Protein Aggregation and Stability in Biopharmaceuticals Banner

 

The phenomenon of protein aggregation is a complex conundrum that impacts biopharmaceutical development at virtually every stage. All mechanisms of aggregation are not conclusively known, but the industry must use every effort to characterize and control these conditions, applying a rapidly changing landscape of assays, instrumentation, formulation strategies and process steps. The PEGS Protein Aggregation and Stability in Biopharmaceutical Products offers important scientific updates and a forum for dialog among the stakeholders in this challenging arena.

Final Agenda

Recommended Short Course(s)*

SC8: Introduction to Biophysical Analysis for Biotherapeutics: Discovery & Development Applications


*Separate registration required.

THURSDAY, MAY 3

AGGREGATION PRECURSORS
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12:00 pm Registration (Commonwealth Hall)

12:35 Luncheon in the Exhibit Hall with Poster Viewing (Commonwealth Hall)

1:40 Chairperson’s Remarks

David Brockwell, PhD, Associate Professor, School of Molecular and Cellular Biology, University of Leeds, United Kingdom

1:50 Emerging Approaches for Understanding Early Aggregation Transients

Arun_IgnatiusArun Alphonse Ignatius, PhD, Principal Scientist, Pfizer

Monoclonal antibodies could be susceptible to aggregation, especially at concentrations relevant to pharmaceutical formulations. (>100 mg/ml). Although conventional low resolution biophysical methods probe conformational stability of mAbs in relation to aggregation, molecular origins of aggregation are largely elusive. Nuclear magnetic resonance (NMR) studies on mAb and mAb fragments provide structural fingerprinting at an atomic resolution that can be potentially correlated to aggregation propensities of mAbs.

2:20 Protein Aggregation and Gelation – Insight from Combining Scattering, Rheology and Computer Simulations

Peter_SchurtenbergerPeter Schurtenberger, PhD, Professor, Physical Chemistry, Lund University, Sweden

Understanding and avoiding protein aggregation and gelation is essential in formulating biopharmaceuticals. We show how we can use a combination of advanced characterization techniques such as small-angle neutron (SANS) and X-ray scattering (SAXS), neutron spin echo measurements and microrheology experiments, combined with the theoretical toolbox from colloid physics and state-of-the-art computer simulations, to assess and predict aggregation and gel formation in concentrated solutions of biopharmaceuticals.


2:50 KEYNOTE PRESENTATION: Complement Activation in Human Serum by Protein Particles Is Influenced by Interactions with Containers

Ted_RandolphTheodore W. Randolph, PhD, Professor, Chemical & Biological Engineering, University of Colorado, Boulder

Acute immune responses to therapeutic proteins include anaphylactic shock, which may result from activation of the complement cascade. Container materials may affect the number of particles formed following freeze-thawing or agitation of protein formulations. Testing of these formulations in human serum shows that levels of activation of Bb, C3a and C5a, all critical intermediates on the complement activation cascade, are strongly correlated with levels of particles within antibody formulations.

 

3:20 Cruise Through Biologics Development with Uncle and Hunky

Dina Finan, Product Manager, Marketing, Unchained Labs

Developing biologics requires picking the best candidates and assessing different formulations to ensure stability and minimize aggregation risk. Whether you’re working with limited amounts of precious sample, or trying to understand even more about your molecules later on, Unchained Labs automates and simplifies these assessments. We’ll discuss how you can use applications like thermal melting, sizing and polydispersity, and chemical denaturation to thoroughly characterize your samples.

3:50 Networking Refreshment Break (Harbor & Mezzanine Level)

4:20 Assessing the Aggregation of Biopharmaceuticals in vitro and in vivo

David_BrockwellDavid Brockwell, PhD, Associate Professor, School of Molecular and Cellular Biology, University of Leeds, United Kingdom

Protein-based biopharmaceuticals are susceptible to unfolding, mis-folding and aggregation by environmental perturbations that include hydrodynamic flow. Aggregation thus poses an enormous challenge to biopharmaceutical development, production, formulation and storage. To address this problem, we describe an in vivo method to rapidly assess candidates for developability and an in vitro method to assess candidate manufacturability or process suitability for the manufacture of aggregation-prone biopharmaceuticals.

