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

Allison L. Hunt, Mariaelena Pierobon, Elisa Baldelli, Julie Oliver, Dave Mitchell, Glenn Gist, Nicholas W. Bateman, G. Larry Maxwell, Emanuel F. Petricoin, Thomas P. Conrads*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

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.

Original languageEnglish
Article number9
JournalClinical Proteomics
Volume17
Issue number1
DOIs
StatePublished - 9 Mar 2020
Externally publishedYes

Keywords

  • Laser capture microdissection
  • Laser microdissection
  • Phosphoprotein
  • Proteomics
  • Reverse phase protein array

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