Cambridge Healthtech Institute’s Eighth Annual
Protein Aggregation and Emerging Analytical Tools
Mechanism, Prediction, Screening, Immunogenicity and Formulation Challenges
January 12-13, 2017 | Hilton San Diego Bayfront | San Diego, CA

The popular Protein Aggregation and Emerging Analytical Tools conference covers latest trends, challenges and solutions in understanding, characterization and mitigation of problems generated by protein aggregation. It features in-depth case studies, new and unpublished data and interactive discussions on mechanisms of aggregation, new tools for detection and quantitation of aggregates, and how the data are used in regulatory filings. It also discusses mechanistic understanding of protein aggregation and presents case studies on prevention of particle formation by engineering and formulation approaches, impact of aggregation on production, aggregates as a factor for immunogenicity, and approaches for improvement of biophysical properties of protein solutions.

THURSDAY, JANUARY 12

7:45 am Conference Registration and Morning Coffee

UNDERSTANDING AND CONTROLLING PROTEIN AGGREGATION DURING PATIENT HANDLING AND USE

8:15 Chairperson’s Opening Remarks

Peter Tessier, Ph.D., Department of Chemical & Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute

Keynote Presentation

8:20 Mishandling of Therapeutic Protein Products by End Users: Particle Formation and Potential Roles in Adverse Immunogenicity

John F. Carpenter, Ph.D., Professor, Pharmaceutical Sciences; Co-Director, Center for Pharmaceutical Biotechnology, University of Colorado Anschutz Medical Center

Subvisible particles can cause adverse immunogenicity, resulting in loss of efficacy of therapeutic proteins in patients. Companies work diligently to control particle levels in products. However, patients can receive high levels of particles due to product mishandling by end users. Mishandling includes exposure of prefilled syringes to temperature extremes, transport of IV bags through pneumatic tube systems in hospitals and repackaging of vialed Avastin into syringes for off-label intraocular use.

9:00 Towards Particle and Silicon-Free Protein Drug Products

Gerhard Winter, Ph.D., Professor, Chair, Pharmaceutical Technology and Biopharmaceutics, LMU Munchen

A more wide-spread bedside filtration and the use of silicon oil free polymer syringes could reduce the amount of particles and silicon oil droplets eventually injected into patients with protein drug products. A critical view on the performance and on open questions associated with the use of the two measures is presented. The quality of filters and the resolution of the oxygen permeability of plastic material are studied in detail.

PREVENTION AND CONTROL OF PROTEIN AGGREGATION

9:30 Understanding and Overcoming Tradeoffs between Antibody Affinity, Specificity and Stability

Peter Tessier, Ph.D., Department of Chemical & Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute

There are many challenges associated with the discovery and development of potent and stable antibody therapeutics. Our lab is addressing some of these challenges, including the design and evolution of antibodies with high affinity, specificity, stability and solubility. We will discuss our findings related to common tradeoffs between these antibody properties as well as directed evolution methods for overcoming these tradeoffs.

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

11:00 Applications of Fluorescence-Detected Analytical Ultracentrifugation (AU-FDS) to High-Concentration Antibody Interactions

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

AU-FDS may be used to characterize the size distributions of antibody complexes. Results will be shown for several systems such as participation of a “foreign” antibody in the self-association complexes of mAb, weak interaction of poly-IgG with mAbs, presence of non-reactive antigen in a poly-IgG binding assay, existence of species cross-reaction in a poly-IgG prep and characterization of Ab:Ag lattice formation ‘overshoot’ kinetics. Also, it can be used to study that the size distribution of Ab:Ag complexes differ in buffer and serum and how AU-FDS uniquely detects large complexes in patient serum.

11:30 Counting & Sizing Protein Aggregates Down to 0.15um in sub-mL Volumes by Novel Focused-Beam Light Scattering Technology

David Nicoli, Ph.D., Vice President, Research & Development, Particle Sizing Systems, LLC

A novel single-particle optical sizing (SPOS) technique collects scattered light from particles passing through a focused laser beam, enabling protein aggregates to be counted/sized down to 0.15 um, at concentrations too high for normal light scattering sensors. Combining this with a second sensor based on traditional light obscuration plus scattering provides an upper size limit of 200 um. Analysis can be made on sub-mL samples, including those of high viscosity, with conservation of the sample.

