Improved workflows for Biopharmaceuticals characterization

Monoclonal antibody (mAb)

Monoclonal antibody (mAb)

Bruker is committed to build on key technologies such as Ultrahigh Resolution (UHR) QTOF and MALDI TOF/TOF to support the development of Biopharmaceuticals.

The use of MALDI in-source decay (ISD) has been widely described as a useful technique to produce protein fragments quite similar to the ones obtained with electron transfer dissociation (ETD) on electrospray instruments; well-known to be useful for protein Top-Down sequencing. The ultrafleXtreme SmartBeam laser enables the formation of these fragments with unmatched speed and their detection with very high sensitivity. Combined with a simple protein cleanup on HPLC, this opens the door to near full length sequence verification of monoclonal antibodies. Similar to ETD, this technique will leave peptide modifications in place to confirm their presence and position. The fact that MALDI spectra only contain singly charged ions simplifies the interpretation of such Top-Down Spectra often containing 100’s of fragments in the case of larger proteins.

Determining the cysteine linkage with disulfide bridges fragmentation

A novel application of this fragmentation technique is to use the selectivity of the method for disulfide bridges (DSB) fragmentation.

Disulfide Bond Analysis

Disulfide Bond Analysis

This allows determining the cysteine linkage in a protein and the heterogeneity of this linkage with a simple peptide map of the unreduced protein.  Software is available to detect patterns characteristics of DSB dipeptides fragmentation and peptide sequence can be verified with a simple MS/MS experiment.

On the other hand, the improvement in UHR QTOF allows for accurate mass measurement of proteins up to 50kDa. For example, it is possible to measure the monoisotopic mass of a reduced antibody light and heavy chain with sub-ppm mass accuracy. We recently demonstrated that even a 5% deamidation can be detected directly at the light chain or IdeS subunit level with a simple LC experiment. This enables the measurement of more quality attributes in upstream development without increasing the workload in terms of sample preparation or data processing.

To read more details about these enhanced workflows, have a look at Bruker’s recent ASMS contributions.

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Mass Spec: Taking UHR (Ultra High Resolution) to a Whole New Level

Average cost to bring a drug to market: $5 Billion

Average time to bring a drug to market: 9 Years

These are truly staggering numbers – that’s $1.5+ million a day!
But what’s that have to do with ultra-high resolution mass spectrometry, you ask?

Simple – data matters, and quality of data is paramount. And the quicker reliable data can be produced and analyzed, the more pharma companies can chip away at that insane hourly rate. Ultra High Resolution (UHR)mass spectrometry enablesfaster, more accurate drug discovery, and the ability to more rapidly analyze compounds with extremely high data confidence.

UHR – Shattering Performance Barriers
The combination of resolution, accuracy and speed is the holy grail of researchers – and modern analytical instrumentation has historically involved tradeoffs. It seems as though researchers are forced to whistle Meatloaf’s “Two Outta Three Ain’t Bad” as they contemplate which feature – speed, accuracy or resolution – will be compromised in the next run. Fact is at $1.5 million a day, compromising anything gets pretty expensive.

UHR redefines our understanding of ‘high resolution,’ breaking past the perceived limitations of mass spec resolution. And it does it without sacrificing accuracy or speed. It delivers extreme sensitivity across the entire mass range – from very small molecules up to large intact proteins.

QTOF-NewsletterApplications for UHR
I’ve been around marketing a while, so I’ve come to dislike tired, generic phrases like ‘quantum leap’ or ‘paradigm shifting.’ Ultra high resolution (UHR), however, is the next evolution of o-TOF mass spectrometry, and its applications range far beyond drug discovery. UHR is perfectly suited to the analysis of unknown compounds in complex matrices, whether they are small molecules or antibodies. It has applications in proteomics, metabolomics, small molecule research, glycomics, screening applications, and more…making it both an incredibly powerful and immensely versatile technology.

