Measurement of osmotic second virial coefficients by zonal size-exclusion chromatography

作者：Winzor Donald J^*

来源：Analytical Biochemistry, 2016, 504: 59-63.

DOI：10.1016/j.ab.2016.04.003

• Thermodynamic nonideality
• Second virial coefficient
• Size-exclusion chromatography
• Bovine serum albumin
• Lysozyme

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 co-efficients (B-22) 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 B-22 magnitudes as the standards against which the experimentally derived values were being tested.

• 出版日期2016-7-1