肺弹性蛋白降解肽段人工抗原的合成与鉴定? |
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引用本文: | 彭方毅,崔玉花,马明炎,李元宽,邬红雨,姜海蓉,周欢.肺弹性蛋白降解肽段人工抗原的合成与鉴定?[J].重庆医学,2015(22):3037-3039. |
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作者姓名: | 彭方毅 崔玉花 马明炎 李元宽 邬红雨 姜海蓉 周欢 |
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作者单位: | 1. 重庆理工大学药学与生物工程学院,重庆 400054; 重庆市垫江县中医院检验科 408300;2. 重庆理工大学药学与生物工程学院,重庆,400054;3. 重庆市垫江县中医院检验科 408300 |
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基金项目: | 国家自然科学基金资助项目,重庆市科委自然科学基金,重庆市卫生局科研项目 |
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摘 要: | 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。
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关 键 词: | 肺弹性蛋白降解肽段 人工抗原 抗体 |
Synthesis and identification of artificial antigens of lung elastin degradation peptide |
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Abstract: | 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. |
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Keywords: | lung elastin degradation peptide artificial antigens antibody |
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