肺弹性蛋白降解肽段人工抗原的合成与鉴定

Synthesis and identification of artificial antigens of lung elastin degradation peptide

目的制备和鉴定肺弹性蛋白降解肽段人工抗原,为进一步研制慢性阻塞性肺疾病(COPD)免疫检测试剂盒打下良好基础。方法以Sulfo-SMCC为偶联剂,钥孔血蓝蛋白(KLH)为载体蛋白制备免疫原;经紫外扫描和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)电泳鉴定偶联情况;以免疫原免疫Balb/c小鼠制备特异性抗体,间接竞争酶联免疫吸附实验(ELISA)法评价抗血清效价及特异性。结果紫外扫描和SDS-PAGE电泳确认了肺弹性蛋白降解肽段人工抗原合成成功,其蛋白浓度为1.181mg/mL,抗血清效价高于1∶64 000,IC50为13.7ng/mL,抗血清与无关肽段无交叉反应。结论本实验成功制备了肺弹性蛋白降解肽段人工抗原,其具有较好的免疫原性,为建立快速准确检测COPD免疫检测方法奠定基础。

Objective To synthesize and identify artificial antigens of lung elastin degradation peptide and for the purpose of preparation of COPD test. Methods The artificial antigens were synthesized by Sulfo-SMCC and KLH. The complete antigens were identified by ultraviolet spectrum and SDS-PAGE. Immunize Balb/c mice was used to prepare antibody. The antiserum activity was evaluated by indirect competitive ELISA, Results The artificial antigens were identified by ultraviolet spectrum and SDS-PAGE. The protein concentration was 1. 181 mg/mL. The titer of antiserum was 1 ： 64 000,and ICS0 was 13.7 ng/mL. The antiserum had no cross-reaction with nonsense peptide. Conclusion The artificial antigens were acquired successfully,which had good immunoge- nicity. The results have laid basis for COPD test.