Deoxynivalenol-mimic nanobody isolated from a naïve phage display nanobody library and its application in immunoassay

Analytica Chimica Acta
Yulou QiuYuan-Yuan Liu

Abstract

In this study, using mycotoxin deoxynivalenol (DON) as a model hapten, we developed a nanobody-based environmental friendly immunoassay for sensitive detection of DON. Two nanobodies (N-28 and N-31) which bind to anti-DON monoclonal antibody (MAb) were isolated from a naive phage display library. These nanobodies are clonable, thermally stable and mycotoxin-free products and can be served as coating antigen mimetics in heterologous immunoassay. The half inhibition concentration (IC50) of the immunoassay developed with N-28 and N-31 was 8.77 ± 0.41 ng mL(-1) and 19.97 ± 0.84 ng mL(-1), respectively, which were 18- and 8-fold more sensitive than the conventional coating antigen (DON-BSA) based immunoassay. In order to better understand the molecular mechanism of antigen mimicry by nanobody, the 3D structure of "nanobody (N-28) - anti-DON MAb" complex was presented and verified by molecular modeling and alanine-scanning mutagenesis. The results showed that hydrogen bond and hydrophobic interaction formed between Thr 102 - Ser 106 of N-28 and CDR H3 residues of anti-DON antibody may contribute to their binding. This novel concept of enhancing sensitivity of immunoassay for DON based on nanobody may provide potential applications in...Continue Reading

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Citations

Sep 30, 2016·Journal of Agricultural and Food Chemistry·Yuanyuan LiuQinghua He
Nov 12, 2017·Analytical and Bioanalytical Chemistry·Riikka PeltomaaMaría C Moreno-Bondi
Jun 4, 2019·Analytical and Bioanalytical Chemistry·Hina MukhtarJia Wang
Nov 8, 2018·International Journal of Molecular Sciences·Hanna J WagnerWilfried Weber
Dec 28, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Li HanZi-Liang Wang
Sep 13, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yanan WangJunjie Chen
Aug 20, 2019·Analytical and Bioanalytical Chemistry·Riikka PeltomaaMaría C Moreno-Bondi
Jun 20, 2021·Molecular Diagnosis & Therapy·Nairo Brilhante-da-SilvaSoraya Dos Santos Pereira
Aug 22, 2021·Analytical and Bioanalytical Chemistry·Riikka PeltomaaMaría C Moreno-Bondi
Jan 25, 2020·Journal of Agricultural and Food Chemistry·Jia-Xiang YanQing-Hua He
Sep 30, 2021·Journal of Agricultural and Food Chemistry·Dan-Tong HuangZhen-Lin Xu

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