Discuss the methodology of radio immune essay and list its advantages.
Radioimmunoassay (RIA) is an in vitro assay that measures the presence of an antigen with very high sensitivity. Basically any biological substance for which a specific antibody exists can be measured, even in minute concentrations. RIA has been the first immunoassay technique developed to analyze nanomolar and picomolar concentrations of hormones in biological fluids.
The target antigen is labeled radioactively and bound to its specific antibodies (a limited and known amount of the specific antibody has to be added). A sample, for example a blood-serum, is then added in order to initiate a competitive reaction of the labeled antigens from the preparation, and the unlabeled antigens from the serum-sample, with the specific antibodies. The competition for the antibodies will release a certain amount of labeled antigen. This amount is proportional to the ratio of labeled to unlabeled antigen. A binding curve can then be generated which allows the amount of antigen in the patient's serum to be derived.
That means that as the concentration of unlabeled antigen is increased, more of it binds to the antibody, displacing the labeled variant. The bound antigens are then separated from the unbound ones, and the radioactivity of the free antigens remaining
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Discuss the methodology of radio immune essay and list its advantages.
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| What are molar conductivity and limited molar conductivity? Calculate the dissociation constant of 0.02 mol dm−3 ethanoic acid, if its molar conductivity is 11.57 × 10−4 S m2 mol−1 and limited molar conductivity is 3.90 × 10−2 S m2 mol−1. |
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