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Development of Ion Mobility Mass Spectrometry Deconvolution Strategies for Use in Data-Independent Acquisition Proteomics
Başlık:
Development of Ion Mobility Mass Spectrometry Deconvolution Strategies for Use in Data-Independent Acquisition Proteomics
Yazar:
Pettit, Michael Edmund, author.
ISBN:
9780438017467
Yazar Ek Girişi:
Fiziksel Tanımlama:
1 electronic resource (234 pages)
Genel Not:
Source: Dissertation Abstracts International, Volume: 79-10(E), Section: B.
Advisors: Touradj Solouki Committee members: C. Kevin Chambliss; Elyssia S. Gallagher; Christopher Kearney; Sung-Joon Kim.
Özet:
Modern mass spectrometers are among the most sensitive and suitable analytical tools for high-throughput protein characterization. However, limitations in ion mobility (IM) and mass spectrometry (MS) resolving powers can limit the success of MS-based proteomics approaches. This work highlights the development of IM-MS deconvolution techniques and their utilization with commercial data processing software to improve results from IM-enhanced data-independent acquisition (HDMSE) proteomics strategies.
Chemometric deconvolution and post-IM/collision-induced dissociation (post-IM/CID) MS were used to extract "pure component" IM profiles and construct CID mass spectra of IM-MS unresolved species. Collision-energy resolved and broadband IM-MS deconvolution strategies were developed and used for successful characterization of simulated complex mixtures and biological samples, respectively. As described in Chapter Two, utilizing m/z-isolation in a novel collision-energy resolved approach, a multicomponent IM unresolved peak was deconvoluted to construct fully resolved individual IM peaks.
A broadband data acquisition approach (i.e., without the use of m/z-isolation) for improved IM-MS deconvolution is presented in Chapter Three where it is demonstrated that broadband IM-MS deconvolution provides time-saving and sensitivity advantages over the scan mode approach. Elimination of the ion isolation step also expands the applicability of IM-MS deconvolution to HDMSE acquisition. Ultra-performance liquid chromatography (UPLC) was used with HDMSE for high-throughput bottom-up proteomics analysis of rat brain tissue. The combined use of broadband IM-MS deconvolution and commercial HDMSE data processing software allowed more efficient precursor and product ion detections and improved proteins' sequence coverages.
The use of data-dependent acquisition (DDA) and HDMSE for high-throughput MS-based investigations of rat brain tissue is presented in Chapter Four. Infrared laser ablation microsampling was used to extract spatially localized rat brain samples for subsequent bottom-up proteomics analysis using both DDA and HDMSE strategies. The use of HDMS E acquisition for bottom-up proteomics of laser ablated rat brain samples provided greater numbers of peptides and proteins detected than DDA. Finally, in Chapter Five, potential future research directions are presented and discussed.
Notlar:
School code: 0014
Konu Başlığı:
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Yer Numarası | Demirbaş Numarası | Shelf Location | Lokasyon / Statüsü / İade Tarihi |
---|---|---|---|
XX(679076.1) | 679076-1001 | Proquest E-Tez Koleksiyonu | Arıyor... |
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