Automated quantification of thyrotropin by radial partition immunoassay

Autor:	S R Dawson, K S Leung, S A Evans, C T Rigl, J A Rugg, C W Flaa
Rok vydání:	1988
Předmět:	Chromatography biology Filter paper medicine.diagnostic_test Chemistry medicine.drug_class Biochemistry (medical) Clinical Biochemistry Monoclonal antibody Fluorescence Fragment crystallizable region Immunoassay Monoclonal medicine biology.protein Antibody Conjugate
Zdroj:	Clinical Chemistry. 34:118-122
ISSN:	1530-8561 0009-9147
DOI:	10.1093/clinchem/34.1.118
Popis:	We describe a radial partition enzyme immunoassay in which fully automated quantification of human thyrotropin (hTSH) takes less than 11 min. This "sandwich"-type assay involves two monoclonal antibodies, both specific for the intact hTSH molecule. The solid phase consists of tabs of glass-fiber filter paper containing a pre-immobilized monoclonal anti-hTSH antibody complexed with a goat antibody specific for the Fc region of mouse IgG. The patient's sample is first applied to the central "reaction zone" of the tab, wherein hTSH binds to the immobilized antibody. Application of a buffered solution containing enzyme-labeled Fab' fragments of the second monoclonal anti-hTSH antibody initiates "sandwich" formation. A wash buffer containing a fluorogenic substrate elutes unbound conjugate to the tab periphery. The bound enzyme conjugate is quantified by measuring the rate of increase in fluorescence in the reaction zone of the tab, then converting the rate to clinical units by comparison with a stored calibration curve. The clinical utility and performance of the present assay compare favorably with those of other sensitive assays for hTSH.
Databáze:	OpenAIRE
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