Stanazolol derived ELISA as a sensitive forensic tool for the detection of multiple 17α-methylated anabolics

Publication date: Available online 5 December 2019Source: SteroidsAuthor(s): Lukáš Huml, Dominika Havlová, Ondřej Longin, Eliška Staňková, Barbora Holubová, Martin Kuchař, Elena Prokudina, Zdeňka Rottnerová, Tomáš Zimmermann, Pavel Drašar, Oldřich Lapčík, Michal JurášekAbstractTwo valuable forensic tools based on enzyme-linked immunoassays (ELISAs) for the analysis of 17α-methylated steroids were developed using haptens of stanazolol and its conjugates with biotin. Haptens containing terminal carboxylic group were conjugated to bovine serum albumin (BSA), rabbit serum albumin (RSA) or ovalbumin (OVA). Eight batches of antisera (RAbs) obtained by immunization of rabbits were tested in an indirect competitive ELISA system using immobilization of RSA conjugate (RSA/hapten) and competitor immobilization of the biotinylated conjugate (AB-ELISA) to avidin (avidin/hapten). The best results were achieved with the RAb 212 antibodies in RSA/ST-3 and avidin/ST-10 assembled variants. For the RSA/ST-3 system, an IC50 of 0.3 ng/mL and a detection limit of 0.02 ng/mL were measured. In case of avidin/ST-10 variant, IC50 was of 3.9 ng/mL and a detection limit of 0.57 ng/mL were obtained. The effect of solvent was tested as well as the stability of coated microtiter plates over four-month period. The cross-reactivity of the developed assays with other anabolic steroids was tested and high sensitivity towards 17α-methylated steroids was observed. RSA/ST-...
Source: Steroids - Category: Drugs & Pharmacology Source Type: research