精子抗原肽P10G和YLP12的合成及在ELISA检测中的意义

目的 人工合成精子肽P10G和YLP12,以此为抗原建立检测抗精子抗体(AsAb)的酶联免疫吸附测定法(ELISA).方法 用PS3全自动合成仪合成精子肽P10G和YLP12,经反相高效液相色谱纯化和质谱鉴定.分别包板制备ELISA试剂盒,用以检测血清中的AsAb.结果 成功合成两种目的肽,质谱结果显示其相对分子质量均与理论相对分子质量相吻合,高效液谱法(HPLC)结果显示其纯度为97.5％和97.8％.以该两种合成肽为抗原,建立AsAb的ELISA,与商品化试剂盒比较均显示极好的一致性(Kappa值为0.77和0.79,＞0.75).结论 全自动固相多肽合成技术可用于合成高纯度的精子抗原肽,所合成的P10G和YLP12均具有较好的抗原活性,可作为包被抗原用于AsAb的ELISA检测.

Synthesis of sperm antigen peptide P10G and YLPI2 and their significance in enzyme-linked immunosorbent assay

Objective To synthesize sperm peptide PI0G and YLP12 and to establish an enzyme-linked immunosorbent assay（ELISA） for detecting antisperm antibodies（AsAb） based on the peptide. Methods The sperm peptide P10G and YLP12 were synthesized by the PS3 automated solid phase peptide synthesizer, purified by RP-HPLC and identified by MS. With the syn- thetic peptide as a coating antigen,the ELISA method was used to detect serum AsAb. Results The molecular weights of two synthesize peptides identified by MS were in good agreement with its calculated molecular weight （MW）. The purity of the synthetic peptides purified by HPLC was 97.5% and 97.8%. AsAb was detected by the indirect ELISA based on the peptide compared with commercial kit, the results showed that both assays were very unanimous （Kappa value 0.77 and 0.79, 〉0.75）. Conclusion Au- tomated solid-phase peptide synthesis could be used to synthesize the high purity sperm antigen peptides. The synthesized peptide P10G and YLP12 show the high antigenic activity and can be used as coating antigen for indirect ELISA to detect AsAb.