S-Trap is an ultrafast sample preparation approach for shotgun proteomics

ABSTRACT: The success of shotgun proteomic analysis depends largely on how samples are prepared. Current approaches such as gel-, solution- or filter-based, although being extensively employed in the field, are time-consuming and less effective with respect to the repetitive sample processing, recovery, and overall yield. As an alternative, suspension trapping (S-Trap) filter is commercially available very recently in the format of single or 96-well filter plate. In contrast to conventional filter aided sample preparation (FASP) approach, which utilizes a molecular weight cutoff (MWCO) membrane as the filter and requires hours of processing before digestion-ready proteins can be obtained, S-Trap employs a three-dimensional porous material as filter media, and traps particulate protein suspension with subsequent depletion of interfering substances and in-filter digestion. Due to the large (sub-micron) pore size, each centrifugation cycle of S-Trap filter only takes around 1 minute, which significantly reduces the total processing time from approximately 3 hours by FASP to less than 15 minutes, suggesting an ultrafast sample preparation approach for shotgun proteomics. Here, for the first time, we comprehensively evaluate the performance of individual S-Trap filter and 96-well filter plate in the context of global protein identification and quantitation using whole cell lysate and clinically relevant sputum samples.

INSTRUMENT(S):

ORGANISM(S): Escherichia Coli

SUBMITTER: Yanbao Yu  

LAB HEAD: Yanbao Yu

PROVIDER: PXD010237 | Pride | 2019-03-28

REPOSITORIES: Pride