A new ELISA method for the rapid quantification of Tamm-Horsfall protein in urine.
The authors present a simplified enzyme-linked immunosorbent assay (ELISA) technique for the quantitative measurement of urinary Tamm-Horsfall protein (THP). Microtiter plates are coated with THP and urine samples at various dilutions without the need for a capture antibody. The bound glycoprotein is then incubated with a monoclonal anti-THP antibody and an alkaline phosphatase conjugated anti-IgG antibody. The assay was validated and gave reproducible results over a wide range of absorbance values. The sensitivity of the assay for THP was 2-5 micrograms/L, the coefficient of variation between assays 7.5%, and the day-to-day variability 11.1% for THP concentrations between 6.25 and 200 micrograms/L. THP excretion was assayed in five volunteers over five days comparing THP concentration in spot urines and in 24-hour urine collections.
Department of Medicine, Wayne State University, Detroit, Michigan.
H H
HH
Reinhart
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N
Obedeanu
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D
Walz
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JD
Sobel
D000328:Adult; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009088:Mucoproteins / Q000652:urine*; D011237:Predictive Value of Tests; D011257:Pregnancy Proteins / Q000652:urine*; D015203:Reproducibility of Results; D058949:Uromodulin
D000911:Antibodies, Monoclonal; D009088:Mucoproteins; D011257:Pregnancy Proteins; C548612:UMOD protein, human; D058949:Uromodulin