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克伦特罗单克隆抗体的纯化及其生物学特性的研究
  • ISSN号:1000-6834
  • 期刊名称:中国应用生理学杂志
  • 时间:2014
  • 页码:413-416
  • 分类:R363[医药卫生—病理学;医药卫生—基础医学]
  • 作者机构:[1]军事医学科学院卫生学环境医学研究所,天津市环境与食品安全风险监控技术重点实验室,天津300050
  • 相关基金:国家自然科学基金(81030052,21177159)
  • 相关项目:几种典型EDCs检测的高性能液相阵列压电传感器研究
中文摘要:

目的:将自制克伦特罗(CL)单克隆抗体纯化并研究其生物学特性,进行性质鉴定并建立检测标准曲线。方法:用ELISA法测定克伦特罗单克隆抗体的亲和常数和抗体活性,ELJSA测定单克隆抗体与BSA的交叉反应及与几种结构和功能类似物的交叉反应,然后采用间接竞争ELISA方法建立检测标准曲线。将制备的含克伦特罗单克隆抗体的小鼠腹水用盐析法和免疫亲和柱层析法进行抗体纯化。结果:经ELISA法测定,单克隆抗体亲和常数为2.90×10mmol/L,抗体效价最高达10^6。单克隆抗体对BSA无反应,对几种结构和功能类似物的交叉反应率均小于0.005%。建立的标准曲线R2=0.9812,最低检测限为1.0ng/ml。结论:建立了间接竞争ELISA检测cL的标准曲线。自制的克伦特罗单克隆抗体亲和力好,特异性高。为以后实际样品的检测及制备CL免疫检测试纸条和试剂盒奠定了基础。

英文摘要:

Objective: To identify the self-preparation monoclonal antibody which target to clenbuterol, and set up the standard curve to clenbuterol (CL) detection. Methods: The affinity constants and activity of the monoclonal antibody which target to CL were determined by ELISA. ELISA was also used to confirm whether the monoclonal antibody had any across-reaction with BSA and CL analogues. The rat ascites which contains the monoclonal antibody target to CL was purified by (NH4)2S04 salt-out method and further by aff'mity colunm. At last, the CL detection standard curve which based on indirect competition EI.ISA was established. Results: The ELISA experiment showed that the antibody fiter was 106 and the monoclonal antibody affinity constants was 2.90 ×1010 L/mol. The result of the indirect competition ELISA confirmed that the monoclonal antibody had no cross-reaction with BSA and a few kind of CL analogue. CL detection standard curve based on indirect compe- tition ELISA was established, which R2 was 0.9812, and the lowest detectable limit was 1.0 ng/ml. Conclusion: The standard curve based on indirectly competitioning ELISA was established. The self-preparation monoclonal antibody which target to CL has high affinity and high specific to CL, which had established the foundation to the advanced develot~nent of the CL immune test paper and CL ELISA kit.

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期刊信息
  • 《中国应用生理学杂志》
  • 北大核心期刊(2008版)
  • 主管单位:
  • 主办单位:中国生理学会 卫生学环境医学研究所 军事医学科学院
  • 主编:汪海
  • 地址:天津市大理道1号
  • 邮编:300050
  • 邮箱:tjzgyish@163.com
  • 电话:022-84655184
  • 国际标准刊号:ISSN:1000-6834
  • 国内统一刊号:ISSN:12-1339/R
  • 邮发代号:6-16
  • 获奖情况:
  • 国家印刷质量二等奖,国防科技期刊一等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),荷兰文摘与引文数据库,美国生物医学检索系统,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2000版)
  • 被引量:9411