The Protein Aggregation & Stability conference at PEGS Europe seeks to highlight the importance in prediction of aggregation propensity affecting stability of solutions and formulations, while exploring new and advanced technologies, strategies and
device design to characterize, control and mitigate aggregation.
Final Agenda
THURSDAY 21 NOVEMBER
13:00 Registration (Foyer A)
13:15 Dessert Break in the Exhibit Hall with Poster Viewing (Rio Pavilion)
14:00 Chairperson’s Opening Remarks
Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire
14:05 KEYNOTE PRESENTATION: Closing the Analytical 0.1-to-2 Micron-Size Gap: Why, When, and How?
Wim Jiskoot, PhD, Professor, BioTherapeutics, Leiden
University
Protein aggregates and other particulate impurities are critical quality attributes of biopharmaceutical drug products. Routine analytical methods, such as size-exclusion chromatography and light obscuration, are blind for aggregates and particles in
the size range between about 0.1 and 2 micrometers. In this presentation, I will explain why it is important to characterize and quantify such “gap range” species, present trends in the development of analytical tools that cover this range,
and discuss how these tools could be applied in product development.
14:35 FEATURED PRESENTATION: Final Container Specifications for Subvisible Particulate Matter in Therapeutic Protein Injections
Ewa Marszal, PhD, Regulatory
Review Scientist, Division of Plasma Protein Therapeutics, Office of Tissues and Advanced Therapies, CBER, FDA
While ICH Q6B guidance recommends setting specifications based on data from preclinical and clinical trials and from lots used for demonstration of manufacturing consistency and product stability, the manufacturers tend to set specifications for subvisible
particulate matter in biologics using as an analytical method light obscuration and the USP <787> and <788> limits. However, these limits were not developed for proteinaceous particulate matter and they are not supported with safety data.
In this presentation, I will remind the history of the USP limits for subvisible particulate matter and will argue that other methods and product-specific limits will provide a better assurance of product safety, efficacy, and manufacturing consistency.
15:05 Are Particulates Hiding in Your Formulation?
Rick Gordon, Vice President, Sales, Halo Labs
See how the HORIZON system from Halo Labs uses Backgrounded Membrane Imaging (BMI) to measure subvisible particles, including translucent protein aggregates to help predict protein drug stability during early stage formulation development. The measurement
is fully automated (up to 96 samples) and uses 1/10th the volume of other techniques.
15:20 Developability Assessment and Property Prediction by pH-Dependent Conformational Sampling
Nels Thorsteinson, PhD, Scientific Services Manager, Biologics, Chemical Computing Group
mAb candidates identified from high-throughput screening or binding affinity optimization often present liabilities for developability, such as aggregation-prone regions or poor solution behavior. In this work, we developed a method for modeling proteins
and performing pH-dependent conformational sampling, which can enhance property calculations such as hydrophobic patches, charge and pI. A retrospective data analysis demonstrates that these 3D descriptors, averaged over conformational sampling and
stochastic titration, can accurately predict pI values, screen candidates and enrich libraries with favorable developability properties for a range of biotherapeutics. The clinical landscape of antibodies is also analyzed and its property profile
and insights thereof are presented.
15:35 Networking Refreshment Break (Foyer D)
16:00 New Design of a Blast Freezer Thawer to Minimize Freeze-Thaw Associated Protein Aggregation
Karoline Bechtold-Peters, PhD, Senior Strategy and Technology Leader, Biologics Technical Development & Manufacturing, Novartis Pharma AG
Freezing and thawing biologics drug substance is an important process step that can lead to protein destabilization and aggregation due to various, phenomena such as freeze concentration and interaction at the large ice-liquid interface. We found that
the commercially available blast or shock freezers for DS bottles are not ideal because they are not working in a sufficiently homogeneous and powerful mode and are also not suitable for thawing. We have developed and further optimized an improved
device to quickly freeze and thaw sensitive therapeutic proteins. The design and process results will be presented in the talk.
16:30 How Industry Handles Aggregate and Particle Issues – What Happens to Your Product Once It’s Out of Your Control?
Christina
Vessely, PhD, RAC, Senior Consultant, Analytical and Formulation Development, Biologics Consulting
Protein aggregates, as well as other particulate matter, have been definitively linked to immunogenicity and adverse outcomes in patients. As formulation and analytical scientists, we work very hard to ensure that the products we are sending out to clinics
are of sufficient quality to maintain safety to patients. But what happens when those materials are no longer in our control? The purpose of this presentation is to inform sponsors as to what can occur between the time that you put your product into
a vial and the time it is actually dosed to patients, as well as how to mitigate the potential issues thereby maintaining patient safety and product quality/efficacy.
17:00 End of Day
17:00 Dinner Short Course Registration*
17:30 – 20:30 Dinner Short Courses
Recommended Short Course*
SC7: Protein Aggregation: Mechanism, Characterization and Consequences - LEARN MORE
*Separate registration required.
