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

被引:45
作者
Wang, Juan [1 ,2 ,5 ]
Wang, Yulian [1 ,2 ,3 ,5 ]
Pan, Yuanhu [1 ,2 ,3 ,5 ]
Chen, Dongmei [1 ,2 ,3 ,4 ,5 ]
Liu, Zhenli [1 ,2 ,3 ,4 ,5 ]
Feng, Liang [1 ,2 ,5 ]
Peng, Dapeng [1 ,2 ,3 ,5 ]
Yuan, Zonghui [1 ,2 ,3 ,4 ,5 ]
机构
[1] Natl Reference Lab Vet Drug Residues HZAU, Wuhan 430070, Hubei, Peoples R China
[2] MAO Key Lab Detect Vet Drug Residues, Wuhan 430070, Hubei, Peoples R China
[3] MOA Lab Risk Assessment Qual & Safety Livestock &, Wuhan 430070, Hubei, Peoples R China
[4] Hubei Collaborat Innovat Ctr Anim Nutr & Feed Saf, Wuhan 430070, Hubei, Peoples R China
[5] Huazhong Agr Univ, Wuhan 430070, Hubei, Peoples R China
关键词
Phenothiazines; Monoclonal antibody; Indirect competitive enzyme linked immunosorbent assay; Feed; LINKED-IMMUNOSORBENT-ASSAY; PERFORMANCE LIQUID-CHROMATOGRAPHY; CHLORPROMAZINE; TRANQUILIZERS; CARAZOLOL; SEDATIVES; RADIOIMMUNOASSAY; THIORIDAZINE; RESIDUES; CHICKEN;
D O I
10.1016/j.foodchem.2016.11.062
中图分类号
O69 [应用化学];
学科分类号
081704 ;
摘要
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 1.1 mu g kg(-1) to 15.3 mu g kg(-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. (C) 2016 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1004 / 1013
页数:10
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