Journal of Chromatography & Separation Techniques
Open Access

Kinetic chromatography: A new approach to study kinetics of reversible protein-drug binding

World Congress on Chromatography

September 21-23, 2016 Amsterdam, Netherlands

Sergey N Krylov

York University, Canada

Keynote: J Chromatogr Sep Tech

Abstract :

Studying the kinetics of reversible binding between a protein and a small-molecule drug is a major challenge. The available approaches require that either the small molecule or the protein modified by labeling or immobilization on a surface. Not only can such modifications be difficult to do but they can also drastically affect the kinetic parameters of the interaction. To solve this problem, we developed kinetic chromatography with mass-spectrometry detection, a solution-based label-free approach. Our novel approach utilizes the ability of some modes of chromatography to separate any small molecule from any protein-small-molecule complex without immobilization and the ability of mass-spectrometry to detect a small molecule without a label. The rate constants of complex formation and dissociation are convoluted in the temporal pattern of small molecule elution measured with mass spectrometry at the exit from the chromatography column. We developed advanced algorithms and software to deconvolute the unknown constants from the temporal pattern by solving an inverse mathematical problem either analytically or numerically. In this lecture, the concept of kinetic chromatography with mass-spectrometry detection will be explained and a examples of its use for studying kinetics for protein-drug interactions will be presented. Prospectives of the utilization of kinetic chromatography with mass-spectrometry detection in the pharmaceutical industry and academic research will be discussed.

Biography :

Sergey Krylov completed his PhD from Moscow State University. He has been Professor of Chemistry at York University in Toronto since 2000. He had held Canada Research Chair Tier II for a maximum term and is now York Research Chair in Bioanalytical Chemistry. He is a Founder and Director of the Centre for Research on Biomolecular Interactions. He is recognized internationally for his pioneering work in the fields of “Chemical cytometry, kinetic analysis of affinity interactions, and selection and applications of oligonucleotide aptamers”. He has authored over 160 peer-reviewed papers and his research contributions have been recognized with a number of distinctions including two awards from the Chemical Society of Canada.

Email: skrylov@yorku.ca