Preparation of a generic monoclonal antibody and development of a highly sensitive indirect competitive ELISA for the detection of phenothiazines in animal feed

Juan Wang; Yulian Wang; Yuanhu Pan; Dongmei Chen; Zhenli Liu; Liang Feng; Dapeng Peng; Zonghui Yuan

doi:10.1016/j.foodchem.2016.11.062

Preparation of a generic monoclonal antibody and development of a highly sensitive indirect competitive ELISA for the detection of phenothiazines in animal feed

Food Chem. 2017 Apr 15:221:1004-1013. doi: 10.1016/j.foodchem.2016.11.062. Epub 2016 Nov 15.

Authors

Juan Wang¹, Yulian Wang², Yuanhu Pan², Dongmei Chen³, Zhenli Liu³, Liang Feng¹, Dapeng Peng⁴, Zonghui Yuan⁵

Affiliations

¹ National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Wuhan, Hubei 430070, China; Huazhong Agricultural University, Wuhan, Hubei 430070, China.
² National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Wuhan, Hubei 430070, China; MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Wuhan, Hubei 430070, China; Huazhong Agricultural University, Wuhan, Hubei 430070, China.
³ National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Wuhan, Hubei 430070, China; MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Wuhan, Hubei 430070, China; Hubei Collaborative Innovation Centre for Animal Nutrition and Feed Safety, Wuhan, Hubei 430070, China; Huazhong Agricultural University, Wuhan, Hubei 430070, China.
⁴ National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Wuhan, Hubei 430070, China; MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Wuhan, Hubei 430070, China; Huazhong Agricultural University, Wuhan, Hubei 430070, China. Electronic address: pengdapeng@mail.hzau.edu.cn.
⁵ National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Wuhan, Hubei 430070, China; MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Wuhan, Hubei 430070, China; Hubei Collaborative Innovation Centre for Animal Nutrition and Feed Safety, Wuhan, Hubei 430070, China; Huazhong Agricultural University, Wuhan, Hubei 430070, China. Electronic address: yuan5802@mail.hzau.edu.cn.

PMID: 27979052
DOI: 10.1016/j.foodchem.2016.11.062

Abstract

In this study, a broadly specific monoclonal antibody was prepared and a sensitive monoclonal-based indirect competitive enzyme linked immunosorbent assay (ic-ELISA) was subsequently developed to determine the phenothiazines in animal feed with a simple sample preparation procedure for the first time. The obtained antibody 3A5 was of the immunoglobulin G1 (IgG1) isotype possessing a kappa light chain, which broadly cross-reacted to nine phenothiazines. The limit of detections of the method ranged from 1.1μgkg^-1 to 15.3μgkg^-1 in the swine feed and the fish feed. The recoveries of the phenothiazines were in the range of 78.2-116.6%. The coefficient of variations were less than 16.7%. A positive correlation (r>0.9249) between the results of the ic-ELISA and the high-performance liquid chromatography were also observed, which indicated that the developed ic-ELISA is reliable and can be used to monitor phenothiazines in animal feed.

Keywords: Feed; Indirect competitive enzyme linked immunosorbent assay; Monoclonal antibody; Phenothiazines.

MeSH terms

Animal Feed / analysis*
Animals
Antibodies, Monoclonal*
Chromatography, High Pressure Liquid / methods
Enzyme-Linked Immunosorbent Assay / methods*
Immunoglobulin G
Phenothiazines / analysis*
Swine

Substances

Antibodies, Monoclonal
Immunoglobulin G
Phenothiazines