Abstract
We have developed a column-based micro-technique for rapid determinations of protein solubilities. While retaining a large crystal surface area, the column dead volume has been reduced to ≤5 μl. The technique was tested with tetragonal lysozyme (pH 4.5, 0.1M acetate, 3.0% NaCl, 5–25°C) and column volumes of ∼ 60, 300, and 900 μl. Identical solubility data were obtained, indicating that equilibration was obtained even in the smallest columns. In addition, solubility data for Br − and I − salts of lysozyme (pH 4.5, 0.1M acetate buffer, 0.5M salt concentrations) were obtained. It appears that the technique can be further miniaturized. The limit in further reducing the crystalline column volume is determined by the minimum solution sample size needed to determine the protein concentration.
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