An enzyme-linked immunoadsorbent assay (ELISA) for measuring human pituitary glycoprotein free α-subunit is described. Pituitary tumours may secrete glycoprotein hormones, their free subunits (α, β) or a combination of these. Non-functioning adenomas often secrete free α-subunit. Assays for free α-subunit have previously used radioimmunoassay or immunoradiometric principles. Some of these methods are time-consuming and lack specificity and sensitivity. In order to overcome these problems, we have developed a two-site ELISA for α-subunit which uses a monoclonal antibody to α-subunit as the capture antibody to provide greater specificity. An affinity-purified polyclonal anti α-subunit conjugated to alkaline phosphatase was the signal antibody. Detection and quantification were based on phenolphthalein monophosphate conversion to phenolphthalein. The ELISA specifically measured glycoprotein free α-subunit in serum or plasma, discriminating between α-subunit and the intact glycoprotein hormones. The assay can be completed in 4 h. The assay sensitivity was 0·03 μg/l, and a normal range of 0·05 to 0·22 μg/l was established. In a retrospective study, elevated circulating glycoprotein α-subunit was detected in 22% of patients with non-functioning pituitary tumours previously treated with surgery and/or radiotherapy.
Journal of Endocrinology (1993) 136, 511–516
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