As new product formats progress through development and into the regulatory process, the role of analytical characterization is taking on new meaning. Regulatory agencies are themselves learning about these new formats, and to support filings, industry companies are being asked to provide ever more complex data across a wide range of analytical methods. The use of platformed analytical programs, developed as a cost and time saver by the industry, is now diminishing as novel molecules and product formats demand more individualized characterization steps. And instrumentation suppliers are striving to support this new era with unique product features, software and feature combinations. The PEGS Characterization of Biotherapeutics meeting will explore these changes in the progression of analytical development, and offer a case study forum for those working in the field to share ideas, experiences and solutions that support the development of exciting new biotherapeutics.
MONDAY, APRIL 25
7:00 am Registration and Morning Coffee
8:30 Chairperson’s Remarks
Hubert Kettenberger, Ph.D., Principal Scientist, Protein Analytics, Roche Pharma Research & Early Development, Roche Innovation Center Penzberg
8:40 KEYNOTE PRESENTATION
Analytics: A View to the Horizon and Applications Advancing Attribute Control
Janet Cheetham, Ph.D., Executive Director, Attribute Sciences Core Technologies, Amgen Inc.
Concurrently, advances in analytical technologies have delivered significant improvements in sensitivity, resolution, speed and precision, as well as increased deployment flexibility through reduced footprints. With a view to the horizon, a strategic roadmap can be shaped and scientific and technology advances and transformation accelerated through a commitment to collaboration between industry peers and vendors leveraging established pre-competitive models, together with a continuing partnership and engagement with the international regulatory agencies.
9:10 Analytics by Design: Tailored Approaches to Detect Product-Related Impurities in Next-Generation Biotherapeutics
Hubert Kettenberger, Ph.D., Senior Principal Scientist, Protein Analytics, Roche Pharma Research & Early Development, Roche Innovation Center Penzberg
Generic analytical methods for the characterization of biotherapeutics save time and resources during early stages of development. However, generic methods may not provide enough information about critical quality attributes for next-generation biotherapeutics such as bispecific antibodies and fusion proteins. Here, we describe tailored approaches for the rapid analytical method development for such proteins. We combine theoretical considerations with analytical data collected during cell line development and developability assessments to draft a control strategy.
9:40 Biophysical and Functional Characterization of an Anti-TNF Polymer
Ivan Correia, Ph.D., Research Fellow; Head, Global Protein Sciences, AbbVie Bioresearch Center
We engineered repeats of the hydrophobic polypeptide “VPGXG” from the ECM protein elastin onto an anti-TNF monoclonal antibody. The engineered molecule retained properties of its parent and in addition showed a temperature driven phase transition (Tt). Insoluble aggregates were formed above the Tt and upon lowering the temperature the molecule re-dissolved and retained properties of the parent molecule. We exploit features of this engineered anti-TNF molecule for novel therapeutic applications.
10:10 Coffee Break
10:50 Selective, Non-Covalent Conjugation of Peptides with Proteins Using Host-Guest Chemistry
Christopher van der Walle, Ph.D., Fellow, MedImmune
In a host-guest chemistry approach, the macrocycle cucurbit[8]uril host is shown to selectively conjugate two guests: a recombinant protein with a synthetic peptide. This non-covalent approach does not require chemical modification of the protein and maintains the biological activity of the conjugated peptide. Calorimetry, fluorescence and light scattering data are used to fully characterize the heteroternary complex. The strategy represents a powerful approach for the development of novel therapeutic biologics.
11:20 All Lights are Changing to LEDs. Do Proteins Like It?
Erinc Sahin, Ph.D., Senior Research Investigator, Drug Product Science & Technology, Bristol-Myers Squibb
Light sources have evolved from incandescent to fluorescent to light emitting diodes (LEDs) at a fast pace within the last two decades. Light sensitivity is a well-known liability for protein therapeutics. Alternate light sources have different spectral signatures (wavelength distribution) with varying degradative potentials. In this study, we have compared effect of fluorescent and LED lighting (with comparable illuminations) on protein therapeutics, in order to investigate if protein therapeutic manufacturing areas can safely switch to LED lighting in their facilities.
11:50 High Throughput Assays for the Determination of the Potency and Comparability of Biosimilars and Innovator Products
Michael G. Tovey, Ph.D., INSERM Director, Research, Laboratory of Biotechnology & Applied Pharmacology, Ecole Normale Supérieure de Cachan
Biosimilar development is dependent upon establishment of validated and standardized assays that allow comparisons of innovator molecules and biosimilars. A validated standardized high throughput assay platform will be described that is applicable to most biopharmaceuticals and that allows direct comparison of drug potency and comparability of innovator molecules and biosimilars in the same assay. Case studies will be presented for biopharmaceuticals ranging from novel forms of human insulin and FGF-21 to structurally diverse TNF-(INSERT SYMBOL) antagonists.