Back on the subject of drug discovery: how does UHR fare chipping away at that daily drug cost referenced above? To be fair, that would really depend on how it’s used, how frequently, and so on, but here’s what I can share:

UHR delivers extremely high resolution detection of low-abundance compounds in sharp UHPLC peaks. Even more impressively, it delivers qualitative and quantitative results from a single LC/MS analysis.

How High Res is Ultra High Res?
At Bruker, we’ve built ultra high resolution technology into both our maXis and impact system lines. The impact HD™ – Bruker’s unique UHR-QqTOF (Ultra-High Resolution Qq-Time-Of-Flight) mass spectrometry system – boasts >40,000 full-sensitivity resolution (FSR). Our maXis HD™ ups the ante even further, with an industry-leading (and record-breaking) full sensitivity resolution of > 75,000.

UHR mass spectrometry enhances analytical performance levels for virtually any application in which trace analysis from complex, high-background matrices is a challenge. This includes fields such as biomarker research, identification of impurities, or residue screening.

Do you have any experience with ultra high resolution mass spectrometry?

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Keeping Honey Safe – Smarter Analytical Techniques & Testing Methods Become the Gold Standard

EVOCThe food industry is constantly facing increasing demands to perform food safety testing to ensure product quality and consumer safety. Development of smarter analytical techniques and testing methodologies aims to improve the accuracy, speed, confidence and ease of use required for efficient food screening.

Today, MRM LC-MS/MS is considered the gold standard for food safety testing and methodologies.

Honey Has its Problems
You may not realize it, but the rules and regulations as to the content, labeling and nature of honey is under continuous debate. With a number of contested environmental, ecological and personal health issues to consider, modern honey manufacturers are faced with multiple challenges.

The treatment of honey with veterinary medicines is a particularly controversial issue and the maximum residue limits of commonly used antibiotics – such as sulfonamides – are commonly set at trace levels. Sulfonamides are an important group of broad spectrum antibacterial drugs, often used in honey production to prevent and treat potential bacterial growth. Sulfonamides also fight disease in honey bees. Consequently, residues of sulfonamides are frequently detected in samples of commercially-available honey products.

Quantitative sulfonamide analysis, however, presents the industry with considerable challenges, since traditional analytical techniques have difficulty achieving the selectivity, specificity and accuracy to perform accurate low level quantitation within complex biological matrices. This means that increasingly sophisticated analytical techniques are needed to meet evolving food safety criteria.

Sulfonamides – Difficult to Detect at Low Levels in Honey
Low-level sulfonamide residues are difficult to analyze. The complex interactions that arise from a biological matrix such as honey aren’t well suited to traditional techniques, which commonly suffer extensive preparation times, limited selectivity and low throughput.

Of the range of MS techniques available, multiple reaction monitoring (MRM) with triple quadrupole LC-MS has emerged as the most effective for food analysis, favored for its high sensitivity and selectivity in complex matrices with minimal sample preparation. MRM methods of mass spectrometry analysis employ a mass filtering function to enhance selectivity during analyte quantitation. Ions of interest are first pre-selected as ‘pre-cursor’ ions before undergoing fragmentation within a collision cell. By then filtering only the sequence-specific fragment ions for analysis, MRM MS enables highly sensitive and easy to perform multiplexing analysis with the specificity required at low analyte levels even within complex biological matrices.

Triple Quadrupole LC-MS with Multiple Reaction Monitoring (MRM)
One of our teams at Bruker used high performance triple quadrupole liquid chromatography mass spectrometry (LC-MS TQ) with multiple reaction monitoring (MRM) methodologies to develop a fast, simple and robust technique for accurate and confident low-level sulfonamide quantitation.

UHPLC & EVOC Elite – Excellent Limits of Detection & Repeatability
Triple Quadrupole MS MRM monitoring is a technology that offers complete analyte selectivity, even for samples within complex matrices. In the case of Bruker’s EVOC Elite – used in the study – we’ve also incorporated a unique Active Exhaust feature that improves triple quadrupole MS sensitivity and noise reduction by eliminating gas recirculation in the system.

