以精子抗原肽为抗原的抗精子抗体ELISA检测方法的建立

目的建立组合多个精子抗原肽为抗原的检测抗精子抗体的酶联免疫吸附测定(ELISA)法。方法用PS3全自动多肽合成仪合成4个特异精子肽,并经反相高效液相色谱(RP-HPLC)纯化和质谱鉴定。用ELISA检测4个合成肽的抗原性,基于对这些合成肽抗原活性的评价,将4个精子抗原肽混合后(组合肽)作为抗精子抗体(AsAb)检测用抗原。结果合成了实验所需要的抗原性肽段,RP-HPLC结果显示纯度达95%以上。以组合肽为抗原建立的检测AsAb的间接ELISA方法,与商品化试剂盒相比符合率为95.6%。结论固相多肽合成技术可用于合成高纯度的精子抗原肽,以组合的精子抗原肽作为包被抗原建立的ELISA方法可提高AsAb检测的灵敏度。

Establishment of an enzyme-linked immunosorbent assay for detecting antisperm antibodies based on sperm antigen peptides

Purpose To establish an enzyme-linked immunosorbent assay for the antisperm antibody detection with combined sperm antigen peptides.Methods Four specific sperm peptides were synthesized by the PS3 automated solid phase peptide synthesizer,purified by RP-HPLC and identified by MS.Based on their antigenicity evaluated by ELISA,the specific sperm peptides were used as coating antigens in the form of combination to detect antisperm antibodies.Results The desired peptides were synthesized,and the purity of these peptides purified by HPLC was over 95%.By comparing the responsibility of the ELISA using combined peptides as coated antigens with commercial kit,a 95.6% coincidence was obtained.Conclusion Solid-phase peptide synthesis could be used to synthesize high purity sperm antigen peptides.The ELISA for the detection of antisperm antibodies using combined sperm antigen peptides as coating antigens can improve the sensitivity.