Cambridge Healthtech Institute’s 11th Annual
Protein Aggregation and Emerging Analytical Tools
Mechanism, Prediction, Immunogenicity, Developability, and Process Challenges
January 23-24, 2020
Part of the Formulation & Stability pipeline
The popular 11th Annual Protein Aggregation and Emerging Analytical Tools conference covers the latest trends, challenges and solutions in understanding, characterization and mitigation of problems generated by protein aggregation in biopharmaceuticals.
This conference will feature in-depth case studies, new and unpublished data and interactive discussions on immunogenicity of aggregates, mechanisms of aggregation, new tools for detection and quantitation of aggregates, and how the data is used in
regulatory filings. It will also discuss mechanistic understanding of protein aggregation and present case studies on prevention of particle formation by engineering and formulation approaches, aggregation in ADCs, bispecifics, impact of aggregation
on production, aggregates as a factor for immunogenicity, and approaches for improvement of biophysical properties of protein solutions.
We invite you to join with colleagues from around the world in this discussion of the key challenges and solutions in protein aggregation in biotherapeutics.
Final Agenda
Tuesday, January 21
5:45 - 8:45 Recommended Dinner Short Course*
SC5: Protein Aggregation: Mechanism, Characterization, and Consequences - Detailed Agenda
Instructors:
Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire
Kevin Mattison, PhD, Principal Scientist, Malvern Pananalytical, Inc.
*Separate registration required
THURSDAY, JANUARY 23
7:45 am Registration (Sapphire West Foyer) and Morning Coffee (Sapphire West & Aqua West Foyer)
8:10 Organizer’s Welcome Remarks
Nandini Kashyap, Conference Director, Cambridge Healthtech Institute
8:15 Chairperson’s Opening Remarks
Peter Schurtenberger, PhD, Professor, Department of Chemistry, Lund University
KEYNOTE PRESENTATION
8:20 Understanding and Predicting Self-Association in High Concentration Antibody Solutions – A Colloid Approach
Peter Schurtenberger, PhD, Professor, Department of Chemistry, Lund University
We address the problem of enhanced self-association in high concentration antibody solutions and the concomitant high viscosities. In order to understand and predict the thermodynamic and flow properties of such formulations, we provide the first
quantitative description of mAb self-association and viscosity as a function of concentration by combining experiments (static and dynamic scattering and microrheology), theory and computer simulations using a model based on analogies to patchy
colloids.
9:00 Estimating Solution Nonideality from Measured Values
Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire
The concentration-dependent chemical potential of a species results from the sum of its repulsive and attractive interactions with neighboring species. The repulsive and attractive terms can be estimated using measured values of size, charge and association
constants from a combination of one- and two-component solution measurements. The underlying concepts will be explained and the need for measured values will be highlighted.
9:30 Detection and Characterization is the First Step to Eliminating Aggregation
Kevin McCowen, Southwest Regional Manager, Wyatt Technology
Size exclusion chromatography (SEC) with UV detection gives limited information on the nature of aggregates. In this presentation, we discuss how multi-angle light scattering in conjunction with SEC as well field flow fractionation and dynamic light
scattering allow the researcher to rapidly assess formulation stability to aid in the elimination of aggregates in the early development phase, detect the presence of large aggregates, and probe aggregate characteristics such as absolute molecular
weight and conformation.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
11:00 Application of an in vitro Skin Model System to Assess Potential Risk of in vivo Aggregation and
Immune Activation of Biotherapeutic Attributes
Josh Tokuda, PhD, Postdoctoral Fellow, Biological Relevance and Characterization, Amgen, Inc.
In this presentation, a new human in vitro skin model system will be presented that can be used to assess the potential of biotherapeutics to aggregate and cause immune activation in vivo.
11:30 PANEL DISCUSSION: Prediction and Control of in vivo Aggregation and Immunogenicity
Discussion Points:
- Current Challenges
- New Predictive Tools
- Data Analysis and Management
- Use of Artifical Intelligence in Better Prediction and Control
Moderator:
Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire
Panelists:
Peter Schurtenberger, PhD, Professor, Department of Chemistry, Lund University
Josh Tokuda, PhD, Postdoctoral Fellow, Biological Relevance and Characterization, Amgen, Inc.
