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Development of an enhanced proteomic method to detect prognostic and diagnostic markers of healing in chronic wound fluid.
Published on Dec 7, 2007in British Journal of Dermatology 6.13
· DOI :10.1111/j.1365-2133.2007.08362.x
Melissa Laura Fernandez4
Estimated H-index: 4
(Queensland University of Technology),
James A. Broadbent7
Estimated H-index: 7
(Queensland University of Technology)
+ 2 AuthorsZee Upton30
Estimated H-index: 30
(Queensland University of Technology)
Abstract
Summary Background  Chronic venous leg ulcers are a significant cause of pain, immobility and decreased quality of life for patients with these wounds. In view of this, research efforts are focused on multiple factors in the wound environment to obtain information regarding the healing of ulcers. Objectives  Chronic wound fluid (CWF), containing a complex mixture of proteins, is an important modulator of the wound environment, and therefore we hypothesized that these proteins may be indicators of the status of wounds and their potential to heal or otherwise. To explore this we developed and validated a proteomic approach to analyse CWF. Methods  In this study, pooled CWF was depleted of high abundant proteins using immunoaffinity chromatography. The flow-through and bound fractions were collected, concentrated, desalted and analysed using a range of techniques. Each fraction was further separated using two-dimensional (2D) gel electrophoresis and 2D liquid chromatography and analysed using mass spectrometry (MS). Results  Western blot analysis against three high abundant proteins confirmed the selective removal of these proteins from CWF. Critically, one-dimensional and 2D gel electrophoresis indicated that subsequent removal of these proteins enhanced the ability to detect proteins in low abundance in CWF. Further, MS demonstrated that depletion of these abundant proteins increased the detection of other proteins in these samples. Conclusions  Results obtained indicate that this approach significantly improves separation of proteins present in low concentrations in CWF. This will facilitate the identification of biomarkers in samples collected from patients with ulcers and lead to improved patient therapies and wound care approaches.
  • References (44)
  • Cited By (34)
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References44
Published on Jan 1, 2000in Ostomy Wound Management 0.99
Staiano-Coico L1
Estimated H-index: 1
,
Higgins Pj1
Estimated H-index: 1
+ 2 AuthorsGoncalves J1
Estimated H-index: 1
Analyzing acute and chronic wound fluids provides an important and intriguing insight into the wound milieu. This review outlines some of the salient features of wound repair and the wound fluid environment. Most studies support the premise that the contents of the wound fluid reflect the status of the wound and can be indicative of whether a wound is on the course of a normal or impaired response to injury. For example, chronic wound fluids often differ from acute wound fluids in their prolifer...
23 Citations
E Krauss1
Estimated H-index: 1
,
C F Polnaszek1
Estimated H-index: 1
+ 3 AuthorsJ L Holtzman1
Estimated H-index: 1
The authors have determined the binding parameters for [14C] lidocaine to several preparations of purified human serum alpha 1-acid glycoprotein (AAG) and human serum albumin (HSA). The authors find that the average Kd for the three different purified AAG preparations was 15.2 +/- 0.5 microM, with an average of 0.567 +/- 0.009 binding sites (N) per AAG molecule, when the concentration of AAG was determined by a standard colorimetric protein assay. The number of binding sites per AAG increased to...
18 Citations
Published on Oct 1, 1996in Wound Repair and Regeneration 2.95
Bruce A. Mast11
Estimated H-index: 11
(University of Florida),
Gregory S. Schultz56
Estimated H-index: 56
(University of Florida)
A healing wound represents a complex series of interactions between cells, soluble mediators, and extracellular matrix. Within this multifaceted environment, there are multiple regulatory points which control the ordered series of events that lead to normal tissue repair. An alteration in this physiologic network can lead to the development of a chronic wound. This article presents an update on the numerous mediators that exist within the wound environment in both acute normal healing and chroni...
