期刊
JOURNAL OF PROTEOME RESEARCH
卷 11, 期 8, 页码 4308-4314出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr3004216
关键词
top-down proteomics; Orbitrap Elite; high-resolution mass spectrometry
资金
- NIH [GM067193, DA018310]
- Chicago Biomedical Consortium
- Searle Funds at The Chicago Community Trust
- NSF-GK12 fellowship Reach For The Stars, Northwestern University
Mass spectrometry based proteomics generally seeks to identify and fully characterize protein species with high accuracy and throughput. Recent improvements in protein separation have greatly expanded the capacity of top-down proteomics (TDP) to identify a large number of intact proteins. To date, TDP has been most tightly associated with Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. Here, we couple the improved separations to a Fourier-transform instrument based not on ICR but using the Orbitrap Elite mass analyzer. Application of this platform to H1299 human lung cancer cells resulted in the unambiguous identification of 690 unique proteins and over 2000 proteoforms identified from proteins with intact masses <50 kDa. This is an early demonstration of high throughput TDP (>500 identifications) in an Orbitrap mass spectrometer and exemplifies an accessible platform for whole protein mass spectrometry.
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