title: Comparison of weak affinity chromatography and surface plasmon resonance in determining affinity of small molecules.
abstract: In this study, we compared affinity data from surface plasmon resonance (SPR) and weak affinity chromatography (WAC), two established techniques for determination of weak affinity (mM-μM) small molecule-protein interactions. In the current comparison, thrombin was used as target protein. In WAC the affinity constant (KD) was determined from retention times, and in SPR it was determined by Langmuir isotherm fitting of steady-state responses. Results indicate a strong correlation between the two methods (R(2)=0.995, P<0.0001).
authors.affiliation: Department of Chemistry and Biomedical Sciences, Linnaeus University, SE-391 82 Kalmar, Sweden.
authors.forename: Minh-Dao
authors.initials: MD
authors.lastname: Duong-Thi
authors.affiliation: Division of Biotechnology/IFM, Linköping University, SE-581 83 Linköping, Sweden.
authors.forename: Gunnar
authors.initials: G
authors.lastname: Bergström
authors.affiliation: Division of Biotechnology/IFM, Linköping University, SE-581 83 Linköping, Sweden.
authors.forename: Carl-Fredrik
authors.initials: CF
authors.lastname: Mandenius
authors.affiliation: Department of Chemistry and Biomedical Sciences, Linnaeus University, SE-391 82 Kalmar, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Bergström
authors.affiliation: Department of Chemistry, Medicinal Chemistry, University of Gothenburg, SE-412 96 Göteborg, Sweden.
authors.forename: Tomas
authors.initials: T
authors.lastname: Fex
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore 637551, Republic of Singapore. Electronic address: sohlson@ntu.edu.sg.
authors.forename: Sten
authors.initials: S
authors.lastname: Ohlson
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D054852:Small Molecule Libraries / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000379:methods*; D013917:Thrombin / Q000378:metabolism*
chemical_list: D054852:Small Molecule Libraries; D013917:Thrombin
keywords: Affinity determination; Fragment-based drug discovery; Surface plasmon resonance; Thrombin; Weak affinity chromatography