Poster Presentations
Day 1, June 10（Wed.） Room P (5F 501+502)
- 1P-04（1D-O1-1502）
Development of Sample Pretreatment StageTips Packed with Restricted Access Media for Simultaneous Analysis of Serum Peptidome and Proteome
(Kyoto Univ.)
oTaiki Asaba, Hiroto Kakiuchi, Eisuke Kanao, Yasushi Ishihama
Blood contains minute amounts of proteins and peptides leaked from organs and tissues, which hold potential as biomarkers. However, detecting these molecules via LC/MS/MS requires overcoming constraints imposed by the need for a concentration dynamic range of approximately 1011 with the presence of highly abundant serum proteins. Here, we have developed disposable tipLC columns for serum proteome pretreatment capable of isolating low-molecular-weight protein fractions (peptidome fractions). This utilizes restricted access media (RAM) as the tipLC stationary phase, which retains only low-molecular-weight molecules accessible to the mesopores, while high-molecular-weight proteins unable to permeate the pores are not retained. Analysis of the peptidome fraction from 1 µL of pooled human serum, without trypsin digestion, identified approximately 800 peptides. Meanwhile, analysis of the flow-through protein fraction using native trypsin digestion without denaturation, reduction, or alkylation identified 624 proteins. These results demonstrate that using RAM-StageTip for serum pretreatment enables simultaneous profiling of the serum peptidome and proteome from a single sample.