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Clinical Chemistry 29: 1777-1780, 1983;
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Clinical Chemistry, Vol 29, 1777-1780, Copyright © 1983 by American Association for Clinical Chemistry

Time-resolved fluoroimmunoassay of human pancreatic phospholipase A2

JU Eskola, TJ Nevalainen and TN Lovgren

We describe an immunofluorometric assay for human pancreatic phospholipase A2 based on time-resolved fluorescence. The labeled antibody technique in combination with the time-resolved 1-s fluorometric detection of the europium label, which essentially eliminates all background fluorescence, resulted in a high sensitivity (20 ng/L) and a wide (5000-fold) linear range. Nonspecific binding was minimized by treating the solid-phase antibody with NaSCN before coating, to remove endogenous antigen, and by immunosorbent purification of the antibody before labeling with europium. This is a one-incubation, multi-site, solid-phase assay on polystyrene microtiter strips, even though a polyclonal antibody was used. As measured by this assay, activity of immunoreactive phospholipase A2 was found to be above normal in sera of patients suffering from acute pancreatitis.




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Copyright © 1983 by the American Association for Clinical Chemistry.