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A qualitative and quantitative proteomic study of human microdialysate and the cutaneous response to injury

A qualitative and quantitative proteomic study of human microdialysate and the cutaneous response to injury
A qualitative and quantitative proteomic study of human microdialysate and the cutaneous response to injury
The extracellular fluid space is the site of intercellular communication and represents an important source of mediators that can shed light on the parenchymal environment. Sampling of this compartment using continuous microdialysis allows assessment of the temporal changes in extracellular mediators involved in tissue homeostasis and disease processes. However, novel biomarker identification is limited by the current need to utilize specific, targeted molecular assays. The aim of our study was to explore the use of qualitative and quantitative proteomic approaches to define the protein content of dermal dialysate. Timed dermal dialysate samples were collected from healthy human volunteers for 5 h following probe insertion, using a 3,000-kDa MWCO membrane perfused at a rate of 3 ?l/min. Dialysate proteins were identified using GeLC–MS/MS and iTRAQ approaches and functions assigned according to the Gene Ontology classification system. More than 80 proteins (size range 11–516 kDa) originating from both extracellular and intracellular fluid space were identified using the qualitative approach of GeLC–MS/MS. Quantitative iTRAQ data were obtained for 27 proteins with relative change ratios between consecutive timed samples showing changes of >1.5-fold. Interstitial proteins can be identified and measured using shotgun proteomic techniques and changes detected during the acute inflammatory response. Our findings provide a platform from which to explore novel protein biomarkers and their modulation in health and disease.
1550-7416
309-317
Gill, Carolyn Anne
33a26d0c-2da6-4823-a356-30a8d9fe35a0
Parkinson, Erika
b7294dcc-43d3-46c4-bd19-7f6795b80fe6
Church, Martin K.
dad189d5-866e-4ae1-b005-0d87f74282b8
Skipp, Paul
1ba7dcf6-9fe7-4b5c-a9d0-e32ed7f42aa5
Scott, D.A.
7d38ada8-f0cf-4f6f-ae35-51e7e4a82a69
White, A.
1bc91e01-fd81-4c14-aaff-dcb3c9a009d3
O'Connor, D.
c4910888-1df5-4be7-8142-af0e919f9100
Clough, G.F.
9f19639e-a929-4976-ac35-259f9011c494
Gill, Carolyn Anne
33a26d0c-2da6-4823-a356-30a8d9fe35a0
Parkinson, Erika
b7294dcc-43d3-46c4-bd19-7f6795b80fe6
Church, Martin K.
dad189d5-866e-4ae1-b005-0d87f74282b8
Skipp, Paul
1ba7dcf6-9fe7-4b5c-a9d0-e32ed7f42aa5
Scott, D.A.
7d38ada8-f0cf-4f6f-ae35-51e7e4a82a69
White, A.
1bc91e01-fd81-4c14-aaff-dcb3c9a009d3
O'Connor, D.
c4910888-1df5-4be7-8142-af0e919f9100
Clough, G.F.
9f19639e-a929-4976-ac35-259f9011c494

Gill, Carolyn Anne, Parkinson, Erika, Church, Martin K., Skipp, Paul, Scott, D.A., White, A., O'Connor, D. and Clough, G.F. (2011) A qualitative and quantitative proteomic study of human microdialysate and the cutaneous response to injury. The AAPS Journal, 13 (2), 309-317. (doi:10.1208/s12248-011-9269-6).

Record type: Article

Abstract

The extracellular fluid space is the site of intercellular communication and represents an important source of mediators that can shed light on the parenchymal environment. Sampling of this compartment using continuous microdialysis allows assessment of the temporal changes in extracellular mediators involved in tissue homeostasis and disease processes. However, novel biomarker identification is limited by the current need to utilize specific, targeted molecular assays. The aim of our study was to explore the use of qualitative and quantitative proteomic approaches to define the protein content of dermal dialysate. Timed dermal dialysate samples were collected from healthy human volunteers for 5 h following probe insertion, using a 3,000-kDa MWCO membrane perfused at a rate of 3 ?l/min. Dialysate proteins were identified using GeLC–MS/MS and iTRAQ approaches and functions assigned according to the Gene Ontology classification system. More than 80 proteins (size range 11–516 kDa) originating from both extracellular and intracellular fluid space were identified using the qualitative approach of GeLC–MS/MS. Quantitative iTRAQ data were obtained for 27 proteins with relative change ratios between consecutive timed samples showing changes of >1.5-fold. Interstitial proteins can be identified and measured using shotgun proteomic techniques and changes detected during the acute inflammatory response. Our findings provide a platform from which to explore novel protein biomarkers and their modulation in health and disease.

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Published date: 13 June 2011
Organisations: Faculty of Medicine

Identifiers

Local EPrints ID: 337395
URI: http://eprints.soton.ac.uk/id/eprint/337395
ISSN: 1550-7416
PURE UUID: 51b21b82-3b1d-44e3-8b45-3ba1ca036053
ORCID for Paul Skipp: ORCID iD orcid.org/0000-0002-2995-2959
ORCID for G.F. Clough: ORCID iD orcid.org/0000-0002-6226-8964

Catalogue record

Date deposited: 25 Apr 2012 11:39
Last modified: 05 Nov 2019 02:06

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Contributors

Author: Carolyn Anne Gill
Author: Erika Parkinson
Author: Martin K. Church
Author: Paul Skipp ORCID iD
Author: D.A. Scott
Author: A. White
Author: D. O'Connor
Author: G.F. Clough ORCID iD

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