12:20 pm Unlock Biologic Stability: Simultaneous Characterization Measurements at Low Volumes
Krista Witte, Ph.D., Vice President, Product Development, Unchained Labs
Early biologic stability decisions are constrained by the sample volume required for multiple analysis methods.This limits the available information on constructs and formulations until later development when more protein is available, which increases risk. We will present methodologies of producing more valuable information on protein and formulation stability, with simultaneous measurements of key analysis parameters.
12:50 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on your Own
1:20 Luncheon Presentation II (Sponsorship Opportunity Available)
1:50 Session Break
2:20 Problem-Solving Breakout Discussions
Product Quality Monitoring-Options and Trends
Christine P. Chan, Ph.D., Principal Scientist and Technical Lead, Global Manufacturing Science and Technology, Genzyme-a SANOFI company
- In-process analytics for process characterization
- Multi-attribute monitoring methods
- On-line, at line assays
- Process linkage studies
Bioprocess Impacts on Drug Product Quality
Shantanu Sule, Ph.D., Senior Engineer, Parenteral Drug Product Development, Biogen
- Impact of high yield drug substance (DS) process on drug product (DP) quality
- Impact of lot-to-lot variability in drug substance (DS) attributes on drug product (DP) stability
- Methods for characterizing process-related impurities
- General understanding about controlling post-translational modifications in DS process
- Learnings about process impurities in diverse biologics: mAbs, ADCs, bispecifics, hormones, cytokines, enzymes
3:20 Refreshment Break in the Exhibit Hall with Poster Viewing
PLENARY KEYNOTE SESSION
4:00 Chairperson’s Remarks
4:10 The Promise of Cancer Immunotherapy: An Overview of Recent Advances and Jounce’s Approach to Delivering the Right Therapy to the Right Patient
Deborah Law, D. Phil. CSO Jounce Therapeutics, Inc.
As immunotherapies become an increasingly important component of cancer treatment the challenge will be to identify ways to provide the best therapy(s) to the individual. This presentation will provide an overview of current cancer immunotherapies as well as highlight some of the challenges ahead including selection of optimal combinations, moving outside of T cell-directed approaches, and will highlight how Jounce Therapeutics is using its Translational Science Platform as an approach to develop and deliver the right therapy to the right patient.
4:50 Antibody as Drugs: Then, Now and Tomorrow
Paul J. Carter, Ph.D., Senior Director and Staff Scientist, Antibody Engineering, Genentech
Antibodies have grown into a clinically and commercially important drug class with more than >45 antibodies marketed for imaging or therapy in the USA and/or Europe and with ~$63 billion in worldwide sales in 2013. This presentation will highlight progress in developing antibody drugs and consider opportunities for future innovation.
5:30 Welcome Reception in the Exhibit Hall with Poster Viewing
6:45 End of Day
TUESDAY, APRIL 26
8:00 am Morning Coffee
8:25 Chairperson’s Remarks
Ivan Correia, Ph.D., Senior Principal Research Scientist; Head, Global Protein Sciences, AbbVie Bioresearch Center
8:30 Use of Molecular Dynamics to Predict Chemical Degradation
Lydia Beasely, Engineer, Early Stage Pharmaceutical Development, Genentech
Liquid formulation of therapeutic monoclonal antibodies requires the protein to be chemically stable during long-term storage. This presentation describes in-silico tools to predict potential for chemical modification. These tools involve building three-dimensional structures using homology modeling and then running molecular dynamics simulations. The potential for both Trp oxidation and Asp isomerization was predicted by extracting structural parameters from molecular dynamics simulations. The in silico tools described allow for rapid screening for mAb candidates during early discovery, enabling selection of molecules with optimal behavior.
9:00 The Role of Analytical Automation in Transforming Drug Product Development
Rahul Rajan, Ph.D., Director and Functional Area Head, Drug Product Formulation, Amgen, Inc.
The industry needs to develop drug products faster, in the face of new modalities that challenge the development status quo. We present case studies where assays and workflows critical to robust drug product development were miniaturized and/or automated. These include attributes such as viscosity and particulate level measurement, as well aspects such as sample preparation and labeling. These advances accelerate product development and create the capability to measure a larger design space.
9:30 Biochemical Analysis and Its Correlation with Functional Assays
Babita Parekh, Ph.D., Director, Bioanalytical Science, Eli Lilly and Company
Therapeutic monoclonal antibodies typically exist as a heterogeneous mixture of product variants. Furthermore proper protein folding is critical to an antibodies biological activity and efficacy. Post translational modifications that impart heterogeneity may change the biological functions of an antibody. Structural characterization helps in pinpointing the change and in establishing structure function correlation. Analytical work performed to understand the molecule and to support product safety, quality and efficacy will be discussed.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing and Poster Awards
10:50 Applications of an Automated and Quantitative CE-Based Size and Charge Western Blot for Therapeutic Proteins and Vaccines
Richard Rustandi, Ph.D., Principal Scientist, Vaccine Analytical Development, Merck Research Labs
Labor intensive SDS-PAGE and IEF slab gels have been replaced with CE-SDS and CE-IEF methods, respectively, in the biopharmaceutical industry. Another traditional slab gel technique is the western blot, which detects proteins using immuno-specific reagents after SDS-PAGE separation. This technique is very laborious, manual, time consuming and only semi-quantitative. Here, we describe the applications of a relatively new CE-based western blot technology that is automated, fast and quantitative.
