Cambridge Healthtech Institute’s Ninth Annual

Protein Aggregation and Emerging Analytical Tools

Mechanism, Prediction, Screening, Immunogenicity and Formulation Challenges

January 11-12, 2018

 

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 in biopharmaceuticals. This conference will feature in-depth case studies, new and unpublished data and interactive discussions on mechanisms of aggregation, new tools for detection and quantitation of aggregates, and how the data is used in regulatory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, bipecifics, 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 present a poster and attend to join with colleagues from around the world in this discussion of the key challenges and solutions in protein aggregation in biotherapeutics.

Final Agenda

THURSDAY, JANUARY 11

7:45 am Registration and Morning Coffee

Mechanism, Prediction and Overcoming Protein Aggregation

8:15 Chairperson’s Opening Remarks

Thomas Laue, Ph.D., Professor Emeritus, Molecular, Cellular and Biomedical Sciences, University of New Hampshire

KEYNOTE PRESENTATION

8:20 Light-Induced Protein Disulfide Degradation: Product Characterization

Christian_SchoneichChristian Schoneich, Ph.D., Distinguished Professor and Chair, Pharmaceutical Chemistry, University of Kansas

Protein biotherapeutics can degrade via a manifold of physical and chemical degradation mechanisms. We will show here, that light exposure of therapeutic disulfide-containing proteins can lead to > 60 different products, generated via novel cross-linking and fragmentation mechanisms. Some of these reactions may be catalyzed by metal impurities such as iron or tungstate, which can be present in pre-filled glass syringes. Mechanistically, product formation can be rationalized by light-induced generation of radicals and reactive oxygen species.

9:00 Protein Solvation: Preventing Aggregation by Forming a Tighter ‘Shield’ around a Protein

Thomas_LaueThomas Laue, Ph.D., Professor Emeritus, Molecular, Cellular and Biomedical Sciences, University of New Hampshire

For proteins to aggregate, they need to come into contact. The hydration/solvation shell around proteins can block protein-protein contacts. This talk will focus on what factors impact the strength of the solvation shell.


9:30 Characterizing and Inhibiting Glucagon Fibrillation

Elizabeth_ToppElizabeth M. Topp, Ph.D., Dane O. Kildsig Chair and Department Head, Department of Industrial and Physical Pharmacy, Purdue University

Glucagon, a peptide hormone, is currently marketed in lyophilized form for treating severe hypoglycemia. The lyophilized form is necessary because glucagon rapidly fibrillates in solution. This presentation summarizes computational and experimental studies of the mechanisms of glucagon fibrillation, and presents novel glucagon derivatives that resist fibrillation.

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

11:00 Design of Stable and Aggregation Resistant Single-Domain Antibody Biotherapeutics

Jamshid_TanhaJamshid Tanha, Ph.D., Senior Research Officer, Human Health Therapeutics Portfolio, National Research Council Canada

Various approaches for improving the stability of VH and VL single-domain antibodies have been described. Here we zoom in on one particular approach, namely disulfide engineering approach, which improves the stability of VHs and VLs. The approach appears to be universally applicable across all VHs and VLs and may also apply to scFvs, Fabs, mAbs and their derivatives.

Malvern Panalytical11:30 Automated, Low Volume Assessment of Stability Parameters in Protein Formulation 

Kevin_MattisonKevin Mattison, Principal Scientist, Bioscience, Malvern PANalytical

Measuring key stability parameters of a protein formulation is critical in the early stage development of biopharmaceutical products. At this early stage the quantity of drug substance available for testing is limited, requiring these tests be performed on very small sample volumes. The Viscosizer TD system is an automated platform to characterize the stability of formulations. The system provides automated, low volume measurement of Viscosity, Hydrodynamic Size, Stokes Radius, and the Diffusion Interaction Parameter (kD).

