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Protein diffusion coefficient measurements by laminar flow analysis: method and applications.

R R Walters, J F Graham, R M Moore

    Analytical Biochemistry
    |July 1, 1984
    PubMed
    Summary
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    This study presents a simple method to determine solute translational diffusion coefficients using band dispersion in laminar flow. This technique accurately measures diffusion for small molecules and proteins, aiding molecular weight estimation.

    Area of Science:

    • Biophysics
    • Analytical Chemistry
    • Physical Chemistry

    Background:

    • Solute band spreading occurs during laminar flow in open tubes.
    • Radial diffusion counteracts this spreading phenomenon.
    • Translational diffusion coefficients are crucial for understanding solute transport and molecular properties.

    Purpose of the Study:

    • To develop and validate a calibration-free method for determining translational diffusion coefficients.
    • To measure diffusion coefficients of small solutes and proteins using a simple apparatus.
    • To assess the utility of diffusion coefficient data for molecular weight estimation.

    Main Methods:

    • Utilizing band dispersion in laminar flow within an open tube.
    • Measuring diffusion coefficients based on band dispersion, tube length, and flow rate.

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  • Employing a simple apparatus for experimental determination.

    • Main Results:

    • The method accurately calculates translational diffusion coefficients without calibration.
    • Diffusion coefficients for small solutes and 18 proteins were measured.
    • Results showed good agreement (+/- 10%) with literature values.
    • Determination times ranged from 7-90 minutes, requiring approximately 25 micrograms of protein.

    Conclusions:

    • The described method provides a straightforward and efficient way to measure translational diffusion coefficients.
    • This technique is applicable to a range of molecules, including proteins.
    • The obtained diffusion data are valuable for estimating molecular weights.