Original Agenda
We are actively working with our speakers to confirm their availability for the virtual event. Initial response from our speakers has been very positive, and we are optimistic we will have the new programs ready to share here soon.

Analytical Characterisation of Biotherapeutics track banner

The Analytical Characterisation of Biotherapeutics at PEGS Europe focuses on the challenges of characterizing novel and complex modalities, while developing a toolbox of advanced techniques, coupled with computational approaches and high-throughput analytics to meet the demands for robustness, sensitivity, accuracy and speed.

Wednesday, 11 November

07:45 Registration and Morning Coffee

CHARACTERISING NOVEL BIOTHERAPEUTICS

08:30

Chairperson's Opening Remarks

Christopher Bravery, PhD, Consulting Regulatory Scientist, Consulting on Advanced Biologicals Ltd.
08:35

Challenges in Characterization and Developability Assessments of Multispecific Antibodies

Melanie Fischer, PhD, Section Head Assays and Analytics, Biologics Research, Sanofi

The complexity of multispecific antibodies requires a comprehensive set of analytical techniques to guide lead discovery and optimization. An overview of these techniques will be presented with a focus on mispairing analysis and functional characterization of multispecific drug candidates. Furthermore, the integrated developability concept at Sanofi Biologics will be presented along with show cases highlighting the challenges in characterization and developability of multispecifics.

09:05

Complementing Biophysical Characterization of Antibodies and ADCs with Mass Spectrometry to Assess Higher-Order Structure

Eef Dirksen, PhD, Group Lead, NBE Extended Characterization, Byondis

The safety and efficacy of complex biopharmaceuticals, such as antibodies and antibody-drug conjugates, not only depend on their purity, but also on their higher-order structure (HOS). Consequently, the characterization of molecular structures is of utmost importance throughout the pharmaceutical development trajectory. To investigate the thermal stability of an antibody and its corresponding drug conjugate, a series of orthogonal analytical techniques, both 'classical' spectroscopic, as well as mass spectrometry-based, was applied. In this way, not only the performance of the methods applied could be studied, but the data obtained also provided insight into degradation pathways of the molecules investigated. In addition to providing a feel for the sensitivity, accuracy, linearity, and precision of biophysical, i.e. spectroscopic, characterization methods, the addition of mass spectrometric data potentially reveals the actual regions of the molecules impacted, which is essential information in the process of assessing whether a change in structure is not only significant, but also scientifically relevant. The results will be presented that together paint a comprehensive picture of: a) the performance and orthogonality of the methods routinely used for HOS characterization in the pharmaceutical industry; and b) the HOS of both antibodies and antibody-drug conjugates, how it differs amongst them, and how it responds to heat.

09:35 Development of Novel Native LC-MS Methods and Solutions for the Characterization of mAbs and Related Products
Shunhai Wang, PhD, Senior Staff Scientist, Analytical Chemistry, Regeneron Pharmaceuticals Inc.

Over the past decade, a wide variety of separation methods have been successfully coupled to native MS to characterize the heterogeneity of mAbs and related products. Here, we report the development of several novel native LC-MS techniques and showcase their applications in drug product characterization. In addition, we introduce an integrated native LC-MS solution that offers large dynamic range, high robustness and great versatility.

10:35 Coffee Break in the Exhibit Hall with Poster Viewing
11:15

In Pursuit of Quality Molecules: Early Biophysical Characterization Tools for the Prediction of in vivo Stability

Sathya Venkataramani, PhD, Associate Director, Biophysics, BDS, Janssen Biotherapeutics

Identifying the "Critical Quality Attributes " of a lead mAb candidate early in the research phase is mandatory for selecting "Right the First Time " molecule and avoiding late-stage failures. Among the various intrinsic properties that require in-depth characterization, the top quality attributes of biotherapeutics, such as conformational and serum stability, non-specific binding, and aggregation propensity, need immediate characterization. Integrated biophysical screening of these attributes, in combination with sequence quality assessment, offers a successful predictive tool for selecting risk-free candidates early on. This talk is designed to provide case studies highlighting the criticality of developing such an early tool kit in the discovery phase. 

