Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1217, Issue 13, Pages 2032-2041Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2010.01.068
Keywords
Surface plasmon resonance; Ion-exchange chromatography; Modeling
Funding
- European Commission [LSHB-2006-037541, LSHB-2006-018933]
- Portuguese Fundacao para a Ciencia a Tecnologia [PTDC/EQU-EQU/71645/2006, SFRH/BD/31257/2006]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/31257/2006, PTDC/EQU-EQU/71645/2006] Funding Source: FCT
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Surface plasmon resonance (SPR) spectroscopy is used as a scaled-down, analytical, pseudo-chromatography tool for analyzing protein binding and elution over an ion-exchange surface under cyclic sorption conditions. A micrometric-scale adsorption surface was produced by immobilizing atypical ion exchange ligand - diethylaminoethyl (DEAE) - onto commercially available planar gold sensor chip surfaces pre-derivatized with a self-assembled monolayer of 11-mercaptoundecanoic acid with known density. An explicit mathematical formulation is provided for the deconvolution and interpretation of the SPR sensorgrams. An adsorption rate model is proposed to describe the SPR sensorgrams for bovine serum albumin, used here as model protein, when the DEAF surface is subjected to a cyclic series of binding and elution steps. Overall, we demonstrate that the adsorption rate model is capable of quantitatively describing BSA binding and elution for protein titers from dilute conditions up to overloaded conditions and a broad range of salt concentrations. (C) 2010 Elsevier B.V. All rights reserved.
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