4:50 Combining Viral Particle Counting, Biological Characterization and Advanced Kinetics to Predict Vaccine Stability

Didier_ClenetDidier Clenet, PhD, Research Scientist, Formulation & Stability, Sanofi Pasteur, France

In-depth characterization of a purified rabies vaccine was performed in term of virus counted, aggregation state, and antigenic and genomic titer. Agreement between results from NTA (nanoparticle tracking analysis) and ELISA was assessed. Additionally, forced degradation study was combined with modern kinetic-based modeling approach to delimitate a time-temperature stability domain into which the vaccine would be kept antigenic. Based on a kinetic model, 3 years of vaccine stability was predicted at 5°C.
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5:20 End of Day

5:20 Registration for Dinner Short Courses (Commonwealth Hall)

Recommended Dinner Short Course(s)*

SC11: Strategic/Modular Bioassay Design and Analysis


*Separate registration required.

FRIDAY, MAY 4

8:00 am Morning Coffee (Harbor Level Lobby)

DEVELOPABILITY EVALUATION
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8:30 Chairperson’s Remarks

Neal Whitaker, PhD, Associate Researcher, Macromolecule and Vaccine Stabilization Center, Pharmaceutical Chemistry, The University of Kansas School of Pharmacy

8:35 Chemical Stability Screening in Early Stage Discovery: New Isomerization and Deamidation Datasets

Yingda_XuYingda Xu, PhD, Director, Protein Analytics, Adimab

Isomerization and deamidation of therapeutic leads can often delay development timelines and provide challenge and risks for downstream process. Sequence-based prediction to scan for NG and DG can often lead to false positive results. Here we report the chemical liability analysis of ~140 clinical stage antibodies, under forced degradation conditions.


9:05 FEATURED PRESENTATION: Methods for Identifying Monoclonal Antibodies with Drug-Like Properties

Peter_TessierPeter M. Tessier, PhD, Professor, Pharmaceutical Sciences and Chemical Engineering, University of Michigan

The success of antibody therapeutics is dependent not only on their specific bioactivities but also on their physiochemical properties. Here we report the development of methods for identifying antibodies with drug-like properties based on their chemical compositions and biophysical properties.

Schrodinger3:15 A Computational Framework for Predicting Protein Liabilities and Improvement of Antibody Developability

Kannan Sankar, PhD, Postdoctoral Associate, Schrödinger

We present a novel algorithm to predict aggregation-prone regions based on 3D structures that uses the distribution of hydrophobic and electrostatic patches on the proteins’ interaction surface. Machine learning techniques are applied to a large set of protein-specific descriptors to enable non-experts to produce statistically sound liability prediction models.

10:05 Networking Coffee Break (Harbor & Mezzanine Level)

10:35 Developing a Screening Platform for Novel Biotherapeutics

Jay_GuoZhi (Jay) Guo, PhD, Senior Scientist, Global Protein Sciences, AbbVie Bioresearch Center

Novel bi-functional biologics with a variety of formats to choose from (scFv, Fab, mAb, etc.) and fused to different payloads (cytokines, growth factors, a different scFv, etc.) is of immense interest for targeted delivery and improved efficacy. Payloads can be fused at different positions and spacer lengths and it is now apparent that payloads dominate production and stability of biologics; thus, confounding drug-like properties (DLPs). To better understand structure-function relationship and rapidly advance candidates with good DLPs we will discuss our throughput screening process.

FORMULATION CONTROL OF PROTEIN AGGREGATION
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11:05 Characterization of Aggregates That Are Concentration-Dependent

Neal_WhitakerNeal Whitaker, PhD, Associate Researcher, Macromolecule and Vaccine Stabilization Center, Pharmaceutical Chemistry, The University of Kansas School of Pharmacy

High protein concentrations in biopharmaceutical drug products (>100 mg/ml) can lead to colloidal instability and elevated levels of aggregation. Numerous analytical techniques are required to characterize these aggregates, from small soluble aggregates, to submicron and subvisible particles to larger visible particles. This talk will present several case studies utilizing these methods with the aim of reducing the propensity of aggregate formation as part of the development of stable protein formulations.

