The phenomenon of protein aggregation has perplexed many scientists. All mechanisms of aggregation are not conclusively known, while sources of aggregation continue to surface. Regulatory authorities put great emphasis on the identification and characterisation of these aggregates, particles and impurities due to their implications on product safety, efficacy and immunogenicity risks; thus, companies are constantly on the lookout for better and more robust ways to detect and mitigate these aggregates.

PEGS Europe’s 5^th Annual Protein Aggregates & Particles brings the latest understanding on mechanisms and sources of aggregation, strategies to mitigate aggregation during engineering and process development, as well as technologies and approaches to characterise and control these impurities.

Final Agenda

Day 1 | Day 2

THURSDAY 15 NOVEMBER

13:00 Registration

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

MECHANISMS OF AND IMPLICATIONS FOR PARTICLE FORMATION

14:00 Chairperson’s Opening Remarks

Christian Schoneich, PhD, Professor, Pharmaceutical Chemistry, University of Kansas

14:05 Regulatory Considerations for Impurity Characterization and Control for Biological Products

Audrey Jia, PhD, Principal Consultant, CMC, DataRevive LLC

Impurity characterization and impact to the product quality is important for both novel biologic and biosimilar development. To what extent the impurity needs to be characterized and analyzed to satisfy the regulatory need will be discussed.

14:35 Factors Influencing Biotherapeutic mAb Aggregation

Linda Yi, PhD, Senior Scientist, Analytical Development, Biogen

Aggregation has been identified as one of the major degradation pathways that may affect safety, quality and efficacy of therapeutic mAbs. Aggregate present in mAb products can be complex, varying by size, type and origin, with underlying mechanisms not always being well-understood. This presentation will provide an overview of the factors that may influence biotherapeutic mAb aggregation. A case study will follow on impact of a chemical modification catalyzed by metals on aggregation of mAbs.

15:05 Presentation to be Announced

15:35 Networking Refreshment Break

16:00 KEYNOTE PRESENTATION: Protein Gelation at Interfaces: Implications for Aggregation and Particle Formation

Theodore W. Randolph, PhD, Kenneth and Genevieve Gillespie Professor, Department of Chemical and Biological Engineering, University of Colorado

16:30 Weak, Promiscuous IgG Self- and Hetero-Association: Implications for Aggregation and Viscosity

Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire

The interactions between seven fluorescently labelled tracer IgG mAbs differing in V and C regions were detected and characterized by analytical ultracentrifugation. All interactions were found to be attractive, variable in strength, affected by both the variable and constant regions, but indiscriminate with respect to IgG subclass. Furthermore, weak attractive interactions were observed for all the mAbs with freshly purified human poly-IgG. The universality of the weak interactions suggests that they may contribute to effector function cooperativity in the normal immune response, and could be important contributors to viscosity.

17:00 End of Day

Day 1 | Day 2

17:00 Dinner Short Course Registration*

17:30 – 20:30 Dinner Short Courses

Recommended Short Course*

SC7: Protein Aggregation: Mechanism, Characterization and Consequences

Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire

Protein aggregation is recognized by regulatory agencies and the biopharmaceutical industry as a key quality attribute of biotherapeutic products. Various aggregates hold the potential for adversely impacting production and patients in a variety of ways. This in-depth workshop reviews the origins and consequences of aggregation in biotherapeutics, and then examines strategies for predicting and quantifying aggregation in biopharmaceuticals. It benefits scientists engaged in development, production, analytical characterization and approval of biotherapeutics and who require a good working knowledge of protein aggregation.

*Separate registration required.

FRIDAY 16 NOVEMBER

08:00 Registration and Morning Coffee

AGGREGATE DETECTION, QUANTIFICATION AND CONTROL

08:30 Chairperson’s Remarks

Joël Richard, PhD, Senior Vice President, Pharmaceutical Development – Peptides, R&D, Ipsen

08:35 Detecting and Separating Sub-Visible Protein Aggregates from Other Particulates

Alain Pluen, PhD, FHEA, Lecturer, Director, BSc Pharma Science, Division of Pharmacy and Optometry, School of Health Sciences, University of Manchester

09:05 A New Alternative to Conventional Aggregate Quantitation and Sizing

Oliver Bannach, PhD, CEO, Attyloid GmbH

According to a recent FDA guideline, subvisible aggregates ranging from 0.1-1 µm are of special concern regarding immunogenicity of biologicals. The FDA further emphasizes the need for quantitative methods covering this size range. sFIDA fills this gap by quantitating aggregates from 10 nm to 50 µm. sFIDA is a novel platform technology for quantitation and sizing of protein aggregates. The technology combines the selectivity of immunological assays with the sensitivity of high-resolution fluorescence microscopy.

09:35 Scale-Down Models for Freezing/Thawing Process to Control Aggregation

Karoline Bechtold-Peters, PhD, Senior Strategy and Technology Leader, Biologics Technical Development & Manufacturing, Novartis Pharma AG

10:05 Networking Coffee Break

10:35 Development Strategy of Fibril-Prone Peptide Therapeutics: Aggregation Kinetics, Predictive Methods, and Detection Methods

Jingtao Zhang, PhD, Principal Scientist, Pharmaceutical Sciences, Merck & Co.

