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Issue 4, 2015
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Pre-equilibration kinetic size-exclusion chromatography with mass spectrometry detection (peKSEC-MS) for label-free solution-based kinetic analysis of protein–small molecule interactions

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Abstract

Here we introduce pre-equilibration kinetic size-exclusion chromatography with mass-spectrometry detection (peKSEC-MS), which is a label-free solution-based kinetic approach for characterizing non-covalent protein–small molecule interactions. In this method, a protein and a small molecule are mixed outside the column and incubated to approach equilibrium. The equilibrium mixture is then introduced into the SEC column to initiate the dissociation process by separating small molecules from the complex inside the column. A numerical model of a 1-dimensional separation was constructed to simulate mass chromatograms of the small molecule for varying rate constants of binding.

Graphical abstract: Pre-equilibration kinetic size-exclusion chromatography with mass spectrometry detection (peKSEC-MS) for label-free solution-based kinetic analysis of protein–small molecule interactions

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Supplementary files

Article information


Submitted
05 Dec 2014
Accepted
06 Jan 2015
First published
07 Jan 2015

Analyst, 2015,140, 990-994
Article type
Communication
Author version available

Pre-equilibration kinetic size-exclusion chromatography with mass spectrometry detection (peKSEC-MS) for label-free solution-based kinetic analysis of protein–small molecule interactions

J. Bao, S. M. Krylova, L. T. Cherney, J. C. Y. Le Blanc, P. Pribil, P. E. Johnson, D. J. Wilson and S. N. Krylov, Analyst, 2015, 140, 990
DOI: 10.1039/C4AN02232G

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