title: An ultra-sensitive monoclonal antibody-based competitive enzyme immunoassay for sterigmatocystin in cereal and oil products.
abstract: Sterigmatocystin (STG), a biosynthesis precursor of aflatoxin B1, is well known for its toxic and carcinogenic effects in humans and animals. STG derivatives and protein conjugates are needed for generation of monoclonal antibodies (mAbs). This work describes a reliable and fast synthesis of novel STG derivatives, based on which novel STG bovine serum albumin conjugates were prepared. With the novel STG bovine serum albumin conjugates, three sensitive and specific mAbs against STG, named VerA 3, VerA 4, and VerA 6, were prepared by semi-solid hypoxanthine/aminopterin/thymidine (HAT) medium using a modified two-step screening procedure. They exhibited high affinity for STG and no cross-reactivity (CR) with aflatoxins B1, B2, G1, G2, and M1. Based on the most sensitive antibody VerA 3, an ultra-sensitive competitive enzyme-linked immunosorbent assay (ELISA) was developed for STG in wheat, maize, and peanuts. Assays were performed in the STG-GA-BSA-coated (0.5 µg · mL(-1)) ELISA format, in which the antibody was diluted to 1:80,000. Several physicochemical factors influencing assay performance, such as pH, ionic strength, blocking solution, and diluting solution, were optimized. The final results showed that the assays had the detection limits of 0.08 ng · g(-1) for wheat, 0.06 ng · g(-1) for maize, and 0.1 ng · g(-1) for peanuts, inter-assay and intra-assay variations of less than 10%, and recoveries ranging from 83% to 110%. These recoveries were in good agreement with those obtained by using HPLC-MS/MS method (90-104%), indicating the importance of the mAb VerA 3 in the study of STG in crude agricultural products.
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, P. R. China; Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, P. R. China; Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, P. R. China; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Peiwu
authors.initials: P
authors.lastname: Li
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wu
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, P. R. China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Fei
authors.initials: F
authors.lastname: Ma
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, P. R. China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Zhaowei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Ding
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, P. R. China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, P. R. China; Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, P. R. China.
authors.forename: Hengling
authors.initials: H
authors.lastname: Wang
mesh_terms: D016604:Aflatoxin B1 / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D002622:Chemistry, Agricultural; D002523:Edible Grain / Q000737:chemistry; D005504:Food Analysis; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D009821:Oils / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology; D013241:Sterigmatocystin / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D009821:Oils; D013241:Sterigmatocystin; D012710:Serum Albumin, Bovine; D016604:Aflatoxin B1