Poster Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

SiTrap: rapid and high throughput clinical multiomics (#915)

John P Wilson 1 , Alexandre Zougman 2 , Darryl J Pappin 3
  1. ProtiFi, LLC, Farmingdale, NEW YORK, United States
  2. University of Leeds, Leeds, UK
  3. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States

Sample processing in omics analyses is obligatory and has traditionally been the largest source of variability in proteomics and metabolomics data. Resultantly, many biomarker “discoveries” are subsequently traced to batch or run order effects. Such errors unfortunately continue to the present day, thus compelling for clinical deployment of omics technologies first serious improvements in reproducibility especially in sample processing. Second, for widespread use, sample throughput should be as high as possible, necessitating extension to an automated platform. Moreover, to enable eventual point of care (POC) deployment of omics techniques, sample processing time should be as rapid as possible. Third, as we do not necessarily have a priori knowledge of which biomolecule will prove to be useful as diagnostic, prognostic, therapeutic or predictive signals for a given biological condition, sample processing should ideally produce from one sample different fractions for multiple omics interrogations. Fourth, such a solution should be affordable enough to be accessible to the majority of labs.

Here, we present SiTrap, a new high-throughput solution to address all necessities for clinical implementation of omics analyses. A new development over the original S-Trap[1-4] sample processing platform, SiTrap yields from the same sample both metabolomic and proteomic fractions with extraordinary inter- and intrarun reproducibility over time. It requires only minutes of processing time per sample and can be executed either in loose spin columns or in an automated 96-well format (4 - 96 samples simultaneously). Importantly, automation and its commensurate high reproducibility can be achieved on an inexpensive Tecan A200 positive pressure workstation, a general automation platform accessible – due to its low cost – to the majority of research and clinical labs. We anticipate that SiTrap will become an essential omics tool in laboratory and clinical settings and will enable novel discoveries, thereby helping to usher in a new era of clinical proteomics.

  1. Zougman A, Selby PJ, Banks RE. Suspension trapping (STrap) sample preparation method for bottom‐up proteomics analysis. Proteomics. 2014 May;14(9):1006-0.
  2. Ludwig KR, Schroll MM, Hummon AB. Comparison of in-solution, FASP, and S-Trap based digestion methods for bottom-up proteomic studies. Journal of proteome research. 2018 May 13;17(7):2480-90.
  3. HaileMariam M, Eguez RV, Singh H, Bekele S, Ameni G, Pieper R, Yu Y. S-Trap, an Ultrafast Sample-Preparation Approach for Shotgun Proteomics. Journal of proteome research. 2018 Aug 16;17(9):2917-24.
  4. Elinger D, Gabashvili A, Levin Y. Suspension trapping (S-trap) is compatible with typical protein extraction buffers and detergents for bottom up proteomics. Journal of proteome research. 2019 Mar 1.