Non-linear elution effects in split-peak chromatography. II. Role of ligand heterogeneity in solute binding to columns with adsorption-limited kinetics

Journal of Chromatography. a
J G Rollag, David S Hage

Abstract

The split-peak effect is a useful phenomenon in studying the kinetic behavior of chromatographic supports. This work examined the combined role of ligand heterogeneity and non-linear elution conditions (i.e., sample load dependence) on the solute free fractions that are measured during split-peak studies. Exact expressions were derived to describe the effects of ligand heterogeneity under linear elution conditions, and simulation models were developed to specifically examine the combined effects of ligand heterogeneity and non-linear elution in systems with adsorption-limited rates for solute binding. The simulations showed that ligand heterogeneity increased the amount of free solute seen at any flow-rate or sample size, with this being most noticeable when using low flow-rates or large samples. One application in which these increases were examined in detail concerned the use of the split-peak effect for association rate constant measurements. It was found that linear extrapolation methods developed for homogeneous systems (as a correction for non-linear elution conditions) could successfully be applied to columns containing heterogeneous ligands. Columns containing immobilized protein A and/or protein G were used as experime...Continue Reading

References

Aug 1, 1987·Analytical Biochemistry·L A Larew, R R Walters
Apr 1, 1983·Analytical Chemistry·J R SportsmanG S Wilson
May 19, 1993·Journal of Chromatography·J P McConnell, D J Anderson

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Citations

Apr 25, 2009·Journal of Separation Science·John E Schiel, David S Hage
Nov 5, 2019·Frontiers in Chemistry·Sazia IftekharDavid S Hage
Jan 19, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Erika L PfaunmillerDavid S Hage
Oct 29, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·A J OosterkampH Irth
Jun 8, 2001·Analytical Chemistry·W ClarkeD S Hage
Apr 15, 2005·Analytical Chemistry·Tao JiangDavid S Hage
Apr 3, 2018·Methods : a Companion to Methods in Enzymology·Jeanethe A AnguizolaDavid S Hage

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