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On-line Access: 2021-04-08

Received: 2020-10-23

Revision Accepted: 2020-11-19

Crosschecked: 2021-02-23

Cited: 0

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Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Weifeng CHEN

https://orcid.org/0000-0001-9948-5826

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Journal of Zhejiang University SCIENCE B 2021 Vol.22 No.4 P.305-309

http://doi.org/10.1631/jzus.B2000661


A tetravalent single-chain variable fragment antibody for the detection of staphylococcal enterotoxin A


Author(s):  Weifeng CHEN, Zhiwei LI, Xingxing DONG, Xiaohong WANG

Affiliation(s):  School of Food and Bioengineering, Henan University of Animal Husbandry and Economy, Zhengzhou 450046, China; more

Corresponding email(s):   wxh@mail.hzau.edu.cn

Key Words: 


Weifeng CHEN, Zhiwei LI, Xingxing DONG, Xiaohong WANG. A tetravalent single-chain variable fragment antibody for the detection of staphylococcal enterotoxin A[J]. Journal of Zhejiang University Science B, 2021, 22(4): 305-309.

@article{title="A tetravalent single-chain variable fragment antibody for the detection of staphylococcal enterotoxin A",
author="Weifeng CHEN, Zhiwei LI, Xingxing DONG, Xiaohong WANG",
journal="Journal of Zhejiang University Science B",
volume="22",
number="4",
pages="305-309",
year="2021",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2000661"
}

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%T A tetravalent single-chain variable fragment antibody for the detection of staphylococcal enterotoxin A
%A Weifeng CHEN
%A Zhiwei LI
%A Xingxing DONG
%A Xiaohong WANG
%J Journal of Zhejiang University SCIENCE B
%V 22
%N 4
%P 305-309
%@ 1673-1581
%D 2021
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2000661

TY - JOUR
T1 - A tetravalent single-chain variable fragment antibody for the detection of staphylococcal enterotoxin A
A1 - Weifeng CHEN
A1 - Zhiwei LI
A1 - Xingxing DONG
A1 - Xiaohong WANG
J0 - Journal of Zhejiang University Science B
VL - 22
IS - 4
SP - 305
EP - 309
%@ 1673-1581
Y1 - 2021
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B2000661


Abstract: 
Staphylococcal enterotoxin A (SEA) synthesized by Staphylococcus aureus is a foodborne and heat-stable toxin, which is a great threat to human health (Pexara et al., 2010). Highly sensitive antibodies are a key factor in the immunological detection of SEA, which is one of the most effective ways to detect SEA because of its accuracy, agility, and efficiency (Nouri et al., 2018). In this study, we constructed a tetravalent anti-SEA antibody gene by linking the tetramerization domain of human p53 to the C-terminus of the anti-SEA single-chain variable fragment (scFv), which was then transformed into Escherichia coli BL21 (DE3) for the production of a SEA-specific tetravalent antibody. Successful expression of the tetravalent antibody was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot. An indirect non-competitive enzyme-linked immunosorbent assay (ELISA) revealed that the tetravalent antibody exhibited SEA-specific binding activity. A sandwich ELISA demonstrated that compared to the scFv monomer, the tetravalent antibody was more sensitive in detecting SEA. Molecular docking analysis revealed that the SEA interacted with the scFv mainly on the opposite side of the residue linked to p53. Thus, this study indicated that genetically engineered tetramerization is a potential way to improve the sensitivity of SEA-specific scFv.

一种用于检测金黄色葡萄球菌A型肠毒素的四聚化单链抗体

概要:免疫学方法可以用于金黄色葡萄球菌A型肠毒素的准确而快速的检测,而快速生产高亲和力抗体是其中的关键步骤。在本研究中,我们通过p53的四聚化功能结构域,在金黄色葡萄球菌A型肠毒素特异性单链抗体的基础上,成功制备了四聚化的单链抗体。酶联免疫吸附试验表明该四聚化抗体对金黄色葡萄球菌A型肠毒素的亲和力显著高于其单体形式,表明基于基因工程的四聚化是提高单链抗体亲和力的一种潜在的可行方法。

关键词:金黄色葡萄球菌A型肠毒素;酶联免疫吸附试验;单链抗体;四聚体

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

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