12:00 pm Session Break

12:15 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing

PREVENTION AND CONTROL OF PROTEIN AGGREGATION (CONT.)

2:00 Chairperson’s Remarks

Jan Jezek, Ph.D., CSO, Research & Development, Arecor, Ltd.

2:05 Control of Protein Aggregation by Unconventional Formulation Parameters

Jan Jezek, Ph.D., CSO, Research & Development, Arecor, Ltd.

A number of formulation parameters, such as pH, ionic strength or surfactant, are traditionally optimized to minimize protein aggregation. The talk will show how unconventional parameters of formulation excipients can be exploited to control aggregation and enable products to be used outside the cold chain. This will be demonstrated on several data driven case studies using relevant therapeutic proteins, describing the specific formulation features employed to achieve superior stability.

2:35 Optimizing Protein Stability through Integration of Cutting-Edge Analytical Tools with Rational, Molecule-Specific Approach to Process and Formulation Development

Danny K. Chou, Pharm.D., Ph.D., President and Founder, Compassion BioSolution; Former Senior Research Scientist, Biologics Development, Gilead Sciences

We are at the dawn of a new era with the emergence of new analytical tools that can enable both prediction and real-time monitoring of protein stability. The author will use real case studies to illustrate how to properly combine some of these new tools with tried and true strategies that incorporate the key factors/forces that impact physical stability (with focus on protein aggregation) of proteins in solution. Emphasis will be placed on high throughput, low sample volume strategies that are useful for industrial application.

3:05 SELECTED POSTER PRESENTATION: Improving ADC Size Exclusion Chromatography Separation with Efficient DoE Based Method Development

David Chiu, Ph.D., Scientist, Analytical Sciences, Seattle Genetics, Inc.

3:20 SELECTED POSTER PRESENTATION: Performance of Capping on Residual Seal Force and Container Closure Integrity

Robert Ovadia, Technical Development Research Associate, Pharmaceutical Processing and Technology Development, Genentech, Inc.

Residual Seal Force (RSF) can be used to assess the "goodness" of a seal and enable non-subjective, consistent setting and validation of cappers across different manufacturing sites. Correlating RSF to Helium leakage (He-leak), the most sensitive container closure integrity (CCI) method, has proven to be challenging. Vials with no measurable RSF (loosely capped) remain integral when measured by He-leak. Our proposed method allows us to independently test each sealing surface. In this study, we explore the relationship between RSF and CCI, and how RSF can be used to ensure crimping robustness across different capping equipment.

3:35 Refreshment Break in the Exhibit Hall with Poster Viewing

4:15 Impact of Interfacial Interactions on the Formation of Particles, Aggregation of Proteins, and Their Prevention: Role of Surface Energetics and Their Application to Container Compatibility and Protein Purification

Jinjiang Li, Ph.D., Senior Principal Scientist, Drug Product Science & Technology, Bristol-Myers Squibb Co.

This talk will discuss the effect of surface energetics on protein adsorption, formation of aggregates, and particle formation, with/without surfactants. Common surfaces encountered in protein purification and storage will be used as examples. Surface energetics were characterized through measuring contact angles. Adsorption of proteins like lyozyme was measured using QCM-D. Particles formed were determined using MFI. For all cases, the protection role of PS 80 and Poloxamer was examined.

4:45 BREAKOUT DISCUSSIONS:

Topic 1: Innovation in Formulation – Novel Approaches versus Established Platforms

Moderator: Jan Jezek, Ph.D., CSO, Research & Development, Arecor, Ltd.

