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.

Protein Stability & Aggregation track banner

The Protein Aggregation & Stability conference at PEGS Europe not only examines the interfaces and protein interactions on aggregation & stability, but also discusses new developments in techniques to predict and study stability and aggregation and their chemical modifications on protein function. The meeting will also include case studies on formulation and purification strategies to lower aggregation propensity and achieve higher purity.

Thursday, 12 November

13:00 Registration
13:15 Dessert Break in the Exhibit Hall with Poster Viewing

STABILITY TESTING FOR AGGREGATES/IMPURITIES CONTROL

14:00

Chairperson's Opening Remarks

Robin Curtis, PhD, Senior Lecturer, University of Manchester
14:05 Identification of HCPs Inducing Particle Formation during Protein Stability Testing
Veronika Reisinger, PhD, Lab Head, Physico Chemical Characterization, Novartis AG

Residual HCPs can impact product quality in different ways. Besides effects on patient safety, product stability might be affected. Here, we present workflows based on LCMS to identify HCPs inducing particle formation. LCMS is the method of choice for identification of unknown HCPs as ELISA is usually not capable to identify individual HCPs. In addition to HCP identification, LCMS allows the relative and absolute identification of single HCPs.

14:35

Measuring Colloidal Stability of Partially Folded scFv Proteins and the Impact on Aggregation

Robin Curtis, PhD, Senior Lecturer, University of Manchester

We present key insights into protein structural factors controlling aggregation behaviour for scFv mutants. Aggregate growth rates are controlled by non-specific electrostatic interactions, while increasing the scFv lysine to arginine content lowers aggregation by preventing partially unfolded regions from associating. We show protein-protein interaction measurements made under chemically denaturing conditions reflect colloidal stability of scFv partially unfolded states, which play key roles in aggregation pathways.

Rick Gordon, Vice President, Sales, Halo Labs

Distinguishing aggregated API from other particle types is important for understanding the root cause of instability. Existing methods are unreliable, too cumbersome and difficult to use in many workflows. With Aura, you can now finally count, size, and characterize aggregates and identify them as proteins, non-proteins, or other molecules.

15:20 Sponsored Presentation (Opportunity Available)
15:35 Networking Refreshment Break
16:00

New Developments in the Use of Synchrotron Techniques for Studying Protein Stability

Ulla Elofsson, PhD, Associate Professor & Senior Scientist, Health and Life Science, Chemical Process and Pharmaceutical Development, RISE Research Institutes of Sweden AB

Synchrotron techniques are increasingly applied in pharmaceutical development. The synchrotron source MAX IV offers new possibilities by means of faster measurements and high brilliance. CoSAXS beamline is designed for studying formulation effects on protein solution structure and interactions using automated sample handling and data analysis. Examples on how synchrotron small angle X-ray techniques can be used, and suggestions on how the new developments at CoSAXS can contribute will be presented.

16:30

Small-Angle Neutron and X-Ray Scattering: Emerging Biophysical Tools for Frozen and Freeze-Dried Biologicals

Evgenyi Y. Shalaev, PhD, Research Investigator, Pharmaceutical Development, Allergan, Inc.

Essentially all biopharmaceutical drug substances and drug product are exposed to freeze-thaw and/or freeze-drying/reconstitution. Recently, frozen and freeze-dried proteins with various excipients (surfactants and lyoprotectors) are studied using small-angle neutron scattering (SANS) and small- and wide-angle X-ray scattering (SAXS/WAXS). Both protein-protein interaction and various crystalline and liquid-crystalline phases of excipients are monitored. SANS and SAXS/WAXS represent valuable orthogonal tools to study mechanisms of protein (de)stabilization during freeze-thaw and freeze-drying.

17:00 End of Day
17:00 Dinner Short Course Registration
17:30 Recommended Dinner Short Course*
SC9: Developing Stable Bioformulations: Predicting and Controlling Protein Aggregation

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

Friday, 13 November

08:00 Registration and Morning Coffee

INTERFACES AND ITS IMPACT ON STABILITY

08:30

Chairperson's Opening Remarks

Karoline B. Bechtold-Peters, PhD, Senior Strategy & Technology Leader, Pharmaceuticals & Biopharma Process, Novartis Pharma AG
08:35 KEYNOTE PRESENTATION: Synergistic Effect of Hydrodynamic Flow and Interfaces on Antibody Aggregation
Paolo Arosio, PhD, Assistant Professor, Chemistry & Applied Biosciences, ETH Zurich

We present our efforts towards the development of small-scale and high-throughput assays for the investigation of the synergistic effect of interfaces and hydrodynamic flow on protein aggregation. Our assays, largely based on microfluidic technology, exhibit an accurate control of interfaces and flow stresses, and pave a way to develop methods for the evaluation of antibody stability against interfaces and hydrodynamic flows, both during early-stage screening and during bioprocessing.

09:05 They Must Be Compatible: Mutual Interaction Phenomena between Protein Formulation and Siliconized/Silicone-Free Syringe Packaging Materials with an Effect on Functionality and Stability
Karoline B. Bechtold-Peters, PhD, Senior Strategy & Technology Leader, Pharmaceuticals & Biopharma Process, Novartis Pharma AG

FIRST DATA/TOP POSTERS HIGHLIGHT

09:35 A Nanoparticle-Based Assay to Evaluate Surface-Mediated Protein Instability in Developability Studies
Marie Kopp, Graduate Student, Biochemical Engineering, ETH Zurich

The physical stability of therapeutic proteins during bioprocessing and formulation is a crucial property to guarantee their safety and efficacy. Air-water and solid-liquid interfaces are well known to potentially trigger protein instability and aggregation. Challenges in the investigation of interface-induced protein aggregation include the control of the amount and type of surfaces, as well as the presence of synergistic effects between interfaces and hydrodynamic flows. Here, we present a newly developed surface stress assay based on polymeric nanoparticles, which could complement developability studies in early-stage development.

