精子肽的固相合成及应用初探

目的 :探讨将多肽固相合成技术用于检测抗精子抗体的ELISA试剂盒制备。方法 :以多肽固相合成法合成特异性精子肽 ,并经高效液相纯化分析及质谱分析。以此合成精子肽包板制备检测抗精子抗体的ELISA试剂盒 ,检测血清标本的AsAb。结果 :HPLC结果显示 ,合成的精子肽纯度达 98.26%;质谱分析结果主峰分子质量与理论值一致。采用合成多肽抗原建立了检测抗精子抗体的酶联免疫吸附测定方法 ;不明原因不育患者组与对照组间AsAb发生率呈非常显著差异(P <0,005 )。结论 :本固相合成法可获得高纯度特异性精子肽 ;该精子肽包板的ELISA试剂盒可靠简便。

Solid-phase Synthesized of the Sperm Peptide and Primary Research on its Using

Objective:To explore the using of solid-phase synthetic peptide in an enzyme linked immunosorbent assay(ELISA) for the detection of anti sperm antibody. Methods:The specific peptide was synthesized by a solid phase method, and then assessed by high pressure liquid chromatography and mass spectrometric analysis. Sera were tested with the assay using synthetic peptide as coated antigen. Results:The synthetic sperm peptide showed a purity of 98 26% as assessed by high pressure liquid chromatography. Good agreement of appearance molecular weight with its calculated M.W. was observed when analysed by Mass spectrometric. An ELISA for the detection of anti sperm antibody using synthetic peptide was established. The positive rate of AsAb between infertility group and unmarried youth group showed significant differences ( P <0 005).Conclusion: The high purity specific peptide was synthesized by our solid phase method. The Kit coated with the sperm peptide is reliable and convenient.;