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Identification of single domain antibodies with insect cytotoxicity using phage-display antibody library screening and Plutella xylostella ATP-binding cassette transporter subfamily C member 2 (ABCC2)-based insect cell expression system

文献类型： 外文期刊

作者： Zhu, Qing 1 ; Hu, Xiaodan 1 ; Liu, Yuan 1 ; Xie, Yajing 1 ; Xu, Chongxin 1 ; Lin, Manman 1 ; Pooe, Ofentse Jacob 2 ; Zhong, Jianfeng 1 ; Gao, Meijing 1 ; Lu, Lina 1 ; Liu, Xianjin 1 ; Zhang, Xiao 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Food Safety & Nutr, Jiangsu Key Lab Food Qual & Safety,Key Lab Contro, State Key Lab Cultivat Base,Minist Agr,Minist Sci, Nanjing 210014, Peoples R China

2.Univ KwaZulu Natal, Coll Agr Engn & Sci, Sch Life Sci, Discipline Biochem, Westville Campus,Private Bag X54001, ZA-4000 Durban, South Africa

3.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang 212013, Jiangsu, Peoples R China

关键词： Phage display; Cell screening; PxABCC2; Cytotoxicity; Molecular docking

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.025； 五年影响因子：7.626 ）

ISSN： 0141-8130

年卷期： 2022 年 209 卷

页码：

收录情况： SCI

摘要： It is extremely imminent to study a new strategy to manage agricultural pest like Plutella xylostella (P. xylostella) which is currently resistant to most of pesticides, including three domain-Cry toxins from Bacillus thuringiensis (Bt). In this study, we reported a phage displayed single domain antibody screening from human domain antibody (DAb) library targeted on Spodoptera frugiperda 9 (Sf9) cells expressed Cry1Ac toxin receptor, ATP dependent binding cassette transporter C2 in P. xylostella (PxABCC2). After three rounds of panning, three cytotoxic antibodies (1D2, 2B7, 3C4) were obtained from thirty-eight antibodies and displayed high binding ability towards PxABCC2-expressed Sf9 cells. Through homology modeling and molecular docking, the interaction mode indicated that the most cytotoxic 1D2 of the three antibodies presented the lowest binding free energy required and had the most hydrogen bond formed with PxABCC2 in molecular docking analysis. Functional assay of key regions in 1D2 via Alanine replacement indicated that complementarity-determining region (CDR) 3 played a crucial role in antibody exerts binding activity and cytotoxicity. This study provides the first trial for discovering of potential cytotoxic antibodies from the human antibody library via specific receptor expressed insect cell system biopanning.

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