Solid-Phase Radioimmunoassay of Steroids
There have been several procedures reported which offer the new investigator interested in developing a radioimmunoassay for steroid hormones a choice of methods for separating the free from bound steroid. Midgley et al. (1969) have described a means for accomplishing separation, based on precipitation of bound antibody-steroid with a second antibody. This separation usually requires incubation for several days. Mikhail et al. (1970) separate free from bound estradiol by polymerizing the antibody, but this adds a high-speed centrifugation step. The recently described solid-phase system for estradiol (Abraham, 1969; Abraham and Odell, 1970) would appear to offer many advantages over the above procedures, particularly the unique feature of accomplishing the equilibration and the separation of free from bound steroid in a single step. It will be the principal aim of this paper to describe in detail the several essential criteria which must be examined for each antiserum, central to its use in the development of new methods for the radioimmunoassay of Steroids. Chemical, immunologic, and endocrinologic aspects of steroid-protein conjugates have been described by Lieberman et al. (1959). Pressman and Grossberg (1968, 1970) have discussed chemical and structural concepts which are involved in antibody-antigen interactions. However, the effects of certain physicochemical factors, such as ionic strength, hydrogen ion concentration, and temperature, on potential radioimmunoassay systems for progesterone and testosterone have not been previously examined in detail and will be described below as they apply to the solid-phase assay of these hormones.
KeywordsNormal Rabbit Serum Immunologic Method Percent Serum Polystyrene Tube Free Hormone
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