title: Generation of monoclonal antibody against trans-resveratrol.
abstract: Monoclonal antibody (MAb) against trans-resveratrol (t-RSV) was obtained from hybridoma clones constructed from splenocytes of BALB/c mice immunized with carrier proteins (bovine serum albumin [BSA] and ovalbumin [OVA]) coupled with synthetic hapten mimicking t-RSV structure. The t-RSV-BSA derivate was more efficient at induction of the immune response than t-RSV-OVA. However, the use of t-RSV-OVA was advantageous during selection of hybridoma clones constructed from splenocytes of t-RSV-BSA-immunized mice. Pre-incubation of immune serum with free t-RSV inhibited the binding of antibody to t-RSV-BSA conjugate, suggesting the specific nature of antibody binding to t-RSV. Splenocytes obtained from the mouse immunized with t-RSV-BSA were used for the hybridoma construction. Expansion of the primary clones, their subsequent screening, and subcloning narrowed our search to allowed isolation of two IgG1a-producing hybridomas designated as 2H9 and 1B1. According to an indirect ELISA assay, the resulting MAb 2H9 had little or no cross-reactivity to cis-RSV. No recognition of trans-RSV-3-O-glucuronide and trans-RSV-3-sulfate was detected. The newly generated MAb against t-RSV may provide a highly valuable and cost-effective tool for the analytical assay for t-RSV in different biological and agricultural specimens.
authors.affiliation: Lycotec Ltd., St John's Innovation Park, Cambridge, United Kingdom.
authors.forename: Ivan M
authors.initials: IM
authors.lastname: Petyaev
authors.forename: Valeriy V
authors.initials: VV
authors.lastname: Tsibezov
authors.forename: Sergey N
authors.initials: SN
authors.lastname: Osipov
authors.forename: Nigel H
authors.initials: NH
authors.lastname: Kyle
authors.forename: Daria V
authors.initials: DV
authors.lastname: Vorobjeva
authors.forename: Yuriy K
authors.initials: YK
authors.lastname: Bashmakov
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis* / Q000737:chemistry; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D005260:Female; D006241:Haptens / Q000737:chemistry; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin / Q000276:immunology; D000077185:Resveratrol; D012710:Serum Albumin, Bovine / Q000276:immunology; D013267:Stilbenes / Q000737:chemistry / Q000276:immunology*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000941:Antigens; D006241:Haptens; D013267:Stilbenes; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D000077185:Resveratrol