John J. Correia, PhD, Professor, Department of Biochemistry, University of Mississippi Medical Center
Reza Esfandiary, Associate Director, Early Stage Formulation Sciences, BioPharmaceutical Development, AstraZeneca
12:00 pm Shear Rate Dependent Viscosity as an Indicator of Protein-Protein Interactions and Cluster Formation
Stacey Elliot, Principal Scientist, R&D, RheoSense, Inc
Protein-protein interactions and the resulting microstructure influence the low shear viscosity and onset of shear thinning, which are both practically relevant to pharmaceutical development. Shear rate dependent viscosity measurements on
concentrated proteins are analyzed using concepts from colloidal systems to facilitate interpretation with respect to attraction strength and cluster size.
12:30 Session Break
12:40 LUNCHEON PRESENTATION: Everything You Wanted to Know About Protein Aggregation but Were Too Afraid To Ask
Kevin Lance, PhD, Product Manager, Marketing, Unchained Labs
Detecting and understanding aggregation is both complex and critical to the successful development of biologics. Thankfully Uncle delivers six powerful tools to ensure there is no shortage of insights on stability, aggregation, and non-specific
interactions. Fluorescence, and static and dynamic light scattering work for your protein, antibody, or viral capsid to understand their willingness to stay folded and solitary, and if you’ve got the right formulation to keep them
that way.
1:10 Ice Cream Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
2:15 Chairperson’s Remarks
Christoph Brandenbusch, PhD, Group Leader, Biochemical and Chemical Engineering, TU Dortmund
2:20 FDS-AUC Analysis of mAb Nonideality and Self-Association in Serum and Formulation Solutions
John J. Correia, PhD, Professor, Department of Biochemistry, University of Mississippi Medical Center
The Aviv Fluorescence Detection System (Aviv-FDS) has allowed the performance of sedimentation velocity experiments on therapeutic antibodies in highly concentrated environments, like serum and formulation buffers. Methods have been implemented
in the software package SEDANAL for the analysis of nonideal, weakly associating AUC data acquired on therapeutic antibodies and proteins using absorbance or FDS optics. This involves determining both hydrodynamic nonideality Ks and thermodynamic
nonideality BM1 plus association constants.
2:50 Mitigation of Reversible Self-Association and Viscosity of Monoclonal Antibodies via Structure-Guided Protein Engineering: Complementing Analytical and
i
n silico Tools
Reza Esfandiary, Associate Director, Early Stage Formulation Sciences, BioPharmaceutical Development, AstraZeneca
High protein concentrations can introduce additional development challenges due to issues such as reversible self-association or high viscosity. Protein engineering can provide a complementary mitigation approach to formulation optimization in
improving high concentration developability properties. Here, case studies utilizing complementary analytical and in silico methods are presented where molecular hotspots in monoclonal antibodies, responsible for
high concentration issues, are systematically identified and engineered to generate variants with improved developability profiles.
3:20 Networking Refreshment Break (Sapphire West & Aqua West Foyer)
3:45 Thermodynamics-Based Approach for Predicting Aggregation Propensity and Beneficial Solution Conditions in Antibody Formulations
Christoph Brandenbusch, PhD, Group Leader, Biochemical and Chemical Engineering, TU Dortmund
Protein aggregation is caused by the molecular interactions of all components in solution. We developed a thermodynamics-based approach to predict beneficial solution conditions taking the competition for water by a specific excipient, as well
as the molecular interactions of the proteins in the presence of excipients into account. This allows predicting a first estimate on aggregation propensity induced by the respective excipients and thus enables a first choice of preferential
excipients with a minimum of experimental effort.
4:15 Discerning the Synergistic Effect of Hydrodynamic Flow and Interfaces on Protein Aggregation
Paolo Arosio, PhD, Professor, Biochemical Engineering, Department of Chemistry and Applied Biosciences, ETH Zurich
Despite being an area of extensive investigation, the effect of hydrodynamic flow and shear on protein aggregation is still controversial. Here, we demonstrate the presence of a synergistic effect of interfaces and hydrodynamic flow in flow-induced
protein aggregation. We propose that hydrodynamic flow and shear stress should be considered in close association with interfaces when discussing sources of protein aggregation.