09:50 Development and Application of Screening Assays to Predict Aggregation upon Long-Term Storage
Fabian Dingfelder, PhD, Industrial Postdoc, Biophysics, Novo Nordisk AS

Protein aggregation remains a challenge for the development of biopharmaceuticals, and currently there are no screening assays validated to predict aggregation upon long-term storage. Identifying predictive assays would be an important advancement to guide the design and selection of druggable candidates in screening campaigns. In this talk, I will show how we probe multiple biophysical properties of different antibody formats to calculate correlations with the aggregation propensity upon long-term storage.

10:05 Networking Coffee Break

IMPLICATIONS OF AGGREGATION AND CHEMICAL MODIFICATIONS ON PROTEIN FUNCTION

10:35 Evolutionary Imprints of Protein Aggregation: Implications for Protein Engineering and Redesign
Frederic Rousseau, PhD, Principal Investigator, Switch Laboratory, VIB-KU Leuven Center for Brain & Disease Research

I will discuss the kinetic and thermodynamic relationship between globular and amyloid protein structure and how this shaped the evolution of protein sequences and their recognition by molecular chaperones. I will also discuss the implications of these findings for protein engineering and redesign.

11:05

Chemical Modifications in Biotherapuetic Proteins and Their Impact on HOS and Function

Sambit R. Kar, PhD, Principal Scientist & Head, Biophysics Center of Excellence, Bristol Myers Squibb Co.
11:35 Sponsored Presentation (Opportunity Available) OR Panel Discussion
12:05 Problem Solving Breakout Discussions with Light Snack*

*Topics to be announced.

PURIFICATION AND FORMULATION DESIGN FOR HIGHER PURITY AND LOWER AGGREGATION PROPENSITY

13:00

Chairperson's Remarks

Michael Siedler, PhD, Section Head, NBE Formulation Sciences & Process Development, Abbvie Deutschland GmbH & Co. KG
13:05 Challenges in Mutein IL2 Purification Process: How to Control the Aggregation and Misfolding Processes
Kathya Rashida de la Luz Hernandez, PhD, Head, Analytical, Center of Molecular Immunology

A mutant of the interleukin-2 molecule without interaction with the alpha chain of the IL-2 receptor was designed at the CIM. This new mutant shows higher antitumor activity and less toxicity than human IL-2, that is why it is a better candidate for human cancer therapy. In order to improve the quality of the final protein some of the purification steps and conditions were modified. The proposed purification process allowed obtaining a consistent purified product, with similar physical-chemical and biological properties to the product, purified with the current process and higher quality.

13:35

Using a High-Throughput Formulation Robustness Screening to Establish the Formulation Design Space for a Monoclonal Antibody – A Case Study

Michael Siedler, PhD, Section Head, NBE Formulation Sciences & Process Development, Abbvie Deutschland GmbH & Co. KG

The presentation will provide an example on how quality by design can be implemented in formulation development especially for demonstrating formulation robustness. Topics explored include the use of down-scale models for identifying critical (formulation) parameters and define the formulation design space; justification of utilized down-scale models and verification of results, and the impact on setting drug product specifications.

14:05 Assessing Refoldability to Select Therapeutic Proteins and Formulations with Lower Aggregation Propensity during Storage
Hristo Svilenov, PhD, Researcher, Pharmacy, Technical University Munich

Protein refoldability is an essential feature of therapeutic protein formulations with lower aggregation tendency during storage. I will introduce you to two approaches that allow the assessment of protein refoldability after unfolding caused by heat or chemical denaturants. Further, I will demonstrate how the presented methods can be useful for the selection of protein molecules with lower aggregation propensity and formulations that impede the formation of aggregates during storage.

14:35

From Formulation Screening to Early Manufacturing

Bernhard Valldorf, PhD, Principal Scientist & Lab Head, Formulation Development, EMD Serono

In the process of developing biotherapeutics early developability and manufacturability assessment is crucial to select the best candidate for the CMC phase. Therefore we focus on miniaturized screening approaches which allow us to predict stability and rank lead candidates for further development. This talk will give an overview of our formulation screening cascade and show its applicability in a case study.

15:05

Long Acting Injectables of Fragile Molecules: Opportunities from New Technologies for the Delivery of Small Bispecific Antibodies

Joel Richard, PhD, Chief Development Officer, MedinCell SA

BEPO® in situ implant forming (ISIF) technology is based on the association of amphiphilic block poly (lactic acid) (PLA) - poly (ethylene glycol) (PEG) copolymers and provides a unique combination for ease of administration, stability of fragile molecules and in vivo control of the drug release rate and duration of action after injection. This talk will provide insight into the many advantages of this new, highly versatile, long-acting injectable formulation technology and present the promising results obtained for the controlled delivery of a small, bispecific short half-life antibody for immunotherapy in prostate cancer. The antibody remains functional and active through formulation and release processes and demonstrates high anti-tumor activity in animal models, while improving subcutaneous bioavailability and half-life of the biomolecule.

15:35 Close of Summit