title: Identification of a high-affinity monoclonal antibody against ochratoxin A and its application in enzyme-linked immunosorbent assay.
abstract: Ochratoxin A (OTA) is one of the most commonly occurring mycotoxins produced by some species of Aspergillus and can contaminate cereal and cereal products. A high-affinity anti-OTA monoclonal antibody (mAb) was generated from a hybridoma cell line 2D8 using splenocytes from a BALB/c mouse immunized with synthesized OTA-bovine serum albumin conjugate. The mAb 2D8 is specific with high affinity (3.75 × 10(9) L/M). An indirect competitive ELISA (ic-ELISA) was then developed using this mAb for quantitative determination of OTA in corn and feed samples. Using the optimized conditions, there was good linearity between OTA concentration and competitive inhibition (y = -0.6076x + 0.2441, R(2) = 0.9923) with the working range from 2.4 to 23.6 μg/kg, IC50 at 7.6 μg/kg and lower limit of detection at 1.4 μg/kg. The recovery rates in spiked samples were 91.2-110.3%. Of the 56 corn and feed samples, this ic-ELISA and a commercial kit both found the same 13 samples positive for OTA with good linear correlation between the two methods in OTA quantification (R(2) = 0.9706). We conclude that this ic-ELISA can be used for rapid and quantitative screening of corn and feed samples for the presence of OTA.
authors.affiliation: Zhejiang University Institute of Preventive Veterinary Medicine, and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China.
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Zhejiang University Institute of Preventive Veterinary Medicine, and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China.
authors.forename: Mengjiao
authors.initials: M
authors.lastname: Sun
authors.affiliation: Zhejiang University Institute of Preventive Veterinary Medicine, and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Kang
authors.affiliation: Zhejiang University Institute of Preventive Veterinary Medicine, and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Xie
authors.affiliation: Zhejiang University Institute of Preventive Veterinary Medicine, and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Zhejiang A&F University, 88 Huanbei Road, Lin'an, Zhejiang, 311300, China.
authors.forename: Houhui
authors.initials: H
authors.lastname: Song
authors.affiliation: Zhejiang University Institute of Preventive Veterinary Medicine, and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China.
authors.forename: Xiaoliang
authors.initials: X
authors.lastname: Li
authors.affiliation: Zhejiang University Institute of Preventive Veterinary Medicine, and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China. Electronic address: whfang@zju.edu.cn.
authors.forename: Weihuan
authors.initials: W
authors.lastname: Fang
mesh_terms: D000821:Animal Feed / Q000382:microbiology / Q000633:toxicity; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000918:Antibody Specificity; D002138:Calibration; D002417:Cattle; D002460:Cell Line; D004784:Environmental Monitoring / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D008807:Mice, Inbred BALB C; D009793:Ochratoxins / Q000737:chemistry / Q000276:immunology*; D010047:Ovalbumin / Q000737:chemistry / Q000276:immunology; D012709:Serum Albumin / Q000737:chemistry / Q000276:immunology; D003313:Zea mays / Q000382:microbiology
chemical_list: D000911:Antibodies, Monoclonal; D009793:Ochratoxins; D012709:Serum Albumin; C025589:ochratoxin A; D010047:Ovalbumin
keywords: Artificial antigen; Monoclonal antibody; Ochratoxin A; Quantification; ic-ELISA