Analytical Characterisation of Biotherapeutics banner

Analytical scientists have their work cut out for them. Multitudes of new, non-platform formats are landing in their labs, requiring them to develop robust methods to characterise these molecules, from which to identify the desired properties and target product profiles.

PEGS Europe’s 5th Annual Analytical Characterisation of Biotherapeutics aims to arm scientists with the tools and knowledge to speed innovation, by showcasing novel technologies, proven techniques or risk-based approaches that can help characterise the new modalities, analyse the structure-function and determine CQAs of the molecules.

Final Agenda


07:45 Registration (Foyer C) and Morning Coffee (Foyer D)

Auditorium VIII

08:30 Chairperson’s Remarks

Bernice Yeung, PhD, Global Head of Characterization, Analytical Development, Shire

08:35 Developing, Qualifying and Validating Analytical Methods for Novel Biologics

Lowney_DeclanDeclan Lowney, MSc, Associate Director, Analytical Development, Janssen R&D

The presentation will cover the analytical strategies deployed for the characterization of novel biologics and the analytical approaches taken for release, stability and characterisation.

09:05 Multi Attribute Method and Native Intact Mass Spectrometry for Characterization of Quality Attributes during Pharmaceutical Development of mAb Mixtures

Kristensen_DanBachDan Bach Kristensen, PhD, Principal Scientist, Analytical Development, Symphogen A/S

Symphogen develops anti-cancer mAb mixtures, which are inherently complex and difficult to characterize using conventional technologies (e.g. methods relying exclusively on chromatographic resolution). To address product complexity and meet regulatory demands, Symphogen is looking increasingly to mass spectrometry for qualitative and quantitative control of critical quality attributes (CQAs). Case studies employing LC MS (Multiple-Attribute Method, MAM) and native mass spectrometry for characterization and control of CQAs during biopharmaceutical development will be presented.

09:35 Identification and Quantitation of Duobody® Bispecific IgG1 Using Mass Spectrometry and Automated Data Processing and Analysis Workflow

vandenBremer_EwaldEwald van den Bremer, PhD, Senior Scientist, Analytical Sciences, Genmab B.V.

The characterization of bispecific antibodies (BsAbs) by mass spectrometry (MS) offers several advantages over traditional chromatographic techniques (e.g. HIC, CEX). MS provides unambiguous identification and relevant quantitative information, and combined with automated data processing and analysis, it can be employed in a high-throughput environment. We present a software solution and the related workflows that enabled us to accelerate BsAb research batch characterization and release, achieving high quality results and significant time and cost savings.

10:05 Reproducible LC/MS in Biopharma: A New Paradigm

John Gebler, PhD, Director, Biopharma Business Development, Pharmaceutical Business, Waters Corporation

10:35 Coffee Break in the Exhibit Hall with Poster Viewing (Pavilion 1)

11:15 KEYNOTE PRESENTATION: Integrating Analytical Strategies into a Comprehensive Development Strategy

Spitznagel_TomThomas Spitznagel, PhD, Senior Vice President, Biopharmaceutical Development & Manufacturing, MacroGenics

Appropriately selected and developed analytics are critical to ensure process development and manufacturing are effectively implemented. Phase appropriate strategies and approaches to developing analytical release, characterization, and in-process methods will be presented that balance speed, risk, and thoroughness. A variety of case studies across different protein platforms will be used to illustrate examples that ensure the overall development strategy is supported by the appropriate set of analytical tools.

11:45 Cutting-Edge Analytical and Structural Methods for the Characterization of Antibodies and Antibody-Drug Conjugates

Wagner_ElsaElsa Wagner-Rousset, PhD, Senior Scientist, NBEs, Analytical Chemistry, Centre d’Immunologie Pierre Fabre

mAbs and ADCs are one of the fastest growing classes of oncology therapeutics. Their development and optimization rely on improving their analytical characterization by assessing critical quality attributes (CQAs) such as sequence liabilities, drug load distribution, drug to antibody ratio and residual small molecular drugs. Therefore, early-developability assessment requires state-of-the-art analytical methods, such as native and ion mobility mass spectrometry (MS), 2D liquid chromatography and capillary electrophoresis coupled to MS.

