Pharmaceutical Analysis

Mass Spectrometry for Discovery and Development Applications

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Mass SpectrometryInnovation with Integrity

Bruker has developed a large portfolio of application based systems for Pharmaceutical Discovery and Development. These systems are capable of analyzing traditional small molecule drugs or biotherapeutics and represent some of the most powerful, yet robust, and easy to use instruments on the market.

Bruker’s Pharmaceutical Analysis Solutions are renowned for:

Market Leading Instrument Performance

Innovative and Easy to Implement Software

Superior Product Robustness and Reliability

Outstanding Expert Service and Support

Whether you are operating in a Discovery operation or a highly regulated Product Development and Manufacturing environment, Bruker can provide a product and service package designed to meet your needs. Our analytical systems are designed for a number of routine and exceptional application areas including:

Discovery Analytical

Chemical Reaction Yield and Purity Determination Compound Library QC

Proteomics/Target Validation

Top down or Bottom up Protein and Peptide Sequencing Biomarker Discovery and Monitoring

ADME-TOX and Pharmacokinetics

Metabolite Identification Drug Distribution

Analytical Development

Stability and Forced Degradation Studies Impurity Identification Formulation and Excipient

Characterization Analytical Method Development

Process Development and Manufacturing QC

Drug Product and Drug Substance Quality Testing Quality Control of Biotherapeutics Sterility Testing

A full Product Portfolio for Pharmaceutical Analysis

The FLEX™ Series of MALDI TOF Mass Spectrometers

Powerful, flexible systems ideal for protein and polymer analysis applications

The maXis™ and micrO-TOF™ Series of TOF and qTOF Mass Spectrometers

Outstanding, UHPLC compatible accurate mass systems for small molecule or biotherapeutics applications

The amaZon™ Series of Ion Trap Mass Spectrometers

Innovative, robust systems ideal for routine or high throughput applications

The solariX™ Series of Fourier Transform Mass Spectrometers

The ultimate system for molecular formula determination

Analytical Solutions Designed for Your Applications

Initial discovery, characterization, and determination of the therapeutic potential of a new compound requires a series of analytical assays. Bruker has utilized several key instrument platforms for molecular characterization applications routinely used in Pharmaceutical Discovery.

Discovery Analytical

Bruker offers several LC-MS options for supporting chemistry operations. Based on either the amaZon Series of ion traps or micrOTOF-Q II or maXis series of qTOFs, these systems can provide detailed information about reaction products, yield and purity.

System options are available for High Throughput, Open Access, or Expert User environments. All of these systems, including very high resolution instruments, are fully compatible with UHPLC methods.

Pharmaceutical Discovery

Analysis of a Synthetic Compound with an amaZon SL ion trap system showing use of MSn fragmentation to identify the compound synthesized.

Sample kindly provided by Dr. M. Rittner, SiChem GmbH, Bremen

Open Access Software used for Reaction Monitoring and QC.

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Proteomics and Target Validation

Validation or Discovery of a putative therapeutic target have always been key steps in both developing a therapeutic and understanding it’s mechanism of action. Bruker has utilized its extensive experience in analyzing proteins by Mass Spectrometry to create a series of solutions for protein identification, quantitation, and sequencing.

Protein identification can be achieved with either our FLEX Series of MALDI-TOF or ESI-TOF instrument range. These systems are fully capable of analyzing a wide variety of proteins isolated by gel or HPLC based methods. Many of these systems also support the sequencing and identification of differentially expressed proteins utilizing labeled or label free bottom up or top down sequencing methodologies.

For analyzing the post translational modifications or proteins and peptides, Bruker offers ion trap based systems which incorporate Electron Transfer Dissociation (ETD) and MSn technologies that greatly enhance the opportunity to identify and analyze glycosylation and phosphorylation patterns on proteins.

Performance and flexibility are the hallmarks of Bruker solutions for Genomics and Proteomics studies. Bruker solutions also offer various levels of analytical capability and performance, and feature specific software (ProteinScape 2) packages developed especially to facilitate large or detailed Proteomics studies.

