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  • 09월 10일 16시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

대한화학회 제124회 학술발표회 및 총회 Efficient and comprehensive proteomic analysis by fully automated DO-NCFC-RP/RPLC

2019년 8월 29일 16시 44분 42초
ANAL.P-168 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
10월 18일 (금요일) 11:00~12:30
Analytical Chemistry
저자 및
Chaewon Kang, Dowoon Nam, Sang-Won Lee*
Department of Chemistry, Korea University, Korea
Recently, human proteome analysis from a single-cell type covering 10,000 proteins is practicable, although some methodological and technical issues remain. There are still significant challenges to identify more than 10,000 proteins, because the identification efficiency of present technology has decreased at this level of identification. Here, we report a dual online reverse-phase/reverse-phase liquid chromatography system, which utilizes an online non-contiguous fractionating and concatenating device (NCFC fractionator). In shotgun proteomics analyses of a human proteome, this system provides considerably improved exploitation of the separation space, resulting in a significant increase in the numbers of proteins identified compared to a general contiguous 2D-RP/RPLC method. Unlike the previously described offline 2D-NCFC-RP/RPLC method, the completely automated online 2D-NCFC-RP/RPLC system bypasses complex manual processes such as offline fractionations, pooling, clean-up, drying, reconstitution, and autosampler fraction injection. As a result, our novel system offers minimal sample loss and highly reproducible 2D-RP/RPLC experiments. With this system, in single shot data-dependent acquisition (DDA), we identified more than 10,000 proteins in one-day data acquisition time. In addition, we performed data independent acquisition (DIA) experiment, which has emerged as an alternative approach to data dependent acquisition (DDA) for comprehensive proteomic analyses. We will discuss the comparison of DIA and DDA data.