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제112회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Micro-scale Depletion of High Abundance Proteins and Ultra-high Pressure Enzymatic Digestion for High-throughput Analyses of Human Bio-fluids

2013년 8월 29일 16시 55분 08초
BIO1-2 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
목 09시 : 30분
생명화학 - Mass Spectrometry for Biological Applications
저자 및
한국표준과학연구원(KRISS) 유기분석표준센터, Korea
Reduction of protein complexity and follow-up sample digestion are necessary for protein identification by mass spectrometry in bottom-up proteomics studies. Bio-fluids such as plasma and cerebrospinal fluid (CSF) have been considered to be good sources for finding biomarker candidates due to the close relations to human diseases. This presentation describes a methodology in terms of depletion of high abundance proteins and high-throughput protein digestion using human bio-fluids. A vast dynamic range of protein concentration in plasma is required to be depleted for high abundance proteins which inhibit the detection of low-abundance proteins by LC-MS/MS. Immunoaffinity is one of the most frequently used protein depletion methods, however, that requires the sacrifice of high expenses for the purchase of depletion column and sometimes, undesired high sample loading as well. As an alternative to those problems, micro-scale depletion column (volume, 346 μL) was prepared using Seppro IgY 14 resin and then the feasibility was demonstrated using human plasma. Further study was carried out with CSF as a limited bio-fluid sample which can be more appropriately applicable to the micro-scale immunoaffinity column. In another study, ultra-high pressure enzymatic digester (UPED, XStreamTM) was developed and demonstrated the effectiveness in respect to digestion time and protein identifications. Moreover, this UPED system was integrated with a custom nano-liquid chromatography system for direct LC-MS/MS analysis upon digestion that can span the automation at a time from protein digestion to LC-MS/MS analysis for high-throughput analysis of proteins.