title: Preparation of highly specific anti-zearalenone antibodies by using the cationic protein conjugate and development of an indirect competitive enzyme-linked immunosorbent assay.
abstract: Although anti-zearalenone (ZEN) antibodies have been widely prepared, these antibodies cross-react with α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), zearalanone (ZAN), α-zearalanol (α-ZAL) and β-zearalanol (β-ZAL). To overcome this problem and improve the specificity of immunoassays, we produced anti-ZEN antibodies based on a ZEN-cationic protein conjugate. In this study, ZEN was coupled with cationic bovine serum albumin (cBSA) via a Mannich reaction. After BALB/c mice were immunized with ZEN-cBSA, an immunological response was rapidly induced. The titers of the polyclonal antisera and monoclonal antibody were 30,000 and 20,000, respectively. Cross-reactivity (CR) values of the anti-ZEN polyclonal antisera and monoclonal antibody with the 5 analogs were <7% and <2%, respectively. An indirect competitive enzyme-linked immunosorbent assay based on the monoclonal anti-ZEN antibody was established. The recovery rates of ZEN in spiked cereal and feed were in the range of 80%-120% with coefficients of variation <15%. The intra-assay variation and inter-assay variation in assay buffer were both <5%. Therefore, the results demonstrated a feasible approach for preparing highly specific, higher titer and more rapidly induced antibodies against ZEN by using a ZEN-cBSA conjugate as the immunogen instead of currently used immunogens.
authors: Institute of Medicinal Plant Development, Chinese Academy of Medical Science & Peking Union Medical College, Beijing, 100193, PR China.
authors: Yuan
authors: Y
authors: Gao
authors: Meihua
authors: M
authors: Yang
authors: Cheng
authors: C
authors: Peng
authors: Xiaohong
authors: X
authors: Li
authors: Runlan
authors: R
authors: Cai
authors: Yun
authors: Y
authors: Qi
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D000918:Antibody Specificity; D002021:Buffers; D002417:Cattle; D003429:Cross Reactions; D002523:Edible Grain / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004968:Estrogens, Non-Steroidal / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D015025:Zearalenone / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D015029:Zeranol / Q000031:analogs & derivatives / Q000032:analysis* / Q000276:immunology / Q000378:metabolism
chemical_list: D000888:Antibodies, Anti-Idiotypic; D002021:Buffers; D004968:Estrogens, Non-Steroidal; D007106:Immune Sera; D012710:Serum Albumin, Bovine; D015025:Zearalenone; D015029:Zeranol