11:45 Original Case Studies in the Toolbox of Biotherapeutics Analytical Characterization
Elsa Wagner-Rousset, PhD, Senior Research Scientist, Analytical Chemistry & Mass Spectrometry, Center of Immunology Pierre Fabre

The early use in the R&D process of mass spectrometry (MS) and hyphenated methods helps to optimize the structure and function of next-generation of biopharmaceuticals, such as monoclonal antibodies (mAbs) and derivates. Recent development in analytics (MS, CE-SDS) and applications to therapeutics will be highlighted within original case studies. Notably, immunoglobulin isotype gives rise to fingerprint when analyzed with such methods. Glyco-profiling is also a key point when dealing with glycoengineered mAbs.

12:15

FTIR Spectroscopy as an Analytical Tool to Compare Glycosylation in Therapeutic Monoclonal Antibodies

Allison Derenne, PhD, Researcher, Chemistry, Structure and Function of Biological Membrane, Université Libre de Bruxelles

Glycosylation is the most common protein post-translational modification (PTM), especially in biopharmaceuticals. It is a critical quality attribute as it impacts product solubility, stability, half-life, pharmacokinetics and pharmacodynamics (PK/PD), bioactivity, and safety (e.g. immunogenicity). Yet, current glycan analysis methods involve multiple and lengthy sample preparation steps, which can affect the robustness of the analyses. The development of orthogonal, direct and simple method is therefore desirable. In this study, we have developed a method based on Fourier Transform Infrared (FTIR) spectroscopy to analyse glycosylation. We show that FTIR spectra of glycoproteins provide a global, but accurate fingerprint of the glycosylation profile. This fingerprint is not only sensitive to large differences, such as the presence or absence of several monosaccharides, but also to smaller modifications of the glycan and monosaccharide content.


Filip Borgions, Global Head, CMC, argenx BV
13:45 Luncheon Presentation II (Sponsorship Opportunity Available)
14:15 Session Break

VECTOR ANALYTICS FOR CELL & GENE THERAPY

14:30

Chairperson's Remarks

Sonya Schermann, PhD, Director, Analytics, Freeline Therapeutics
14:35 KEYNOTE PRESENTATION: ATMP Potency: Metrological Considerations
Christopher Bravery, PhD, Consulting Regulatory Scientist, Consulting on Advanced Biologicals Ltd.

Establishing suitable release assays for gene, and especially, cell therapies presents a number of unusual challenges. This talk will highlight a few common issues and provide some examples of how these have been addressed so far.

15:05 Analytical Technologies for Determining the Proportion of AAV Capsids Containing Full-Length Vector Genomes
Sonya Schermann, PhD, Director, Analytics, Freeline Therapeutics

A critical quality parameter for all AAV products is the proportion of capsids, which contain the gene of interest in its full length, the so-called ‘full-to-empty ratio’. It will be shown that no single currently available analytical method allows an accurate determination of this parameter. As such, a panel of assays should be used to fully characterise the AAV product. This talk will discuss several of these assays, along with their advantages and possible pitfalls.

15:35 Refreshment Break in the Exhibit Hall with Poster Viewing
16:15 AUC Characterization of Empty, Partial, and Full AAV Particles
Chris Rogers, Senior Scientist, Allergan Biologics
17:15

Development, Transfer and Validation of Analytical Methods for Cell and Gene Therapy Products: Learnings & Challenges

Francesca Rossetti, Analytical Methods Development Manager, MolMed S.P.A.

MolMed has developed an analytical panel, lentiviral platform based, in order to release and characterize vectors and LV genetically modified cell products. All the assays have been studied and optimized in order to achieve the best optimization in term of volume of sample used, time for the analysis, applicability of different projects and GMP suitability according ICH Q2 (R1) guidelines. Each test is developed in order to be suitable for clinical trial application and to be completely validated in case of marketing submission. Increase robustness, accuracy and throughput by automation and decreasing method variability are goals, which are in continuous improvement.

17:45 Networking Reception in the Exhibit Hall with Poster Viewing
18:45 Problem Solving Breakout Discussions*

*Topics to be announced.

