S-Trap, an Ultrafast Sample-Preparation Approach for Shotgun Proteomics

Milkessa Hailemariam, Rodrigo Vargas Eguez, Harinder Singh, Shiferaw Bekele, Gobena Ameni, Rembert Pieper, Yanbao Yu

Research output: Contribution to journalArticlepeer-review

106 Citations (Scopus)

Abstract

The success of shotgun proteomic analysis depends largely on how samples are prepared. Current approaches (such as those that are 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, the suspension trapping (S-Trap) filter has been commercially available very recently in the format of a single or 96-well filter plate. In contrast to the conventional filter-aided sample preparation (FASP) approach, which utilizes a molecular weight cut-off (MWCO) membrane as the filter and requires hours of processing before digestion-ready proteins can be obtained, the S-Trap employs a three-dimensional porous material as filter media and traps particulate protein suspensions with the subsequent depletion of interfering substances and in-filter digestion. Due to the large (submicron) pore size, each centrifugation cycle of the S-Trap filter only takes 1 min, which significantly reduces the total processing time from approximately 3 h by FASP to less than 15 min, suggesting an ultrafast sample-preparation approach for shotgun proteomics. Here, we comprehensively evaluate the performance of the 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.

Original languageEnglish
Pages (from-to)2917-2924
Number of pages8
JournalJournal of Proteome Research
Volume17
Issue number9
DOIs
Publication statusPublished - Sep 7 2018
Externally publishedYes

Keywords

  • FASP
  • Klebsiella pneumoniae
  • Proteomics
  • Sample preparation
  • Sputum
  • Suspension trapping
  • Tuberculosis

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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