重组人ALR单克隆抗体的制备、鉴定和初步应用  

Preparation and preliminary application of monoclonal antibody against recombinant human augmentor of liver regeneration

作　　者：王彦芳[1] 焦艳晴 董爱华[1] 耿娅[1] 贾茜[1] 

机构地区：[1]华北制药集团新药研究开发有限责任公司,河北石家庄050015

出　　处：《免疫学杂志》2004年第1期70-73,共4页Immunological Journal

摘　　要：目的　制备抗重组人肝再生增强因子 (ALR)的单克隆抗体 (McAb)。方法　以基因重组人ALR二聚体(drhALR)为抗原 ,免疫Balb c小鼠 ,制备单克隆抗体。用常规法制备抗ALR单体 (mrhALR)多抗 ,建立夹心ELISA法测定ALR ,初步确定ALR在小鼠组织的分布 ;用荧光免疫方法对胎鼠各组织进行ALR检测。结果　筛选出 3株稳定分泌抗drhALR的单抗杂交瘤细胞株 ,免疫球蛋白亚类分别为IgG1、IgG2a、IgG2b,特异性高 ,用于rhALR的免疫印迹及ELISA检测 ,可同时识别rhALR单体 (mrhALR)和二聚体 (drhALR) ;夹心ELISA法检测小鼠肝、肾、脾提取液ALR含量与鼠龄无关 ,各脏器的ALR含量不同 ,分别为肝 0 .36 μg g、肾 0 .11μg g、脾 0 .0 5 μg g ;免疫荧光检测胎鼠 ,各器官有不同程度ALR的存在 ,无器官特异性。 结论　用drhALR为抗原制备的抗rhALR单克隆抗体 ,能够用于ALR的体内外免疫学检测 。Objective To obtain monoclonal antibodies(McAb) against recombinant human augmentor of liver regeneration(rhALR).Methods Recombinant human augmentor of liver regeneration dimer(drhALR) was used as antigen to immune Balb/c mice. Monoclonal anti bodies against ALR were prepared by normal hybridoma technology. Polyclonal antiserum against ALR was produced by immunizing rabbit with mrhALR. A sandwich ELISA was established to detect ALR of different tissues of adult and fetal mice. Its distribution in mice tissue was determined by immunofluorescence method. Results Three hybridmas producing antibodies against drhALR were obtained. IgG isotypes of three McAbs were IgG1,IgG2a and IgG2b.The antibodies were proved to be specific for mrhALR. Either drhALR or ALR western blotting have the same result. Investigation the AlR content of the mice liver, spleen and kidney extract did not show any difference in respect to the animal age, but the level between each organ differed significantly, i.e. 0.36 μg/g (liver),0.11 μg/g(kidney) and 0.05 μg/g (spleen). Immunofluorescent assay indicated that ALR is not organ specific. Conclusion The rhALR McAb which using drhALR as antigen prepared in this paper can be used for ALR immunoassay in vitro and in vivo , and provide a basis for research on pharmacology and quality criteria of rhALR and ALR from animal source.

关 键 词：肝再生增强因子 基因重组人ALR的二聚体 单克隆抗体 夹心ELISA ALR的组织定位 

分 类 号：R392[医药卫生—免疫学]