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Top-down sequencing of proteins based on MALDI-TOF MS to simultaneously obtain N- and C- terminal information.

Analysis of an IgG Glycopeptide Tryptic Digest with an amaZon ETD system.

Biomarker Discovery and Monitoring

Many groups have been working towards the identification of small molecule or protein/peptide biomarkers as indicators of disease or to measure the effectiveness of therapeutic intervention.

Bruker offers a number of system options based on the FLEX series of MALDI TOF instruments or the micrOTOF-Q II and maXis UHR-TOF for protein/peptide or small molecule biomarker identification. These systems have been specifically designed with the necessary statistical data analysis software packages (ProfileAnalyis, MALDI Molecular Imager, Metabolite Tools) to measure and compare differences in biological samples such as (cell or tissue lysates, serum, urine, etc.).

Analysis of peptides from tryptic digest call lysates

Pharmaceutical Discovery

60,000 Ions 12,000 Peptides

ProteinScape Functionality: Protein Database Search and Interactive Viewers

Automatic result assessment

SequenceViewer

Spectrum Viewer LC MS Survey

Support of different fragmentation types

Molecular Tissue Imaging

One of the innovative and exciting uses of Mass Spectrometry has been its application to in situ Tissue Imaging. In this application, Mass Spectrometers are used for molecular histology.

Bruker has been a pioneer in the applica-tion of MALDI-TOF Mass Spectrometers for molecular imaging applications. To facilitate sample preparation for Imaging, Bruker has developed the ImagePrep system for evenly and completely prepa-ring tissue sections with chemical matrix for subsequent Imaging measurements by Mass Spec.

Bruker molecular imaging Mass Spec-trometry solutions combine the market leading performance of the FLEX Series MALDI-TOF systems with powerful Flex-Imaging software for easy and definitive data and statistical analysis.

Molecular Imaging mass spectrometry

Brain section imaged at 200 khz and 1,000 khz.

Pharmaceutical Discovery

Sample Preparation

Statistics

Data Evaluation

Data Acquisition

Determining the metabolic fate of a potential therapeutic by identifying and quantitating its metabolites is a key step in developing a therapeutic. To facilitate these operations, Bruker has utilized its micrOTOF- Q II and maXis systems for metabolite identification and quantitation and the solariX FTMS platform for drug distribution studies.

Metabolite identification

Bruker LC-MS solutions for metabolite identification utilize a combination of powerful qTOF LC-MS systems (micrOTOF-Q II and maXis) and purposefully designed software packages. Bruker solutions are able to calculate difference spectra and apply mass defect filtering to detect potential drug metabolites. Metabolites can by identified by determining the molecular formula and correlating predicted metabolite structures with MS/MS information. All knowledge gained is subsequently used to generate an automated report. Incorporating programs such as Metabolite Tools and SmartFormula 3D, Bruker systems are fully capable of identifying even low level metabolites in complex matrices such as plasma or serum.

ADME-TOX and Pharmacokinetics

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Identification of a drug metabolites in plasma by mass defect filtering.

Mass defect chromatogram

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Metabolite Identification in human plasma-calculating the difference chromatogram of sample and refenence by the superior eXopseTM algorithm reveals even minor metabolites.

Calculate the difference between sample and reference

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C15 H16 NO4S2: Parent + hydroxylation + sulfation

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In vivo Drug Distribution

Typically, in vivo distribution studies are done late in the DMPK process and utilize expensive radiolabelled compounds. While informative, these studies are costly and often can‘t distinguish between parent drug and metabolite distribution in tissues. Bruker has developed the solariX based FTMS system to detect both parent compounds and metabolites in tissues at therapeutic doses without the need for any label. Incorporating an accessible FTMS platform with specially designed image analysis software tools, the solariX is the most versatile MS system capable of providing in situ spatial small molecule drug and metabolite distribution data at therapeutic dosages.

The innovative FAST-SRM on FLEX series MALDI-TOF instruments facilitates measurement of the distribution of a single drug or metabolite.

