[1]俞华齐,王 娟,姜忍忍,等.重金属铬单克隆抗体特异性ELISA免疫检测方法的建立[J].江苏农业学报,2016,(02):460-466.[doi:10.3969/j.issn.1000-4440.2016.02.034]
　YU Hua-qi,WANG Juan,JIANG Ren-ren,et al.Development of specific ELISA for the detection of monoclonal antibody against heavy metal Cr[J].,2016,(02):460-466.[doi:10.3969/j.issn.1000-4440.2016.02.034]
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重金属铬单克隆抗体特异性ELISA免疫检测方法的建立()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年02期
页码:
460-466
栏目:
加工贮藏·质量安全
出版日期:
2016-03-20

文章信息/Info

Title:
Development of specific ELISA for the detection of monoclonal antibody against heavy metal Cr
作者:
俞华齐1;  王 娟2;  姜忍忍2;  姚 刚2;  刘少伟1
1.华东理工大学生物工程学院国家重点实验室,上海 200237; 2.中国科学院上海生命科学研究院营养科学研究所食品安全研究中心,上海 200031
Author(s):
YU Hua-qi1;  WANG Juan2;  JIANG Ren-ren2;  YAO Gang2;  LIU Shao-wei1
1.The State Key Laboratory of Bioreactor Engineering,Department of Biological Engineering, East China University of Science and Technology,Shanghai 200237,China; 2.Key Laboratory for Food Safety Research, Institute of Nutritional Sciences, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031,China
关键词:
重金属Cr;  纳米抗体;  噬菌体展示;  间接竞争性ELISA
Keywords:
heavy metal Cr;  nanobody;  phage display;  indirect competitive ELISA
分类号:
X832
DOI:
10.3969/j.issn.1000-4440.2016.02.034
文献标志码:
A
摘要:
为了利用抗体库筛选技术制备检测重金属铬的单克隆抗体,本研究利用螯合剂DTPA成功制备了重金属Cr的完全抗原,并利用制备的完全抗原结合噬菌体展示技术成功筛选到了特异性识别Cr-DTPA的噬菌体抗体Cr-DTPA-VHH 4-63。 结果显示,获得的抗体亲和常数为 1.279×109 L/mol,与载体蛋白BSA/OVA及无金属抗原DTPA-BSA/OVA均无交叉反应,与其他金属离子也没有交叉反应,特异性强。利用制备的Cr-DTPA噬菌体抗体初步建立了ELISA免疫检测法,其方法最低检测限达到1 μg/L,水样加标回收试验平均回收率 90%～114%,达到国家饮用水检测标准,可用于农业灌溉用水及生活饮用水中的重金属铬的检测。
Abstract:
A complete antigen Cr-DTPA for heavy metal Cr was prepared using chelator p-SCN-Bn-DTPA, and the phage antibody Cr-DTPA-VHH 4-63 specifically identifying Cr-DTPA was selected by phage display technique. The nanobody showed a strong specificity, with no cross reactions with carrier protein BSA/OVA, non-metallic antigen DTPA-BSA/OVA, and other metallic ions. The indirect competitive ELISA method developed based on the nanobody could detect as low as 1 μg/L Cr in the water sample. The average recovery of Cr spiked in the water sample reached 90%-114%, meeting the national standard for drinking water detection. The method established in this study could be applied for heavy metal detection in irrigation water and drinking water.

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备注/Memo

备注/Memo:
收稿日期:2016-01-27 基金项目:科技部“十二五”国家科技支撑计划项目(2011BAZ0319814、2012BAK01B00) 作者简介:俞华齐(1990-),男,江苏淮安人,硕士研究生,主要从事生物技术和食品安全方面的研究。(E-mail)jsjhyhq@163.com 通讯作者:刘少伟,(E-mail)swliu@ecust.edu.cn
更新日期/Last Update: 2016-03-20