CAJ | 학술논문

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1.181 mg/mL,抗血清效价高于1∶64000,IC50为13.7 ng/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。
목적：제비화감정폐탄성단백강해태단인공항원,위진일보연제만성조새성폐질병(COPD)면역검측시제합타하량호기출。방법이 Sulfo-SMCC 위우련제,약공혈람단백(KLH)위재체단백제비면역원；경자외소묘화십이완기류산납-취병희선알응효전영법(SDS-PAGE)전영감정우련정황；이면역원면역 Balb/c 소서제비특이성항체,간접경쟁매련면역흡부실험(ELISA)법평개항혈청효개급특이성。결과자외소묘화 SDS-PAGE 전영학인료폐탄성단백강해태단인공항원합성성공,기단백농도위1.181 mg/mL,항혈청효개고우1∶64000,IC50위13.7 ng/mL,항혈청여무관태단무교차반응。결론본실험성공제비료폐탄성단백강해태단인공항원,기구유교호적면역원성,위건립쾌속준학검측 COPD 면역검측방법전정기출。
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 IC50 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.