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Kinetics of Acetone Precipitation: Optimization of conditions to efficiently concentrate and purify protein samples in minutes with the ProTrap XG

Jessica L. Nicherson, Alan A. Doucette, Department of Chemistry, Dalhousie University

Introduction

Acetone precipitation is a widely used method of concentrating and purifying complex protein samples in top-down proteomic workflows. Our group previously established the necessity of incorporating salt to provide consistently high protein recoveries (>95%) [1]. We later developed a two-stage filtration cartridge (ProTrap XG), which automates the process of protein precipitation [2]. The current bottleneck involved with acetone precipitation relates to the lengthy (overnight) incubation at -20°C. This study examines the kinetics of protein precipitation, with emphasis on optimizing the variables leading to the most efficient recovery.