12:15 Product and Process Characterisation of Novel High-Potency Neurotoxin Therapeutics

Spencer_DavidDavid Spencer, MSc, Characterization Manager, Ipsen

Distinct molecular attributes can critically modulate the physicochemical and pharmacological properties of protein biotherapeutics. Due to their highly potent nature, and resultant low dose, toxin-based therapeutics present some unique analytical challenges. Case studies will be presented where insight into quality attributes and key degradation pathways has been gained using novel, highly sensitive analytical characterisation techniques. Application of this knowledge to determine toxin manufacturing critical process parameters will also be discussed.

12:45 Array-Based SPR Imaging as a Powerful Tool at Multiple Stages in Candidate Selection Cascades

Alex van der Kooi, Manager Interaction Laboratory, IBIS Technologies

An immunization campaign often results in hundreds to thousands of lead candidates. A number of tests is applied to narrow down the best possible candidate(s). In this presentation we demonstrate that the IBIS MX96, an array-based SPR imaging platform, can be used at multiple stages in such a screening cascade.

13:00 Recent Advances in the Use of Capillary Electrophoresis for Biopharmaceutical Analysis

Jim Thorn, Senior Manager, Marketing & Applications Separations, SCIEX
We will present a case study of the rapid and complete characterization of the NIST mAb. Then, we will present the identification of biopharmaceutical modifications by the connection of CE to MS, and we will preview the acceleration of cell line development and process control through high throughput glycan screening.

NanotemperTechnologies 13:15 Luncheon Presentation I: A Quick Check of Protein Quality that will Improve Biotherapeutic Candidate Purification/Characterization Workflow

Peter Fung, Senior Manager, Product Marketing, Marketing, NanoTemper Technologies

Starting with material of questionable quality for protein purifcation and characterization leads to irreproducible or ambiguous results.Transitioning between upstream and downstream workflows can be challenging for bioprocessing researchers, especially when the quality of the sample material is not known.We offer a new platform that identifies sample quality and relative functionality in minutes complementing and guiding bioprocessing workflows-making go/no go decisions easy and quick-saving time, effort and producing more consistent results.

13:45 Luncheon Presentation II (Sponsorship Opportunity Available)

14:15 Session Break

Auditorium VIII

14:30 Chairperson’s Remarks

Mario Lebendiker, PhD, Head, Protein Purification Facility, Wolfson Centre for Applied Structural Biology, Hebrew University of Jerusalem

14:35 Structure-Function Characterization of Sulfated Glycans in Recombinant Idursulfase

Yeung_BerniceBernice Yeung, PhD, Global Head of Characterization, Analytical Development, Shire

Recombinant Idursulfase is used in enzyme replacement therapy for treatment of Hunter Syndrome. Characterization of Idursulfase has been performed to understand its structure-function relationships. In this highly glycosylated protein, it has been recognized that phosphorylated glycans are critical for facilitating cellular uptake of Idursulfase, while sialylated glycans are needed for pharmacokinetic effects. The discovery of sulfated glycans in Idursulfase has led to unexpected understanding of the role of glycosylation on its function.

15:05 HOS in QC Environment - Could Native Peptide Mapping Support/Replace Bioassay for Stability Monitoring of Biopharmaceuticals?

Michel DegueldreMichel Degueldre, Scientist, Analytical Sciences Biologicals, UCB Pharma SA

Higher Order Structure is one of the critical quality attributes to be controlled for therapeutic proteins. Current HOS techniques are hardly amenable to QC. The potential of a novel peptide mapping method for routine monitoring of HOS will be presented. This presentation will show how promising this QC-friendly method is to understand the link between structure (HOS) and function (biological activity) and the degradation pathways under different stress conditions.

