Streamlined SDS-based Workflows in the ProTrap XG for Top-down or Bottom-up Proteomics

Alan Doucette, with Jessica Nickerson, Ziheng Dang, Venus Baghalabadi, Kate-Lynn Smith, Shayna Phonchareon
Dalhousie University, Halifax, Nova Scotia

Introduction

SDS is beneficial for front-end proteome sample preparation, though detrimental to MS analysis. Solvent precipitation is a proven approach to remove SDS ahead of analysis. However, achieving optimal protein recovery following solvent precipitation can be challenging. High yields are possible, assuming the proper conditions are met, including incorporation of sufficient ionic strength [1], as well as avoiding accidental pipetting of the sample pellet. The ProTrap XG is a disposable, two-stage filtration and extraction cartridge, designed to overcome the latter issue by isolating the protein pellet on a porous PTFE membrane [2]. Within this device, a complete SDS-based workflow for top-down and bottom-up sample processing is achievable.