title: Selection and application of broad-specificity human domain antibody for simultaneous detection of Bt Cry toxins.
abstract: Bt Cry toxin is a kind of bio-toxins that used for genetically modified crops (GMC) transformation widely. In this study, total 15 positive clones could bind the Bt Cry toxins which isolated from a human domain antibody library by 5 rounds affinity selection. According to analyzing of PCR amplification and enzyme-linked immunosorbent assay (ELISA), the most positive phage domain antibody (named F5) gene was cloned into the pET26b vector and expressed in E. coli BL21. The purified antibody was used to develop an indirect competitive ELISA (IC-ELISA) for Cry1Ab, Cry1Ac, Cry1B, Cry1C and Cry1F toxins, respectively. The working range of detection for standard curves in IC-ELISA were 0.258-1.407 μg/mL, the medium inhibition concentration (IC50) were 0.727-0.892 μg/mL and detection limit (IC10) were 0.029-0.074 μg/mL for those Bt Cry toxins. The affinity of F5 domain antibody with Cry1Ab, Cry1Ac, Cry1B, Cry1C and Cry1F toxins were 1.21-5.94 × 10(7) M(-1). The average recoveries of the 5 kinds of Bt Cry toxins from spiked wheat samples were ranged from 81.2%-100.8% with a CV at 2.5%-9.4%. The results showed that we successfully obtained the broad-specificity human domain antibody for simultaneous detection of Bt Cry toxins in agricultural product samples.
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Chongxin
authors.initials: C
authors.lastname: Xu
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Huaihua Vocational and Technical College, Huaihua 418007, China.
authors.forename: Xiaoqin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Xiaodan
authors.initials: X
authors.lastname: Hu
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Zhong
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Cunzheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu/Province-State Key Laboratory Breeding Base, Institute of Food Quality Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China. Electronic address: hhxyxcx@163.com.
authors.forename: Xianjin
authors.initials: X
authors.lastname: Liu
mesh_terms: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000032:analysis*; D004731:Endotoxins / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D006460:Hemolysin Proteins / Q000032:analysis*; D006801:Humans; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D057127:Single-Chain Antibodies; C030956:insecticidal crystal protein, Bacillus Thuringiensis
keywords: Bt Cry toxins; ELISA; Human domain antibody; Soluble expression