From assessing a molecule’s developability to determining the structure-function relationships, analytical techniques play an important role to identify and characterize the physico-chemical properties of the molecules, allowing scientists to better understand their molecules and design strategies to overcome challenges and optimize their druggability. These techniques include but are not limited to: microscopy, spectroscopy, light scattering, crystallography, calorimetry, chromatography etc.
At the 2nd Annual Advances in Characterization Methods & Approaches conference, we invite scientists to share their current thinking and latest techniques and tools they have used to elucidate protein structure and function, assess developability, evaluate particles and impurities, and to characterize the new and emerging biomolecules.
Final Agenda
Tuesday, March 21, 2017
12:20 pm Registration
1:45 Chairperson’s Opening Remarks
Danny Chou, Ph.D., President & Founder, Protein Formulation and Characterization, Compassion BioSolution, LLC
1:50 Featured Presentation: Cutting-Edge LC and CE-MS Methods for mAbs, Biosimilars, ADCs and Bispecific Antibodies
Alain Beck, Ph.D., Senior Director, Analytical Chemistry, NBEs, Center d’Immunology Pierre Fabre
The development and optimization of mAbs and related products rely on improving their analytical and bioanalytical characterization by assessing multiple critical quality attributes (CQAs). Case studies on progresses of multi-level (top, middle, bottom, SMDs) state-of-the-art MS methods will be presented. These methods include native MS, Ion Mobility-MS, CE-MS, 2D-LC-MS, Extended Bottom Up and Top Down Sequencing combined with chromatographic and electrophoretic techniques.
2:20 Solving Protein Development Challenges with Hydrogen Exchange Mass Spectrometry
David D. Weis, Ph.D., Associate Professor, Chemistry and Pharmaceutical Chemistry, University of Kansas
Hydrogen exchange mass spectrometry (HX-MS) is gaining acceptance for its ability to monitor higher order structure in proteins, in particular in antibody-based therapeutics. This talk will highlight two areas of recent work focused on solving challenges in development. First, work aimed at developing a molecular level understanding of how protein-excipient interactions alter physical stability will be presented. Second, the use of HX-MS to map sites of reversible self-association in IgG1 mAbs at protein concentrations > 50 mg/mL will be presented.
2:50 Integrated Methods for Fingerprint Analysis of Recombinant Proteins
Darryl Davies, Ph.D., Associate Director, Analytical Development, Janssen R&D
3:20 Automated Data Processing for Quality Monitoring of Biotherapeutics by Multi-Attribute Methods (MAMs)
Joe Shambaugh, Ph.D., Senior Scientific Account Manager, Expressionist, Genedata
MS-based methodologies enable measurement at the molecular level of many quality attributes of biomolecules. Applying these multi-attribute methods (MAMs) can increase product quality while reducing development and manufacturing costs. We present an implementation of MAMs using Genedata Expressionist®, the software platform for processing, analysis, and management of MS data. Fully automated workflows are employed as part of a bioprocess control strategy to measure critical quality attributes, while searching for impurities and checking the system suitability.
3:50 Refreshment Break in the Exhibit Hall with Poster Viewing
4:30 Leveraging the Multi-Attribute Method (MAM) to Improve Biotherapeutic Characterization
Richard Rogers, Ph.D., Scientist 4, Just Biotherapeutics
Characterization of complex biotherapeutics using highly resolving mass spectrometry has resulted in a better understanding of the post-translational modifications that are crucial for safety and efficacy. These modifications are used to guide the manufacturing process and the release strategy for biotherapeutics. We have developed and implemented a mass spectrometry based multi-attribute method (MAM) that monitors known modifications but also has the ability to identify new modifications on the biotherapeutics.
5:00 Considerations for Optimization of Multi-Attribute Peptide Mapping Protocols
Trina Formolo, Ph.D., Research Chemist, Bioanalytical Sciences Division, Material Measurement Laboratory, National Institute of Standards and Technology (NIST)
While a powerful, streamlined data analysis workflow is important in building a successful multi-attribute method (MAM) platform, the platform must also incorporate optimized sample preparation and analysis protocols. The generation of reliable, meaningful results from any MAM platform begins with quality sample preparation and analysis. This includes developing robust, reproducible tryptic digestion protocols and optimizing chromatographic separation methods to facilitate the detection and quantitation of critical quality attributes (CQA).
