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The impact of ultraviolet- and infrared-based laser microdissection technology on phosphoprotein detection in the laser microdissection-reverse phase protein array workflow

Authors
  • Hunt, Allison L.1, 2
  • Pierobon, Mariaelena3
  • Baldelli, Elisa3
  • Oliver, Julie2, 4
  • Mitchell, Dave2, 4
  • Gist, Glenn2, 4
  • Bateman, Nicholas W.2, 4
  • Larry Maxwell, G.1, 2
  • Petricoin, Emanuel F.3
  • Conrads, Thomas P.1, 2,
  • 1 Inova Health System, 3300 Gallows Rd., Falls Church, VA, 22042, USA , Falls Church (United States)
  • 2 Uniformed Services University and Walter Reed National Military Medical Center, 8901 Wisconsin Avenue, Bethesda, MD, 20889, USA , Bethesda (United States)
  • 3 George Mason University, Manassas, VA, USA , Manassas (United States)
  • 4 The Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., 720A Rockledge Drive, Suite 100, Bethesda, MD, 20817, USA , Bethesda (United States)
Type
Published Article
Journal
Clinical Proteomics
Publisher
BioMed Central
Publication Date
Mar 09, 2020
Volume
17
Issue
1
Identifiers
DOI: 10.1186/s12014-020-09272-z
Source
Springer Nature
Keywords
License
Green

Abstract

Reversible protein phosphorylation represents a key mechanism by which signals are transduced in eukaryotic cells. Dysregulated phosphorylation is also a hallmark of carcinogenesis and represents key drug targets in the precision medicine space. Thus, methods that preserve phosphoprotein integrity in the context of clinical tissue analyses are crucially important in cancer research. Here we investigated the impact of UV laser microdissection (UV LMD) and IR laser capture microdissection (IR LCM) on phosphoprotein abundance of key cancer signaling protein targets assessed by reverse-phase protein microarray (RPPA). Tumor epithelial cells from consecutive thin sections obtained from four high-grade serous ovarian cancers were harvested using either UV LMD or IR LCM methods. Phosphoprotein abundances for ten phosphoproteins that represent important drug targets were assessed by RPPA and revealed no significant differences in phosphoprotein integrity from those obtained using higher-energy UV versus the lower-energy IR laser methods.

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