A sensitive and specific ELISA using a monoclonal capture antibody for detection of Tamm-Horsfall urinary glycoprotein in serum.
An enzyme-linked immunosorbent assay (ELISA) has been established using Nunc polystyrene immunoplates coated with a monoclonal antibody to human Tamm-Horsfall urinary glycoprotein (THP) to detect and measure THP in human serum. Optimal reaction conditions for both the monoclonal capture antibody and the affinity-purified rabbit anti-human THP second antibody were established to produce standard curves which showed linearity between 20-90 ng/ml with a sensitivity of 2-3 ng/ml. The plate-to-plate standard curve mean coefficient of variation (CV) was 5.9 +/- 2.9% on assays performed on the same day while day to day mean CV was 13.3 +/- 2.4%. The specificity of the ELISA was demonstrated by inhibition of binding after preincubation of both urinary THP standards and serum with monoclonal anti-THP antibody. Sera from 195 blood donors tested by the ELISA had a mean concentration of THP antigenic determinants of 260 +/- 105 ng/ml. Results from three control sera run on all plates used in the survey showed mean CV less than 7.6% while no binding was observed with sera from an anephric patient.
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D008970:Molecular Weight; D009088:Mucoproteins / Q000097:blood*; D013696:Temperature; D013997:Time Factors; D058949:Uromodulin
D000911:Antibodies, Monoclonal; D009088:Mucoproteins; C548612:UMOD protein, human; D058949:Uromodulin