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Diffusive Interfacial Transport:  A New Approach to Concentrated Protein Solution Studies

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Miami Valley Laboratories, The Procter and Gamble Company, Cincinnati, Ohio 45247
Cite this: Langmuir 2001, 17, 16, 4693–4696
Publication Date (Web):July 3, 2001
https://doi.org/10.1021/la010226v
Copyright © 2001 American Chemical Society

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    Abstract

    A spatial gradient in the concentration of binary protein−water systems is deliberately created by slow evaporation of a solution of solute. Such samples are scanned by optical interferometry to record the refractive index profile and by near-infrared microspectroscopy to determine the water composition along the spatial gradient. Combination of the two data sets allows the change in refractive index with concentration and its increment dn/dc to be determined beyond the usual report limited to low protein−solvent volume fractions. Further, phase behavior of protein solutions can be studied. To illustrate the method, cytochrome c is chosen; clean, bubble-free samples of extremely concentrated (ca. 80%) metastable solutions can be produced in this way, and in time, protein crystals separate from these solutions.

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    Cited By

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