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 IC-50 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 IC-50 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 IC-50 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.