Biophysical analysis is now playing a significant role in the research and early development for a new generation of complex protein therapeutics. The combination of improved understandings of the structure and function of proteins with advanced, higher
resolution analytical methods helps researchers identify and fine-tune candidate molecules before advancing them into development. As protein engineers and analytical scientists increase their reliance on biophysical analysis, they are driving a move
to instruments with higher throughput and resolution – and working to quantify analytical results previously used only for qualitative assessments. Biophysical Analysis of Biotherapeutics will bring together an international
audience of protein scientists and analytical specialists to explore the latest technologies and techniques used for problem solving in this dynamic field.
WEDNESDAY, MAY 6
7:00 am Registration and Morning Coffee
8:00 Chairperson’s Remarks
Hubert Kettenberger, Ph.D., Principal Scientist, Protein Analytics, Roche Pharma Research & Early Development, Roche Innovation Center Penzberg
8:10 KEYNOTE PRESENTATION
Competing Effects for Optimizing Protein Behavior at Low and High Concentrations
Christopher J. Roberts, Ph.D., Associate Professor, Chemical & Biomolecular Engineering, University of Delaware
Multiple factors are considered when selecting protein product formulation and manufacturing conditions, as there may be competing effects that require optimization. This presentation focuses on examples from therapeutic proteins and model proteins
that highlight the balance between different behaviors (e.g., aggregation, solubility, viscosity) and how to reasonably account for or predict these in at least a semi-quantitative manner based on biophysical measurements.
8:40 Beauty or Beast? In Silico Molecule Assessment to Support Lead Selection
Hubert Kettenberger,
Ph.D., Principal Scientist, Protein Analytics, Roche Pharma Research & Early Development, Roche Innovation Center Penzberg
Successful new biotherapeutics should possess – besides the desired biological activity – high biochemical and biophysical stability. In silico prediction of chemical degradation “hotspots”, charge distribution, molecular dynamics
and other features can support the lead selection and protein engineering process.
9:10 Structure-Based Predictive Tools for Engineering Biophysical Characteristics of Biotherapeutics
Sudharsan (Sid) Sridharan, Ph.D., Scientist, Antibody Discovery and Protein Engineering, MedImmune
Structural data of antibodies and other biotherapeutics provide key knowledge for understanding their biophysical characteristics which impacts both research and development of these molecules. Predictive tools using structures complement experimental
analyses and can enable rational engineering of biotherapeutics. In this talk examples of these tools and their applications in engineering favourable biophysical properties of biotherapeutics will be highlighted.
9:40 Structural Flexibility and Allosteric Responses are Widely Redistributed Within Antibody Fragments Upon Mutation
Dennis Livesay, Ph.D., Professor,
Bioinformatics and Genomics, University of North Carolina
Recombinant antibody fragments have emerged as credible alternatives to full therapeutic antibodies. Unfortunately, reduced thermostability is frequently observed, limiting their broad utility. In response, screening for mutants that increase stability
without compromising affinity is commonly employed. Little is known about how the uncovered mutations affect dynamical properties. In this talk, I will discuss the frequency and scale of changes in structural flexibility and allostery across a number
of different antibody fragment systems.
10:10 Analyzing Varying Biochemical and Biophysical Data in Discovery Biologics
Timothy Fenn, Ph.D., Principal Scientist,
Boehringer Ingelheim
In discovery biologics, simple and information rich biochemical/biophysical methods are required to provide the wide variety of information necessary to evaluate antigens, multispecifics, and potential antibody leads. This talk will focus on some common
analytical methods and how the information derived from each can be used in a complementary manner. Examples will be provided in a variety of contexts, including bispecific scaffold selection, antibody purification and protein reagent analysis.
10:40 Coffee Break in the Exhibit Hall with Poster Viewing
11:25 Combining Biophysical Tools and Transgenic Mouse Models to Understand the Immunogenic Potential of Subvisible Particles
Emilien Folzer, Scientist,
Pharma Technical Development Europe (Biologics) Analytics, Roche
Theoretical concerns regarding the potential immunogenicity of proteinaceous sub-visible particles in protein therapeutics have being widely debated in literature with very limited experimental data available to date. This talk will focus on describing
two methods for particle fractionation that have been developed to isolate defined particle species, as well as the detailed physicochemical and biological characterization of these species.
