Q-TOF Newsletter - Bruker · • POLYWAX LP; A: 90% ACN, 0.1% acetic acid; B: ... Q-TOF Newsletter 13 Edition Sample: 5 µg Hela digest ... Data measured with impact UHR-TOF prior

impact HD™ – sensational new capabilities in shotgun proteomics

Identification of more than 4500-5000 proteins in a single LC-MS/MS run, more than 8400 from a combined 2D experiment.

(Minneapolis, June 2013) At the ASMS 2013 conference Bruker introduces exciting new High-Definition capa-bilities in complex-sample proteomics and biomarker research. Latest generation ion optics, record high-speed digitization and patented nano-flow ionization technology work in concert to define your sample more sharply than ever before. Analyzing your HD data using Bruker‘s exceptional proprietary Protein and Glycoprotein Characterization software gives you unique insights into your research challenges.

Figure 1

• 50 cm LC column• Separation with 4 hour gradient• impact HD UHR-TOF with CaptiveSpray™ nanoBooster™• Instant Expertise™ software• ProteinScape™

Figure 2

• POLYWAX LP; A: 90% ACN, 0.1% acetic acid; B: 30% ACN, 0.1% formic acid• MS: impact HD• Source: CaptiveSpray nanoBooster• 8444 proteins identified with FDR of < 1.0%

See poster MP24, 491 and MP24, 486 for more details.

Q-TOF Newsletter

ASMS 2013 Edition

Sample: 5 µg Hela digest

4,587 proteins

39,549 peptides

Figu

re 1

ERLIC separation into 6 fractions

Figu

re 2

45554830

50204695

4374 4486

8444

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1 2 3 4 5 6 7

Pep

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ERLIC fractions 1-6 plus total (7)

proteins peptide hits

Innovation with IntegrityLC-MS/MS

New sensational

performance level

impact HD enhanced dynamic range 50 GBit/sec technology

CaptiveSpray nanoBooster™ extreme sensitivity supercharging capabilities

Instant Expertise™ Software IDAS RT2

Introducing High-Definition, Instant Expertise™ and CaptiveSpray nanoBooster™ in ultra-high resolution LC-MS/MS in a research-performance bench-top Q-TOF

• 50 GBit/sec sampling technology maintaining full specification resolution even during ultrafast chromatography. 5 orders of magnitude dynamic range in one second for definitive trace analysis from complex, high-background matrices

• CaptiveSpray nanoBooster with enhanced peptide sensitivity matching best shot-gun proteomics identifications

• Intelligent self-optimizing MSMS routines for expert-caliber results first time

5 orders of magnitude dynamic range @ 1 Hz speed LC timescale

195.0878

217.0696

233.0436

+MS, 4.2-4.3min #247-256

0

2

4

6

6x10Intens.

0 25 50 75 100 125 150 175 200 225 m/z

Caffeine 0.6 ppmI = 7,155,285 counts

90.0550

+MS, 4.2-4.3min #247-256

0

10

20

30

40

50

Intens.

86 88 90 92 94 96 m/z

Alanine 0.5 ppm I = 46 counts

1.6 x 10 5

• 3 - 4 x higher dynamic range from 10 bit ADC technology

• Provides 50 GBit/sec highest sampling speeds and maintains high resolution

• Much higher dynamic range in protein concentration measurable

Figure 3: Five orders of magnitude dynamic range @ 1 Hz speed LC timescale

Dynamic range at real LC speed

CaptiveSpray nanoBooster™ for ultimate proteomics performance

Figure 4: Optimized ionization condition

Smart dopant enrichment of nitrogen

Figure 5: Significant increase in signal intensities for low sample amounts

Figure 6: Data measured with impact UHR-TOF prior to HD improvement

CaptiveSpray nanoBooster

• Boost nanoflow sensitivity

• Enable glycoanalysis

• Supercharging capability

• Push up ID rates

• Reduce noise

• Patented technology

Performance and ease of use

• Direct benefit for quantitation / glycosylation detection

• One method for all concentrations

without CaptiveSpray nanoBooster

CaptiveSpray nanoBooster

Hela cell digest,100 ng lysate

Enhance signal intensity and charge state for enhanced Protein ID capabilities. CaptiveSpray nanoBooster is particularly sensitive for low-abundance peptides,lower quantitation limits, and detection of glycosylation sites. Data measured with impact „classic“ prior to HD improvement.

1027

2654

1286

3018

100 ng HeLa lysate 1000 ng HeLa lysate

w/o w/nanoBooster

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Bruker Daltonik GmbH

Bremen · GermanyPhone +49 (0)421-2205-0 Fax +49 (0)421-2205-103 [email protected]

Bruker Daltonics Inc.

Billerica, MA · USA Phone +1 (978) 663-3660 Fax +1 (978) 667-5993 [email protected]

www.bruker.com

Fremont, CA · USA Phone +1 (510) 683-4300 Fax +1 (510) 490-6586 [email protected]

Instant Expertise™: Method for optimal quant and ID results in one acquisition

Issue: Classical Auto MS/MS strategies use basically the same pre-set conditions for fragmentation; independent of the peptide amount and chroma-tographic resolution – meaning that dynamic range and MS/MS quality are compromised .

Solution: Instant Expertise Software with new autoMSMS strategies:

• IDAS (Intensity Dependent Acquisition Speed) MSMS acquisition time automatically derived from precursor intensity

• RT2 (RealTime Re-Think) RealTime Re-Think reconsiders an additional MS/MS at a higher precursor signal intensity before activating dynamic exclusion

See Application Note LCMS-81 for more details.

• Higher productivity – one acquisition method for all concentration levels

• Higher yield (> 50%) of good MSMS => higher ID rates

• High accuracy for quantitation

• Always obtain optimal results, independent of sample amount or complexity

Figure 7: Sampling rate automatically derived from precursor intensity

MSMS acquisition time automatically derived from precursor intensity

2Hz 20HzMSMS spectra rate:

Aut

oMS

MS

cyc

le t

ime

[s]

AutoMSMS cycles

0.00

0.50

1.00

1.50

2.00

2.50

1 2 3 4

MS MS MS MS

30 MS/MS 29 MS/MS 16 MS/MS 21 MS/MS

18 19

# of MS/MS spectra within 2.5 sec

Always obtain optimal results, independent of sample amount or complexity