In the study, an experiment was carried out into the simultaneous determination of sulfonamide residues in honey using Advance UHPLC and the EVOQ Elite triple quadrupole LC-MS/MS system with MRM-based sulfonamide analysis. The system demonstrated excellent detection limits and repeatability for all nine sulfonamides in the study, making it a useful method for the screening and quantification of medicinal sulfonamides commonly used in honey production, to the level required by the EU and other nations.

Which analytical tool do you most frequently use to perform your most challenging analyses?

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Having an impact…II

impactIIAt the annual ASMS conference down in Baltimore in mid-June, Bruker introduced the impact II – our follow-on to the industry changing impact QTOF platform.

The impact II is our new ‘omics knowledge generation platform, delivering best-in-class quantitative proteomics, intact protein analysis and metabolomics capabilities.

The impact II retains many of the features that made the original impact an essential part of the proteomics toolbox, such as the CaptiveSpray nanoBooster. (One user shared that their impact was capable of delivering “untouched performances for over 6-8 weeks of 24/7 use” for routine shotgun bottom up proteomics).

The impact II raises the stakes with triple-quadropole-like sensitivity – benefits of the system’s latest UHR TOF technology: a dual ion funnel, an IonBooster Source, the CaptiveSpray Nanobooster and an HDC collision cell. The sum of these parts? Unrivaled performance.ImpactIIb


Taking Performance Beyond Known Limits
The impact II delivers:

  • 3-4X higher dynamic range
  • 50GB/sec sampling speeds deliver high definition resolution on an LC timescale
  • One-shot plug-and-play acquisition with triple-quad sensitivity
  • Much higher dynamic range in analyte concentration

From Data to Knowledge
Delivering high quality data is a must for any analytical instrument. With mass spec and proteomics, however, it isn’t enough. Data must be parsed, analyzed, and transformed into knowledge. ProteinScape – Bruker’s central knowledge generation database for proteomics and glycomics – transforms the impact II data into relevant biological knowledge. Data from the impact II can be combined with data from other MS systems, and can be linked to knowledge databases (e.g., 3D structures, Gene Ontology, Pride).

proteinscapeBiology’s Diversity Demands Versatility
The impact II’s unique spectral qualities (resolution, sensitivity, mass accuracy), wide dynamic range (5 orders of magnitude), benchtop design and ability to deliver results in a single LC run make it an incredibly versatile system. Yes – it provides best-in-class quantitative proteomics, intact protein analysis and metabolomics capabilities. It’s also a speed demon when it comes to ultra-fast shotgun proteomics experiments. But it’s also the perfect system for:

  • drug metabolite, degradant and impurity identification and quantitation
  • synthetic chemistry support
  • forensic and doping control
  • food and water testing

Learn more about the Bruker impact II. (LINK TO PRESS RELEASE OR PRODUCT PAGE, WHEN LIVE)

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Bruker @ ASMS2014

ASMS kicks off down in Baltimore on Sunday, June 15, and hopefully you’ll be getting a chance to enjoy spectacular views of the Inner Harbor like this one:

Bruker has tons of events, posters and oral sessions on the agenda this year, and a rocking hospitality suite open on Monday, Tuesday and Wednesday (with a live band) nights.  Everything kicks off Sunday morning with our User Meeting, followed by our customer appreciation party at the Hard Rock Café.

