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目的:以已经制备的抗肌糖蛋白C(TN-C)的单克隆抗体(mAb)为基础,建立能定量检测肌糖蛋白C浓度的夹心ELISA方法,并初步于临床血清标本检测。方法:将3株mAb(克隆号分别为:1A8、3H7和4D6)制备腹水后纯化,分别与辣根过氧化物酶交联后,两两配对,以重组肌糖蛋白C蛋白为检测抗原,分析抗体之间最佳组合;利用建立的夹心ELISA方法检测收集的临床骨肉瘤患者和正常人血清标本。结果:包被1A8 mAb,HRP-4D6配对敏感性最高;骨肉瘤患者血清中肌糖蛋白C浓度明显高于正常人。结论:成功建立检测肌糖蛋白C的夹心ELISA方法,并利用其检测到肌糖蛋白C在骨肉瘤患者中的浓度异于正常人。
OBJECTIVE: To establish a sandwich ELISA method for the quantitative determination of muscle glycoprotein C based on monoclonal antibody (mAb) against muscle glycoprotein C (TN-C). The sandwich ELISA was initially detected in clinical serum samples. Methods: Three mAbs (clone numbers: 1A8, 3H7, and 4D6, respectively) were prepared and purified ascites fluid. After being crosslinked with horseradish peroxidase, the three mAbs were paired and the recombinant protein was used as detection antigen. The optimal combination of antibodies was analyzed. Serum samples from patients with clinical osteosarcomas and normal controls were tested using the established sandwich ELISA. Results: 1A8 mAb and HRP-4D6 had the highest sensitivity. Osteosarcoma in patients with serum osteogonosin C concentration was significantly higher than normal. CONCLUSIONS: Sandwich ELISA for detecting muscle glycoprotein C was successfully established and its concentration of muscle glycogen C in osteosarcoma was detected to be different from that of normal human.