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Ultra high resolution linear ion trap Orbitrap mass spectrometer (Orbitrap Elite) facilitates top down LC MS/MS and versatile peptide fragmentation modes.

Michalski A, Damoc E, Lange O, Denisov E, Nolting D, Müller M, Viner R, Schwartz J, Remes P, Belford M, Dunyach JJ, Cox J, Horning S, Mann M, Makarov A - Mol. Cell Proteomics (2011)

Bottom Line: Here we describe a novel version of this instrument family, the Orbitrap Elite, which is improved in three main areas.For bottom-up proteomics, we combined survey scans at 240,000 resolving power with data-dependent collision-induced dissociation of the 20 most abundant precursors in a total cycle time of 2.5 s-increasing protein identifications in complex mixtures by about 30%.The speed of the Orbitrap Elite furthermore allows scan modes in which complementary dissociation mechanisms are routinely obtained of all fragmented peptides.

View Article: PubMed Central - PubMed

Affiliation: Department of Proteomics and Signal Transduction, Max-Planck Institute for Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany.

ABSTRACT
Although only a few years old, the combination of a linear ion trap with an Orbitrap analyzer has become one of the standard mass spectrometers to characterize proteins and proteomes. Here we describe a novel version of this instrument family, the Orbitrap Elite, which is improved in three main areas. The ion transfer optics has an ion path that blocks the line of sight to achieve more robust operation. The tandem MS acquisition speed of the dual cell linear ion trap now exceeds 12 Hz. Most importantly, the resolving power of the Orbitrap analyzer has been increased twofold for the same transient length by employing a compact, high-field Orbitrap analyzer that almost doubles the observed frequencies. An enhanced Fourier Transform algorithm-incorporating phase information-further doubles the resolving power to 240,000 at m/z 400 for a 768 ms transient. For top-down experiments, we combine a survey scan with a selected ion monitoring scan of the charge state of the protein to be fragmented and with several HCD microscans. Despite the 120,000 resolving power for SIM and HCD scans, the total cycle time is within several seconds and therefore suitable for liquid chromatography tandem MS. For bottom-up proteomics, we combined survey scans at 240,000 resolving power with data-dependent collision-induced dissociation of the 20 most abundant precursors in a total cycle time of 2.5 s-increasing protein identifications in complex mixtures by about 30%. The speed of the Orbitrap Elite furthermore allows scan modes in which complementary dissociation mechanisms are routinely obtained of all fragmented peptides.

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Related in: MedlinePlus

Isotope resolved spectrum of enolase with Orbitrap Elite and LTQ Orbitrap Velos instruments. Spectra were acquired with transients of A, 768 ms on the Orbitrap Elite and B, 1536 ms on the LTQ Orbitrap Velos. In these conditions, the superior resolution of the Orbitrap Elite instrument (>2 × higher in 2 × shorter transient time) helps to baseline resolve the 47+ charge state of the intact yeast enolase.
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Figure 3: Isotope resolved spectrum of enolase with Orbitrap Elite and LTQ Orbitrap Velos instruments. Spectra were acquired with transients of A, 768 ms on the Orbitrap Elite and B, 1536 ms on the LTQ Orbitrap Velos. In these conditions, the superior resolution of the Orbitrap Elite instrument (>2 × higher in 2 × shorter transient time) helps to baseline resolve the 47+ charge state of the intact yeast enolase.

Mentions: The implementation of the compact Orbitrap analyzer and eFT signal processing allows the Orbitrap Elite to reach a comparably high resolution as a 17 Tesla FT ICR instruments at m/z 400 Th and with standard signal processing methods. In contrast to the inversely linear dependence of FT ICR resolving power on m/z, however, the Orbitrap resolving power is inversely proportional to the square root of m/z (1). Therefore, for subsecond acquisition on an LC-MS time scale its resolving power at m/z 1000 already corresponds to a 25 Tesla FT-ICR instrument, and it should be particularly suitable for top down experiments of larger proteins such as BSA (66.4 kDa) and enolase (46.64 kDa). Using static electrospray conditions we found that charge state 47+ of intact yeast enolase could readily be baseline resolved at a resolution setting of 240,000 corresponding to 768 ms transients (Fig. 3A). To reach this resolution, a vacuum better than 10−10 Torr was necessary. On the predecessor instrument, partial resolution of intact yeast enolase could occasionally be observed (Fig. 3B).