Peptide aggregation such as fibrillation presents significant challenges for DS and DP development of peptide therapeutics. Different development criteria and control strategy is required for fibril development in contrast to protein aggregation. Approaches to close gaps in these areas will be shared in the presentation, which includes the investigation on the aggregation kinetics of a fibril-prone peptide, the projection of physical stability shelf-life, and the development of highly sensitive characterization methods for fibrils.

11:05 Capreomycin Inhibits the Initiation of Amyloid Fibrillation and Suppresses Amyloid Induced Cell Toxicity

Rizwan Khan, PhD, Professor, Interdisciplinary Biotechnology Unit, Aligarh Muslim University

Protein aggregation and amyloid fibrillation are responsible for several serious pathological conditions (like type II diabetes, Alzheimer’s and Parkinson’s diseases, etc.) and protein drugs ineffectiveness. Therefore, a molecule that can inhibit the amyloid fibrillation and potentially clear amyloid fibrils is of great therapeutic value. In this manuscript, we investigated the antiamyloidogenic, fibril disaggregating, as well as cell protective effect of an anti-tuberculosis drug, Capreomycin (CN).

11:35 Sponsored Presentation (Opportunity Available)

11:50 New Tools to Anticipate and Prevent Protein Aggregation Undergoing Freeze-Thaw

Miguel Rodrigues, Professor, Co-Founder, SmartFreez

12:05 Problem-Solving Breakout Discussions with a Light Snack

PREDICTIONS OF AGGREGATION PROPENSITY

13:00 Chairperson’s Remarks

Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire

13:05 Using Chemical Denaturants to Improve Predictions of Biopharmaceutical Aggregation during Storage

Robin Curtis, PhD, Senior Lecturer, Chemical Engineering and Analytical Science, University of Manchester

Here we show how to account for the high concentration of denaturants in the data analysis, which if unaccounted for, could lead to data misinterpretation. We then correlate light scattering measurements of five monoclonal antibodies at high denaturant concentration with their shelf stability. The results indicate there is a window of denaturant concentration over which the self association behaviour provides an indicator for the aggregation propensity upon storage.

13:35 Prediction of Aggregation Propensity at Early Development Stage Using Orthogonal Characterization Methods

Joël Richard, PhD, Senior Vice President, Pharmaceutical Development – Peptides, R&D, Ipsen

In the perspective of accelerating early stage protein formulation development, it has become key to anticipate and predict formulation, storage and processing conditions that will lead to aggregation. To anticipate aggregation propensity, evaluate aggregation rate under critical conditions and mitigate associated immunogenicity risks, it is proposed to focus on the very early steps of aggregation, often involving higher order structure (HOS) alterations and loss of colloidal stability, using a set of orthogonal characterization techniques.

14:05 Using Physics-Based Simulations to Understand the Determinants of Viscosity in Concentrated Antibody Solutions

Saeed Izadi, PhD, Scientist, Early Stage Pharmaceutical Development, Genentech, Inc.

We have developed a physics-based multi-scale coarse-grained approach to explore how high viscosity can emerge from weak self-interactions. Key developments that enabled this approach are discussed, including our novel strategies to preserve electrostatic fields and hydrophobicity features. Our method improves on currently available approaches, evidenced by its accuracy when compared against experimental data, along with providing unexpected insights into the problem that enable consideration of mitigation strategies before material is available for testing.

DEGRADATION MONITORING FOR STABILITY AND FORMULATION DEVELOPMENT

14:35 Stability of Protein Disulfides: Sensitive Targets for Oxidation and Light-Induced Degradation

Christian Schoneich, PhD, Professor, Pharmaceutical Chemistry, University of Kansas

Protein disulfides are important for the structural integrity of proteins. Nevertheless, they are sensitive targets for multiple pathways of protein degradation, where specific degradation products have been detected in immunogenic protein samples. Hence, a thorough characterization of disulfide degradation pathways of biotherapeutics presents an important task. Recently, we detected about 60 different degradation products resulting from the light-induced degradation of a small protein, human growth hormone, and significantly more products may be expected from the degradation of monoclonal antibodies.

15:05 Quantification and Degradation Monitoring of PS80 in One Single Analysis Using QDa Mass Detector

Pierre Guibal, PhD, Deputy Head, Analytical Development, BioAnalytics, Sanofi

Different methods already exist for PS80 monitoring, but to our knowledge, no method is able to quantify intact PS80 and monitor its potential degradation – oxidation or hydrolysis – in one single analysis. We present here a new chromatographic method based on ion source versatility of QDa, a single quadrupole mass detector. Using specific single ion recording signals, this is the first method allowing quantification of intact PS80 and covering all known degradation in one single analysis.

15:35 Protein-Excipient Interactions Evaluated via NMR Studies in Polysorbate-Based Multi-Dose Protein Formulations: Influence on Antimicrobial Efficacy and Potential Study Approach

Riccardo Torosantucci, PhD, Lab Head, Formulation Development, Pharmaceutical Development Biologics, Sanofi-Aventis Deutschland GmbH

Preservatives are excipients needed in biopharmaceutical multi-dose formulations to prevent microbial growth; however, they are known to interact with non-ionic surfactants like polysorbate and potentially with the active pharmaceutical ingredient (API). In the current study those interactions were successfully quantified via NMR and correlated to the antimicrobial activity of the formulations. NMR represents therefore a powerful tool to support formulation development of multi-dose formulations.

16:05 End of Summit

Day 1 | Day 2

* The program is subject to change without notice, due to unforeseen reason.