5:30 Subunit Analysis Leads to Simultaneous Measurement of Multiple Attributes of Monoclonal Antibodies
Linda Yi, Ph.D., Scientist II, Anaytical Development, Biogen
Heterogeneity of therapeutic antibody products as a result of enzymatic/chemical modifications during bioprocessing and storage is well-documented. Routine monitoring of the modifications is essential to ensure product quality. IdeS, an endopeptidase, cleaves antibody at heavy chains below the hinge region, producing F(ab’)2 and Fc/2 fragments. LC-MS analyses of the IdeS digestS, with or without further reduction, enables simultaneous characterization of domain-specific modifications and probing higher order interactions such as high molecular weight species.
6:00 Welcome Reception in the Exhibit Hall with Poster Viewing
7:00 End of Day
Wednesday, March 22, 2017
8:00 am Morning Coffee
8:30 Chairperson’s Opening Remarks
Alain Beck, Ph.D., Senior Director, Analytical Chemistry, NBEs, Center d’Immunology Pierre Fabre
8:40 Developability Assessment and Epitope Characterization at the Early Stage of Therapeutic Antibody Discovery
Sam Wu, Ph.D., Principal Scientist, Biologics Research, Janssen R&D
Biophysical and chemical properties are critical factors that can dictate success or failure in the development of therapeutic antibodies. Early quality control and risk assessment help prevent failure in later stages of antibody development. An early-stage developability workflow and high-throughput epitope selection were designed to set stage gate for molecules progressing to late-stage development. The collective information identify the best candidates to move forward through the development process.
9:10 Developability Assessment of Bispecific Antibodies: Analytical Platform and Stability Studies
Sagit Hindi-Jacoel, Ph.D., Principal Scientist, BioAnalytical Sciences, Eli Lilly and Company
The development of bispecific antibodies can present unique challenges for manufacturing and characterization. Attributes such as product purity, biophysical properties, stability and solubility are critical for the development of bispecific antibodies and need to be investigated at early stage of development. This presentation will discuss the analytical platform and stability studies that enable efficient developability assessment of IgG-like bi-specific antibodies.
9:40 Roundtable Breakout Session
Table 1: Technologies and Epitope Mapping Strategies for Antibodies
Moderator: Sam Wu, Ph.D., Principal Scientist, Biologics Research, Janssen R&D
- Antibody epitopes: A significant factor in therapeutic antibody selection
- Evaluation of biophysical technologies applied in epitope mapping
- Discussion on complementary strategies in mapping antigen-antibody interactions
Table 2: Rational Selection of Assays for Early Stage Characterization
Moderator: Liangyi Zhang, Ph.D., Senior Scientist III, Analytical Development, Abbvie
- Assays for discovery, candidate selection and developability assessment
- Throughput vs quality of analytical assays
- Adoption of novel technologies
- Bridging assays between Discovery and Development
Table 3: Developability Assessment for Complex Antibodies Platforms
Moderator: Sagit Hindi-Jacoel, Ph.D., Principal Scientist, BioAnalytical Sciences, Eli Lilly and Company
- Critical attributes for manufacturing risk assessment
- Challenges in characterization of complex therapeutics
- Choosing the right analytical platform for your molecule
- Using orthogonal methods for analytical characterization
- Essential stability studies for therapeutics development
Table 4: Analytical Tools and Techniques for Stability Monitoring and Formulation Development
Moderator: Danny Chou, PharmD, Ph.D., President, Compassion BioSolution, LLC.
10:40 Coffee Break in the Exhibit Hall with Poster Viewing
11:20 Selection of High Quality Biotherapeutics: A Staged Approach to Biophysical Assessment of Early Leads
Amy King, Senior Scientist, Biomedicine Design, Pfizer
A two-staged approach is used to identify lead molecules with superior biophysical properties and qualities that support ease of developability. Stage I assessment is designed to triage and de-select proteins that may have low thermal stability, high aggregation propensity, or a tendency toward self-association. Stage II assessment is performed on the prioritized leads to explore behavior at high concentration, such as solubility, viscosity, and colloidal stability. Another key selection criterion is performance in the forced degradation assay to assess sequence hot spot liabilities.