11:55 Biophysical Tools for Molecular Assessment of mAbs
Arun Alphonse Ignatius, Ph.D.,
Principal Scientist, Biotherapeutics Pharmaceutical Science, Pfizer, Inc.
Molecular assessment is a key component of the candidate selection process. Previously, candidate selection has focused mainly on the biological properties of the protein and developability/manufacturability aspects were not considered. Many programs
required additional resources and non-platform approaches during development due to lack of early developability assessment data. In this presentation, biophysical characterization tools optimized to probe into the conformational and colloidal
stability of mAbs will be used on a set of Pfizer candidates to enable de-selection of risky candidates and ranking during molecular assessment stages.
12:25 pm Molar Mass, Size, Charge and Conformation: Light Scattering
Tools for Biophysical Characterization of Macromolecules
John Champagne, Ph.D., Senior Applications Scientist & Northeast Regional Manager, Wyatt Technology Corporation
This seminar describes a comprehensive suite of characterization tools based on static and dynamic light scattering, which work together with size-based separation, to provide first principles biophysical characterization of macromolecules. Some
of the key applications of the light scattering toolbox include analyses of molar mass and size distributions, aggregation, branching and other measures of conformation, and the composition of complex protein systems and other conjugated macromolecules.
12:55 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:55 Session Break
2:10 Chairperson’s Remarks
Sudharsan (Sid) Sridharan, Ph.D., Scientist, Antibody Discovery and Protein Engineering, MedImmune
2:15 Optimizing Selection of Aggregation-Resistant Antibodies Using Self-Interaction Nanoparticle Spectroscopy
Jiemin (Jimmie) Wu, Research Associate,
Chemical and Biological Engineering, Rensselaer Polytechnic Institute
Conventional methods of measuring mAb self-association are difficult to employ during antibody discovery due to the low purities, limited quantities, and large numbers of mAb candidates. To address this challenge, we are developing a robust method
(affinity-capture self-interaction nanoparticle spectroscopy, AC-SINS) capable of identifying mAbs with low self-association propensity at extremely low antibody concentrations and in the presence of contaminants.
2:45 Catching the Seeds of Destruction: Probing Structurally Altered and Aggregated States of Therapeutically Relevant Proteins Using GroEL Coupled to Bio-Layer Interferometry
Mark T. Fisher, Ph.D., Professor,
Biochemistry and Molecular Biology, University of Kansas Medical School
Nature employs a wide array of chaperone proteins to prevent large scale aggregation in vivo by interacting with partially folded protein species. This chaperone network actively or passively reverses misfolding to prevent and/or reverse the accumulation
of aggregation prone species. Taking this tact, we demonstrate that a GroEL chaperonin-based biolayer interferometry platform can successfully detect initial “seed” preaggregate species in therapeutic protein formulations prior
to large aggregate formation in vitro.
3:15
Simple Approaches to Monitoring Bio Pharmaceutical Purification with Flow Imaging
Aaron Noyes, Ph.D., Process Engineer, Purification Process
Development, Pfizer
Sub-visible particle analysis using micro-flow imaging is critical to monitor biopharmaceutical product quality and safety. Compared to older techniques which only provide particle size and count and can undercount, MFI also provides particle
morphology. As a result, MFI can resolve complex particle types such as protein aggregates from silicone oil and other contaminants. During purification of biologicals, complex particle environments are common. In this webinar, Aaron Noyes,
Scientist at Pfizer, will show how MFI presents a fresh approach for characterizing biopharmaceutical production processes of mAbs and vaccines. Case studies focused on flocculation, precipitation, and filtration will demonstrate how measuring
particle size, count and morphology can enhance process understanding, improve productivity, and ultimately, increase the robustness, of biopharmaceutical production processes.
3:45 Refreshment Break in the Exhibit Hall with Poster Viewing
4:45 Microspectroscopic Analysis of Free Fatty Acid Particles in Protein Formulations
Xiaolin Cao, Ph.D., Principal Scientist,
Amgen
Free fatty acid particles were identified in a number of protein formulations containing the surfactant polysorbate 20 using multiple microspectroscopic methods. These fatty acid particles were numerous in number, granular or sand-like in morphology
and were several microns in size. In addition, protein particles and particles containing a mixture of protein and fatty acids were also identified. The techniques and observations described in this case study will be useful for the identification
of microparticles in pharmaceutical products.
