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

doi:10.1016/j.jchromb.2016.04.041

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 journal › Article › Academic › peer-review

9 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 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	https://doi.org/10.1016/j.jchromb.2016.04.041
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

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Quantitative antibody-free LC-MS-MS analysis of sTRAIL in sputum and
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Wilffert, D., Donzelli, R., Asselman, A., Hermans, J., Govorukhina, N., ten Hacken, N. H. T., Quax, W. J., Merbel, van de, N., & Bischoff, R. (2016). Quantitative antibody-free LC-MS/MS analysis of sTRAIL in sputum and saliva at the sub-ng/mL level. Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 1032, 205-210. https://doi.org/10.1016/j.jchromb.2016.04.041

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

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.",

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",

author = "Daniel Wilffert and Riccardo Donzelli and Angela Asselman and Jos Hermans and Natalia Govorukhina and {ten Hacken}, {Nick H. T.} and Quax, {Wim J.} and {Merbel, van de}, Nico and Rainer Bischoff",

year = "2016",

month = oct,

day = "1",

doi = "10.1016/j.jchromb.2016.04.041",

language = "English",

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Wilffert, D, Donzelli, R, Asselman, A, Hermans, J, Govorukhina, N, ten Hacken, NHT, Quax, WJ , Merbel, van de, N & Bischoff, R 2016, 'Quantitative antibody-free LC-MS/MS analysis of sTRAIL in sputum and saliva at the sub-ng/mL level', Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, vol. 1032, pp. 205-210. https://doi.org/10.1016/j.jchromb.2016.04.041

Quantitative antibody-free LC-MS/MS analysis of sTRAIL in sputum and saliva at the sub-ng/mL level. / Wilffert, Daniel; Donzelli, Riccardo; Asselman, Angela et al.
In: Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, Vol. 1032, 01.10.2016, p. 205-210.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - Quantitative antibody-free LC-MS/MS analysis of sTRAIL in sputum and saliva at the sub-ng/mL level

AU - Wilffert, Daniel

AU - Donzelli, Riccardo

AU - Asselman, Angela

AU - Hermans, Jos

AU - Govorukhina, Natalia

AU - ten Hacken, Nick H. T.

AU - Quax, Wim J.

AU - Merbel, van de, Nico

AU - Bischoff, Rainer

PY - 2016/10/1

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N2 - 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.

AB - 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.

KW - Soluble tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL)

KW - Saliva

KW - Induced sputum

KW - Antibody-free LC-MS/MS

KW - Immobilized metal affinity (IMAC) enrichment

KW - N-15-Metabolically labeled internal standard

KW - MULTIPLE-SCLEROSIS

KW - BIOLOGICAL SAMPLES

KW - MASS-SPECTROMETRY

KW - TRAIL

KW - APOPTOSIS

KW - QUANTIFICATION

KW - EXPRESSION

KW - CANCER

KW - APO2L/TRAIL

KW - PROTEOMICS

U2 - 10.1016/j.jchromb.2016.04.041

DO - 10.1016/j.jchromb.2016.04.041

M3 - Article

C2 - 27250581

SN - 1570-0232

VL - 1032

SP - 205

EP - 210

JO - Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences

ER -

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

Abstract

Keywords

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