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Isolation and Proteomic Analysis of Platelets by SELDI-TOF MS

  • Sean R. DowningEmail author
  • Giannoula L. Klement
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 818)

Abstract

Many growth factors, leukotrines, and biological ligands are not circulating free in plasma or serum, except in the case of late or disseminated disease. During early tumor growth and angiogenesis, platelets actively and selectively sequester regulators of angiogenesis and, as such, the platelet protein content can be used as a marker of early tumor growth or angiogenesis. With the recent increase in the clinical use of biologic modifiers in cancer and chronic disease therapy, the search for markers of early disease, therapeutic response, and/or recurrence has suggested that analysis of platelet proteins may be more relevant and accurate. We provide a guideline for the proteomic analysis of platelet proteome, placing specific emphasis on angiogenesis regulators, even though other platelet proteins may serve as markers of disease in the future. The analysis of serum/plasma has been fraught with difficulties because of the extraordinarily large number of proteins and because some of the proteins are contained in extraordinarily large amounts, masking the less abundant proteins. Thus, platelets may provide a much more biologically relevant analyte for biomarker discovery.

Key words

Platelet Platelet proteomics Angiogenesis Markers of tumor growth Vascular endothelial growth factor (VEGF) Basic fibroblast growth factor (bFGF) Platelet-derived growth factor (PDGF) 

Abbreviations

Coag

Coagulation, referring to a test used to measure coagulation time of whole blood

EGF

Epidermal growth factor

Fbgn

Fibrinogen

bFGF

Basic fibroblast growth factor

PGE

Prostaglandin

PRP

Platelet rich plasma

PDGF

Platelet derived growth factor

PPP

Platelet poor plasma

SELDI-TOF MS

Surface enhanced laser desorption/ionization – time-of-flight mass spectrometry

SPA

Sinapinic acid

TFA

Trifluoroacetic acid

VEGF

Vascular endothelial growth factor

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Vascular Biology ProgramChildren’s Hospital BostonBostonUSA
  2. 2.Children’s HospitalBostonUSA

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