Abstract
A novel method for studying the impurity distribution coefficient in protein crystals was proposed. We observed the impurity distribution in a crystal by a fluorescence method. Conventional electrophoresis can only provide the average effective distribution coefficient (K e f f ) of the whole crystal, but the fluorescence method can reveal the impurity incorporation process during crystal growth. As a model experiment, we examined the incorporation of lysozyme dimers into tetragonal lysozyme crystals at various NaCl concentrations. Using the electrophoresis method, we found that the K e f f value was less than 1 at low NaCl concentrations and exceeded 1 at high NaCl concentrations. The fluorescence method clearly indicated increases or decreases in the impurity distribution, depending on the NaCl concentrations in the solution. The characteristics of the impurity distribution obtained by the two methods were the same.
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