Abstract
Sex hormone binding globulin (SHBG) is a hormone binding protein which plays an important role in regulating the transport and availability of biologically active androgens and estradiol to target cells and used to calculate free testosterone concentrations. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed, featuring an albumin removal step followed by a tryptic digestion. After a reduction step with dithiothreitol and alkylation with iodoacetamide three signature peptides were used for the quantification of SHBG. The method enables the quantification of serum and plasma SHBG over the clinically relevant range of 200-20,000ng/mL and was validated according to the mostrecent guidelines. The LC-MS/MS method correlates wellwith the Abbott Alinity immunoassay (R2>0.95), but theLC-MS/MS results are on average 16-17% lower than the immunoassay results, which is consistent for all three signature peptides. The LC-MS/MS method which includes an albumin depletion step allows quantification of SHBG in serum and plasma without an immunocapture step at clinically relevant SHBG levels, thus contributing to better lab-to-lab consistency of results.
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