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 language | English |
---|---|
Pages (from-to) | 205-210 |
Number of pages | 6 |
Journal | Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences |
Volume | 1032 |
DOIs | |
Publication status | Published - 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