2017 Archived Content

Protein Aggregates and Particles Track Banner


Aggregation impacts biopharmaceutical development at every stage from discovery to development to processing. The mechanisms and kinetics of aggregation, particularly of the novel and complex biomolecules, are not well-defined, thus regulators are highly concerned about its impact on drug product safety and immunogenicity. Companies hence expend great efforts to better understand, predict, identify, characterize, and minimize the occurrence of aggregates and particles. The 4th Annual Protein Aggregates & Particles conference invites scientists to showcase their latest techniques and approaches in this ever challenging field.

Final Agenda

THURSDAY 16 NOVEMBER

12:30 Registration

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

SOURCES AND MECHANISMS OF PROTEIN AGGREGATION

13:30 Chairperson’s Opening Remarks

Paul Dalby, MA, Ph.D., FRSC, AMIChemE, Deputy Head of Department (Research),

Biochemical Engineering, University College London

13:35 Microscopic Mechanisms Underlying the Effect of Surfaces and Mechanical Agitation on Protein Aggregation

Paolo_ArosioPaolo Arosio, Ph.D., Professor, Institute of Chemical and Bioengineering, ETH Zurich

Despite the fact that empirical effects of surfaces and mechanical forces on protein aggregation are well documented, the molecular details of the mechanisms underlying this behaviour are still largely elusive. In this work, we design an experimental assay based on nanoparticles to investigate independently the effect of surfaces and mechanical forces on the formation of amyloid fibrils from human insulin, and we apply a chemical kinetic platform to analyse the molecular mechanisms at the origin of this effect.

14:05 Review on Silicone as Source for Subvisible/Visible Particles and Potential Protein Aggregation Promoter

Karoline_Bechtold-PetersKaroline Bechtold-Peters, Ph.D., Senior Strategy & Technology Leader, Pharmaceutics, Biologics Technical Development & Manufacturing, Novartis

While silicone is toxicologically safe, the use of silicone as a lubricant in packaging materials for injection purposes is associated with the increase of subvisible as well as visible particles and is also described as a source of protein aggregation for biologics. The talk will discuss various published studies and describe analytical methods to characterize silicone layers, which can be used within the context of a control strategy.

14:35 See Stability in Hi-Def with Hunky

Greg Manley, Senior Applications Specialist, Unchained Labs

Developing biologics requires identifying ideal constructs and assessing a wide range of formulation space to ensure stability and minimize aggregation. Assessing ΔG is a powerful approach for the quantitative assessment of conformational stability and aggregation. Unchained Labs automates the tedious and manual task of determining ΔG, allowing for quantitative stability and aggregation assessment throughout biologic development. We'll discuss how automated chemical denaturation can be used with more traditional approaches to assess stability.



15:05 Refreshment Break in the Exhibit Hall with Poster Viewing

QUANTIFYING AND CHARACTERISING AGGREGATES

15:50 KEYNOTE PRESENTATION: Regulatory Perspective in New Analytical Methods for Protein Aggregates Quantification and Characterization

Ewa_MarszalEwa Marszal, Ph.D., CMC Reviewer, Plasma Derivatives Branch, Division of Plasma Protein Therapeutics, Office of Tissues and Advanced Therapies, Center for Biologics Evaluation and Research, US FDA




16:20 A Semi-Quantitative Method for Visible Particles in Biotechnological Solutions – Why Do We Care?

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Pat_CashPatricia Winter Cash, Ph.D., Independent Consultant

Presence of particulates is a common reason for Product Recalls. Formation of protein particles is recognized as an inherent quality attribute. All proteins contain particles. There is an industry-wide need for particle standards so biopharmaceutical manufacturers can monitor and control the presence of particles in their protein products. This talk will explain the need for particle standards, describe one method for estimating particles and introduce new particle standards in development.

