Objective: To synthesize sperm peptides for establishment of the enzyme - linked immunosorbent assay ( ELJSA ) to detect antisperm - antibodies ( AsAb) . Methods; The sperm peptide P10G, YLP12 and SP17 were synthesized by the PS3 automated solid phase peptide synthesizer, purified by RP - HPLC and identified by mass spectrum (MS). With the synthetic peptide as a coating antigen, the ELISA method was used to detect serum concentration of AsAb. Results; The molecular weights of three synthesize peptides identified by MS were in good agreement with calculated molecular weights. The purity of the synthetic peptides purified by HPLC was 97.5% , 97.78% and 95.5%. AsAb was detected by the indirect ELISA based on the peptide compared with the commercial kit, and results showed that both assays were unanimous with Kappa value more than 0.75. Conclusion; Automated solid - phase peptide synthesis could be used to synthesize high purity sperm antigen peptides. Hie synthesized peptide P10G, YLP12 and SP17 show high antigenic activities and can be used as a coating antigen for indirect ELISA to detect AsAb.%目的:以合成精子抗原肽为抗原建立酶联免疫吸附测定法( ELISA)检测抗精子抗体(AsAb).方法:通过PS3全自动合成仪合成精于肽P10G、YLP12和SP17抗原肽,经反相高效液相色谱(HPLC)纯化和质谱鉴定,分别包板制备ELISA试剂盒,并检测血清标本.结果:成功合成3种目的肽,质谱分析结果主峰分子量与理论值基本一致,HPLC结果显示其纯度为97.5％、97.78％和95.5％.以3种合成肽为抗原建立ELISA检测AsAb,与商品化试剂盒比较,均显示极好的一致性(Kappa值＞0.75).结论:全自动固相多肽合成技术可用于高纯度的精子抗原肽合成,所合成的P10G、YLP12和SP17抗原肽均具有较好的抗原活性,可作为ELISA的包被抗原用于AsAb检测.
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