title: Determination of aculeatisides based on immunoassay using a polyclonal antibody against aculeatiside A.
abstract: An enzyme-linked immunosorbent assay (ELISA) was developed for determination of aculeatisides. Aculeatiside A was conjugated with bovine serum albumin (BSA) for immunization. The ratio of hapten in an antigen conjugate was determined by matrix-assisted laser desorption/ionization TOF mass spectrometry. Polyclonal antibody was developed in rabbits against an aculeatiside A-BSA conjugate. The antibody was specific for aculeatiside A and aculeatiside B. The range of the immunoassay extended from 100 ng ml(-1) to 5 pg ml(-1) of aculeatisides. Good correlation between ELISA and HPLC methods was obtained when crude extracts of plant samples were analyzed. The optimized ELISA was found to be applicable to the determination of total aculeatisides in various plant samples.
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Thailand.
authors.forename: Waraporn
authors.initials: W
authors.lastname: Putalun
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Toshiya
authors.initials: T
authors.lastname: Muranaka
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Shoyama
mesh_terms: D000906:Antibodies; D004144:Diosgenin / Q000031:analogs & derivatives* / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D010047:Ovalbumin; D012503:Saponins / Q000032:analysis* / Q000276:immunology; D012680:Sensitivity and Specificity; D032322:Solanum / Q000737:chemistry*
chemical_list: D000906:Antibodies; D012503:Saponins; C115560:aculeoside A; C115561:aculeoside B; D010047:Ovalbumin; D004144:Diosgenin