Analytical characterization of chiral drug-protein interactions: comparison between the optical biosensor (surface plasmon resonance) assay and the HPLC perturbation method

Analytical Chemistry
Robert ArnellTorgny Fornstedt

Abstract

Two modern, fundamentally different methods were used for a detailed investigation of enantioselective drug-protein interactions, a surface plasmon resonance (SPR)-based Biacore 2000 biosensor assay and the previously validated HPLC perturbation method (HPLC-PM). This is the first time SPR has been used for this purpose. The fundamental features of the two methods were investigated, and the consequences for operation and data evaluation were addressed. With HPLC-PM, chiral data could be obtained directly from the racemic mixture, whereas a separate analysis of each pure enantiomer was required to obtain chiral data with SPR. It was shown that if chirality is not attributed in the SPR analysis, misleading average racemic binding constants will be obtained. Both drug and protein consumption were considerably higher with HPLC-PM. HPLC-PM was found to be best suited for measurements of weak affinity interactions, whereas the SPR method was best for strong interactions. With both methods, the presence of DMSO in the samples severely affected the interactions, introducing errors. The binding of the beta-blockers alprenolol and propranolol to Cel7a cellulase was used as a model system. These methods gave results that agreed quite well...Continue Reading

References

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Citations

Aug 10, 2013·Journal of Chromatography. a·Víctor Agmo HernándezTorgny Fornstedt
Jan 1, 2010·Sensors·Katsuhiko ArigaJonathan P Hill
Dec 13, 2007·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Apr 4, 2008·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Bo ZhouYi Liu
May 23, 2009·Molecular BioSystems·Jianjun SuiWei Ning Chen
Mar 4, 2020·Journal of Separation Science·Laurence MuhrLaetitia Canabady-Rochelle
Apr 8, 2021·Analytical Chemistry·Yingqian WangHaibing Li
Jul 24, 2012·Analytical Chemistry·Nicola MarchettiAlberto Cavazzini
Jun 7, 2015·Analytical Chemistry·Martina CataniAlberto Cavazzini

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