title: Development and validation of an enzyme-linked immunosorbent assay for the quantification of gelonin in mouse plasma.
abstract: This article details the development and validation of an enzyme-linked immunosorbent assay (ELISA) for the quantification of gelonin in mouse plasma. The ELISA was validated for intra- and inter-day variability and for accuracy over a standard curve range of 7.5-100 ng/mL. The assay was then applied to assess gelonin pharmacokinetics in mice. Results from the ELISA were compared to data obtained from a parallel study conducted with (125)Iodine-labeled gelonin, with quantification via gamma counting. The ELISA demonstrated good precision, as the percent coefficient of variation of quality control samples in intra-day and inter-day validation ranged from 5.4-9.3% and 2.9-7.3%, respectively. Sample recoveries ranged from 98.3-105% of nominal values. The ELISA method yielded lower plasma concentrations of gelonin than found from the less-specific gamma counting method. Consequently, pharmacokinetic analyses yielded significantly higher estimates for volume of distribution (106 ± 31 vs. 55.8 ± 13 mL/kg) and plasma clearance (34.7 ± 6.6 vs. 10.9 ± 2.1 mL/min/kg) for data determined by ELISA vs. by gamma counting.
authors.affiliation: a Department of Pharmaceutical Sciences , School of Pharmacy and Pharmaceutical Sciences, University at Buffalo - The State University of New York , Buffalo , New York , USA.
authors.forename: Frank A
authors.initials: FA
authors.lastname: Engler
authors.affiliation: a Department of Pharmaceutical Sciences , School of Pharmacy and Pharmaceutical Sciences, University at Buffalo - The State University of New York , Buffalo , New York , USA.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Balthasar
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008297:Male; D051379:Mice; D054789:Ribosome Inactivating Proteins, Type 1 / Q000097:blood* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D054789:Ribosome Inactivating Proteins, Type 1; C025674:GEL protein, Gelonium multiflorum
keywords: ELISA; assay; gelonin; pharmacokinetics; protein toxin; ribosome-inactivating protein