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Poster Presentations
Day 4, June 25(Wed.)
Room P (Maesato East, Foyer, Ocean Wing)
- 4P-PM-26
Developing OrthoCell Matrix to Enhance Absolute Protein Quantification Performance in LC-MS/MS: Demonstration of EGFR Mutation in Lung Cancer
(1NTU, 2IOC)
oChia-Yen Wang1,2, Huan-Chi Chiu2, Li-Yu Chen2, Ching-Yi Wan2, Yu-Ju Chen1,2
MS-based targeted proteomics is the clinical gold standard but struggles with regulatory compliance and sensitivity. Here, we developed the “OrthoCell matrix" strategy, combined with PRM-MS, for absolute quantitation of low-input druggable mutation peptides. Unlike the conventional matrix effect, which typically causes ion suppression, we observed that adding small amounts of peptides as matrices enhances detection sensitivity and signal stability in LC-MS/MS by preventing non-specific adsorption and boosting signal intensity.
The term "Ortho" refers to "orthogonal," as the matrix is composed of completely different peptide sequences from the identical proteome of the sample. Its compositional similarity to the sample minimizes potential sample loss while preserving complexity without containing endogenous peptides identical to the analytes, making it an ideal surrogate matrix for absolute quantification.
We validated this strategy by quantifying eight EGFR peptides, achieving a 4.6-fold increase in signal intensity and a 7.1-fold reduction in CV%, enabling exclusive detection of the EGFR Del-19 peptide. Notably, more hydrophilic and complex matrices improved sensitivity and stability. The ease and versatility of the OrthoCell matrix make it applicable to a wide range of sample types and experimental conditions in MS-based targeted proteomics