FRIDAY 22 NOVEMBER
08:00 Registration (Foyer A) and Morning Coffee (Foyer D)
08:30 Chairperson’s Remarks
Karoline Bechtold-Peters, PhD, Senior Strategy and Technology Leader, Biologics Technical Development & Manufacturing, Novartis Pharma AG
08:35 Progress in High Throughput Biophysical Characterization Approaches to Predict Long-Term Physical Stability of Protein Drugs in Pharmaceutical Formulations
Gerhard Winter,
PhD, Professor, Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-Universität
Finding the most suitable conditions which allow to minimize protein degradation during long-term storage is still a challenging task. We discuss the application of DSC 2, DSF 4, MST 8, ICD 3,5,6 and a new technique called TOPC 7, and the usefulness of
determining the apparent protein-melting temperatures to select protein formulations with high physical stability. Finally, we show how the assessment of the aggregation of partially folded species during refolding can provide additional information
for the selection of protein formulations with high physical stability during storage. The talk will conclude with general suggestions on how to select solution conditions that impede aggregation.
09:05 Different Grades of Polysorbate for Biopharmaceutical Products – Comparison of Their Degradation Propensity and Evaluation of Their Functional Properties
Klaus Wuchner, PhD,
Scientific Director, DPDS/BioTD-Analytical Development, Janssen R&D, Cilag AG
Polysorbate 20 (PS20) and polysorbate 80 (PS80) grades compliant with major pharmacopeias (EP, USP, JP) are the most widely used surfactants in biopharmaceutical products. Chinese pharmacopoeia (ChP) requires an oleic acid content of ≥98.0%. However,
still little is known about the stability and functional properties of these “higher” purity-grade PS in biopharmaceutical formulations. This talk will provide some insight into degradation behavior of different PS grades, present novel
markers for oxidation, and compare the functional properties with respect to stabilizing protein against interfacial stress.
09:35 Thermodynamic Prediction of the Concentration-Dependence of Protein Solutions
Thomas Laue, PhD, Professor
Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire
Prediction of the high-concentration behavior of biotherapeutics is of interest to their development and formulation. Approximations of solution thermodynamics are shown that allow a first-order prediction of protein behavior at high-concentrations. Though
not exact, the predictions provide a useful guide to how protein and solvent characteristics impact solution behavior.
10:05 Networking Coffee Break (Foyer D)
10:35 A Mechanistic Approach for the Assessment of Protein Aggregation Propensity in Therapeutic Proteins: Practical Application in Biopharmaceutical Drug Candidate Selection and Pre-Formulation Development
Danny K. Chou, PharmD, PhD, President, Compassion BioSolution, LLC
In recent years, our understanding of the fundamental mechanisms of protein aggregation has increased significantly. To identify the most stable and manufacturable drug candidates and develop the most robust formulation for such molecules, the logical
approach is to take advantage of the current knowledge and make it the foundation of a stability assessment/formulation development plan. This presentation will show how one can implement such an approach to successfully evaluate stability of protein
pharmaceuticals and develop a suitable formulation in a rapid manner.
11:05 Understanding Protein Aggregation via Computational Means
Sandeep Kumar, PhD, Senior Research Fellow, Biotherapeutics, Boehringer-Ingelheim Pharmaceuticals
- Protein folding and aggregation
- Aggregation process
- Aggregation prone regions in thermophiles, human proteome and biotherapeutics
- Benefits of aggregation mitigation
- Prediction of change in aggregation rate upon mutation
- Overall trends
11:35 Prediction and Measurement in Viscosity
NEW: Virginie Le Brun, PhD, Senior Principal Scientist, Drug Product Services, Lonza Pharma & Biotech
12:05 Problem-Solving Breakout Discussions with a Light Snack*
TABLE 29 Particle Analytics – Advanced Instrumentation and Data Modeling
Moderators:
Jonas Hoeg Thygesen, PhD, Area Specialist, R&D Microanalysis Centre, Novo Nordisk Pharmatech A/S, and
Anacelia Rios Quiroz, PhD, Scientist, Group Leader, Particle Lab, Pharma Technical Development Biologics Europe, Hoffmann-La Roche
- What are the current analytical methods available for identification and characterization of particles in biopharmaceuticals?
- Orthogonal control strategies for visible particles
- What are the advantages and challenges with the methods? Possibilities for automation/high-throughput?
- What are the challenges with analysis of sub-visual vs. visual particles
- What data analytical tools may be used?
- May deep learning and machine learning be used for particle identification? If so what are the challenges and advantages by these methods?
TABLE 30: The New Challenges of Developing Formulations for Advanced Therapy Medicinal Products (ATMPs)
Moderator: Danny K Chou, PharmD, PhD, President, Compassion BioSolution, LLC
- What are some of the new types of therapeutic modalities that just arrived on the market or are on the horizon that are different from monoclonal antibodies?
- What types of challenges will we face when developing stable/efficacious dosage forms for these new types of therapy?
