title: Development of a competitive indirect enzyme-linked immunosorbent assay based on monoclonal antibodies for the detection of 2-dodecylcyclobutanone in irradiated beef.
abstract: A highly sensitive and specific competitive indirect enzyme-linked immunosorbent assay (ciELISA) based on monoclonal antibodies was developed for the detection of 2-dodecylcyclobutanone (2-DCB), a chemical marker for irradiated lipid-containing foods. 2-Oxocyclobutane undecanoic acid was used as an alternative to 2-DCB and conjugated to BSA and OVA via a conventional carbodiimide condensation reaction to prepare the immunogen and the coating antigen for 2-DCB. The monoclonal antibody against 2-DCB was obtained using the hybridoma technique, with a high specificity and low cross-reactivity for 2-tetradecylcyclobutanone (2-TCB; <8%) and other structurally related compounds (<0.1%). The ciELISA method was applicable at optimal experimental conditions of 0.001-100 μg/mL 2-DCB in a buffer solution, with an IC50 value of 0.25 μg/mL and a limit of detection (defined as the IC20) of approximately 0.004 μg/mL. The recovery efficiency of 2-DCB from ground beef patties ranged from 84.4 to 109.8%. The intra-assay and interassay coefficients of variation were <10.0 and <12.0%, respectively. The proposed method was validated by gas chromatography-mass spectrometry with high correlation. The same method was used to detect 2-DCB in ground beef patties irradiated at 0.5, 1.0, 3.0, 5.0, and 7.0 kGy; the 2-DCB concentration linearly increased with the radiation dose.
authors.affiliation: Institute of Agro-food Science and Technology, Chinese Academy of Agricultural Sciences /Key Opening Laboratory of Agricultural Products Processing and Quality Control, Ministry of Agriculture, Beijing 100193, China.
authors.forename: Yueliang
authors.initials: Y
authors.lastname: Zhao
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.forename: Weiming
authors.initials: W
authors.lastname: Li
authors.forename: Yiming
authors.initials: Y
authors.lastname: Ha
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D002417:Cattle; D003503:Cyclobutanes / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005514:Food Irradiation; D008460:Meat / Q000032:analysis* / Q000528:radiation effects*; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000528:radiation effects
chemical_list: D000911:Antibodies, Monoclonal; D003503:Cyclobutanes; C485436:2-tetradecylcyclobutanone