【作者】 王娜；

【导师】 刘杞；

【作者基本信息】 重庆医科大学 ， 内科学， 2006， 硕士

【摘要】 目的:肝再生增强因子(Augmenter of liver regeneration, ALR)是Hagiya等于1994年对肝刺激因子(Hepatic stimulatory substance,HSS)的研究中发现的一种非特异性、具有热稳定性、不同于肝细胞生长因子(Hepatocyte growth factor , HGF)的促肝细胞再生因子。为进一步了解血清中人肝再生增强因子( Human augmenter of liver regeneration,hALR)的含量,研究hALR与不同程度肝病,尤其是与乙型肝炎不同阶段的关系,依据经典免疫学理论,我们拟建立一种不同于以往双抗体夹心法或间接ELISA方法的、适用于检测hALR这种小分子蛋白质的可靠检测方法。本研究拟采用直接竞争的反应模式来建立hALR的血清检测方法。通过制备hALR的原核表达纯品,用辣根过氧化物酶(Horseradish peroxidase,HRP)及铕元素(Europium,Eu)标记,为竞争抑制检测方法的建立提供抗原;利用抗hALR的单克隆杂交瘤细胞株,制备并扩增小鼠抗hALR的单克隆抗体(Monoclonal antibody,McAb);进而建立血清hALR直接竞争的检测方法,了解不同肝病患者血清中hALR的含量及意义。方法:1.抗原及标记抗原的制备更多还原

【Abstract】 Objective: Augmenter of liver regeneration (ALR), a heat-stable hepatotrophic growth factor, was first discovered and cloned from neonatal rat livers. Although its action is similar to hepatocyte growth factor (HGF) in promoting liver regeneration, its molecular structure is different from HGF. Because of the low molecular weight protein—human augmenter of liver regeneration (hALR), it’s necessary for us to establish a newly detectable method different from the past ones for the purpose to investigate the serum levels of hALR in various liver diseases and the association between them, which was different from the present detect methods. Our objective in this study was to establish an assay of direct competitive reaction to observe the serum levels of hALR. Recombinant prokaryotic expression plasmid of hALR was expressed in Escherichia coli in this study and the target protein was purified by affinity chromatography technique as antigen in the competitive reaction. The selected hybridomas secreting anti-hALR monoclonal antibody (McAb) were used to prepare McAb to human augmenter of liver regeneration as the antibody in the更多还原