目的 制备和鉴定抗肌红蛋白（Mb）单克隆抗体，为人肌红蛋白检测试剂的研制奠定基础。方法 用人肌红蛋白抗原免疫BALB/c小鼠，采用杂交瘤技术进行细胞融合，用ELISA方法筛选，有限稀释法进行克隆化培养阳性杂交瘤细胞株。ELISA法测定小鼠腹水滴度，CLIA法对抗体进行配对实验与特异性鉴定。结果 筛选培养出9株分泌抗肌红蛋白单克隆抗体的杂交瘤细胞株，腹水滴度为1:5×104～1:2×105。通过配对实验，有5株抗体6种组合可以成功配对。这5株抗体的亲和常数为6.46×108～3.68×109L/mol。用5株抗体所建立的6种化学免疫分析方法与人Hb交叉反应率为7.434×10-3%～2.904×10-2%，与人ALB交叉反应率为4.980×10-7%～3.830×10-5%。用Mb17B6与Mb21E8两株抗体建立的CLIA标准曲线的线性系数为0.997，灵敏度为6.02ng/mL。结论 成功制备了可以应用于免疫分析的抗肌红蛋白单克隆抗体。

Objective To prepare and characteriz the monoclonal antibody against Mb to provide basis for the development of Mb diagnostic reagent. Methods BALB/c mice were immunized with human myoglobin (Mb) antigen, the positive hybridoma were obtained by using hybridoma cell technique. The characterization of antibody were determined by ELISA. Results Nine monoclonal antibodies against Mb were prepared. The titers of ascites were 1:5×104～1:2×105. Five monoclonal antibodies successfully formed six anatibody pairs. The affinity constants of five monoclonal antibodies were in the range of 6.46×108 to 3.68×109 L/mol. The cross-reaction ratios with HB and ALB were 7.434×10-3%～2.904×10-2% and 4.980×10-7%～3.830×10-5% respectively. Two monoclonal antibodies (Mb17B6 and Mb21E8) were used to develop the chemiluminescence immunoassay. The assay sensitivity was 6.02 ng/mL. Conclusion The monoclonal antibodies against Mb are prepared successfully for the development of immunoassay.