These assays rely on the ability of physical chemical methods to predict in vivo activity. This is extremely difficult to accomplish due to the inherent complexity of a macromolecule consisting of significant secondary, tertiary and quaternary structures. It is generally the case that biological function is much more sensitive to minor changes to such secondary structures than a typical physical chemical technique. Therefore, validation of such a physical chemical technique to be a true indicator of potency requires extensive validation against reliable whole animal and/or cell culture based bioassays. Although difficult to develop, upon demonstrable relevance and indication of biological activity, they are highly desirable due to their high precision, potential for automation, cost and rapid analysis times. An example of such an assay is the LC
assay of human insulin which has been extensively validated to be a good predictor of biological activity [60, 61].
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