4:45 Computer Simulations of Aggregation of Proteins and Peptides
Andrzej Kloczkowski, PhD, Professor, Pediatrics, Nationwide Children’s Hospital and The Ohio State University
Aggregation of proteins and peptides is an important biological phenomenon often related to protein misfolding and correlated with various diseases, such as Alzheimer’s or Parkinson’s; recently it has been shown that preeclampsia
has similar molecular mechanism as Alzheimer’s. Computer simulations are excellent tools to study the molecular mechanism, structural features and dynamics of protein aggregation, and formation of amyloid filaments and fibrils. Results
of our recent computational simulations studies focused on specific diseases such as Alzheimer’s, and preeclampsia will be revealed.
5:15 Close of Day
FRIDAY, JANUARY 24
8:00 am Registration (Sapphire West Foyer)
8:00 BuzZ Sessions with Continental Breakfast
Protein therapeutics is a fast-growing global market. As the science improves, so does the complexity of the R&D organization. Ensuring product quality plus speed to market requires insights from stakeholders working across the stages
of protein science R&D. Join experts representing this PepTalk pipeline, peers, and colleagues for an interactive roundtable discussion. Topics include highlights from the week’s presentations, new technologies and strategies,
challenges, and future trends.
Click here for more details
9:00 Chairperson’s Remarks
Gerhard Winter, PhD, Professor, Chair, Pharmaceutical Technology and Biopharmaceutics, LMU Munchen
9:05 Qualifying a New Method for Submicron Particle Counting and Why It Matters
Gerhard Winter, PhD, Professor, Chair, Pharmaceutical Technology and Biopharmaceutics, LMU Munchen
We have tested and critically evaluated TRPS (Tunable Resistive Pulse Sensing) by using a rather affordable analytical equipment (IZON) and providing information on how to collect data, how to analyze and how to critically assess them.
Comparisons with other techniques like RMM and NTA are made, and examples from very different primary packaging materials on what to expect from submicron particle counting and how to reduce particle burden are provided.
9:35 Challenges in Characterization and Developability Assessments of Multispecific Antibodies
Christian Lange, PhD, R&D Biologics Research, Assays and Analytics, Protein Therapeutics, Sanofi-Aventis Deutschland GmbH
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 showcases highlighting potential challenges in characterization
and developability of multispecifics.
10:05 Controlling Aggregation of a Range of Novel Biopharmaceutical Product Modalities
Jan Jezek, PhD, CSO, Arecor Ltd.
Whilst controlling aggregation of monoclonal antibodies has become a routine task through smart candidate screening and platform formulations, there are numerous novel modalities, such as multi-specific antibodies, ADCs, or gene
therapy products, where aggregation remains a key problem. This talk will present case studies showing novel formulation approaches to reduce aggregation in these products and enable user-friendly formats.
10:35 Networking Coffee Break (Sapphire West & Aqua West Foyer)
11:00 Fill/Finish Strategies to Prevent and Overcome Aggregation Challenges
Marcel Tigges, PhD, Associate Director, The Janssen Pharmaceutical Companies of Johnson & Johnson
Fill & Finish processes for large molecule parenterals require a quality control toolbox that allows for efficient monitoring of stress factors potentially impacting drug product quality. New technologies and PAT (Process
Analytical Technology) tools allow for real-time monitoring of protein concentration (FlowVPE) and low volume protein stability analysis (nanoDSF). Identification of critical process parameters (CPPs) and process steps
that potentially cause protein aggregation guides the design of robust processes towards optimal mixing, filtration and filling conditions for highest drug product quality and stability standards.
11:30 Selected Poster Presentation: Detection and Characterization of
Antibody Aggregates by Light Scattering and Fluorescence
Cornelia S. Ziegler, PhD, Scientist, Biologics Development-Bioanalytics, Sanofi France
Therapeutic antibodies are prone to aggregation like all proteins. A prime aspect of their development is the monitoring of their aggregation state during their shelf-life studies. Dynamic light scattering (DLS) is widely used
to describe the colloidal status of the samples in the submicron size range. Currently, we are developing an orthogonal fluorescence-based technique, which will allow the specific detection and quantification of denatured
state and native state protein aggregates.
12:00 pm Conference Wrap-Up
Jan Jezek, PhD, CSO, Arecor Ltd.
Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire
12:30 Close of Conference