兴奋剂促红细胞生成素免疫检测方法的研究

A Monoclonal Antibody－ based Immuoassay for Doping of Erythropoietin in Sports

摘要：目的 探索兴奋剂促红细胞生成素 (EPO)的检测方法。方法 制备了抗重组 EPO(rHuEPO)的两株单克隆抗体： McAbF8和 McAbF11,同时纯化了天然螺旋藻中的藻胆蛋白。以 McAbF8作包被抗体，建立了检测血清中 EPO浓度的竞争 ELISA法；并将藻胆蛋白作为荧光探针标记 McAbF8，建立了检测血清中 EPO浓度的荧光免疫检测法，其检测线性范围为 10－ 10～ 10－ 12mol/L。结果 用于同样样本的血清检测，与竞争酶联免疫吸附法所测结果无显著性差异 (P >0.05)。结论 荧光免疫检测法在灵敏度和特异性等方面，并不亚于传统的酶联免疫吸附法 (ELISA)，而在有些方面还要优于 ELISA，它在检测中将获得更大的应用。

Objective To find the detecting method of erythropoietin(EPO).Methods A kind of doping,two hybridoma cell lines,McAbF8 and McAbF11,were set up and phycobiliprotein was purified using McAbF8 as the coating antibody,the method of the competitive ELISA to detect EPO in serum was set up.At the same time,McAbF8 was labeled with phycobiliprotein and a new type of fluroscent probe was developed.The linear scope ranges from 10－ 12 to 10－ 10 mmol/L.Results The probe was used in detecting EPO concentration in the same samples. No significant difference was found between the two methods.(P >0.05).Conclusion The imunofluorescent assays is not inferior to the traditional enzyem－ linked immunoadsorbent assay(ELISA)in some respects,it has even advantages over ELISA.So the method may be put into more utilization in the detection.