11:20 The Neonatal Fc Receptor (FcRn) Binds Independently to Both Sites of the IgG Homodimer with Identical Affinity mAbs
Yasmina Abdiche, Ph.D., Research Fellow; Head, Bioanalytical Group, Rinat-Pfizer
FcRn plays a central role in extending an antibody’s half-life, yet published studies on the molecular binding mechanism of the FcRn/IgG interaction have been confounded by avidity, often due to an earlier hypothesis of disparate FcRn-binding sites. We demonstrate that two FcRn molecules bind an IgG homodimer with identical affinities at independent sites. Our in vivo studies highlight the biological importance of avidity in extending an IgG’s serum half-life.
11:50 Deciphering Factors that Have Impacts on Glycosylation of mAb and its Biophysical Properties
Zhimei Du, Ph.D., Senior Principal Scientist, Merck
Consistent and reproducible generation of mAb glycoform profiles still remains a considerable challenge in biopharmaceutical industry. To assess the mechanism of immature glycan process, and to minimize the environment-mediated lot-to-lot variations, we identified a broad range of factors that impact on the glycosylation maturation of mAbs. Our results indicate that high mannose may lead to changes in biophysical properties including protein conformation, thermal stability, colloidal stability, and aggregation propensity.
1:20 Ice Cream Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Remarks
Babita Parekh, Ph.D., Director, Eli Lilly and Company
2:05 Molecular Interaction Analysis of a Non-Equivalent Two-Site Protein-Protein Interaction with Multiple Biophysical Methods
Michael Doyle, Ph.D., Senior Principal Scientist, Protein Science, Bristol-Myers Squibb
Quantitative analysis of protein interactions with biophysical methods plays a key role in discovery of drug candidates. The accuracies of different biophysical methods is influenced by different caveats and assumptions. We used a bivalent form of barnase and barstar as a model system to test the abilities of ITC, SPR and AUC to resolve both binding equilibrium constants. The strengths, weaknesses, and synergies of these technologies will be discussed.
2:35 Optimization of the Sensitivity and Selectivity of an Immunoassay for an ADC mAb in Cancer Patients
Josh Albert, Bioanalyst, Bioanalysis, Immunogenicity and Biomarkers, GlaxoSmithKline R&D
Therapeutic target receptor up-regulation and shedding in disease populations can be problematic for adequate immunoassay sensitivity and selectivity. Competition between the up-regulated/shed target and immunoassay capture mechanisms require careful optimization to ensure the most robust tolerance to circulating target concentrations. Strategies for proper optimization of the capture reagent, sample diluent and wash steps to allow suitable clinical support will be discussed in detail.
3:05 Meet Maurice, The Advanced CE System
Alpana Prasad, Product Manager, iCE Marketing, ProteinSimple
Need cIEF and CE-SDS data for your biologics? In this presentation, we introduce the advanced CE system, Maurice™ that enables fast development of cIEF and CE-SDS methods for IgG Identity, Purity and Heterogeneity analysis with excellent reproducibility and precision. Just pop in one of his ready-to-go cartridges, drop in your sample vials or a 96-well plate, and hit start. The new data acquisition and analysis software reduces time to generate and review results. Additionally, the simple and streamlined workflow with automated shutdown processes minimize the hands-on time.
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing
4:25 Hydrogen Exchange Mass Spectrometry to Address Questions of Higher Order Structure for Biotherapeutics
Roxana Iacob, Ph.D., Research Assistant Professor, Northeastern University
The higher-order structure of proteins is responsible for their uniqueness and dictates their function. Higher-order structure can be interrogated with mass spectrometry and in particular by hydrogen exchange mass spectrometry (HDX MS). HDX MS is a widely used tool for the structural characterization of biotherapeutics and plays an important role in the biopharmaceutical industry. Here, recent advances in HDX MS and its applicability in biotherapeutics characterization will be presented.
4:55 A Customized HDX Workflow for Locating Protein Binding Interactions and Conformational Differences
Kristopher Truncali, Scientist, Mass Spectrometry, Boehringer Ingelheim Pharmaceuticals
Through changes in deuterium uptake, Hydrogen-Deuterium Exchange (HDX) mass spectrometry can identify differences in solvent accessibility. A customized HDX workflow was developed utilizing a Leap H/D-X PAL, an Orbitrap Fusion, and an in-house software platform. This lecture will provide an overview of the implementation, optimization, and limitations of this customized HDX workflow. Case studies will then be presented where protein binding interactions and conformational differences were identified by the HDX workflow.
5:25 End of Characterization of Biotherapeutics
5:30 Registration for Dinner Short Courses
Add these programs to your agenda for a full week of comprehensive coverage!
Biophysical Analysis of Biotherapeutics
Protein Aggregation and Stability in Biopharmaceuticals