11:35 Optimizing Cryoprotectant to mAb Ratios to Mitigate Aggregation

Tom_CrowleyTom Crowley, PhD, Principal Scientist, Pharmaceutical R&D, Pfizer, Inc.

The use of disaccharides such as sucrose and trehalose as cryoprotectants in monoclonal antibody formulations is a well-established strategy in protecting against freeze thaw induced aggregation. This work explores the impact of varying the ratio of cryoprotectant to antibody to meet the challenges of ensuring protein stability, controlling viscosity, and maintaining isotonicity for high concentration dosage forms.

12:05 pm Controlled Nucleation During Lyophilization - Comparison of Nucleation Techniques and their Impact on Protein Aggregation

Andrea_AllmendingerAndrea Allmendinger, PhD, Senior Scientist, Late-Stage Pharmaceutical and Processing Development, Biologics, F.Hoffmann-La Roche, Switzerland

Stabilizing proteins in liquid state can be challenging. Lyophilization offers a gentle way to increase storage stability, which is dependent on product attributes like residual moisture and cake structure directly impacted by the design of the lyophilization cycle. There is growing interest in controlling the ice nucleation event in order to reduce variability across samples. Roche/Genentech has evaluated the comparability of cakes produced with different technologies and findings are presented in this talk.

12:35 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:05 Networking Refreshment Break (Harbor & Mezzanine Level)

PROCESS CONTROL OF PROTEIN AGGREGATION
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1:35 Chairperson’s Remarks

Zhi (Jay) Guo, PhD, Senior Scientist, Global Protein Sciences, AbbVie Bioresearch Center

1:40 Rapid, High-Resolution Analysis of Monoclonal Antibody Aggregates Using Chip LFMC

Raja_GhoshRaja Ghosh, PhD, Professor, Chemical Engineering, McMaster University, Canada

Laterally-fed membrane chromatography (LFMC) has been shown to be suitable for high-speed, high-resolution separation and analysis of biologicals such as monoclonal antibodies. A scaled-down Chip LFMC device suitable to rapid and high-resolution analysis of monoclonal antibody aggregates will be discussed. The typical separation time with Chip LFMC is of the order of a couple of minutes, which is significantly faster than UPLC. Also, the separation can be carried out using inexpensive low-pressure liquid chromatography systems.

2:10 Product Quality Control Strategy Development for Non-mAb Complex Modalities by Using Combinatorial Cell Engineering and OMICS Screening Tools

Zhimei_DuZhimei Du, PhD, Director, Bioprocess, Merck & Company, Inc.

New product-related impurities have been found to accompany non-mAb complex modalities, which usually don’t exist in standard mAb production. Many of these PQAs are related to protein folding and assembly efficiency inside the cell, which impact post-translational modifications directly or indirectly. Our presentation will illustrate the importance of selecting appropriate cell/upstream conditions through screening and/or engineering, as part of quality control strategy to obtain the desired recombinant protein PQA profile.

2:40 Process Control Challenges in Controlling Aggregation

Jason_FernandezJason Fernandez, Scientist, Protein Pharmaceutical Development, Biogen

Control of aggregation is a common goal across biopharmaceutical manufacturing processes, from the bioreactor through fill finish operations. As process controls vary along the manufacturing process, with each processing operation having unique challenges to aggregation control, an end-to-end process control strategy is important in minimizing the ultimate aggregate level in finished products. This talk will highlight the concept and challenges of end-to-end process control of aggregation.

3:10 Evaluation of Sub-Visible Particle Technologies Used to Characterize Particles from Packaging Components

John_RechJohn Rech, Technology Manager, Particle Testing, Analytical Laboratory, West Pharmaceutical Services

Measurement and control of sub-visible particles (SVP) in injectables is a growing concern in pharma because of potential to cause patient harm. Global compendia have specifications in the final drug product for particles >100µm and >10µm, but current methods can measure below 10µm and into the submicron range. To better understand SVP contribution, particularly from silicone-treated stoppers, a study was designed to determine optimal methods to measure and identify SVP.

3:40 End of Conference


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