Molecular imaging of olanzapine in kidney. Dosage – 5 mg/kg, 6h post dosage analysis. For detailed information, please read Application Note FTMS-37.

with CASI full mass

Degradent and Impurity Identification

Identification of degradents and impurities are two very important tasks in the process of bringing a therapeutic to market. Increasingly stringent regulatory requirements have led to the need for an increase in the analytical capabilities of many Pharmaceutical Development groups.

To assist in the identification and measurement of low level components in drug products and drug substances, Bruker has developed a series of solutions based on the micrOTOF Q-II and maxis series of LC-MS systems.

These systems incorporate the SmartFormula software packagefor rapid identification of unknowns. They also possess the dynamic range to easily identify impurities well below the important 0.1% level. Development of Biotherapeutics

Today, many companies have made significant investments in the development of Biotherapeutics. Bruker offers a range of qTOF LC-MS (micrOTOF Q-II and maXis) or ion trap (amaZon ETD) and MALDI-TOF based (FLEX series) solutions to analyze either the intact or digested protein, nucleic acid core, including modifications such as phosphorylation, glycosylation, or PEGylation of a potential Biotherapeutic.

High Intra-spectra dynamic range of mass accuracy allows for correct simultaneous detection of high and low abundant components.

Analysis of a recoombinant protein biotherapeutic

Analytical and Process Development

LC-MS of a reduced monoclonal antibody

Intra-spectra dynamic range

Process Development and Manufacturing QC

As a potential therapeutic moves closer to the market, the scale up and development of a quality, cost effective product becomes very important. For these later stage development applications, Bruker offers a range of MALDI-TOF (microflex and autoflex speed), ion trap (amaZon SL) and ESI-TOF based LC-MS solutions that can answer many basic, but important questions in a quick, easy to interpret fashion.

Many of the Bruker solutions for Analytical Development detailed above can be utilized for QC operations, particularly in the area of Biotherapeutic analysis where the detection and identification of a protein truncation, point mutation, or glycosylation are common analytical challenges. Pharmaceutical Microbiology

When a positive culture is detected in a sample from a production environment, identification of the cultured microbe is required. Further production may be shut down and/or product shipments quarantined until the identification is completed. The MALDI Biotyper is a revolutionary new technique for microbial identification based on microbial protein fingerprints measured on the Bruker Daltonics microflex MALDI-TOF bench-top mass spectrometer. The MALDI Biotyper ready-to-use reference library comprises thousands of individual strains and covers microorganisms from clinical microbiology, food and feed safety, environmental analysis, as well as industrial quality control. Thus, even fastidious microorganisms like anaerobic and non-fermenting bacteria as well as multicellular fungi can be identified with one approach. No expertise in mass spectrometry is required, and the identification can be obtained in-house

within minutes after a positive culture is detected, with very low consumables costs. This time savings can directly translate into enormous cost savings in terms of saving lost product shipments and production line up time. The implementation of the MALDI Biotyper is assisted by IQ/OQ documentation and 21 CFR Part 11 supporting software.

Edmass analysis of PEGylated therapeutic peptides. MALDI-TOF of protein. Yoo et al. 2008, JASMS.

The MALDI Biotyper; It takes only minutes to identify microorganisms – in an easy, wizard-driven workflow.

Identify microorganisms in minutes

Edmass analysis of unknown by-products in recombiant proteins

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Sample kindly provided by Prof. M. Novak, Dr. B. Kovachech, Institute of Neu-roimmunology, Slovak Academy of Sciences, Bratislava, Slovakia.

Characterization of minor components X and Y.

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For research use only. Not for use in diagnostic procedures.

Bruker Daltonik GmbH

Bremen · GermanyPhone +49 (0)421-2205-0 Fax +49 (0)421-2205-103 sales@bdal.de

Bruker Daltonics Inc.

Billerica, MA · USAPhone +1 (978) 663-3660 Fax +1 (978) 667-5993 ms-sales@bdal.com

www.bruker.com Bru

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© B

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L 12

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0, #

2732

97