A Dialysis Technique for Determining Aggregate Concentrations in Crystallizing Protein Solutions

Document Type

Article

Publication Date

8-14-1993

Description

We have adapted a dialysis technique which provides aggregate concentrations of protein molecules in solutions which lead to crystal growth. In dialysis, the flux across a semipermeable membrane is directly proportional to the concentration of the diffusible solute inside the bag provided that the solute concentration in the bulk solution is infinitely dilute. Using membranes of varying porosity, the concentrations of different size solutes can be measured by measuring the flux rate across the membrane. We have used this technique to independently measure the concentrations of monomers, dimers, trimers and higher aggregates of lysozyme in both 1% NaCI and 3% NaCI (0.1 M NaAc, pH4) using 25 K and 50 K molecular weight cut-off membranes. We compare these concentration profiles with (110) face growth rate data under the same conditions.

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