Quantitative antibody-free LC-MS/MS analysis of sTRAIL in sputum and saliva at the sub-ng/mL level

Daniel Wilffert, Riccardo Donzelli, Angela Asselman, Jos Hermans, Natalia Govorukhina, Nick H. T. ten Hacken, Wim J. Quax, Nico Merbel, van de, Rainer Bischoff*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

8 Citations (Scopus)

Abstract

Soluble tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL) induces apoptosis via the extrinsic death receptor pathway and maybe a biomarker in the pathogenesis of a broad range of diseases. To investigate the role of sTRAIL in asthma, we developed a quantitative LC-MS/MS method with a lower limit of quantitation (LLOQ) of approximate to 3 pM in induced sputum (174 pg/mL) and saliva (198 pg/mL) without the use of antibodies. sTRAIL was enriched by immobilized metal affinity chromatography (IMAC) solid phase extraction (SPE) followed by tryptic digestion and subsequent enrichment of a signature peptide by strong cation exchange (SCX) SPE. The method was validated with respect to stability, accuracy and precision using the standard addition approach and fully metabolically N-15-labelled hrTRAIL as internal standard. Our results indicate that it is possible to quantify cytokines like sTRAIL at the pM level by LC-MS/MS without the use of antibodies, which has, to our knowledge, never been shown before. (C) 2016 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)205-210
Number of pages6
JournalJournal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
Volume1032
DOIs
Publication statusPublished - 1-Oct-2016

Keywords

  • Soluble tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL)
  • Saliva
  • Induced sputum
  • Antibody-free LC-MS/MS
  • Immobilized metal affinity (IMAC) enrichment
  • N-15-Metabolically labeled internal standard
  • MULTIPLE-SCLEROSIS
  • BIOLOGICAL SAMPLES
  • MASS-SPECTROMETRY
  • TRAIL
  • APOPTOSIS
  • QUANTIFICATION
  • EXPRESSION
  • CANCER
  • APO2L/TRAIL
  • PROTEOMICS

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