Clinical Chemistry
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Clinical Chemistry 32: 45-49, 1986;
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Clinical Chemistry, Vol 32, 45-49, Copyright © 1986 by American Association for Clinical Chemistry

One-step immunoassays for free (unbound) hormones: the effect of tracer binding by serum proteins

D Geiseler, P Chodha and R Ekins

In binding assays for determination of free (non-protein-bound) hormones in serum or plasma, the influence of the measuring system on the original analyte concentration in the sample must be considered. In one- or single-step free-hormone immunoassays, the labeled analyte or analog-tracer not only is bound to the antibody, it also is bound, to some extent, to serum proteins. The dependence of the assay response on two unknown variables--the concentration of free analyte and the binding potential of serum for the tracer--introduces a bias between the actual (original) and measured hormone concentrations. The significance of this protein effect is described by mathematical modeling of the analyte-protein distribution in the assay system. The theoretical consideration is validated by a clearly defined one-step assay system for measurement of free-thyroxin concentration, with a labeled thyroxin-immunoglobulin conjugate used as tracer.




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Copyright © 1986 by the American Association for Clinical Chemistry.