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An enzyme-linked immunoadsorbent assay (ELISA) for measuring human pituitary glycoprotein free -subunit is described. Pituitary tumours may secrete glycoprotein hormones, their free subunits (, β) or a combination of these. Non-functioning adenomas often secrete free -subunit. Assays for free -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 -subunit which uses a monoclonal antibody to -subunit as the capture antibody to provide greater specificity. An affinity-purified polyclonal anti -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 -subunit in serum or plasma, discriminating between -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 -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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