To see our full ASMS 2014 schedule of events, head over to our events site at:

Here’s a quick rundown of what’s on tap this week at ASMS:

Sunday, June 15
8:00am – 12:30pm, User Meeting, Marriott Inner Harbor at Camden Yards, Grand Ball Room, 110 S Eutaw Street
12:30 – 4:30pm, Bruker Customer Appreciation Event – Hard Rock Cafe, 601 E Pratt Street

Monday, June 16
7:00am – 8:15am, Bruker Breakfast Workshop, Baltimore Convention Center – Room 341/342
8:00am – 9:00am, Bruker Media Event, Hilton Hotel, Key Ballroom 6
9:00am – 5:00pm, Poster Session, Exhibits, Booth 49 • Baltimore Convention Center
8:00pm – 11:00pm, Bruker Hospitality Suite, Key Ballroom, Hilton Hotel

Tuesday, June 17
7:00am – 8:15am, Bruker Breakfast Workshop, Baltimore Convention Center – Room 341/342
9:00am – 5:00pm, Poster Session, Exhibits, Booth 49 • Baltimore Convention Center
8:00pm – 11:00pm, Bruker Hospitality Suite, Key Ballroom, Hilton Hotel

Wednesday, June 18
7:00am – 8:15am, Bruker Breakfast Workshop, Baltimore Convention Center – Room 341/342
9:00am-5:00pm, Poster Session, Exhibits, Booth 49 • Baltimore Convention Center
8:00pm – 11:00pm, Bruker Hospitality Suite, Key Ballroom, Hilton Hotel – Live band

Thursday, June 19
9:00am -2:30pm, Poster Session, Exhibits, Booth 49 • Baltimore Convention Center

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Out and About: Bruker Upcoming Events

Quickly see which shows Bruker will be attending at Bdalevents.comFor a complete list of events and tradeshows at which Bruker will be exhibiting, presenting, speaking, hosting hospitality suites, running demos and more, visit Bruker’s Event Calendar at

This Month: June 2014

06/01/2014 – Forum LABO & BIOTECH 2014
Port de Versailles, France

06/01/2014 – College Canada Conference
Vancouver, BC, CA – College Chemistry Canada (C3) Conference

06/04/2014 – EMIM 2014
Antwerp, Belgium – 9th European Molecular Imaging Meeting

06/15/2014 – ASMS 2014
Baltimore, Maryland
Booth – 49; Hospitality Suite – Key Ballroom 6
Check out our events site for in-depth coverage.

06/19/2014 – Annual Congress of the Swiss Society for Microbiology
Fribourg, Switzerland

06/21/2014 – IFT 2014
New Orleans, LA / US – Institute of Food Technologies

06/23/2014 – Metabolomics 2014
Tsuruoka, Japan – 10th International Conference of the Metabolomics Society
Register for the free Bruker Luncheon Event at-the-metabolomics-2014.html

Next Month: July 2014

07/01/2014 – Microscience
Manchester, UK

07/09/2014 – New Frontiers in Biophysics
Warwick, UK – British Biophysical Society, biennial meeting

07/12/2014 – AAIC 2014
Copenhagen, Denmark – Alzheimer´s Associations International Conference

07/21/2014 – BCWGC 2014
Penticton, Canada – 14th Annual Enology & Viticulture Conference

07/21/2014 – Foodtechnology 2014
Las Vegas, NV, US – 3rd International Conference and Exhibition on Food Processing & Technology

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Impact HD – Proteomics, Forensics, Drug Discovery, Impurity Detection, Food Testing and More

impacthdOur new brochure for the impact HD went live a few weeks back, and we’ve gotten some feedback from scientists who were surprised at the breadth of functionality the HD offers.

Often, we run across users of a mass spec instrument who are pretty cloistered in terms of their research. Proteomics scientists tend to study…well….proteins. Food safety scientists typically screen for contaminants.

Working for an analytical instrument company involved in many disciplines allows us to see the ‘bigger capabilities picture’ more often, so the system’s broad usefulness comes as less of a surprise.

The impact HD is ideal for:

  • Biomarker discovery and validation in proteomics and metabolomics
  • Drug metabolite, degradant and impurity identification and quantitation
  • Synthetic chemistry support
  • Intact protein analysis and characterization of biopharmaceuticals
  • Forensic and doping control
  • Food and water testing

It’s a pretty amazing system, and the brochure is worth a read to see how the impact HD solves difficult analytical challenges and delivers results in these diverse applications.

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