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The free hormone hypothesis postulates that only the nonbound fraction the free fraction of hormones that otherwise circulate in blood bound to their carrier proteins is able to enter cells and exert biologic effects. In this review, I will examine four hormone groups-vitamin D metabolites especially 25OHD , thyroid hormones especially thyroxine [T4] , sex steroids especially testosterone , and glucocorticoids especially cortisol -that are bound to various degrees to their respective binding proteins-vitamin D-binding protein DBP , thyroid-binding globulin TBG , sex hormone-binding globulin SHBG , and cortisol-binding globulin CBG -for which a strong case can be made that measurement of the free hormone level provides a better assessment of hormonal status than the measurement of total hormonal levels under conditions in which the binding proteins are affected in levels or affinities for the hormones to which they bind. I will discuss the rationale for this argument based on the free hormone hypothesis, discuss potential exceptions to the free hormone hypothesis, and review functions of the binding proteins that may be independent of their transport role. I will then review the complications involved with measuring the free hormone levels and the efforts to calculate those levels based on estimates of binding constants and levels of both total hormone and total binding protein. In this review, the major focus will be on DBP and free 25OHD, but the parallels and differences with the other binding proteins and hormones will be highlighted. Vitamin D and its metabolites, thyroid hormones, sex steroids, and glucocorticoids are transported in blood bound to serum proteins. The tightness of binding varies depending on the hormone and the binding protein such that the percent free varies from 0. Although the major function of the primary carrier proteins DBP, TBG, SHBG, and CBG may be to transport their respective lipophilic hormones within the aqueous media that is plasma, these proteins may have other functions independent of their transport function.
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