title: Optimization of surface plasmon resonance experiments: Case of high mobility group box 1 (HMGB1) interactions.
abstract: Surface plasmon resonance (SPR) is a powerful technique for evaluating protein-protein interactions in real time. However, inappropriately optimized experiments can often lead to problems in the interpretation of data, leading to unreliable kinetic constants and binding models. Optimization of SPR experiments involving "sticky" proteins, or proteins that tend to aggregate, represents a typical scenario where it is important to minimize errors in the data and the kinetic analysis of those data. This is the case of High Mobility Group Box 1 and the receptor of advanced glycation end products. A number of improvements in protein purification, buffer composition, immobilization conditions, and the choice of flow rate are shown to result in substantial improvements in the accurate characterization of the interactions of these proteins and the derivation of the corresponding kinetic constants.
authors: School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845, Australia. Electronic address: rain_1904@hotmail.com.
authors: Wresti L
authors: WL
authors: Anggayasti
authors: School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845, Australia.
authors: Ricardo L
authors: RL
authors: Mancera
authors: School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845, Australia.
authors: Steven
authors: S
authors: Bottomley
authors: School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845, Australia.
authors: Erik
authors: E
authors: Helmerhorst
mesh_terms: D017127:Glycation End Products, Advanced / Q000737:chemistry* / Q000302:isolation & purification; D024243:HMGB1 Protein / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D020349:Surface Plasmon Resonance
chemical_list: D017127:Glycation End Products, Advanced; D024243:HMGB1 Protein; C585491:HMGB1 protein, human
keywords: Affinity constants; Experimental design; HMGB1; Protein–protein interaction; RAGE; SPR