Thermo Scientific Solutions for Intact-Protein Analysis

Biotherapeutic DevelopmentBetter, Faster Decisions for

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Therapeutic proteins and monoclonal antibodies (MAb) have transformed biotechnology

and the pharmaceutical industry. Together they form the largest part of the rapidly growing

biologics drug market. Protein-based drugs include blood factors, thrombolytic agents,

hormones, hematopoietic growth factors, interferons, interleukins, tumor necrosis factor,

and therapeutic enzymes.

Essential to the development of biotherapeutics is ability to quickly and accurately

assess product quality and safety—including factors such as sequence integrity,

glycan heterogeneity, and purity—at each step. Thermo Scientific workflows for

intact-protein analysis enable these assessments early in the development process

without the effort of complete characterization. Our workflows can provide insights

that allow you to make better, faster decisions about how to proceed with your

biotherapeutic development.

Assess Product Quality and SafetyQuickly and Accurately

For biotherapeutics, fast accurate analysis at each step of the development process is essential

Develop recombinant cell line

Develop drugproduct

Develop purificationprocess

Develop bioprocess

Traditional workflow for assessing MAb purity

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High-quality, high-throughput analyses are key to development of monoclonal antibodies and

other biotherapeutics. A typical intact-protein workflow involves MAb separation and fraction

collection followed by fast cleanup, desalting, and separation by reverse-phase HPLC prior

to mass spectrometric analysis. Intelligent Thermo Scientific HPLC platforms and innovative

column chemistries give users the flexibility to adopt exactly the workflows needed for specific

therapeutic development challenges.

With Thermo Scientific Dionex UltiMate 3000 HPLC, UHPLC+, and RSLCnano systems

and Thermo Scientific EASY-nLC systems, the right combination of chromatographic

performance and capabilities is always available. Thermo Scientific LCs offer:

• Superb accuracy and precision at flow ranges from mL down to nL

• Robust, reliable operation for greater productivity

• Biocompatible flow paths to maintain protein integrity and preserve labile PTMs

• Operating pressures up to 1000 bar for faster analyses

• Configurations for many 1D and 2D workflows

• Available faction collection for automated re-injection in multidimensional or multistep analyses

All of these LC systems can be seamlessly integrated with Thermo Scientific MS systems for fast,

sensitive and accurate analyses.

better results and higher throughputSuperior separations for

MAb samples Desalting on RP column MS of intact protein for protein and glycoform ID

Optional top-downsequencing

Capture by Protein A

Aggregate analysisby SEC

Charge variantanalysis by IEC

Hydrophobicityanalysis by RP/HIC

Optional on-linefraction collection

LC-MS/MS capabilities for greater confidence and additional information

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Fast, accurate mass spectrometry is essential to analysis of intact proteins like monoclonal antibodies.

Thermo Scientific Orbitrap technology offers an unequalled combination of high resolution and

accurate mass (HR/AM), sensitivity, and dynamic range. It enables fast confirmation of mass

and primary sequence, heterogeneity, and purity, allowing quicker, more confident decisions about

how to proceed with biotherapeutic development.

For researchers who plan to dedicate

an instrument primarily to intact protein

screening, the Thermo Scientific Q Exactive

hybrid quadrupole-Orbitrap instrument is

an excellent option, providing:

• High-quality spectra

• Robustness for high throughput

• Compact bench-top design

For researchers who plan to follow screening with complete top-down characterization

or glycan and PTM structural analysis, the Thermo Scientific Orbitrap Elite hybrid

ion trap-Orbitrap mass spectrometer offers the ultimate in performance and flexibility.

It features:

• Stunning 240K mass resolution

• MSn for complete top-down analysis

• Optional electron transfer dissociation (ETD) for fast, accurate location of PTM and glycosylation sites

• 140K mass resolution, low-ppm mass accuracy, and outstanding sensitivity

• 2X– 4X better maximum mass resolution than TOF technology

• Consistent performance for high- and low-abundance samples

• No tradeoff of sensitivity or spectral quality to achieve high mass resolution

• Isotopic resolution for proteins <50K Da

accurate-mass MS/MS provides greater confidenceUncompromising high-resolution,

MAb samples Desalting on RP column MS of intact protein for protein and glycoform ID

Optional top-downsequencing

Capture by Protein A

Aggregate analysisby SEC

Charge variantanalysis by IEC

Hydrophobicityanalysis by RP/HIC

Optional on-linefraction collection

High-resolution, accurate-mass performance of Orbitrap technology clearly resolves IgG glycoforms.

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full advantage of high-quality dataData processing that takes

Accurate deconvolution is essential for analysis of the multi-charge-state data from intact proteins.

Thermo Scientific Protein Deconvolution software maintains the integrity of the high mass accuracy

provided by Orbitrap™ technology. The software employs not one, but two deconvolution algorithms.

The Xtract algorithm is specifically engineered to provide superb deconvolution results on data from

smaller proteins (< ~50K Da) that can be isotopically resolved by Orbitrap technology. The ReSpect™

algorithm provides accurate deconvolution of unresolved data from larger proteins. It is dramatically

faster than other common deconvolution algorithms. This deconvolution power is made accessible

through an entirely new, simplified interface that guides users through the protein deconvolution

workflow and produces clean, easy-to-interpret reports.

For researchers who need to follow up screening with complete top-down characterization or glycan and

PTM structural analysis, we have the specialized software necessary to simplify those tasks, including:

• Thermo Scientific ProSightPC software for powerful top-down analysis with high-resolution, accurate-mass data

• SimGlycan® software from PREMIER Biosoft for glycan structural elucidation

Simplified interface of the Protein Deconvolution software guides users through the deconvolution workflow and produces clean, easy-to-interpret reports

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