Production of Antibodies for the Quantitative Detection of the Anabolically Active Androgens 17β‐Boldenone and Methylboldenone

Abstract

New polyclonal (pAb) and monoclonal antibodies (mAb) for the immunochemical detection of the anabolic steroids 17β‐boldenone and methylboldenone have been developed. Enzyme‐linked immunosorbent assays (ELISA) were established for the effective control of these illegally applied growth promoters. The IC₅₀ for methylboldenone of pAb 53 was determined 1.69 ng/l providing a linear measuring range in the ELISA between 0.23 and 12.47 ng/l. The IC₅₀ for methylboldenone of pAb 52 was calculated to be 0.54 µg/l with a linear measuring range between 0.03 and 9.54 µg/l. The mAb Bak detects a minimum concentration of 0.13 µg/l 17β‐boldenone. The IC₅₀ corresponds to 1.6 µg/l. The lower detection limit of mAb Zak was determined at 0.3 µg/l 17β‐boldenone. The IC₅₀ for this antibody is about 1.2 µg/l. Sample measurements in urine matrices spiked with 2, 3, and 10 µg/l steroid revealed appropriate precision (average CV for pAb of 4.3%, and for mAb of 10.7%) and recovery rates for the developed ELISAs.

Keywords:

• Enzyme‐linked immunosorbent assay
• 17β‐boldenone
• methylboldenone
• monoclonal antibody
• polyclonal antibody

The authors would like to thank Dr. Reiner Stephany from CRL Bilthoven, Netherlands for providing his valuable expertise on steroid analysis. Also we like to thank Ms Helena Geltl for technical assistance and the Euoropean Community (QRLT‐2000‐01670), the Spanish Ministry of Education (Contract Number. NAN2004‐09415‐C05‐02) and the CSIC‐PIF (BIPTOMAG) for financial support. The AMR group is a Grup de Recerca de la Generalitat de Catalunya and has support from the Departament d'Universitats, Recerca i Societat de la Informació la Generalitat de Catalunya (expedient 2005SGR 00207). This publication is dedicated to Prof. Dr. Bertold Hock, Chair of Cell Biology, Technical University of München.