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Research Article | Open Access

Selection of specific nanobodies against peanut allergen through unbiased immunization strategy and the developed immuno-assay

Yaozhong Hua,1, Chuan Zhangb,1, Jing Lina, Yi Wanga, Sihao Wua, Ying Sunb, Bowei Zhanga, Huan Lüa, Xuemeng Jia, Yang Lub, Shuo Wanga( )
Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China
College of Food Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China

1 These authors contribute equally.Peer review under responsibility of KeAi Communications Co., Ltd.]]>

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Abstract

Peanut allergy is considered to be a major health issue with global effects. To date, no effective curative approach has been applied for the therapy of the anaphylaxis resulting from the peanut allergens. The accurate and effective detection methods for the surveillance of allergens in food are still the primary strategies to avoid allergic diseases. In this study, nanobodies (Nbs) derived from the Heavy-Chain only Antibodies (HCAbs) were selected against the general peanut protein extract through the unbiased strategy to facilitate the development of the sandwich ELISA for the detection and surveillance of peanut allergen contamination. The target antigen of the selected Nb was identified as peanut allergen Ara h 3, and a cross-reaction was observed with the member of Gly 1 from the Ara h 3 family. The applicability of the self-paired Nb P43 on the establishment of the immuno-assay was verified. A sandwich ELISA against peanut allergen was developed, which reached a linear range of 0.2－10.6 μg/mL, and a limit of detection of 53.13 ng/mL.

Keywords

NanobodyPeanut allergenAra h 3Sandwich ELISAGly 1

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Food Science and Human Wellness
Volume 12 Issue 3,
May 2023
Pages 745-754
DOI: 10.1016/j.fshw.2022.09.008
Cite this article:
Hu Y, Zhang C, Lin J, et al. Selection of specific nanobodies against peanut allergen through unbiased immunization strategy and the developed immuno-assay. Food Science and Human Wellness, 2023, 12(3): 745-754. https://doi.org/10.1016/j.fshw.2022.09.008

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Received: 29 December 2020
Revised: 26 January 2021
Accepted: 18 February 2021
Published: 15 October 2022
© 2023 Beijing Academy of Food Sciences. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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