CG1 belonged to Ab2β anti-idiotypic antibody,IgG2a subclass.The results confirmed that CG1 may activate growth hormone receptor throught dimerization and signal transduction protein JAK2.【Conclusion】 The anti-body CG1 may bind to growth hormone receptor expressed on the surface of rat liver cell and trigger signal transduction protein JAK2.

关 键 词：猪生长激素 抗独特型抗体 信号转导 

分 类 号：S852.43[农业科学—基础兽医学]