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Year : 2023, Volume : 12, Issue : suppl
First page : ( 138) Last page : ( 138)
Print ISSN : 2320-1193. Online ISSN : 2394-448X. Published online : 2023 December 12.
Article DOI : 10.5958/2394-448X.2023.00057.3

Synthesis of Nanobody Against Botulinum Type A for Toxin Enrichment Application

Phan Thanh-Thuy Thi1,a, Nguyen Duc-Hai Le1,a, Nguyen Anh-Nguyet Thi1, Luong Trung-Hieu, Nguyen Thanh-Trung3, Le Linh Thi Phuong1, Nguyen Loan Thi Hong1,2, Pham Yen Bao2, Phan Tuan-Nghia1,2, Le Nhung Thi Hong1,2,*

1Faculty of Biology, VNU University Of Science, Vietnam National University, Vietnam

2The Key Laboratory of Enzyme and Protein Technology, VNU University of Science, Vietnam

3National Institute for Food Control, Vietnam

*Corresponding Author : nhungle@hus.edu.vn

a: co-first author

Online Published on 12 December, 2023.

Received:  12  October,  2023; Accepted:  10  November,  2023.

Keywords

Nanobody, Botulinum type A, Toxin, Enrichment.

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Nanobodies are recombinantly synthesized single-domain antibodies based on sequences of mini-antibodies containing only heavy chains, with no light chains, isolated from Illamas and sharks. Nanobody has currently being used as a solution to enhance the detection of toxins in food (Lam et al. 2020; McNutt et al. 2021; Miyashita et al. 2021).

In this study, our objective is to recombinantly synthesize the Alc-B8 (A8) nanobody molecule that specifically binds to the light chain of Botulinum toxin type A (BoNT/A) for toxin enrichment applications. Based on the published data, our research team has identify suitable conditions for the expression of nanobody A8 using pET-28a vector in E.coli BL21 (DE3) RIL strain. Subsequently, nanobody A8 was purified using Ni-Sepharose and tested for its specificity against BoNT/A. Our initial results demonstrated that the synthesized Nanobody A8 is capable of recognizing BoNT/A antigen at the concentration of 100 mg/ml. We are applying this nanobody on magnetic beads for BoNT/A enrichment.

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Acknowledgements

We would like to sincerely thank Dr. Nguyen Van Sang, Faculty of Biology, VNU University Of Science and Dr. Bui Le Minh, Nguyen Tat Thanh University for kindly giving us thrombin enzyme for protein digestion. This research was conducted within the framework of the KLEPT.22.03 funded by Vienam National University.

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