Construction of a reusable Gel-Eluted Liquid Fraction Entrapment Electrophoresis (GELFrEE) using 3D printing

Abstract

Gel-Eluted Liquid Fraction Entrapment (GELFrEE) is commonly utilized in proteomics research for size-based separation and fractionation of proteins. Commercial GELFrEE cartridges are wasteful due to their single-use application and, furthermore, are being discontinued for leaving researchers that utilize the technique without needed resources. We have designed a 3D-printed, reusable GELFrEE cartridge that can be utilized with existing GELFrEE instruments. This cartridge allows the user to clean and repeatedly refill the chambers with new gels eliminating unnecessary plastic waste. Additionally, we have optimized the construction of the cartridge with respect to the type of plastic as well as coating materials and sealing glues to avoid interaction with buffers, solvents, and proteins. The 3D-printed cartridge will further allow us to optimize the composition of the separating gel according to the experimental purposes. Lastly, the cartridge has been refined to successfully separate complex E. coli lysate utilizing twelve and fifteen percent acrylamide gel.