title: Modeling protein binding and elution over a chromatographic surface probed by surface plasmon resonance.
abstract: 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 a typical 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 DEAE 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.
authors.affiliation: IBET, Apartado 12, P-2781-901 Oeiras, Portugal.
authors.forename: Tiago
authors.initials: T
authors.lastname: Vicente
authors.forename: José P B
authors.initials: JP
authors.lastname: Mota
authors.forename: Cristina
authors.initials: C
authors.lastname: Peixoto
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.forename: Manuel J T
authors.initials: MJ
authors.lastname: Carrondo
mesh_terms: D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange; D007700:Kinetics; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D012710:Serum Albumin, Bovine