Nanobody-based electrochemical immunoassay for Bacillus thuringiensis Cry1Ab toxin by detecting the enzymatic formation of polyaniline

doi:10.1007/s00604-015-1602-9

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	Nanobody-based electrochemical immunoassay for Bacillus thuringiensis Cry1Ab toxin by detecting the enzymatic formation of polyaniline
	Zhu, Min 1,6; Li, Min 6; Li, Guanghui 1,6; Zhou, Zikai 6; Liu, Hong5 ; Lei, Hongtao 4; Shen, Yanfei 3; Wan, Yakun 1,2,6
刊名	MICROCHIMICA ACTA
	2015-11
卷号	182 期号:15-16 页码:2451-2459
关键词	Nanobody Polyaniline Cry1Ab toxin Sandwich immunoassay Immunosensor Differential pulse voltammetry
ISSN号	0026-3672
DOI	10.1007/s00604-015-1602-9
文献子类	Article
英文摘要	We describe an electrochemical immunoassay for the Cry1Ab toxin that is produced by Bacillus thuringiensis. It is making use of a nanobody (a heavy-chain only antibody) that was selected from an immune phage displayed library. A biotinylated primary nanobody and a HRP-conjugated secondary nanobody were applied in a sandwich immunoassay where horseradish peroxidase (HRP) is used to produce polyaniline (PANI) from aniline. PANI can be easily detected by differential pulse voltammetry at a working voltage as low as 40 mV (vs. Ag/AgCl) which makes the assay fairly selective. This immunoassay for Cry1Ab has an analytical range from 0.1 to 1000 nga (TM) mL(-1) and a 0.07 nga (TM) mL(-1) lower limit of detection. The average recoveries of the toxin from spiked samples are in the range from 102 to 114 %, with a relative standard deviation of < 7.5 %. The results demonstrated that the assay represented an attractive alternative to existing immunoassays in enabling affordable, sensitive, robust and specific determination of this toxin.
资助项目	Jiangsu Nanobody Engineering and Research Center of China[2015-06] ; Program for New Century Excellent Talents in University[NCET-20130127] ; National Natural Science Foundation of China[31271365] ; National Natural Science Foundation of China[31471216] ; National Natural Science Foundation of China[21305065] ; Key Program from the Natural Science Foundation of Jiangsu province[BK20130788] ; Fundamental Research Funds for the Central Universities[00000000] ; Natural Science Foundation of China[U1301214] ; Guangdong Natural Science Foundation[S2013030013338]
WOS关键词	PESTICIDAL CRYSTAL PROTEINS ; SENSITIVE DETECTION ; CHAIN ANTIBODIES ; CARBON NANOTUBES
WOS研究方向	Chemistry
语种	英语
出版者	SPRINGER WIEN
WOS记录号	WOS:000363038800011
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/276349]
专题	药理学第一研究室
通讯作者	Shen, Yanfei
作者单位	1.Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China; 2.Jiangsu Nanobody Engn & Res Ctr, Nantong 226010, Peoples R China; 3.Southeast Univ, Sch Med, Nanjing 210009, Jiangsu, Peoples R China; 4.South China Agr Univ, Guangdong Prov Key Lab Food Qual & Safety, Guangzhou 510642, Guangdong, Peoples R China 5.Plexera LLC, Woodinville, WA 98072 USA; 6.Southeast Univ, Inst Life Sci, Nanjing 210096, Jiangsu, Peoples R China;
推荐引用方式 GB/T 7714	Zhu, Min,Li, Min,Li, Guanghui,et al. Nanobody-based electrochemical immunoassay for Bacillus thuringiensis Cry1Ab toxin by detecting the enzymatic formation of polyaniline[J]. MICROCHIMICA ACTA,2015,182(15-16):2451-2459.
APA	Zhu, Min.,Li, Min.,Li, Guanghui.,Zhou, Zikai.,Liu, Hong.,...&Wan, Yakun.(2015).Nanobody-based electrochemical immunoassay for Bacillus thuringiensis Cry1Ab toxin by detecting the enzymatic formation of polyaniline.MICROCHIMICA ACTA,182(15-16),2451-2459.
MLA	Zhu, Min,et al."Nanobody-based electrochemical immunoassay for Bacillus thuringiensis Cry1Ab toxin by detecting the enzymatic formation of polyaniline".MICROCHIMICA ACTA 182.15-16(2015):2451-2459.