抗人肌红蛋白单克隆抗体的制备和初步鉴定

Preparation and Characterization of Monoclonal Antibody to Human Myoglobin

用纯化的人肌红蛋白（Ｍｂ）抗原免疫Ｂａｌｂ／ｃ小鼠通过细胞杂交技术。获得１０株稳定分泌单克隆抗体的杂交瘤细胞株．其中３株分泌ＩｇＧ_１亚类抗体，２株为ＩｇＧ_（２ａ），５株为ＩｇＧ_（２ｂ）．间接ＥＬＩＳＡ法证明单抗腹水与Ｍｂ起特异性反应，与人血红蛋白（Ｈｂ）．人白蛋白（Ａｌｂ）无交叉反应．免疫印迹检测结果表明，６株单抗腹水能特异性识别分子量１７ＫＤ的Ｍｂ，抗Ｍｂ单克隆抗体的制备，对于急性心肌梗塞早期诊断很有应用价值．

Purified human myoglobin (Mb) was used as antigen for immunization of BAIB/C mice. Hybridoma technique was employed to prepare monoclonal antibody against Mb. Ten cell strains capable of steady production of monoclonal antibody were established. The monoclonal antibodies Ig subclass were secreted three IgG_1, two IgG_(2a), and five IgG_(2b). ELISA results showed that all these monoclonal antibodies reacted only with Mb and had no cross-reaction with human hemoglobin and albumin, In immunoblotting test the monoclonal antibody revealed a band of 17 KD corresponding to the molecular weight of Mb. The preparation of specific monoclonal antibody to Mb has potential utility in the establishment of serum Mb determination for the early diagnosis of acute myocardial infarction.