Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (5): 191-200.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0991

Previous Articles     Next Articles

Screening and Identification of Humanized Genetically Engineered Antibody Targeting to Simulate the Anti-insect Function of Bt Cry1C Protein

XU Chong-xin(), ZHANG Xiao, LIU Yuan(), ZHONG Jian-feng, XIE Ya-jing, LU Li-na, GAO Mei-jing, LIU Xian-jin()   

  1. Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and technology,Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Sciences,Nanjing 210014
  • Received:2021-08-05 Online:2022-05-26 Published:2022-06-10
  • Contact: LIU Yuan,LIU Xian-jin E-mail:hhxyxcx@163.com;liuyuan@jaas.ac.cn;jaasliu@jaas.ac.cn

Abstract:

It has become a hot spot to explore new safe insect-resistant materials,as the pest resistance problem being increasingly prominent alongside the promotion and application of Bt protein and its transgenic crops. The anti-idiotype antibody technology based on antibody immune network theory has been proved to be useful for antigenic bioactive analogue research,and is expected to be a new approach for the targeted creation of the materials simulating the anti-insect function of Bt protein. Using Bt Cry1C protien polyclonal antibody as coated antigen,there were nine positive monoclone obtained from a humanized phage single-chain antibody library. Among them,the E8-Anti-Id scFv was identified as a β type anti-idiotypic genetically engineered single-chain antibody. The half-maximum inhibition concentrations(IC50)of Bt Cry1C polyclonal antibodies(pAbs),Plutella xylostella Brush border membrane vesicles(BBMV)and Cnaphalocrocis medinalis BBMV to E8-Anti-Id scFv were 2.625,3.638 and 4.605 μg/mL,respectively. Laboratory bioassay showed that the corrected mortality of C. medinalis BBMV and P. xylostella BBMV at 72 h were 58.69% and 52.82% at the phage-displayed E8-Anti-Id scFv of titer 1.2×108 CFU/mL,respectively. Its anti-insect activity reached 77.87% and 73.21% of original Bt Cry1C protein(20 μg/mL),respectively. These results indicate that the E8-Anti-Id scFv has preliminary characteristics of mimics the insect-resistant function of Bt Cry1C protein,which lays a technical and material foundation for further research and application.

Key words: Bt Cry1C protein, anti-idiotype antibody, phage antibody library, genetically engineered antibody