16:50 End of Day

17:00 Dinner Short Course Registration

17:30-20:30 Recommended Dinner Short Course*

SC9: Protein Aggregation: Mechanism, Characterisation and Consequences

*Separate registration required

FRIDAY 17 NOVEMBER

08:00 Registration and Morning Coffee

IMPACT OF AGGREGATES ON PRODUCT PROPERTIES

08:30 Chairperson’s Remarks

Vasco Filipe, Ph.D., Unit Head, Formulation Development of Biopharmaceuticals, Sanofi


08:35 KEYNOTE PRESENTATION: Approaches to Predict Protein-Protein Interactions, and Their Impact on Product Properties

Christopher_RobertsChristopher J. Roberts, Ph.D., Professor, Chemical & Biomolecular Engineering, University of Delaware

There is increasing interest in the development of experimental approaches and computational models to predict product properties such as high solution viscosity, solubility limits, and aggregation rates from low- to high-concentration conditions. This presentation focuses on a combination of experimental and modeling approaches to predict protein-protein interactions and their impact on stability and viscosity for monoclonal antibody products, illustrating both successes and outstanding challenges.

09:05 Combining Stability and Surface Aggregation Predictions to Design Protein Solubility

Salvador_VenturaSalvador Ventura, Ph.D., Chair, Professor of Biochemistry & Molecular Biology, Institute of Biotechnology and Biomedicine, Universitat Autonoma de Barcelona

Mutations that increase protein stability usually result in reduced aggregation-propensity. The same effect can be attained with amino acid changes that increase the solubility of structural surfaces. Very often these changes result in decreased protein stability or even a gain in protein aggregation-propensity. AGGRESCAN3D is a software that is able to forecast simultaneously these two effects and to select protein variants with higher probabilities to be evolved into soluble drugs.

09:35 A New Strategy to Assess Ligand or Solution-Based Stability of Aggregation-Prone Proteins Using an Automated Chaperonin Biolayer Interferometry Platform

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Mark_FisherMark T. Fisher, Ph.D., Professor, University of Kansas Medical Center

Stabilizing the folded state of metastable and/or aggregation-prone proteins through ligand binding is an appealing strategy to decrease disease pathologies brought on by deleterious protein folding defects. This presentation describes an automated GroEL chaperonin-based biolayer interferometry (BLI) denaturant-pulse platform for assessing the kinetic stability of aggregation prone proteins and allows us to validate the effects of ligand stabilization or destabilization of these kinetically unstable proteins.

10:05 Coffee Break with Poster Viewing

AGGREGATE-INDUCED IMMUNOGENICITY RISKS

10:35 Probing Protein Aggregation and Immunogenicity by Epitope Profiling

Timothy_EyesTim Eyes, Ph.D., Research Associate, Immunology and Biotechnology, University of Manchester

Aggregation is thought to be a significant risk factor in promoting the unwanted immunogenicity of therapeutic protein products. Our work has examined the relationship between oxidation, aggregation and the influence of heat shock proteins, as prominent HCPs, on the quality and vigor of the immune response to selected model biotherapeutic proteins. More recently we have demonstrated how B-cell epitope profiling can shed light on how antibodies specifically target protein aggregates.

11:05 Aggregation of Monoclonal Antibodies: Causes, Bioavailability and Immunogenicity

Vasco_FilipeVasco Filipe, Ph.D., Unit Head, Formulation Development of Biopharmaceuticals, Sanofi

The mechanisms and factors influencing protein aggregation will be reviewed. Some examples and interesting cases from Sanofi and literature will be shown. A review of the current knowledge and ongoing initiatives on aggregate-induced immunogenicity will be presented.

11:35 Mechanisms Behind Immunogenicity Risk due to Aggregates and Impurities during Biologics Development

Vibha_JawaVibha Jawa, Ph.D., Director, Biologics & Vaccines Bioanalytics, MSD

Human immune system is not exposed to aggregated proteins endogenously. There is an uncertainty around potential risk of aggregates associated with biologics in clinic as there are confounding factors like dose, frequency, route that can also contribute to risk. To address this uncertainty, it is important to identify attributes that can cause a clinically relevant response, establish the threshold of activation of such a response that will support establishing ranges and understand the extent and nature of the response and how close it is to physiological relevance.

12:05 Late Breaking Presentation


12:35 Problem-Solving Breakout Discussions with a Light Snack in the Foyer* (View All Breakout Discussions)

Concerns about Surfactant Application in Protein Drug Products

Moderator: Christoph Grapentin, Ph.D., Postdoctoral Fellow, Pharmaceutical Development & Supplies, PTD Biologics - Europe, F.Hoffmann-La Roche, Ltd.