Ultra high resolution linear ion trap Orbitrap mass spectrometer (Orbitrap Elite) facilitates top down LC MS/MS and versatile peptide fragmentation modes.

Michalski A, Damoc E, Lange O, Denisov E, Nolting D, Müller M, Viner R, Schwartz J, Remes P, Belford M, Dunyach JJ, Cox J, Horning S, Mann M, Makarov A - Mol. Cell Proteomics (2011)

Isotope resolved spectrum of enolase with Orbitrap Elite and LTQ Orbitrap Velos instruments. Spectra were acquired with transients of A, 768 ms on the Orbitrap Elite and B, 1536 ms on the LTQ Orbitrap Velos. In these conditions, the superior resolution of the Orbitrap Elite instrument (>2 × higher in 2 × shorter transient time) helps to baseline resolve the 47+ charge state of the intact yeast enolase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3316736&req=5

Figure 3: Isotope resolved spectrum of enolase with Orbitrap Elite and LTQ Orbitrap Velos instruments. Spectra were acquired with transients of A, 768 ms on the Orbitrap Elite and B, 1536 ms on the LTQ Orbitrap Velos. In these conditions, the superior resolution of the Orbitrap Elite instrument (>2 × higher in 2 × shorter transient time) helps to baseline resolve the 47+ charge state of the intact yeast enolase.
Mentions: The implementation of the compact Orbitrap analyzer and eFT signal processing allows the Orbitrap Elite to reach a comparably high resolution as a 17 Tesla FT ICR instruments at m/z 400 Th and with standard signal processing methods. In contrast to the inversely linear dependence of FT ICR resolving power on m/z, however, the Orbitrap resolving power is inversely proportional to the square root of m/z (1). Therefore, for subsecond acquisition on an LC-MS time scale its resolving power at m/z 1000 already corresponds to a 25 Tesla FT-ICR instrument, and it should be particularly suitable for top down experiments of larger proteins such as BSA (66.4 kDa) and enolase (46.64 kDa). Using static electrospray conditions we found that charge state 47+ of intact yeast enolase could readily be baseline resolved at a resolution setting of 240,000 corresponding to 768 ms transients (Fig. 3A). To reach this resolution, a vacuum better than 10−10 Torr was necessary. On the predecessor instrument, partial resolution of intact yeast enolase could occasionally be observed (Fig. 3B).

Bottom Line: Here we describe a novel version of this instrument family, the Orbitrap Elite, which is improved in three main areas.For bottom-up proteomics, we combined survey scans at 240,000 resolving power with data-dependent collision-induced dissociation of the 20 most abundant precursors in a total cycle time of 2.5 s-increasing protein identifications in complex mixtures by about 30%.The speed of the Orbitrap Elite furthermore allows scan modes in which complementary dissociation mechanisms are routinely obtained of all fragmented peptides.

View Article: PubMed Central - PubMed

Affiliation: Department of Proteomics and Signal Transduction, Max-Planck Institute for Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany.

ABSTRACT
Although only a few years old, the combination of a linear ion trap with an Orbitrap analyzer has become one of the standard mass spectrometers to characterize proteins and proteomes. Here we describe a novel version of this instrument family, the Orbitrap Elite, which is improved in three main areas. The ion transfer optics has an ion path that blocks the line of sight to achieve more robust operation. The tandem MS acquisition speed of the dual cell linear ion trap now exceeds 12 Hz. Most importantly, the resolving power of the Orbitrap analyzer has been increased twofold for the same transient length by employing a compact, high-field Orbitrap analyzer that almost doubles the observed frequencies. An enhanced Fourier Transform algorithm-incorporating phase information-further doubles the resolving power to 240,000 at m/z 400 for a 768 ms transient. For top-down experiments, we combine a survey scan with a selected ion monitoring scan of the charge state of the protein to be fragmented and with several HCD microscans. Despite the 120,000 resolving power for SIM and HCD scans, the total cycle time is within several seconds and therefore suitable for liquid chromatography tandem MS. For bottom-up proteomics, we combined survey scans at 240,000 resolving power with data-dependent collision-induced dissociation of the 20 most abundant precursors in a total cycle time of 2.5 s-increasing protein identifications in complex mixtures by about 30%. The speed of the Orbitrap Elite furthermore allows scan modes in which complementary dissociation mechanisms are routinely obtained of all fragmented peptides.

Show MeSH
Related in: MedlinePlus