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[目的]生产抗癌胚抗原(CEA)的单克隆抗体(McAb),制备检测血清CEA含量的酶联免疫吸附分析(ELISA)试剂盒。[方法]用亲和层析法纯化的CEA抗原免疫BALB/C小鼠,经细胞融合挑选出两株适用于CEAELISA试剂盒的McAbs进行重点鉴定。用ELISA法测定亲和力常数和免疫球蛋白亚类,用中和抑制法和免疫印迹电泳法鉴定特异性。[结果]两株McAbs均为鼠IgG1亚类,其亲和力常数分别为1.03×10-9M和2.25×10-9M,只有CEA阳性血清能抑制McAbs与包被CEA的反应,免疫印迹电泳显示两株McAbs只与200kDa的CEA形成一条带,所组成的试剂盒与放射免疫分析试剂盒比较,其灵敏度为96.60%,特异性为99.11%,总符合率为98.52%。[结论]两株McAbs具有较好的特异性和亲和力,用该两株McAbs组成的ELISA试剂盒可以应用于临床检测CEA。
[Objective] Monoclonal antibody (McAb) against carcinoembryonic antigen (CEA) was produced to prepare ELISA kit for detecting serum CEA content. [Method] BALB / C mice were immunized with CEA antigen purified by affinity chromatography. Two McAbs suitable for CEA ELISA kit were selected by cell fusion for identification. Affinity constants and immunoglobulin subclasses were determined by ELISA, neutralization inhibition and Western blot electrophoresis to identify specificity. [Results] The two McAbs were all mouse IgG1 subclasses with affinity constants of 1.03 × 10-9 M and 2.25 × 10-9 M, respectively. Only CEA-positive serum could inhibit the reaction of McAbs and CEA-coated cells. Immunoblot electrophoresis showed that two strains McAbs only formed a band with 200kDa CEA. The kit consisted of 96.60% sensitivity, 99.11% specificity and 98.52% compliance with radioimmunoassay kit. [Conclusion] The two McAbs have good specificity and affinity. The ELISA kit composed of the two McAbs can be used in the clinical detection of CEA.