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

第一作者： Lin, Manman

作者： Lin, Manman;Liu, Yuan;Zhang, Xiao;Xu, Chongxin;Hu, Xiaodan;Zhu, Qing;Xie, Yajing;Chen, Wei;Liu, Xianjin;Lin, Manman;Hu, Xiaodan;Xie, Yajing;Lin, Johnson;Liu, Yuan;Meng, Meng;Jin, Jiafeng;Shen, Cheng

作者机构：

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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