  • Polysorbate quality, enzymatic degradation and particles from free fatty acids
  • Utility of poloxamer
  • Future alternative surfactants
  • Stabilizing mechanisms of surfactants
  • Interplay of surfactants, proteins and interfaces

Strategies to Evaluate Aggregate-Induced Immunogenicity

Moderator: Vasco Filipe, Ph.D., Unit Head, Formulation Development of Biopharmaceuticals, Sanofi

  • Predictive approaches (in vitro, in vivo, in silico)
  • Which aggregates should we worry about?
  • How many aggregates are too much?
  • Relevance of accelerated stress studies

Reviving Otherwise Hard-to-Stabilize Biopharmaceuticals through Protein Co-Formulation

Moderator: Elenora Cerasoli, Ph.D., Senior Research Scientist, Bioprocess Characterisation , Albumedix

  • What novel formulation strategies and stabilization agents are emerging?
  • Advantages and disadvantages with new stabilization and solubilization agents?
  • Regulatory and clincial challenges with using proteinous excipients in drug formulation?

13:35 Session Break

FORMULATION TO IMPROVE STABILITY AND REDUCE AGGREGATION

14:00 Chairperson’s Remarks

Christopher J. Roberts, Ph.D., Professor, Chemical & Biomolecular Engineering, University of Delaware

14:05 Impact of Protein Mutational Variability upon Formulation Designs

Paul_DalbyPaul Dalby, MA, Ph.D., FRSC, AMIChemE, Deputy Head of Department (Research), Biochemical Engineering, University College London

Protein stability is a critical factor for the successful development of non-aggregating biopharmaceuticals. Routes to predictably engineer protein stability are therefore crucial. We have used a wide range of biophysical analyses to characterise the aggregation landscape of proteins, and used this understanding to inform both better formulation design strategies, and improved protein engineering strategies. The relationships between the conformational stabilities imparted by protein engineering and formulation excipients will be discussed.

Albumedix  14:35 When Standard Biological Drug Formulation Strategies Fail?

Eleonora_CerasoliEleonora Cerasoli, Ph.D., Senior Research Scientist, Bioprocess Characterisation, Albumedix

The expanding field of peptide and protein therapeutics gives promise for improvement of treatments against several diseases. Many of the biopharmaceuticals found to be efficacious, however continue to face ex vivo instability challenges despite advancements in protein and peptide engineering. What novel formulation strategies and stabilization agents are emerging? Advantages and disadvantages with new stabilization and solubilization agents? Regulatory and clincial challenges with using proteinous excipients in drug formulation?

15:05 The Role of Poloxamer 188 in Protecting Protein Formulations

Christoph_GrapetinChristoph Grapentin, Ph.D., Postdoctoral Fellow, Pharmaceutical Development & Supplies, PTD Biologics Europe, (PTDE-P), F. Hoffmann-La Roche, Ltd.

The liabilities of polysorbates have become a hot industry topic recently. Attention has been enhanced to the use of alternative surfactants like poloxamer 188 (P188). The presentation will evaluate a range of state-of-the-art techniques to study the interplay of P188 and proteins at interfaces (air, silicone oil) plus the characteristics of P188 in solution (i.a. dilation rheology, small angle x-ray scattering). Similarities and differences of P188 to polysorbates will be discussed.

15:35 Tackling “Tornado-Like” Particle Formation Issues with an IgG1 Molecule by Formulation Optimisation

Peng_KePeng Ke, Ph.D., Scientist I, Formulation Sciences, MedImmune, a member of the AstraZeneca Group

During formulation development of an IgG1 molecule, “tornado-like” particles were observed in the solutions after storage for a certain period of time. The onset of particle formation was variable and difficult to predict using common analytical techniques. Therefore, significant formulation development challenges were presented and in order to solve the particle issues, systematic investigational works and formulation optimisation were conducted.

16:05 A Rational Strategy for the Reduction of Protein Aggregation in Mammalian Cell Cultures

Friedemann_HesseFriedemann Hesse, Ph.D., Professor, Institute for Applied Biotechnology, Biberach University of Applied Sciences

Although protein aggregation is a well-known quality issue in the production of recombinant biopharmaceuticals which can already occur in early stages of the manufacturing process, only few studies address protein aggregation during upstream processing. Therefore, this study was performed in order to establish a rational approach for the optimization of upstream bioprocesses to reduce protein aggregation in mammalian cell cultures.

16:35 End of Conference