Effective and reliable protease digestion of biological samples is critical to the success in bottom-up proteomics analysis. Various filter-based approaches using different types of membranes have been developed in the past several years and largely implemented in sample preparations for modern proteomics. However, these approaches rely heavily on commercial filter products, which are not only costly but also limited in membrane options. Here, we present a plug-and-play device for filter assembly and protease digestion. The device can accommodate a variety of membrane types, can be packed in-house with minimal difficulty, and is extremely cost-effective and reliable. Our protocol offers a versatile platform for general proteome analyses and clinical mass spectrometry.
Keywords: Filter-aided sample preparation (FASP); Glass fiber; Membrane; Polyethersulfone (PES); Polyvinylidene fluoride (PVDF); Saliva; Shotgun proteomics; Suspension trapping (STrap); Tissue; Urine.