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Updated: Mar 22, 2026

Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity
Published on: March 31, 2021
Measurement of osmotic second virial coefficients by zonal size-exclusion chromatography
1School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland 4072, Australia.
Abstract:
Numerical simulation of protein migration reflecting linear concentration dependence of the partition isotherm has been used to invalidate a published procedure for measuring osmotic second virial coefficients (B22) by zonal exclusion chromatography. Failure of the zonal procedure to emulate its frontal chromatographic counterpart reflects ambiguity about the solute concentration that should be used to replace the applied concentration in the rigorous quantitative expression for frontal migration; the recommended use of the peak concentration in the eluted zone is incorrect on theoretical grounds. Furthermore, the claim for its validation on empirical grounds has been traced to the use of inappropriate B22 magnitudes as the standards against which the experimentally derived values were being tested.
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