| 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。 |
| 关键词: 猪生长激素 抗独特型抗体 信号转导 |
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| 基金项目:国家自然科学基金项目（30871842） |
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| Preparation of porcine growth hormone anti-idiotypic antibody and detection of its biological activity |
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| Abstract: |
| 【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. |
| Key words: porcine growth hormone anti-idiotypic antibody signal transduction |
