Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major Trauma

Antibody mediated strategies for protein biomarker detection are common, but may limit discovery. We hypothesized that the use of antibody-free proteomics is feasible for detecting protein biomarkers in plasma of patients sustaining major trauma. A subset of subjects with major trauma from a prospec...

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Format: Article
Language:English
Published: Mary Ann Liebert 2021-07-01
Series:Neurotrauma Reports
Online Access:https://www.liebertpub.com/doi/full/10.1089/NEUR.2021.0007
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spelling doaj-4b09d5b4a9f448efab9c82900b40d9402021-07-02T03:09:13ZengMary Ann LiebertNeurotrauma Reports2689-288X2021-07-0110.1089/NEUR.2021.0007Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major TraumaAntibody mediated strategies for protein biomarker detection are common, but may limit discovery. We hypothesized that the use of antibody-free proteomics is feasible for detecting protein biomarkers in plasma of patients sustaining major trauma. A subset of subjects with major trauma from a prospective observational trial were analyzed. Patients were assigned to one of four groups based on their presenting Abbreviated Injury Severity Score (AIS). Sensitive, antibody-free selective reaction monitoring (SRM) mass spectrometry (MS), with spiked-in isotopically labeled synthetic peptides, was used for targeted protein quantification of a panel of 10 prospective targets. An overall tiered sensitivity analytical approach was used for peptide detection and quantification based upon plasma immunoaffinity depletion and PRISM fractionation. Forty-four patients were included in the analysis, of which 82% were men with a mean age of 50 (?19) years. Half had isolated head injury (n?=?22), with the remaining patients experiencing multiple injuries or polytrauma (n?=?14), isolated body injury (n?=?2), or minor injury (n?=?6). Peptides from 3 proteins (vascular adhesion molecule 1 [VCAM1], intercellular adhesion molecule 1 [ICAM1], and matrix metalloproteinase 9 [MMP9]) were detected and quantified in non-depleted processed plasma. Peptides from 2 proteins (angiopoietin 2 [Ang2] and plasminogen activator inhibitor-1 [PAI1]) were detected and quantification in depleted plasma, whereas the remaining 5 of the 10 prospective targets were undetected. VCAM1 (p?=?0.02) and MMP9 (p?=?0.03) were significantly upregulated in in the major trauma groups (1?3) versus mild injury group (4), whereas the others were not. There were no differences in protein expression between patients with traumatic brain injury (TBI; groups 1 and 2) versus those without TBI (groups 3 and 4). We detected non-specific upregulation of proteins reflecting blood?brain barrier breakdown in severely injured patients, indicating label-free MS techniques are feasible and may be informative.https://www.liebertpub.com/doi/full/10.1089/NEUR.2021.0007
collection DOAJ
language English
format Article
sources DOAJ
title Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major Trauma
spellingShingle Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major Trauma
Neurotrauma Reports
title_short Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major Trauma
title_full Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major Trauma
title_fullStr Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major Trauma
title_full_unstemmed Antibody-Free Mass Spectrometry Identification of Vascular Integrity Markers in Major Trauma
title_sort antibody-free mass spectrometry identification of vascular integrity markers in major trauma
publisher Mary Ann Liebert
series Neurotrauma Reports
issn 2689-288X
publishDate 2021-07-01
description Antibody mediated strategies for protein biomarker detection are common, but may limit discovery. We hypothesized that the use of antibody-free proteomics is feasible for detecting protein biomarkers in plasma of patients sustaining major trauma. A subset of subjects with major trauma from a prospective observational trial were analyzed. Patients were assigned to one of four groups based on their presenting Abbreviated Injury Severity Score (AIS). Sensitive, antibody-free selective reaction monitoring (SRM) mass spectrometry (MS), with spiked-in isotopically labeled synthetic peptides, was used for targeted protein quantification of a panel of 10 prospective targets. An overall tiered sensitivity analytical approach was used for peptide detection and quantification based upon plasma immunoaffinity depletion and PRISM fractionation. Forty-four patients were included in the analysis, of which 82% were men with a mean age of 50 (?19) years. Half had isolated head injury (n?=?22), with the remaining patients experiencing multiple injuries or polytrauma (n?=?14), isolated body injury (n?=?2), or minor injury (n?=?6). Peptides from 3 proteins (vascular adhesion molecule 1 [VCAM1], intercellular adhesion molecule 1 [ICAM1], and matrix metalloproteinase 9 [MMP9]) were detected and quantified in non-depleted processed plasma. Peptides from 2 proteins (angiopoietin 2 [Ang2] and plasminogen activator inhibitor-1 [PAI1]) were detected and quantification in depleted plasma, whereas the remaining 5 of the 10 prospective targets were undetected. VCAM1 (p?=?0.02) and MMP9 (p?=?0.03) were significantly upregulated in in the major trauma groups (1?3) versus mild injury group (4), whereas the others were not. There were no differences in protein expression between patients with traumatic brain injury (TBI; groups 1 and 2) versus those without TBI (groups 3 and 4). We detected non-specific upregulation of proteins reflecting blood?brain barrier breakdown in severely injured patients, indicating label-free MS techniques are feasible and may be informative.
url https://www.liebertpub.com/doi/full/10.1089/NEUR.2021.0007
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