摘要
为了制备重组人肝再生增强因子（rh ALR）单克隆抗体（Mc Ab）,并鉴定其特异性,从而研发一种特异性较好的检测试剂,试验以纯化的rh ALR为抗原免疫Balb/c小鼠,通过细胞融合建立能稳定分泌抗rh ALR单克隆抗体的杂交瘤细胞株,制备单克隆抗体,用免疫荧光法对自行制备的单克隆抗体与购买的商品化多克隆抗体进行比较。结果表明：筛选出的5株稳定分泌抗rh ALR的单克隆抗体酶联免疫吸附方法（ELISA）鉴定其培养上清液和腹水中效价分别为1×10~3、1×10~3、1×10~3、2×10~3、2×10~3和1×10~4、1×10~5、1×10~3、1×10~4、5×10~5。说明与商品化多克隆抗体相比,自行制备的单克隆抗体具有较好的敏感性。
To prepare monoclonal antibodies （McAbs） against recombinant human augmenter of liver regeneration （rhALR） and identiiy their sensitivity, thus developing a detection reagent with better specificity, the Batb/c mice were immuned using rhALR as an antigen in the test. The McAbs were prepared through the establishment of monoclonal hybridoma cell lines secreting anti rhALR by cell fusion, and then the self - prepared McAbs were compared with commercial polyclonal antibodies purchased using an immunofluorescent method. The results showed that the McAbs from five strains of monoclonal hybridoma cell lines secreting anti rhALR were obtained. The titers of culture supernatants and ascites identified by ELISA method, were 1×10~3、1×10~3、1×10~3、2×10~3、2×10~3 and 1×10~4、1×10~5、1×10~3、1×10~4、5×10~5, respectively. The results indicate that the self - prepared monoelonal antibodies were better than commercial polyclonal antibodies in the sensitivity.
基金
国家自然科学基金项目(81301499
81460324)