365 Citations Source Cite
Published on Aug 1, 2005in Proteomics 3.53
Alex J. Rai8
Estimated H-index: 8
(Johns Hopkins University School of Medicine),
Craig A. Gelfand1
Estimated H-index: 1
+ 13 AuthorsHarald Tammen16
Estimated H-index: 16
There is a substantial list of pre-analytical variables that can alter the analysis of blood-derived samples. We have undertaken studies on some of these issues including choice of sample type, stability during storage, use of protease inhibitors, and clinical standardization. As there is a wide range of sample variables and a broad spectrum of analytical techniques in the HUPO PPP effort, it is not possible to define a single list of pre-analytical standards for samples or their processing. We ...
411 Citations Source Cite
Published on Oct 1, 2005in Journal of Proteome Research 3.95
James Martosella7
Estimated H-index: 7
,
Nina Zolotarjova8
Estimated H-index: 8
+ 2 AuthorsBarry E. Boyes6
Estimated H-index: 6
Serum analysis represents an extreme challenge due to the dynamic range of the proteins of interest, and the high structural complexity of the constituent proteins. In serum, the quantities of proteins and peptides of interest range from those considered “high abundance”, present at 2−70% by mass of total protein, to those considered “low abundance”, present at 10-12 M or less. This range of analytical target molecules is outside the realm of available technologies for proteomic analysis. Theref...
105 Citations Source Cite
Published on Aug 1, 2005in Proteomics 3.53
Sang Yun Cho19
Estimated H-index: 19
(Yonsei University),
Eun Young Lee26
Estimated H-index: 26
(Yonsei University)
+ 12 AuthorsJin Young Kim19
Estimated H-index: 19
Human plasma is the most clinically valuable specimen, containing not only a dynamic concentration range of protein components, but also several groups of high-abundance proteins that seriously interfere with the detection of low-abundance potential biomarker proteins. To establish a high-throughput method for efficient depletion of high-abundance proteins and subsequent fractionation, prior to molecular analysis of proteins, we explored how coupled immunoaffinity columns, commercially available...
108 Citations Source Cite
Published on Jan 1, 2005in Proteomics 3.53
Karin Björhall2
Estimated H-index: 2
(AstraZeneca),
Tasso Miliotis12
Estimated H-index: 12
(AstraZeneca),
Pia Davidsson49
Estimated H-index: 49
(AstraZeneca)
Serum proteins may often serve as indicators of disease and is a rich source for biomarker discovery. However, the large dynamic range of proteins in serum makes the analysis very challenging because high-abundant proteins tend to mask those of lower abundance. A prefractionation step, such as depletion of a few high-abundant proteins before protein profiling, can assist in the discovery and detection of less abundant proteins that may prove to be informative biomarkers. In the present study, fi...
299 Citations Source Cite
Published on Jun 24, 1999in Acta Dermato-venereologica
Artur Schmidtchen40
Estimated H-index: 40
There is growing evidence that chronic wounds show specific temporospatial patterns with respect to the expression of various molecules such as proteinases and their inhibitors. A careful analysis of molecules in the exudate of chronic wounds can be expected to provide useful information regarding events in the underlying tissue. In this connection there is a need for a method applicable to large amounts of patient material for collecting and analysing wound fluid from leg ulcers. A method of th...
18 Citations Source Cite
S.R. Baker3
Estimated H-index: 3
(Fremantle Hospital),
Michael Stacey25
Estimated H-index: 25
(Fremantle Hospital)
The aim of this study was to determine the point prevalence of ulceration of the leg in an Australian community. This was a population-based epidemiological study. Two hundred and fifty-nine patients with leg ulceration were identified by health care professionals of all types working in hospitals and private practices, as well as by self referral from patients. The base population in the study area was 238000. The following data were recorded: age, sex, site of ulceration. duration of ulceratio...
