Accurate Radioimmunoassay of Human Growth Hormone with Separation on Polyacrylamide Gel Electrophoresis of Free Antigen; Antigen-Anti-Body Complex and Damaged Labelled Antigen: A Study Is Performed on This Last One for the Purpose

摘要

The objective of this porject was to develope of a radioimmunoassay method accurate and precise enough to furnish a suitable way for determining human growth hormone (HGH) in extracts or in physiological fluids, useful more for specific research purposes than for routine clinical assays, and where the labelled products could be used as long as possible. Only one technique was found that these requirements, though in some respects more laborious than others: Polyacrylamide Gel Electrophoresis (PAGE). This was used introducing some modifications to the original method of Davis, and it was possible, using tubes 11 cm long, to separate on the same gel, the free, the antibody bound, and the damaged labelled antigen. The method, being able to detect separately and independently these three components and to give a better control on the analytically dangerous 'damaged', furnished accurate and reproducible curves. An example of determination is the one on KABI-Crescormon, that compares the results obatined with the present technique to those presented by another laboratory. Thanks to this method the labelled antigen could be used up to one month of time. After this period a re-purification on Sephadex was introduced so that the same labelled product was profitable for two more months. Parallel to this work a study has been performed on the various components originating in this so called process of 'damaging', and a particular importance has also been given to a more precise knowledge of the amount of antigen, in terms of mass, present in an assay. (Atomindex citation 09:386216)