19:45 End of Day

Thursday, 12 November

08:00 Registration and Morning Coffee

CUTTING-EDGE TECHNOLOGIES AND NEW APPLICATIONS

08:30

Chairperson's Opening Remarks

Alexander Bepperling, PhD, Lab Head, Biophysical Characterisation II, Technical R&D/Technical Development Biosimilars, Novartis Global Drug Development, c/o Hexal AG
08:35

Exploring the Non-Ideality of High-Concentration Biopharmaceutical Formulations by AUC

Alexander Bepperling, PhD, Lab Head, Biophysical Characterisation II, Technical R&D/Technical Development Biosimilars, Novartis Global Drug Development, c/o Hexal AG

AUC is widely used as an orthogonal technique to confirm the aggregation content routinely measured by size-exclusion chromatography. In order to optimize accuracy and precision, it is typically employed at moderate concentrations around 0.5 mg per ml in a simple PBS-like buffer. The aim is to optimize the sample composition in order to meet the requirements of the technique best. However, the advent of high-concentration formulations of biopharmaceuticals at 100 mg per ml or even higher, and many of them self-buffering, renders this approach questionable. Furthermore, predictions on long-term stability and aggregation propensity are only valid if the intact system, the undiluted protein in its native formulation, is analyzed. The recent development of new fitting algorithms for non-ideal sedimentation in Sedanal and sedfit allows the determination of self- and cross-sedimentation terms, as well as the non-ideality parameters, kS and kD. In combination with 3D printed centerpieces, this allows the accurate description of protein solutions up to 100 mg per ml, and even the discovery of previously unrecognized self-association (Chaturvedi et al. 2019). We have explored the capacity of AUC to accurately quantify the aggregation content of a marketed antibody formulation (Humira) using a model protein mimicking an antibody dimer. We extended this approach on a nano-particle drug, using 1 mm centerpieces to a concentration of 50 mg/ml.

09:05 Development and Application of High-Throughput MAM Approach to Support Process Development of Site-Specific ADC Intermediates
Nick Bond, PhD, Senior Scientist, Analytical & CMC Team Lead, AstraZeneca
09:35

A Novel Approach in Middle-Down Biologics Characterization

Francois Griaud, PhD, Functional Lead & Principal Scientist, Analytical Development & Characterization, Novartis Pharma AG

Top and middle-down biologics characterization approaches aim at capturing a direct snapshot of all proteoforms with their combinatorial distribution. This presentation will focus on a new data analysis workflow to reveal relevant diagnostic information in middle-down MS spectra, missed by the classical middle-down approach. This new workflow enabled the localization of a deamidation event and a sequence variant site directly from the gas-phase fragmentation of a mAb light chain.

Jana Hersch, PhD, Scientific Consultant, Biologics, Genedata

Molecular design, systematic cloning, expression, characterization and the eventual selection of the most promising candidate involve generating and analyzing large amounts of data, which serve as a foundation for informed decision-making. We will present new trends and current approaches in digital data management in the context of bispecific, TCR and CAR-T modalities with a focus on immuno-oncology applications.

10:35 Coffee Break in the Exhibit Hall with Poster Viewing
11:15 From Intact Native MS to Peptide Mapping in Biopharmaceutical Development: Platform Diversity & Synergy, Robustness, Automation of Sample Preparation, and Case Studies
Dan Bach Kristensen, PhD, Principal Scientist, Symphogen

Topics that will be covered in the presentation include: 1) native MS platform robustness, including impact of transferring from one MS platform to another; 2) streamlining of sample preparation, with a focus on simplicity, robustness, and a hands-off approach; and 3) case studies from stability, lead selection, and clone selection studies.

11:45 Exploring the Heterogeneity of Intact Biopharmaceuticals Using Charge-Sensitive Separation Modes Directly Interfaced to Mass Spectrometry
Florian Fuessl, PhD, Postdoc Researcher, Bones Lab, National Institute for Bioprocessing Research & Training (NIBRT)

Recent years have seen great advances in interfacing formerly MS-incompatible separation modes to mass spectrometric detection. In this course, we developed MS-friendly, pH-gradient-based cation- and anion-exchange chromatography methods, which have proven exceptionally powerful for the characterisation of monoclonal antibodies and other protein formats. Additionally, the CE-MS-based ZipChip platform was explored and found to be a highly potent and complementary technique for the sensitive and comprehensive characterisation of complex biopharmaceuticals.

12:15 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
13:15 Dessert Break in the Exhibit Hall with Poster Viewing
14:00 Close of Analytical Characterisation of Biotherapeutics
17:00 Dinner Short Course Registration
17:30 Recommended Dinner Short Course*
SC10: Potency Assay Development for Cell and Gene Therapy Products

*Separate registration required. See short course page for details.