Expression and Functional Properties of an Anti-Triazophos High-Affinity Single-Chain Variable Fragment Antibody with Specific Lambda Light Chain

文献类型： 外文期刊

第一作者： Liu, Rui

作者： Liu, Rui;Liang, Xiao;Xiang, Dandan;Guo, Yirong;Zhu, Guonian;Liang, Xiao;Liu, Yihua

作者机构：

关键词： triazophos;scFv;soluble expression;ELISA;SPR

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN： 1422-0067

年卷期： 2016 年 17 卷 6 期

页码：

收录情况： SCI

摘要： Triazophos is a widely used organophosphorous insecticide that has potentially adverse effects to organisms. In the present study, a high-affinity single-chain variable fragment (scFv) antibody with specific lambda light chain was developed for residue monitoring. First, the specific variable regions were correctly amplified from a hybridoma cell line 8C10 that secreted monoclonal antibody (mAb) against triazophos. The regions were then assembled as scFv via splicing by overlap extension polymerase chain reaction. Subsequently, the recombinant anti-triazophos scFv-8C10 was successfully expressed in Escherichia coli strain HB2151 in soluble form, purified through immobilized metal ion affinity chromatography, and verified via Western blot and peptide mass fingerprinting analyses. Afterward, an indirect competitive enzyme-linked immunosorbent assay was established based on the purified anti-triazophos scFv-8C10 antibody. The assay exhibited properties similar to those based on the parent mAb, with a high sensitivity (IC50 of 1.73 ng/mL) to triazophos and no cross reaction for other organophosphorus pesticides; it was reliable in detecting triazophos residues in spiked water samples. Moreover, kinetic measurement using a surface plasmon resonance biosensor indicated that the purified scFv-8C10 antibody had a high affinity of 1.8 x 10(-10) M and exhibited good binding stability. Results indicated that the recombinant high-affinity scFv-8C10 antibody was an effective detection material that would be promising for monitoring triazophos residues in environment samples.

分类号：

  • 相关文献

[1]Selection and application of broad-specificity human domain antibody for simultaneous detection of Bt Cry toxins. Xu, Chongxin,Zhang, Xiao,Liu, Yuan,Hu, Xiaodan,Zhong, Jianfeng,Zhang, Cunzheng,Liu, Xianjin,Liu, Xiaoqin.

[2]Functional and binding characterization of a single chain Fv antibody to abscisic acid and conjugated abscisic acid. Chen, Xiaojiao,Tan, Guiyu,Liu, Wei,Wang, Baomin,Zhang, Mingcai,Li, Zhaohu,Lu, Yao,Cao, Zhen.

[3]Quantitative investigation of the direct interaction between Hemagglutinin and fusion proteins of Peste des petits ruminant virus using surface Plasmon resonance. Meng, Xuelian,Deng, Ruixue,Zhu, Xueliang,Zhang, Zhidong. 2018

[4]Enhancing the soluble expression of an amylase in Escherichia coli by the mutations related to its domain interactions. Wang, Peili,Qin, Weitong,Xu, Jiangtao,Yan, Yaru,Tian, Jian,Wu, Ningfeng,Wang, Peili,Yao, Bin.

[5]Interaction of Hsp90 with phospholipid model membranes. Zhang, Muhan,Wang, Daoying,Li, Pengpeng,Sun, Chong,Xu, Rong,Geng, Zhiming,Xu, Weimin,Dai, Zhaoqi,Xu, Weimin. 2018

[6]Construction of scFv phage display library with hapten-specific repertories and characterization of anti-ivermectin fragment isolated from the library. Zhang, Xiao,Zhang, Cunzheng,Liu, Yuan,Yu, Xiangyang,Liu, Xianjin. 2010

[7]Established a new double antibodies Sandwich enzyme-linked immunosorbent assay for detecting Bacillus thuringiensis (Bt) Cry1Ab toxin based single-chain variable fragments from a naive mouse phage displayed library. Zhang, Xiao,Xu, Chongxin,Zhang, Cunzheng,Liu, Yuan,Xie, Yajing,Liu, Xianjin. 2014

[8]Rapid isolation of single-chain antibodies from a human synthetic phage display library for detection of Bacillus thuringiensis (Bt) Cry1B toxin. Zhang, Xiao,Liu, Yuan,Zhang, Cunzheng,Wang, Yun,Liu, Xianjin,Zhang, Xiao,Wang, Yun,Xu, Chongxin. 2012

[9]Selection of a Single Chain Variable Fragment Antibody against Ivermectin from a Phage Displayed Library. Wen, Shuang,Zhang, Xiao,Liu, Yuan,Zhang, Qingqing,Liu, Xianjin,Wen, Shuang,Liang, Jiansheng.

