人精子蛋白17抗原肽的合成及其在抗精子抗体检测中的应用研究

SYNTHESIS OF THE ANTIGEN PEPTIDE OF HUMAN SPERM PROTEIN 17 AND ITS APPLIED RESEARCH IN ANTISPERM ANTIBODIES DETECTION

目的:合成人精子蛋白17的线性B细胞表位肽(118-127aa),并以此为抗原建立抗精子抗体(AsAb)检测的ELISA方法。方法:用PS3全自动多肽合成仪合成人精子蛋白17抗原肽,并经反相高效液相色谱纯化和质谱鉴定。该合成肽作为ELISA法的包被抗原来检测血清中的AsAb(IgG)。结果:合成了实验所需要的抗原性肽段,高效液相色谱法(HPLC)结果显示其纯度为95.5%。以该合成肽为抗原建立的检测AsAb(IgG)的间接ELISA方法,与商品化试剂盒相比符合率为93.3%。结论:所合成的人SP17抗原肽具有较好的抗原活性,可作为间接ELISA法的包被抗原用于抗精子抗体的检测。

Objective：To synthesize linear B-cell epitope peptide of human sperm protein 17（118-127aa） and establish an enzyme-linked immunosorbent assay （ELISA） to detect antisperm antibodies（AsAb） based on the peptide. Methods： Antigen peptide of human sperm protein 17 was synthesized by the PS3 automated solid phase peptide synthesizer, purified by RP-HPLC and identified by MS. With the synthetic peptide as a coating antigen, the ELISA method was used to detect serum AsAb （IgG）. Results： The purity of the synthesized human sperm protein 17 purified by HPLC was 95.5%. AsAb（IgG）was detected by the indirect ELISA based on the peptide compared with commercial kit, the result showed that the coincidence between them was 93.3%. Conclusion：The synthesized human sperm protein 17 shows a high antigenic activity and can be used as coating antigen for indirect ELISA to detect AsAb.