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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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Publication details

The article was received on 05 Dec 2014, accepted on 06 Jan 2015 and first published on 07 Jan 2015


Article type: Communication
DOI: 10.1039/C4AN02232G
Author version available: Download Author version (PDF)
Citation: Analyst, 2015,140, 990-994
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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

    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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