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The development of an antibody affinity enrichment and mass spectrometry-based assay (iMALDI) for the characterization of EGFR and EGFR isoforms from human brain cancer tissue

EGFR (Epidermal Growth Factor Receptor) is a protein that is ubiquitous in the human
body. Aberrant activity of EGFR or its isoforms is implicated in a number of cancers,
notably brain cancer. An isoform of EGFR, EGFRvIII (EGFR variant III), is particularly
relevant to brain cancer since it is only naturally found in brain tumour tissue. However,
the presence and activity of EGFRvIII is not well characterized. I hypothesize that the
different levels of EGFRvIII expression and its expression relative to wild type EGFR in
human brain tumour tissue can be used to diagnose the different stages and progression of
disease in the glioblastoma multiforme (GM) type of brain cancer.
The work presented in this thesis is an attempt to develop a method for the accurate and
absolute quantitation of EGFRvIII from brain tumour tissue. Using iMALDI
(immunoMALDI), which combines the high-specificity of MALDI mass spectrometry
with antibody immunoaffinity enrichment, I have optimized and developed a highthroughput
technique for sensitive, specific and quantitative detection and differentiation
of EGFR and EGFRvIII. I have also demonstrated a proof-of-concept by applying this
assay to the isolation and detection of these proteins from brain tumour tissue. / Graduate

Identiferoai:union.ndltd.org:uvic.ca/oai:dspace.library.uvic.ca:1828/3278
Date02 May 2011
CreatorsShah, Brinda
ContributorsBorchers, Christoph H.
Source SetsUniversity of Victoria
LanguageEnglish, English
Detected LanguageEnglish
TypeThesis
RightsAvailable to the World Wide Web

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