Pre-equilibration kinetic size-exclusion chromatography with mass spectrometry detection (peKSEC-MS) for label-free solution-based kinetic analysis of protein–small molecule interactions. Issue 4 (22nd January 2015)

• Record Type: Journal Article
• Authors: Bao, Jiayin; Krylova, Svetlana M.; Cherney, Leonid T.; Le Blanc, J. C. Yves; Pribil, Patrick; Johnson, Philip E.; Wilson, Derek J.; Krylov, Sergey N.
• Is Part Of: Analyst. Volume 140:Issue 4(2015)
• Page Start: 990
• Page End: 994
• Publication Date: 2015-01-22
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/c4an02232g
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : Label-free solution-based kinetic analysis of reversible binding between protein and small molecule. 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.