11:50 Analytical Characterization of Small Molecule Impurities in Biologics
Liangyi Zhang, Ph.D., Senior Scientist III, Analytical Development, AbbVie
12:20 Luncheon Presentation: Exposing Character of a Protein with Advanced Analytics
Gayathri Vasudevan, Ph.D., Principal Scientist Group Leader, Analytical and Formulations Development, FUJIFILM Diosynth Biotechnologies
The emergence of more complex biotherapeutics has diminished the use of platformed analytics and increased the demand for more advanced analytics and individualized characterization steps for development of these products. How we evaluate the "character of a protein" along the process development trajectory is a critical component in developing robust processes and overcoming challenges associated with these new, unique and difficult molecules.
1:20 Dessert Break in the Exhibit Hall with Poster Viewing
1:55 Chairperson’s Remarks
Sam Wu, Ph.D., Principal Scientist, Biologics Research, Janssen R&D
2:00 Optimizing Protein Stability through Integration of Cutting-Edge and Rapid Analytical Tools with Rational, Molecule-Specific Approach to Process Development
Danny Chou, Ph.D., President & Founder, Protein Formulation and Characterization, Compassion BioSolution, LLC
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. This presentation will illustrate how to properly combine some of these new tools with tried and true strategies that consider the key factors/forces that impact physical stability of proteins in solution. Emphasis will be placed on high-throughput, low sample requirement strategies that are useful for industrial application.
2:30 Excipient Quantification and Stability Monitoring Are Critical for the Formulation Development of Biologics
Joseph Liu, Ph.D., Principal Research Scientist, Formulation, Eli Lilly and Company
The quantity and stability of excipients have direct impact on formulations. They may affect stability and efficacy of API as well as stability and effectiveness of other excipients. Here, we display a method for monitoring the quantity and stability of a commonly used excipient in biologics formulation. Other excipients existing in the same formulation also changed the stability of this critical excipient. Real case studies and their outcomes are presented.
3:00 Recent Advancements in Differential Scanning Calorimetry for Stability Profiling of Biotherapeutics
Verna Frasca, Product Marketing Manager, Sales and Marketing, Malvern Instruments
Microcalorimetry is the gold standard technique for thermal stability characterization of protein drugs. DSC measures the protein’s thermal transition temperature (TM), and heat (ΔH) of unfolding. DSC data are used to optimize formulations, choose “developable” candidates, and comparability and biosimilarity studies. The new MicroCal PEAQ-DSC introduces advancements in DSC function, speed, and accessibility, including tools to support regulatory requirements. Protein stability characterization by DSC complements data from Dynamic Light Scattering and Taylor Dispersion Analysis.
3:30 Refreshment Break in the Exhibit Hall with Poster Viewing
4:10 Regulatory Perspectives in New Analytical Methods for Protein Aggregates Quantification and Characterization
Ewa Marszal, Ph.D., CMC Reviewer, Hematology Research and Review, Center for Biologics Evaluation and Research, US FDA
Characterization of the size, structural and chemical heterogeneity of protein aggregates in biotherapeutics requires the use of multiple analytical methods. To facilitate and enhance aggregates characterization new methods are being developed, current methods are being improved and established methods are being adapted from other fields. Understanding the advantages and limitations of these techniques by biotherapeutics manufacturers, analytical service labs and regulators is critical to the improvement of the product quality control and determining the links between the presence of protein aggregates and product safety.
4:40 SEC-MS Analysis: A Method for Fast Identification of Causes of Covalent Aggregate of Antibody
Chongfeng Xu, Ph.D., Scientist II, Analytical Development, Biogen
Middle-down subunit analysis of mAb using IdeS digestion has gained popularity for analyzing post-translation modifications, such as glycosylation. Here it was also demonstrated to be valuable for analyzing mAb aggregation. A reducing SEC-MS method with the use of mass spectrometry-compatible running buffer was developed to study highly-aggregated samples of a human IgG1 mAb, and the subunits involved in aggregation were confidently identified.
5:10 Investigation of IgG Monoclonal Antibody Aggregation by HDX, Cross-Linking, and Biophysical Tools
Joomi Ahn, Ph.D., Scientist II, Analytical Sciences, MedImmune
Aggregation in recombinant human monoclonal antibodies is one of the major concerns for the biopharmaceutical industry and regulators. In this study, we utilized various higher order structural analytics such as hydrogen deuterium exchange mass spectrometry, cross-linking mass spectrometry, and conventional biophysical tools to investigate the structural differences and the interaction of IgG aggregated forms.
5:40 Close of Advances in Characterization Methods & Approaches