title: The Preparation and Identification of a Monoclonal Antibody against Citrinin and the Development of Detection via Indirect Competitive ELISA.
abstract: Citrinin (CTN) is a hepato-nephrotoxic mycotoxin produced by fungi genera ofAspergillus,Monauscus, andPenicillium.CTN contaminates grains, fruits, juices and vegetables, and causes various toxic effects to humans and animals. It has small molecular weight, which is non-immunogenic to animals. Thus, CTN was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA), respectively, by amide bonds using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) andN-hydroxysuccinimide (NHS). Mice were immunized with CTN-BSA conjugates, and spleen cells of the immunized mice were fused with Sp2/0 myeloma cells to obtain 21H27 hybriodoma cell. Ascitic fluid of hybridoma cell was produced in mice abdomen, and purified using caprylic/ammonium sulfate precipitation method. The 21H27 anti-CTN mcAb was the IgG2a antibody subclass, and cross-reactivity results indicated that anti-CTN mcAb is specific to CTN with high affinity (2.0 × 10⁸ L/mol). Indirect competitive ELISA (ic-ELISA) results showed that the linear range of detection was 0.01-5.96 ng/mL and the IC50was 0.28 ng/mL with a lower detection limit (LOD) of 0.01 ng/mL. The average recovery was 93.8% ± 1.6% with a coefficient variation of 1.0%-4.3%. Hence, anti-CTN mcAb secreted by 21H27 hybridoma cell was successfully produced and can be used to detect CTN contaminated feed and foodstuffs.
authors.affiliation: Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Key Laboratory of Biopesticide and Chemical Biology of The Education Minister, School of Life Science, Fujian Agriculture And Forestry University, Fuzhou 35002, China. kalayu.yirga@yahoo.com.
authors.forename: Shimuye
authors.initials: S
authors.lastname: Kalayu Yirga
authors.affiliation: Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Key Laboratory of Biopesticide and Chemical Biology of The Education Minister, School of Life Science, Fujian Agriculture And Forestry University, Fuzhou 35002, China. Lsmpu2008@163.com.
authors.forename: Sumei
authors.initials: S
authors.lastname: Ling
authors.affiliation: Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Key Laboratory of Biopesticide and Chemical Biology of The Education Minister, School of Life Science, Fujian Agriculture And Forestry University, Fuzhou 35002, China. yylwsh@sina.com.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Key Laboratory of Biopesticide and Chemical Biology of The Education Minister, School of Life Science, Fujian Agriculture And Forestry University, Fuzhou 35002, China. yjmail2008@126.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Key Laboratory of Biopesticide and Chemical Biology of The Education Minister, School of Life Science, Fujian Agriculture And Forestry University, Fuzhou 35002, China. wshyyl@sina.com.
authors.forename: Shihua
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002953:Citrinin / Q000032:analysis / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005433:Flour / Q000032:analysis; D005506:Food Contamination / Q000032:analysis; D006825:Hybridomas; D018796:Immunoconjugates / Q000276:immunology; D008807:Mice, Inbred BALB C; D012275:Oryza / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; D012710:Serum Albumin, Bovine; D002953:Citrinin
keywords: cell fusion; citrinin; conjugation; ic-ELISA; monoclonal antibody