[10]Isolation of single chain variable fragment (scFv) specific for Cry1C toxin from human single fold scFv libraries. Wang, Yun,Zhang, Xiao,Liu, Xianjin,Wang, Yun,Zhang, Xiao,Zhang, Cunzheng,Liu, Yuan,Liu, Xianjin.

[11]A simple and sensitive competitive bio-barcode immunoassay for triazophos based on multi-modified gold nanoparticles and fluorescent signal amplification. Zhang, Chan,Jiang, Zejun,Jin, Maojun,Chen, Ge,Cao, Xiaolin,Cui, Xueyan,Zhang, Yudan,Li, Ruixing,Wang, Jing,Zhang, Chan,Jiang, Zejun,Jin, Maojun,Chen, Ge,Cao, Xiaolin,Cui, Xueyan,Zhang, Yudan,Li, Ruixing,Wang, Jing,Du, Pengfei,Du, Pengfei,Du, Pengfei,Abd El-Aty, A. M.,Abd El-Aty, A. M.. 2018

[12]Highly sensitive detection of triazophos pesticide using a novel bio-bar-code amplification competitive immunoassay in a micro well plate-based platform. Du, Pengfei,Jin, Maojun,Zhang, Chan,Chen, Ge,Cui, Xueyan,Zhang, Yudan,Zhang, Yanxin,Zou, Pan,Jiang, Zejun,Cao, Xiaolin,She, Yongxin,Jin, Fen,Wang, Jing. 2018

[13]Gender-specific metabolic responses in gonad of mussel Perna viridis to triazophos. Zhang, Linbao,Cai, Wengui. 2017

[14]Enhanced Competitive Chemiluminescent Enzyme Immunoassay for the Trace Detection of Insecticide Triazophos. Jin, Maojun,Shao, Hua,Jin, Fen,Shi, Xiaomei,Wang, Jing,Gui, Wenjun. 2012

[15]Competitive colorimetric triazophos immunoassay employing magnetic microspheres and multi-labeled gold nanoparticles along with enzymatic signal enhancement. Du, Pengfei,Jin, Maojun,Chen, Ge,Zhang, Chan,Cui, Xueyan,Zhang, Yudan,Zhang, Yanxin,Zou, Pan,Jiang, Zejun,Cao, Xiaolin,She, Yongxin,Jin, Fen,Wang, Jing.

[16]Mutation in acetylcholinesterase1 associated with triazophos resistance in rice stem borer, Chilo suppressalis (Lepidoptera: Pyralidae). Jiang, Xiaojing,Jiang, Weihua,Han, Zhaojun,Qu, Mingjing,Denholm, Ian,Fang, Jichao. 2009

[17]Cloning of Two Acetylcholinesterase Genes and Analysis of Point Mutations Putatively Associated with Triazophos Resistance in Chilo auricilius (Lepidoptera: Pyralidae). Li, Xiao-Huan,Zhang, Zhi-Chun,Liu, Bao-Sheng,Fang, Ji-Chao,Li, Xiao-Huan,Fang, Ji-Chao,Huang, Shui-Jin.

[18]Polyclonal antibodies to grain gliadins and their application in wheat quality prediction. Li, Q. Y.,Song, X. Y.,Zhang, E. Y.,Pei, Y. H.,Li, Q. Y.,Ji, K. M.,Yan, Y. M.. 2008

[19]Quantification of toxins in a Cry1Ac+CpTI cotton cultivar and its potential effects on the honey bee Apis mellifera L.. Peng Han,Chang-Ying Niu,Chao-Liang Lei,Jin-Jie Cui,Nicolas Desneux. 2010

[20]Serological Survey of Canine Leptospirosis in Southern China. Liu, Mingchao,Liu, Jianzhu,Wang, Lin,Wang, Zhenyong,Wang, Shujing,Yang, Dubao,Chai, Tongjie,Shi, Dayou,Guo, Shining,Liao, Shenquan,Sun, Mingfei. 2012

作者其他论文 更多>>

微信小程序