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




Shah, Brinda

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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.



brain cancer, tumors