95 Citations Source Cite
Published on Oct 1, 2004in Drug Discovery Today: Technologies
Xiangming Fang7
Estimated H-index: 7
,
Lei Huang4
Estimated H-index: 4
+ 1 AuthorsWei-Wei Zhang6
Estimated H-index: 6
A major challenge in protein target discovery and validation is how to specifically dissect complex protein mixtures and measure trace targets. Immunoaffinity-based protein capture, separation and detection have proven to be one of the most effective approaches. Avian IgY antibody microbeads (Seppro™), representing a type of novel and specific protein sorbent, have several distinct advantages over IgG. Their utility and applications are compared with those of IgG and other affinity reagents.
22 Citations Source Cite
Cited By34
Published on Jan 1, 2009
Laura E. Edsberg13
Estimated H-index: 13
(Daemen College)
Proteome level information is necessary to understand the function of specific cell types and their roles in health and disease. Proteomics is a rapidly developing field with a wide range of applications in wound healing. The ability to use proteomics to assess the wound healing process would have many benefits, including earlier evidence of healing and better understanding of how different treatments affect the wound at the protein level. The basis of what is known about the chronic wound prote...
2 Citations Source Cite
Published on Aug 1, 2015in Rechtsmedizin 0.64
M. Fattouh , C.T. Buschmann7
Estimated H-index: 7
(Charité)
+ 3 AuthorsMichael Tsokos30
Estimated H-index: 30
(Charité)
Hintergrund Die Expression leukozytarer und endothelialer Adhasionsmolekule aus der Gruppe der Selektin-, Integrin- und der Immunglobulinsuperfamilie in primar heilenden Hautwunden ist tierexperimentell sowie an humanem Operations- und Autopsiematerial untersucht worden. Fur die Dekubituswunde existieren bezuglich der Gewebeexpression leukozytarer und endothelialer Adhasionsmolekule bisher allerdings keine gesicherten Erkenntnisse.
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Markus W. Löffler8
Estimated H-index: 8
(University of Tübingen),
Heiko Schuster12
Estimated H-index: 12
(University of Tübingen)
+ 1 AuthorsStefan Beckert16
Estimated H-index: 16
(University of Tübingen)
Valid and reproducible sampling techniques as well as processing protocols are required for the assessment of biomarkers and mediators contained in wound exudate. Moreover, the ideal technique should be easy to use even in daily clinical routine. This is challenging since wound fluid represents an inhomogeneous mixture of different exogenous and endogenous sources. Analyzing wound fluid, however, may facilitate clinical decision making. Many techniques for obtaining wound fluid have been describ...
8 Citations Source Cite
Published on Aug 1, 2008in Journal of Proteomics 3.72
Xiangming Fang1
Estimated H-index: 1
,
Wei-Wei Zhang1
Estimated H-index: 1
Abstract Protein separation or enrichment is one of the rate-limiting steps in proteomic studies. Specific capture and removal of highly-abundant proteins (HAP) with large sample-handling capacities are in great demand for enabling detection and analysis of low-abundant proteins (LAP). How to grasp and enrich these specific proteins or LAP in complex protein mixtures is also an outstanding challenge for biomarker discovery and validation. In response to these needs, various approaches for remova...
114 Citations Source Cite
Published on Feb 1, 2010in Proteomics Clinical Applications 3.57
James A. Broadbent7
Estimated H-index: 7
(Queensland University of Technology),
Terry Walsh8
Estimated H-index: 8
(Queensland University of Technology),
Zee Upton30
Estimated H-index: 30
(Queensland University of Technology)
Chronic wounds, such as venous and diabetic leg ulcers, represent a significant health and financial burden to individuals and healthcare systems. In worst-case scenarios this condition may require the amputation of an affected limb, with significant impact on patient quality of life and health. Presently, there are no clinical biochemical analyses used in the diagnosis and management of this condition; moreover few biochemical therapies are accessible to patients. This presents a significant ch...
