若手研究者セッション
- 第2日 5月16日（火） 09:30～09:45 C会場（会議室1009）
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2C-O-0930（2P-15） PDF
スポンジ状高分子基材を用いたプロテオーム計測の簡便化・高感度化
Shotgun proteome analysis based on nano-liquid chromatography/tandem mass spectrometry (nanoLC/MS/MS) is an effective method for comprehensive and quantitative analysis of proteins in living organisms. The workflow includes protein extraction, enzymatic digestion, and nanoLC/MS/MS measurement of the fragment peptides. Especially due to the remarkable advancements in nanoLC/MS/MS, researchers can easily identify and quantify 4,000-5,000 proteins in just a few hours. However, pre-treatment agents have not shown much innovation in the last decade and there are still several challenges to be addressed, including reducing sample loss during desalting and improving the efficiency of enzyme reactions.
Herein, we synthesized two kinds of spongy-like hyperpermeable polymers (spongy-monolith) consisting of poly(ethylene-co-vinyl acetate) or poly(ethylene-co-glycidyl methacrylate), respectively. The former has hydrophobicity equivalent to the commonly used C18 particles and the latter can immobilize enzymes via the epoxy groups on their own surface. Spongy-monolith has the flexibility to be easily filled into any format and can be used at high flow rates without increasing the back pressure due to their large through-pores (10-100 μm). Notably, they are cost-effective polymers. We report on the application of spongy-monolith for desalting via hydrophobic interaction and high-throughput tryptic digestion of proteome samples, showing their performance in shotgun proteomics.