Generation of anti-idiotypic antibodies mimicking Cry2Aa toxin from an immunized mouse phage display library as potential insecticidal agents against Plutella xylostella
文献类型： 外文期刊
作者： Lin, Manman 1 ; Liu, Yuan 1 ; Shen, Cheng 4 ; Meng, Meng 3 ; Zhang, Xiao 1 ; Xu, Chongxin 1 ; Jin, Jiafeng 3 ; Hu, Xiaodan 1 ; Zhu, Qing 1 ; Xie, Yajing 1 ; Chen, Wei 1 ; Liu, Xianjin 1 ; Lin, Johnson 2 ;
作者机构： 1.Jiangsu Acad Agr Sci, Inst Food Safety & Nutr, Jiangsu Key Lab Food Qual & Safety State Key Lab C, Minist Sci & Technol,Key Lab Control Technol & Sta, Nanjing 210014, Peoples R China
2.Univ KwaZulu Natal, Sch Life Sci, Coll Agr Engn & Sci, Discipline Microbiol, Westville Campus,Private Bag X54001, ZA-4000 Durban, South Africa
3.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang 212013, Peoples R China
4.Nanjing Agr Univ, Coll Plant Protect, Nanjing 210095, Peoples R China
关键词： Anti-idiotypic antibody; Single-chain variable fragment; Cry2Aa toxin; Insecticidal protein; Plutella xylostella
期刊名称：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS （ 影响因子：3.1； 五年影响因子：3.2 ）
ISSN： 0006-291X
年卷期： 2024 年 691 卷
页码：
收录情况： SCI
摘要： This study tried to generate anti-idiotypic antibodies (Ab2s) which mimic Cry2Aa toxin using a phage-display antibody library (2.8 x 107 CFU/mL). The latter was constructed from a mouse immunized with F (ab')2 fragments digested from anti-Cry2Aa polyclonal antibodies. The F (ab')2 fragments and Plutella xylostella (P. xylostella) brush border membrane vesicles (BBMV) were utilized as targets for selection. Eight mouse phagedisplay single-chain variable fragments (scFvs) were isolated and identified by enzyme-linked immunoassay (ELISA), PCR and DNA sequencing after four rounds of biopanning. Among them, M3 exhibited the highest binding affinity with F (ab')2, while M4 bound the best with the toxin binding region of cadherin of P. xylostella (PxCad-TBR). Both of these two fragments were chosen for prokaryotic expression. The expressed M3 and M4 proteins with molecular weights of 30 kDa were purified. The M4 showed a binding affinity of 29.9 +/- 2.4 nM with the PxCad-TBR and resulted in 27.8 +/- 4.3 % larvae mortality against P. xylostella. Computer-assisted molecular modeling and docking analysis showed that mouse scFv M4 mimicked some Cry2Aa toxin binding sites when interacting with PxCad-TBR. Therefore, anti-idiotypic antibodies generated by BBMV-based screening could be useful for the development of new bio-insecticides as an alternative to Cry2Aa toxin for pest control.
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