Buffers used to formulate proteins should not serve as substrates or inhibitors. They should exhibit little or no change in pH with temperature, show insignificant penetration through biological membranes, and have maximum buffer capacity at a pH where the protein exhibits optimal stability. In conformity with the proposition that “Nature designs the optimum molecules,” buffers should mimic the antidenaturant properties of nature exhibited by osmolytes (1–5) that are independent of the evolutionary history of the proteins (6, 7). Such properties may include preferential exclusion from the protein domain (8–11) and stabilization without changing the denaturation Gibbs energy (
Gd) (12).

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