An ionic liquid-assisted sample preparation method for sensitive integral-membrane proteome analysis

Abstract

Abstract Many ion channels and receptorproteins are potential targets for new drugs. However, standard methods for profiling these integral membrane proteins (IMPs) have not been fully established, especially when applied to rare and quantity-limited biological samples. In this study, we present a combined ion liquid (IL)-assisted proteomic sample preparation platform (termed pTRUST) which is compatible with starting materials in the submicrogram range, using our recently developed IL-based sample preparation strategy (iBOPs) and an in-StageTip technique. This novel and straightforward approach allows for the rapid solubilization and processing of a variety of IMPs from human samplesto support highly sensitive mass spectrometry analysis. We also demonstrated that the performance of this technology surpasses that of conventional methods such as filter-aided sample preparation methods, FASP and i-FASP. The convenience and availability of pTRUST technology have great potential for proteomic identification and characterization of novel drug targets and novel disease biology in research and clinical settings.