Exploring ECD on a Benchtop Q Exactive Orbitrap Mass Spectrometer

Kyle L Fort; Christian N. Cramer; Valery G Voinov; Yury V Vasil'ev; Nathan I Lopez; Joseph S Beckman; Albert J R Heck

doi:10.1021/acs.jproteome.7b00622

Exploring ECD on a Benchtop Q Exactive Orbitrap Mass Spectrometer

Kyle L Fort, Christian N. Cramer, Valery G Voinov, Yury V Vasil'ev, Nathan I Lopez, Joseph S Beckman, Albert J R Heck

29 Citationer (Scopus)

46 Downloads (Pure)

Abstract

As the application of mass spectrometry intensifies in scope and diversity, the need for advanced instrumentation addressing a wide variety of analytical needs also increases. To this end, many modern, top-end mass spectrometers are designed or modified to include a wider range of fragmentation technologies, for example, ECD, ETD, EThcD, and UVPD. Still, the majority of instrument platforms are limited to more conventional methods, such as CID and HCD. While these latter methods have performed well, the less conventional fragmentation methods have been shown to lead to increased information in many applications including middle-down proteomics, top-down proteomics, glycoproteomics, and disulfide bond mapping. We describe the modification of the popular Q Exactive Orbitrap mass spectrometer to extend its fragmentation capabilities to include ECD. We show that this modification allows ≥85% matched ion intensity to originate from ECD fragment ion types as well as provides high sequence coverage (≥60%) of intact proteins and high fragment identification rates with ∼70% of ion signals matched. Finally, the ECD implementation promotes selective disulfide bond dissociation, facilitating the identification of disulfide-linked peptide conjugates. Collectively, this modification extends the capabilities of the Q Exactive Orbitrap mass spectrometer to a range of new applications.

Originalsprog	Engelsk
Tidsskrift	Journal of Proteome Research
Vol/bind	17
Udgave nummer	2
Sider (fra-til)	926-933
Antal sider	8
ISSN	1535-3893
DOI	https://doi.org/10.1021/acs.jproteome.7b00622
Status	Udgivet - 2 feb. 2018

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10.1021/acs.jproteome.7b00622Licens: CC BY-NC-ND

Exploring ECD on a Benchtop Q Exactive Orbitrap Mass SpectrometerForlagets udgivne version, 2,37 MBLicens: CC BY-NC-ND

Citationsformater

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abstract = "As the application of mass spectrometry intensifies in scope and diversity, the need for advanced instrumentation addressing a wide variety of analytical needs also increases. To this end, many modern, top-end mass spectrometers are designed or modified to include a wider range of fragmentation technologies, for example, ECD, ETD, EThcD, and UVPD. Still, the majority of instrument platforms are limited to more conventional methods, such as CID and HCD. While these latter methods have performed well, the less conventional fragmentation methods have been shown to lead to increased information in many applications including middle-down proteomics, top-down proteomics, glycoproteomics, and disulfide bond mapping. We describe the modification of the popular Q Exactive Orbitrap mass spectrometer to extend its fragmentation capabilities to include ECD. We show that this modification allows ≥85% matched ion intensity to originate from ECD fragment ion types as well as provides high sequence coverage (≥60%) of intact proteins and high fragment identification rates with ∼70% of ion signals matched. Finally, the ECD implementation promotes selective disulfide bond dissociation, facilitating the identification of disulfide-linked peptide conjugates. Collectively, this modification extends the capabilities of the Q Exactive Orbitrap mass spectrometer to a range of new applications.",

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AU - Fort, Kyle L

AU - Cramer, Christian N.

AU - Voinov, Valery G

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AU - Lopez, Nathan I

AU - Beckman, Joseph S

AU - Heck, Albert J R

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AB - As the application of mass spectrometry intensifies in scope and diversity, the need for advanced instrumentation addressing a wide variety of analytical needs also increases. To this end, many modern, top-end mass spectrometers are designed or modified to include a wider range of fragmentation technologies, for example, ECD, ETD, EThcD, and UVPD. Still, the majority of instrument platforms are limited to more conventional methods, such as CID and HCD. While these latter methods have performed well, the less conventional fragmentation methods have been shown to lead to increased information in many applications including middle-down proteomics, top-down proteomics, glycoproteomics, and disulfide bond mapping. We describe the modification of the popular Q Exactive Orbitrap mass spectrometer to extend its fragmentation capabilities to include ECD. We show that this modification allows ≥85% matched ion intensity to originate from ECD fragment ion types as well as provides high sequence coverage (≥60%) of intact proteins and high fragment identification rates with ∼70% of ion signals matched. Finally, the ECD implementation promotes selective disulfide bond dissociation, facilitating the identification of disulfide-linked peptide conjugates. Collectively, this modification extends the capabilities of the Q Exactive Orbitrap mass spectrometer to a range of new applications.

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JO - Journal of Proteome Research

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Exploring ECD on a Benchtop Q Exactive Orbitrap Mass Spectrometer

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