A biotin-avidin labeled enzyme immunoassay for the quantitation of serum TSH using protein-layered solid phase |
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Authors: | Myung Ja Choi Eun-Young Song Tai-Wha Chung |
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Institution: | (1) Korea Research Institute of Bioscience & Biotechnology, KIST, Taejon, Korea;(2) Bioanalysis & Biotransformation Research Center, KIST, Cheongryang, P. O Box 131, 130-650 Seoul, Korea |
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Abstract: | A sensitive enzyme immunoassay for serum TSH has been developed utilizing the tight binding between biotin and avidin, and
three layered protein polystyrene beads as solid phase. To increase binding capacity of TSH and sensitivity of the assay,
the polystyrene beads were coated sequentially with mouse immunoglobulin as first layer, rabbit antimouse immunoglobulin as
second layer and monoclonal anti-TSH as third layer. A serum sample was incubated simultaneously with a monoclonal anti-TSH
immobilized polystyrene beads and a second monoclonal anti-TSH covalently attached to biotin. After washing, the antibody
bound serum TSH-anti-TSH-biotin complex is reacted with horseradish peroxidase (HRP)-labeled avidin. Following a second wash,
the bound HRP activity was measured colorimetrically. Reproducible results were obtained within 4 hours for serum TSH in the
range between 0 μIU/ml and 50 μIU/ml with detection limit of 0.1 μIU per test. |
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Keywords: | Biotin-avidin EIA TSH Trilayered solid phase |
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