12:00 pm Enjoy Lunch on Your Own

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

NEW TOOLS AND STRATEGIES FOR HTS SCREENING

2:00 Chairperson’s Remarks

Wayne F. Reed, Ph.D., Professor, Physics, Tulane University

2:05 Recent Progress in Light Scattering Determination of Aggregation Rates with Small Samples in Parallel Format

Wayne_ReedWayne F. Reed, Ph.D., Professor, Physics, Tulane University

Monitoring protein aggregation via simultaneous multiple sample light scattering in real time allows rapid, parallel determination of aggregation rates (AR), which are directly related to protein stability for different formulations. Recent progress beyond AR includes: interpretation of non-linear light scattering signatures in terms of mechanisms, relationship of AR to GPC data, distinguishing small populations of large aggregates from large populations of small aggregates, and considerations of reproducibility and predictability of aggregation.

Halo Labs 2:35 High Throughput, Low Volume Subvisible Particle Screening

Bernardo_CordovezBernardo Cordovez, Ph.D., President, Halo Labs

Halo labs will present a subvisible particle screening tool, the HORIZON, with detailed explanation of its Backgrounded Membrane Imaging (BMI) technology. A comparative analysis between HORIZON and flow imaging will be presented and key performance indicators including sample volume, throughput, dynamic range, instrument repeatability will be evaluated.

2:50 Protein (In)stability and Analytical Tools for Monitoring Aggregation

Vasco_FilipeVasco Filipe, Ph.D., Lab Head, Pharmaceutical Development Biologics, Sanofi

Drug product development of therapeutic proteins involves a complex selection and optimization process aimed at making proteins manufacturable, stable and deliverable. Avoiding protein aggregation is one of the main concerns. Choosing the right formulation and setting up good strategies to monitor, predict and avoid protein aggregation during the various steps of the process is crucial. Formulation development considerations and analytical tools used to monitor protein aggregation will be presented.

Wyatt 3:05 Sifting through the Noise-Utilizing Light Scattering for Rapid Formulation Development

Lynette_SchroeterLynette Schroeter, Crystalomics, Group Lead, Formulations Development, Althea CMO

Delivering high concentration, low viscosity biotherapeutics to patients is an attractive option for innovator companies. Althea's Crystalomics® technology delivers formulated protein crystal suspensions subcutaneously, whereby the crystals dissolve readily with no adverse effect. Light scattering analysis, particularly rapid screening utilizing the Wyatt Technology Dynapro® Plate Reader II, is critical to rapidly optimizing formulation conditions and characterizing the protein pre and post crystallization..

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

FORMULATION, PROCESS AND MANUFACTURING STRATEGIES TO OVERCOME AGGREGATION

4:15 Formulation, Process and Manufacturing Strategies to Prevent and Overcome Aggregation Challenges in Early and Late Stage Development

Sreedhara_AlavattamSreedhara Alavattam, Ph.D., Principal Scientist and Senior Group Leader, Genentech, Inc.

Aggregation remains a challenge during protein process development. Aggregation during production, purification and product handling can have potential impacts on immunogenicity. The talk will cover various aspects of decreasing aggregates and potentially remediate the aggregation during handling of drug products in the clinical setting.

4:45 Particulate Formation during Fill & Finish Operations

Cheng_HerCheng Her, Ph.D., Postdoctoral Research Fellow, Carpenter Lab, Pharmaceutical Sciences, University of Colorado-Denver, Anschutz Medical Campus

As the final step before the drug product reaches the patient, it is vital that a fill & finish operation mitigate both particulate formation and aggregation. It was the aim of this study to have a more comprehensive look at particulate formation during fill & finish operations, from the tubing and pumps used, to the storage and handling of the drug product after fill & finish operations.

5:15 PANEL DISCUSSION: Preventive and Analytical Approaches for Reduction and Removal of Aggregates that Work

Moderator:

Wayne F. Reed, Ph.D., Professor, Physics, Tulane University


Panelists:

Bernardo Cordovez, President, Halo Labs

Cheng Her, Ph.D., Postdoctoral Research Fellow, Carpenter Lab, Pharmaceutical Sciences, University of Colorado-Denver, Anschutz Medical Campus

Sophia Kenrick, Ph.D., Senior Applications Scientist, Wyatt Technology

Sreedhara Alavattam, Ph.D., Principal Scientist and Senior Group Leader, Genentech

5:45 Close of Day

FRIDAY, JANUARY 12

8:00 am Registration

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.