21 Citations Source Cite
Published on Apr 2, 2013in Analytical Chemistry 6.04
Kelly M. Hines6
Estimated H-index: 6
,
Samir Ashfaq1
Estimated H-index: 1
+ 3 AuthorsJohn A. McLean36
Estimated H-index: 36
Wound fluid is a complex biological sample containing byproducts associated with the wound repair process. Contemporary techniques, such as immunoblotting and enzyme immunoassays, require extensive sample manipulation and do not permit the simultaneous analysis of multiple classes of biomolecular species. Structural mass spectrometry, implemented as ion mobility-mass spectrometry (IM-MS), comprises two sequential, gas-phase dispersion techniques well suited for the study of complex biological sa...
11 Citations Source Cite
Published on Oct 1, 2011in Archives of Dermatological Research 2.15
Rikke Zillmer1
Estimated H-index: 1
(University of Copenhagen),
Hannah Trøstrup2
Estimated H-index: 2
(University of Copenhagen)
+ 2 AuthorsMagnus S. Ågren22
Estimated H-index: 22
(University of Copenhagen)
Wound fluid collected from chronic wounds may be used as a simple gauge of the processes taking place in the tissue. There is lack of information on the optimal conditions for wound fluid procurement. We have studied possible diurnal variations and duration of wound fluid accumulation using retentive hydrophobic foam on the levels of prototypic cytokines [interleukin (IL)-1α, IL-1β], a chemokine (IL-8) and proteinases [matrix metalloproteinase (MMP)-9] in 23 chronic venous leg ulcer patients. Bi...
13 Citations Source Cite
Published on Jan 1, 2010in BMC Research Notes
Lars Steinsträßer5
Estimated H-index: 5
(Ruhr University Bochum),
Frank Jacobsen20
Estimated H-index: 20
(Ruhr University Bochum)
+ 4 AuthorsDirk Wolters28
Estimated H-index: 28
(Ruhr University Bochum)
Background The process of wound healing consists of several well distinguishable and finely tuned phases. For most of these phases specific proteins have been characterized, although the underlying mechanisms of regulation are not yet fully understood. It is an open question as to whether deficits in wound healing can be traced back to chronic illnesses such as diabetes mellitus. Previous research efforts in this field focus largely on a restricted set of marker proteins due to the limitations d...
9 Citations Source Cite
Published on May 18, 2010in International Wound Journal 2.38
Jennifer T. Wyffels8
Estimated H-index: 8
(Daemen College),
Kristin M. Fries6
Estimated H-index: 6
(Daemen College)
+ 5 AuthorsLaura E. Edsberg13
Estimated H-index: 13
(Daemen College)
The incidence rate of pressure ulcers in the USA ranges from 0·4% to 38% in acute care settings and from 2·2% to 23·9% in long-term care settings, and their treatment costs are in the billions of dollars yearly. The proteome of wound fluid may contain early indicators or biomarkers associated with healing in pressure ulcers that would enable treatment regimes to be optimised for each individual. Wound fluid was collected from the interior and periphery of 19 chronic pressure ulcers at 15 time po...
13 Citations Source Cite
Published on Feb 1, 2015in Biomaterials 8.81
Mark D. Swartzlander5
Estimated H-index: 5
(University of Colorado Boulder),
Christopher A. Barnes6
Estimated H-index: 6
(ETH Zurich)
+ 3 AuthorsStephanie J. Bryant37
Estimated H-index: 37
(University of Colorado Boulder)
Abstract Poly(ethylene glycol) (PEG) hydrogels with their highly tunable properties are promising implantable materials, but as with all non-biological materials, they elicit a foreign body response (FBR). Recent studies, however, have shown that incorporating the oligopeptide RGD into PEG hydrogels reduces the FBR. To better understand the mechanisms involved and the role of RGD in mediating the FBR, PEG, PEG-RGD and PEG-RDG hydrogels were investigated. After a 28-day subcutaneous implantation ...
30 Citations Source Cite
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