- What types of analytical challenges will ATMPs bring to the industry?
- What are some of the known critical quality attributes associated with each class of APMTs (i.e., Gene therapy, MRNA, etc).
- How can one prepare for the arrival of ATMPs from the pharmaceutical development point of view?
TABLE 31: Polysorbate in Bipharmaceutical Products – Current Concerns and Future State
Moderator: Klaus Wuchner, PhD, Scientific Director, DPDS/BioTD-Analytical Development, Janssen R&D, Cilag AG
- Experience with different grades/purities of polysorbate (Do you set specific requirements for PS as raw material for biothech products)?
- Chinese pharmacopoeia (ChP) requires an oleic acid content of ≥98.0% in PS80; do you consider switching to this grade or to other higher purity PS (e.g., all laureate PS20)? Any experience with respect to their stability, impact on protein stability?
- Do you consider alternatives to PS?
- How do you handle Polysorbate from receipt to processing?
- What are currently your main issues related to Polysorbate in biopharmaceutical products? (e.g., particle formation, induction of oxidation of protein….)
- Did you develop/implement successful mitigation strategies for PS degradation?
- Which heath authority requirements/questions/concerns related to PS did you face?
13:00 Chairperson’s Remarks
Anacelia Rios Quiroz, PhD, Scientist, Group Leader, Particle Lab, Pharma Technical Development Biologics Europe, Hoffmann-La Roche
13:05 Chemometrics and Advanced Data Analytics for Particle Analysis
Jonas
Hoeg Thygesen, PhD, Principal Scientist, R&D Microanalysis, Novo Nordisk Pharmatech A/S
Regulatory agencies call for identification and characterization of any intrinsic, inherent, or extrinsic particles present in pharmaceuticals. Of the many tools in the analytical toolbox for particulate/foreign material identification, the methods of
Fourier-Transform Infrared (FTIR) microscopy and Energy Dispersive X-ray Spectroscopy (EDS) have developed into industry standard workhorses. This presentation will highlight how chemometrics and advanced data analytics may be used to gain more insight
from such analytical data collected during particle analysis.
13:35 Monitoring and Characterizing Aggregation Variants in Co-Formulated Biologic Products: Utilizing 2D Chromatography to Assess Aggregation
Mark Anthony Haverick,
Associate Principal Scientist, Biologics Anaytical R&D, MSD
Co-formulated drug products are currently being developed though the combination of multiple mAbs in a single vial. Coformulation of mAbs enables simplified dosing, better production and easier handling. Monitoring aggregation in co-formulated mAbs poses
several challenges due to the similar size and shape of the individual mAbs. This requires a more sophisticated “analytical toolkit” to characterize the aggregation. During this presentation, we discuss the analytical control strategy
for characterization of aggregation in co-formulated drug products, and the importance of understanding method performance for purity. Additionally, we will also discuss the application of two-dimensional liquid chromatography to understand aggregation
and improve product knowledge.
14:05 NEW: sFIDA: A General Method for Detection and Quantitation of Protein Aggregates with Single-Particle Sensitivity for Quality Control of Biologics
Oliver Bannach, PhD, Senior Scientist, Structural Biochemistry, Forschungszentrum Jülich
We have developed sFIDA as a general tool for detection and quantitation of protein aggregates with single-particle sensitivity and total insensitivity to non-aggregated protein. Thus, it allows the determination of the concentration and sizes of protein
aggregate particles in any medium without pre-treatment. The method is fully automated and adaptable to any specific protein. We will demonstrate the technology and give some examples for its successful application.
14:35 Solution NMR Assessments of Therapeutic Protein Behavior
Mark McCoy, PhD, Principal Scientist, Discovery Chemistry – Screening, Target and Compound Profiling, MSD
Solution NMR spectroscopy provides detailed assessments of therapeutic peptide and protein behavior. Structural fingerprints capture solution structure, conformation, and probe for site-specific interactions at atomic resolution. Additionally, diffusion
and dynamic profiling methods are used to understand self-association, assembly, aggregation, and the impact of sequence and formulation on molecular motions and interactions. Examples will be drawn from discovery and development applications that
include higher-order structure characterization, developability assessments, formulation, and co-formulation studies.
15:05 Characterization of Subvisible Particles: Old Challenges and Newest Improvements
Anacelia Rios Quiroz, PhD, Scientist, Group Leader, Particle Lab, Pharma Technical Development Biologics Europe, Hoffmann-La Roche
The talk will give an overview on commercially-available counting methodologies for detection of sub visible particles (SbVP). This species, ubiquitously present in protein formulations, had been in focus due to immunogenicity and quality attributes of
biotechnological products. Thus, the analytical toolbox to characterize them undergoes constant renewals and innovations. Their applicability towards the assessment of a meaningful array for particle-counting characterization will be discussed, including
examples of their use in the frame of immunogenicity studies.
15:35 End of Protein Aggregation & Stability