title: Isolation of broad-specificity domain antibody from phage library for development of pyrethroid immunoassay.
abstract: An antibody to phenoxybenzoic acid (PBA), the conserved chemical region of pyrethroids, was developed using a domain antibody (DAB) library to enable pyrethroid detection in agricultural products. The DAB library, constructed without animal immunization and based on a human VH framework, displayed repertoires on filamentous bacteriophage. After four rounds of panning, we obtained five domain antibodies that are capable of binding to PBA. Antibody A3 has strong identification capability to cypermethrin, β-cypermethrin, and fenvalerate. The antibody A3 was used to develop an enzyme-linked immunosorbent assay (ELISA). The IC50 values were 2.586, 1.814, and 2.251 μg/ml for cypermethrin, β-cypermethrin, and fenvalerate, respectively. The assay shows weak competition with flucythrinate but shows no competition with fenpropathrin, deltamethrin, and permethrin. The developed ELISA process was successfully applied to fortified Chinese cabbage samples, with the recoveries of cypermethrin, β-cypermethrin, and fenvalerate ranging from 84.4 to 112.3%. We developed an immunoassay to detect pyrethroids depending on the domain antibody library, which overcomes the limitation of requiring protein antigen to immunize animals raising antibody.
authors.affiliation: College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Xiaodan
authors.initials: X
authors.lastname: Hu
authors.affiliation: School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia.
authors.forename: Sicong
authors.initials: S
authors.lastname: Tu
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Aihua
authors.initials: A
authors.lastname: Wu
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Cunzheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Zhong
authors.affiliation: College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.
authors.forename: Shengming
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Food Quality and Safety of Jiangsu Province, Jiangsu Academy of Agricultural Science, Nanjing, 210014, People's Republic of China; Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture, Nanjing, 210014, People's Republic of China.
authors.forename: Xianjin
authors.initials: X
authors.lastname: Liu
authors.affiliation: College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China. Electronic address: kangtu@njau.edu.cn.
authors.forename: Kang
authors.initials: K
authors.lastname: Tu
mesh_terms: D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015394:Molecular Structure; D019151:Peptide Library; D011722:Pyrethrins / Q000032:analysis* / Q000276:immunology*
chemical_list: D000906:Antibodies; D019151:Peptide Library; D011722:Pyrethrins
keywords: Domain antibody; Enzyme-linked immunosorbent assay; Phenoxybenzoic acid; Pyrethroid