Oral Sessions
（Day1, Day2, Day3, Day4）
Poster Presentations
（Day1, Day2, Day3, Day4）
Luncheon Seminars
（Day1, Day2, Day3, Day4）
Poster Presentations
- Day 4, May 18（Fri.） Poster
-
4P-09 PDF
Development of Fe3+-IMAC phosphopeptide enrichment for high throughput large scale phosphoproteomics
The techniques of phosphoproteomics allow identification and quantification of several tens of thousands of protein phosphorylation sites by using large amount of proteins (e.g., milligram scale), which can give valuable insights for understanding biological phenomena at system level. In such a large scale study, sample preparation usually requires laborious work that can disturb the analysis of large number of samples, which is often required in clinical studies and so on. Therefore we developed the phosphopeptide enrichment by Fe-ion immobilized metal affinity chromatography (Fe-IMAC). In our laboratory, 0.4 mL of IMAC resin was used for 1 mg of tryptic digest. This volume can disturb downsizing experimental system in order to simplify the operation of IMAC treatment. Then we study the capacity in order to simplify the IMAC operation. As a result, we found the IMAC resin can be reduced by up to 1/8 in comparison with our laboratory protocol. These results suggest the large scale IMAC treatment can be performed in a smaller space such as 1mL size of the solid phase extraction (SPE) cartridge and 96-well format SPE plate, facilitating the throughput of phosphopepitde enrichment by Fe-IMAC.