15:35 Refreshment Break in the Exhibit Hall with Poster Viewing (Pavilion 1)

16:15 Characterization of Complex Glycosylation Patterns for Biopharmaceuticals

Urs Lohrig, PhD, Labhead AL1, Global Drug Development / Technical, Research & Development, Novartis

Glycosylation remains a major challenge when probing the molecular structure of biopharmaceuticals. Tools for the in-depth characterization of N- and O-glycosylations have been developed over the years and were employed on various levels within the industry. Here, we present approaches including hyphenated LC-MS/MS on various levels to tackle the complexity of glycan modifications within biosimilar characterization workflows. This enables a suitable depth of knowledge for manufacturing process development as well as supporting final biosimilarity evaluation.

16:45 Characterization of Variants of a Clinical Antibody Candidate Using Alternative and Novel Separation Methods

Eisen_DavidDavid Eisen, PhD, Scientist, Technical R&D, Technical Development Biosimilars, Novartis

This talk focuses on the characterization of charged variants of an analytically challenging monoclonal antibody candidate. We enriched the charged protein variants for analytical testing using OFFGEL electrophoresis. By applying specific RP-UPLC-MS and wide-pore HILIC-UPLC-MS methods, we could characterize these variants on intact, subunit and peptide level with enhanced resolution. These analyses helped to elucidate the degradation mechanism and criticality of an aspartate isomerization site in the variable domain of the antibody.

17:15 Poster Highlight: Rapid Online and Inprocess Characterization of Cetuximab Fd-Glycoprofile by Direct Monitoring and Control

Eisen_DavidMartin Hedstrom, PhD, Head, Bioanalysis, Division of Biotechnology, Lund University

An automated analytical platform is developed for real time in-process monitoring of monoclonal antibody quality. The system was adapted to continuously monitor a simulated mammalian cell cultivation for the profiling of the cetuximab Fd-glycosylation profile using HR-QTOF-MS as detection method. With this novel technique, the Fc and F(ab')2 subunits are completely separated and sequentially eluted and analysed, allowing for a reliable assignment and profiling of minor glycoforms and other PTMs.

17:45 Networking Reception in the Exhibit Hall with Poster Viewing (Pavilion 1)

18:45 Problem-Solving Breakout Discussions (Foyer E&F)

Stability Strategies for Large Molecules

Moderator: Declan Lowney, MSc, Associate Director, Analytical Development, Janssen R&D

  • Design of studies
  • Influence of Tg’ on stability strategy
  • Stability strategy for self-administered products
  • Statistical modelling and extrapolation

Multi Attribute Methods (MAM): Applications in Discovery, Development and CMC

Dan Bach Kristensen, PhD, Principal Scientist, Analytical Development, Symphogen A/S

  • How to prioritize optimization of the MAM workflow for robustness?
  • Is (high) accuracy and reproducibility even obtainable across different platforms and between laboratories?
  • Knowledge sharing regarding MAM analysis of challenging degradants (deamidation, isomerization, succinimide)
  • Do you view MAM as a stand-alone analytical powerhouse or an addition to your existing analytical platform?
  • How do you handle new peak detection in a QC environment (for release and stability testing of GMP batches)?



19:45 End of Day


08:00 Registration (Foyer C) and Morning Coffee (Foyer D)

Auditorium VIII

08:30 Chairperson’s Remarks

Dan Bach Kristensen, PhD, Principal Scientist, Analytical Development, Symphogen A/S

08:35 A New Application of Multi Angle Light Scattering Coupled to Ion Exchange Chromatography (IEX-MALS) for Protein Characterization

Lebendiker_MarioMario Lebendiker, PhD, Head, Protein Purification Facility, Wolfson Centre for Applied Structural Biology, Hebrew University of Jerusalem

We present here a new analytical tool for protein characterization that combines the high resolution ion exchange (IEX) chromatography method with multi-angle light scattering (MALS). The limited resolution of SEC interferes in some cases with the accurate analysis that can be achieved by MALS. Here we show that combining MALS with the higher resolution separation technique IEX (IEX-MALS) allows a precise analysis of samples that cannot be resolved by SEC-MALS. We conclude that IEX-MALS is a valuable technique for proteins that have limited analysis achieved with SEC-MALS.

