Proteomic Analyses using an Accurate Mass and Time Tag Strategy
An accurate mass and time (AMT) tag approach for proteomic analyses has been developed over the last several years to facilitate comprehensive high throughput proteomic measurements. An AMT tag database for an organism, tissue or cell line is established by initially performing standard shotgun proteomic alalysis and, most importantly, by validating peptide identifications using the mass measurement accuracy of Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). Creation of an AMT tag database largely obviates the need for subseqent MS/MS analyses, and thus facilitates high throughput analyses. The strength of this technology resides in the ability to achieve highly efficient and reproducible 1D reversed-phased LC separations in conjunction with highly accurate mass measurements using FTICR MS. Recent improvements allow analysis of as little as picrogram amounts of proteome samples by minimizing sample handling and maximizing peptide recovery. Nanoproteomics platform has also demonstrated the ability to detect over 10 9 differences in protein abundance in human plasma and identify more abundant proteins from sub-picogram amounts of samples. The AMT tag approach is poised to become a new standard technique for the in-depth and high throughput analysis of complex organisms, clinical samples, with potential to extend the analysis to single mammalian cell.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 15010720
- Report Number(s):
- PNNL-SA-42437; BTNQDO; 400412000; TRN: US200501%%187
- Journal Information:
- BioTechniques, 37(4):621-636, Vol. 37, Issue 4; ISSN 0736-6205
- Country of Publication:
- United States
- Language:
- English
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