Table Moderator: Thomas Laue, Ph.D., Professor Emeritus, Molecular, Cellular and Biomedical Sciences, University of New Hampshire

 

IMMUNOGENICITY, DEVELOPABILITY, EXCIPIENTS, AND STABILITY

9:00 Chairperson’s Remarks

Nathan H. Joh, Ph.D., Scientist, Attribute Sciences, Amgen

9:05 High Molecular Weight Species of a Therapeutic Antibody is Chemically Modified, Lacks Distinct Structure, and Shows No Increased Risk of Immunogenicity in Model Systems

Nathan_JohNathan H. Joh, Ph.D., Scientist, Attribute Sciences, Amgen

High-molecular-weight (HMW) species from monoclonal antibody drug substance was investigated to elucidate structure, chemical modifications, and potential risk of immunogenicity. Higher levels of oxidized methionine and tryptophan were observed in HMW compared to monomer. HMW species lacked a well-defined molecular structure. Little to no risk of immunogenicity was observed for HMW in multiple immune model systems, including heterozygous transgenic mice, human peripheral blood mononuclear cells, and engineered immune cells.


9:35 Evaluation and Development of Screening Methods for Antibody Developability Assessment

Nikolai_LorenzenNikolai Lorenzen, Ph.D., Large Protein Biophysics, Novo Nordisk A/S

Biophysical screening is widely used in early phase development of monoclonal antibodies to guide selection of molecules with a high potential to reach clinical testing. Using the in silico solubility predictor CamSol we have designed a modelsystem of antibody variants displaying a range of solubilities. I will present how selected biophysical measures as e.g. AC-SINS, CIC, PEG precipitation and diffusion interaction parameter predict for formulation relevant measures for this modelsystem.

Partical Sizing Systems 10:05 Counting and Sizing Protein Aggregates Down to 0.15 um at High Concentrations by Focused-Beam SPOS

David_NicoliDavid Nicoli, Ph.D., Vice-President R&D, Particle Sizing Systems LLC

Protein aggregates as small as 0.15-um can be counted/sized at concentrations 100-1000X higher than is possible with light scattering sensors of conventional design, using a novel focused-beam single-particle optical sizing (SPOS) technique. Adding a second sensor that combines light obscuration and scattering extends the upper particle size limit to 200 microns. Analysis can be made on sub-mL samples, including those of high viscosity, and the sample is conserved following analysis.

 10:35 Coffee Break with a Poster Pavilion

PepTalk is proud to support and recognize the protein scientists of tomorrow during the Poster Pavilion. This time has been set aside to view the Student Fellowship posters and interact with presenters one on one. This opportunity gives job seekers the chance to share their expertise with future/potential employers or develop contacts to further their research.

11:15 PANEL DISCUSSION: Protein Aggregation, Degradation Mechanisms, Characterization and Prevention

• Understanding physical and chemical degradation mechanisms 

• Light-Induced protein disulfide degradation 

• Product characterization techniques 

• Protein solvation and preventing aggregation 

 • Managing aggregates during the various product life cycle

Moderator:

Sreedhara Alavattam, Ph.D., Principal Scientist and Senior Group Leader, Genentech


Panelists:

Bernardo Cordovez, President, Halo Labs

David Nicoli, Ph.D., Vice-President R&D, Particle Sizing Systems

Jamshid Tanha, Ph.D., Senior Research Officer, Human Health Therapeutics Portfolio, National Research Council Canada

Lynette Schroeter, Crystalomics, Group Lead, Formulations Development, Althea CMO

Nathan Joh, Ph.D., Scientist, Attribute Sciences, Amgen

Thomas Laue, Ph.D., Professor Emeritus, Molecular, Cellular and Biomedical Sciences, University of New Hampshire

12:15 pm Conference Wrap-Up

Thomas Laue, Ph.D., Professor Emeritus, Molecular, Cellular and Biomedical Sciences, University of New Hampshire

 

12:45 Close of Conference