Reproducible Quantification of Cancer-Associated Proteins in Body Fluids Using Targeted Proteomics

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Abstract

The rigorous testing of hypotheses on suitable sample cohorts is a major limitation in translational research. This is particularly the case for the validation of protein biomarkers; the lack of accurate, reproducible, and sensitive assays for most proteins has precluded the systematic assessment of hundreds of potential marker proteins described in the literature. Here, we describe a high-throughput method for the development and refinement of selected reaction monitoring (SRM) assays for human proteins. The method was applied to generate such assays for more than 1000 cancer-associated proteins, which are functionally related to candidate cancer driver mutations. We used the assays to determine the detectability of the target proteins in two clinically relevant samples: plasma and urine. One hundred eighty-two proteins were detected in depleted plasma, spanning five orders of magnitude in abundance and reaching below a concentration of 10 ng/ml. The narrower concentration range of proteins in urine allowed the detection of 408 proteins. Moreover, we demonstrate that these SRM assays allow reproducible quantification by monitoring 34 biomarker candidates across 83 patient plasma samples. Through public access to the entire assay library, researchers will be able to target their cancer-associated proteins of interest in any sample type using the detectability information in plasma and urine as a guide. The generated expandable reference map of SRM assays for cancer-associated proteins will be a valuable resource for accelerating and planning biomarker verification studies.

Detaljer

Författare
  • Ruth Huttenhain
  • Martin Soste
  • Nathalie Selevsek
  • Hannes Roest
  • Atul Sethi
  • Christine Carapito
  • Terry Farrah
  • Eric W. Deutsch
  • Ulrike Kusebauch
  • Robert L. Moritz
  • Emma Niméus
  • Oliver Rinner
  • Ruedi Aebersold
Enheter & grupper
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Cancer och onkologi
Originalspråkengelska
Sidor (från-till)142ra94
TidskriftScience Translational Medicine
Volym4
Utgivningsnummer142
StatusPublished - 2012
PublikationskategoriForskning
Peer review utfördJa