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Clinical Chemistry 34: 98-102, 1988;
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Clinical Chemistry, Vol 34, 98-102, Copyright © 1988 by American Association for Clinical Chemistry

Highly specific enzyme immunoassay for the beta-subunit of human thyrotropin with use of monoclonal antibodies

Y Iijima, Y Endo, N Hata, H Fujita, M Unoki and K Miyal
Department of Laboratory Medicine, Osaka University Medical School, Japan.

A highly specific enzyme-linked "sandwich" immunoassay is described for determining free human thyrotropin (hTSH) beta-subunit in serum by using a anti-hTSH beta-subunit monoclonal antibody conjugated with beta- D-galactosidase (EC 3.2.1.23) and a solid phase consisting of silicone rods coated with another monoclonal antibody. We could detect as little as 0.04 ng of beta-subunit per assay. The measurable range of hTSH beta- subunit concentrations in serum was 0.4 to 50 micrograms/L. The assay demonstrated little or no cross reactivity with intact hTSH, hTSH alpha- subunit, or human choriogonadotropin. The mean CVs were 12.2% within assay, 13.9% between assay. The hTSH beta-subunit was not detectable in sera from healthy subjects, patients with hyperthyroidism, or two patients with pituitary tumors producing TSH. It was measurable (at concentrations of 0.65 to 2.70 micrograms/L) in sera from eight of 23 hypothyroid patients. In five of the hypothyroid patients examined, the concentration of hTSH beta-subunit in serum increased after administration of thyroliberin. This method may be useful in elucidating the physiological and pathological significance of the hTSH beta-subunit and in examining the function of the hypothalamus- pituitary-thyroid axis.




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Copyright © 1988 by the American Association for Clinical Chemistry.