Topic 2: In silico Tools for Predicting Biotherapeutic Aggregation

Moderator: Russ Lehrman, Ph.D., President/Founder, R&D Consultation, BioSuperior Technology

Topic 3: Applications of AU-FDS to High-Concentration Antibody Interactions

Moderator: Thomas Laue, Ph.D., Professor, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire

5:15 End of Breakout Discussions and Discussion Report Out

5:45 Close of Day

FRIDAY, JANUARY 13

8:00 am Conference Registration and Morning Coffee

RAPID TOOLS FOR PREDICTION, MEASUREMENT OF AGGREGATION, VISCOSITY AND STABILITY

8:30 Chairperson’s Remarks

Danny K. Chou, Pharm.D., Ph.D., President and Founder, Compassion BioSolution; Former Senior Research Scientist, Biologics Development, Gilead Sciences

8:35 Application of in silico Predictions to the Mitigation of Biotherapeutic Aggregation

Russ Lehrman, Ph.D., President/Founder, R&D Consultation, BioSuperior Technology

Aggregation of biotherapeutic candidates affects drug safety, efficacy, and manufacturability. This degradation pathway occurs largely due to the presence of aggregation prone regions (APRs). Computer tools that identify APRs are available, but since each program is based on different properties, they often detect distinct protein regions. Effective use of these tools depends on an understanding of protein structure and aggregation mechanisms. Case studies that help demonstrate how to effectively use these programs for the identification of APRs will be presented.

9:05 A Rapid Real-Time Biosensor to Detect Preaggregates and Screen Formulations of Therapeutic Proteins

Subhashchandra Naik, Ph.D., Postdoctoral Researcher, Chemical and Biomolecular Engineering, University of Delaware

A biosensor for monitoring the structural integrity and physical stability of therapeutic proteins was developed by combining a bacterial protein GroEL with real-time biolayer interferometry. This biosensor detects structurally altered pre-aggregates within a minute in real-time, is sample sparing and is non-destructive. The data was supported and validated by orthogonal biophysical techniques. The biosensor can also be used to screen for formulations/excipients that stabilize proteins and suppress aggregate formation.

9:35 Integrating Novel Tools into Development Workflow of Biologics: nanoDSF and MST for Discovery, Development and QC

Alexey Rak, Ph.D., Director, Bio Structure & Biophysics, Sanofi R&D

Biophysical approaches are routinely used to assess biologics activities, stability and quality. Modern drug discovery operations require characterization of biomolecular interactions to be both time- and cost-effective as well as to be highly precise and reproducible. Here we report applications of two novel methods, nano-Differential Scanning Fluorimetry (nanoDSF) and MicroScale Thermophoresis (MST), that we are applying in our biologics discovery and characterization operations. The examples of the demonstrated effectiveness of the nanoDSF and MST will be presented and discussed.

10:05 Coffee Break with a Poster Pavilion

11:00 Advancements and Comparability Assessments of Amgen Automated Visual Inspection System for Therapeutic Molecules

David Le, MSc, Scientist, Drug Product Technologies, Amgen

Amgen has developed a robotic system that fully automates visual inspections of liquid formulations. The system provides versatility in handling several container configurations that includes glass and plastic pre-filled syringes and vials. The instrument utilizes high resolution imaging photo optics and proprietary algorithms to capture and quantify the levels of both visible and sub-visible particles. Results for precision, linearity, and comparability to manual inspections will be discussed.

11:30 Detection and Mitigation of Antibody Aggregation in Intravenous Infusion Solutions

Yin Lai, Ph.D., Senior Scientist, Formulation Development, Eli Lilly and Company

Administration of therapeutic monoclonal antibodies through intravenous route involves dilution of drug product using intravenous infusion solutions such as saline or 5% dextrose. As a result, the original drug product is diluted, which will potentially change physical and chemical stability of the API. This presentation will discuss structural characterization, protein-protein interaction, concentration dependent aggregation, and methods to monitor time-dependent self-association in intravenous infusion solutions containing therapeutic antibodies.

12:00 pm IT’S A WRAP: PEPTALK 2017 CLOSING PLENARY PANEL DISCUSSION

1:15 Close of Conference