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


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.