5:15 HELM: Setting the Standard for Biomolecular Data Exchange
Sergio Rotstein, Ph.D.,
Director, Research Business Technology, Pfizer, Inc.
The Hierarchical Editing Language for Macromolecules (HELM) enables the representation of complex biologic entities such as oligonucleotides, peptides, proteins, antibodies and bioconjugates in a flexible and compact fashion. The technology, originally
developed at Pfizer Incorporated, was released into the public domain through the Pistoia Alliance and is well on its way to becoming the industry standard for the exchange and manipulation of complex biomolecule structures and their associated
data.
5:45 Networking Reception in the Exhibit Hall with Poster Viewing
7:00 End of Day
THURSDAY, MAY 7
8:00 am Morning Coffee
8:30 Chairperson’s Remarks
Wim Jiskoot, Ph.D., Professor of Drug Delivery Technology, Leiden University, The Netherlands
8:35 Leveraging the Power of Mass Spectrometry to Study ADME Properties of Therapeutic Biologics
Ji Ma, Ph.D., Principal Scientist,
Amgen
Benefiting from rapid advances in protein engineering field, discovery of therapeutic biologics has evolved from primarily monoclonal antibodies to different molecular types, e.g. ADCs, huFc and HSA fusions. Using several case studies, cutting
edge mass spectroscopic technologies have been applied to understand ADME properties of therapeutic biologics in discovery stage.
9:05 Using Hydrogen-Deuterium Exchange Mass Spectrometry to Characterize Protein Higher-Order Structure and Dynamics
George Bou-Assaf,
Ph.D., Scientist, Biogen Idec
Hydrogen/Deuterium exchange mass spectrometry provides superior assessment of protein higher-order structure. Here, we discuss the applications of HDX-MS in various aspects of drug discovery and development. We showcase how its use in comparability
and biosimilarity studies enables stronger regulatory filings. We discuss how the technique has been instrumental for epitope mapping and in characterizing protein-protein or protein-ligand interactions. Finally, we explore how it could be
used in protein formulation development.
9:35 Terahertz Spectroscopy of mAb Formulations
Christopher
van der Walle, Ph.D., Principal Scientist, Development, MedImmune
Terahertz time domain spectroscopy (THz-TDS) provides insight into the interaction between proteins and water, or ‘hydration shell’, by analyzing the nonlinear relationship between protein concentration and THz absorption. Distinct
changes in THz absorption were observed for mAbs formulated up to 150 mg/ml in different excipients: NaCl, sucrose, proline and arginine. Relating these changes to key formulation parameters such as viscosity will improve our understanding
of mAb behavior at high concentrations.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing
11:05 Light Obscuration Measurements of Highly Viscous Solutions: Sample Pressurization Overcomes Underestimation of Subvisible Particle Counts
Wim Jiskoot, Ph.D., Professor of
Drug Delivery Technology, Leiden University
Light obscuration (LO) is a standard technique for subvisible particle analysis of therapeutic protein products. Some of those, however, exhibit high viscosities, which can lead to an underestimation of subvisible particle concentrations. To solve
this problem, we evaluated the application of sample pressurization during analysis, which turned out to be an elegant way to restore the reliability of LO analysis of highly viscous products without the need for dilution.
11:35 Protein Intermolecular Organization within Clusters in High Concentration Solutions Effectively Differentiates Crystallization, Liquid-Liquid Phase Separation, Gelation and Viscous Rheology Processes
Wenhua Wang, Ph.D., Postdoctoral
Fellow, Late Stage Pharmaceutical Development Department, Genentech, Inc.
Protein self-association, particularly at high concentrations, often leads to manufacturing and administration problems such as aggregation, high viscosity, opalescence, and liquid-liquid phase separation (LLPS). Here, the physical nature of phase
behavior in concentrated monoclonal antibody solutions is addressed by characterizing protein interactions and cluster structures. A molecular-level understanding of protein self-assembly of different high concentration phase behaviors helps
to control opalescence/LLPS and mitigate viscosities in drug product formulations.
12:05 pm Adopting Imaging and Other Analytical Techniques to Better Characterize and Study Novel Therapeutic Modes Such As the DVD-IgTM Molecule
Ivan Correia, MBA, Ph.D.,Senior Principal Research Scientist,cHead, Global Protein Sciences,
AbbVie Bioresearch Center (ABC)
12:35 End of Conference
5:15 Registration for Dinner Short Courses