2012 Fiscal Year Final Research Report
Development of molecular diagnosis system of cancer drug resistance using absolute quantitative proteomics
| Project/Area Number |
23701084
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Tumor diagnosis
|
|---|
| Research Institution | Ehime University |
|---|
Principal Investigator |
TAKEMORI Nobuaki 愛媛大学, プロテオ医学研究センター, 講師 (40533047)
|
|---|
| Project Period (FY) |
2011 – 2012
|
| Keywords | 薬剤耐性 / 絶対定量プロテオミクス / 定量MRM法 / 小麦無細胞タンパク質合成系 |
|---|
| Research Abstract |
We have established a molecular diagnosis system for quantifying proteomic alterations of cancer drug resistance using wheat germ cell-free protein synthesis system and quantitative mass spectrometry. Stable isotope (SI) labeled recombinant proteins as an internal standard for protein quantification were in vitro synthesized using wheat cell-free synthesis system. We selected a set of digested peptides for each protein based on their ionization efficiency and established multiple reaction monitoring (MRM) assay using triple quadrupole mass spectrometer. We are currently constructing an absolute quantitative platform of human biopsy samples by combining MRM assay and synthesized SI-labeled protein library, which allows an accurate measurement of the absolute amount of targeted proteins from a limited amount of tissue samples.
|