09:05 Multiple Attribute Monitoring – Optimization of Automated Sample Preparation

Pfenninger_AnjaAnja Pfenninger, PhD, Lab Head, Mass Spectrometry, Bioanalytics/Biopharmaceutical Development, Sanofi

Automated sample preparation of tryptic digests using a liquid-handling roboter is key for successful multiple-attribute monitoring in medium-throughput mode (100-200 samples per week). The optimization procedure will be discussed with regards to different aspects such as digest quality, costs of goods, speed and reproducibility.

09:35 Energetic Epitope Mapping of an Antibody/Interleukin-23 Interaction

Schneeweis_LumelleLumelle A. Schneeweis, PhD, Protein Science, Molecular Discovery Technologies, Bristol-Myers Squibb

Identification of the energetic epitope of the hot-spot residues which dominate the binding affinity of an antibody supports mechanistic interpretation, antibody optimization, and intellectual property claims. Complementary mass spectrometry, computational analysis, mutagenesis, and binding analytics provide a clear picture of the discontinuous interfacial hot-spot epitope on interleukin-23 (IL-23) that dominates its binding affinity for an anti-IL-23 antibody.

10:05 Next Steps in Biophysical Characterization and Screening: RPC/IEX-MALS and HT-SLS

Daniel Some, PhD, Principal Scientist, Marketing, Wyatt Technology Corp

SEC-MALS and high-throughput DLS (HT-DLS) are widely implemented across biopharma to characterize molar mass, aggregation, oligomerization and fragmentation, and to screen candidates and formulations for aggregation and stability. Recent extensions of light scattering will be presented: a light-scattering plate reader that measures both dynamic and static light scattering, to determine size, molar mass, kD, A2, thermal stability and viscosity; and the use of multi-angle light scattering with reversed-phase and ion-exchange chromatography.

10:35 Coffee Break in the Exhibit Hall with Poster Viewing (Pavilion 1)

11:15 Highly Glycosylated Therapeutic Proteins Neorecormon® and Mircera®: Novel Methods for Sample Generation, Structural and Functional Characterization

Buettner_AlexanderAlexander Buettner, PhD, Scientist, Analytical Development and Quality Control, Pharma Technical Development Europe, Roche 

NeoRecormon® and Mircera® are therapeutic proteins for the treatment of anaemia and contain highly complex glycans. Glycan variety in combination with presence of multiple glycosylation sites makes CQA assessment cumbersome. We have been developing and applying sample generation techniques as well as structural and functional analysis methods to examine structure function relationships in this complex situation. The talk will address selected topics from these fields and present results of structure function case studies.

11:45 Establishing High Throughput, Low Protein Consuming Biophysical Platform for Biologics Characterization

Villinger_SaskiaSaskia Villinger, PhD, Scientist, Formulation & Process Development / Biologics Drug Product Development, Sanofi R&D

The presentation will cover new biophysical applications for biologics characterization using 1) Second Harmonic Generation (SHG); 2) Nano Differential Scanning Fluorimetry (nano-DSF); and 3) Micro Scale Thermophoresis (MST). It will also discuss the kinetics for thermal stability and activation energy barrier determination for biologics stability assessment, as well as present long-term stability prediction by computational biophysical approaches.

12:15 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

13:15 Dessert Break in the Exhibit Hall with Poster Viewing (Pavilion 1)

14:00 End of Analytical Characterisation of Biotherapeutics

17:00 Dinner Short Course Registration* (Foyer C)

17:3020:30 Dinner Short Courses