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 requiring industry companies to provide ever more complex data across a
wide range of analytical methods. And instrumentation suppliers are striving to support this new era with unique product features, software and feature combinations. The PEGS Characterization of Biotherapeutics meeting explores
these changes in the progression of analytical development, and offers a case study forum for those working in the field to share ideas, experiences and solutions that support the development of exciting new biotherapeutics.
Sunday, April 30
SC2: Translational Considerations for Development of Monoclonal Antibodies and Emerging Constructs: Part I: Focus on Construct Design
SC7: Translational Considerations for Development of Monoclonal Antibodies and Emerging Constructs: Part 2: Focus on Preclinical Development
*Separate registration required.
MONDAY, APRIL 30
7:00 am Registration (Commonwealth Hall) and Morning Coffee (Harbor Level)
8:30 Chairperson’s Remarks
Jennifer F. Nemeth, PhD, SCPM, Director, Biophysics, Structural Characterization, Biologics Discovery Sciences, Janssen Research & Development
8:40 Development of a Cell-Based Assay Measuring Bispecific Antibody Binding to Predict PK-PD Responses
Lynn Kamen, PhD, Scientist, Bioanalytical Sciences, Genentech
PK-PD modeling has traditionally relied upon drug concentration in serum along with affinity generated by SPR to predict efficacy and exposure. However, the advent of novel drug formats such as bispecific antibodies has heightened the need to
develop biological characterization assays that are able to better predict PK-PD responses. This presentation will highlight a case study in which a novel cell-based binding assay was used to predict the PK-PD effects of a bispecific therapy.
9:10 Characterization of CAR Ts and Cell Therapies
Eric S. Alonzo, PhD, Scientist, Cellular Analytics, bluebird bio
Clinical-grade CAR T cell drug products contain a heterogenous mixture of phenotypically and functionally distinct cells. Such heterogeneity necessitates innovative strategies to define biomarkers that may predict responses to CAR T cell therapy.
We improved biomarker characterization of our CAR T cell drug products by combining high dimensional mass cytometry with global gene expression analysis. These strategies identified multiple distinct memory T cell populations that may be associated
with positive outcomes in CAR T cell therapy.
9:40 Characterization of Gene Therapy Products
Zhenhong Li, PhD, Senior Director, Analytics, RegenxBio
Thanks to the pioneers’ perseverance and innovations, after greater than 20 years of development, the gene therapy industry is taking serious steps to advance therapeutics, leveraging knowledge and understanding from well-characterized biologics.
The presentation will share our approach in selecting analytical methods to assess product characteristics with regard to product identity, content, purity/impurity, potency, and structural integrity in support of early phase clinical trials,
including product release, stability and comparability.
10:10 Networking Coffee Break (Harbor & Mezzanine Level)
10:50 Top-Down Mass Spectrometry Strategy for Novel Drug Characterization
Zhe Zhang, PhD, Senior Scientist, Integrated Biologics Profiling, Novartis
Mass spectrometry has shown to be a powerful tool to characterize different therapeutic protein formats. With peptide mapping, site-specific characterization can be achieved. Top-down mass spectrometry, however, can provide added value, for example
avoiding digestion and artifact generation, sequence coverage on special regions, etc. Two case studies focusing on post-translation modification and clipping will be presented using top-down mass spectrometry. Integration of this technology
can facilitate drug candidates developability-assessment.
11:20 Analytical Characterization and Control of mAb Combination Products
Michael Adamo, Senior Scientist, Bristol-Myers Squibb
Immuno-Oncology (I-O) may require targeting multiple pathways in the immune system leading to the emergence of combination I-O products and significant analytical challenges. This talk will discuss the analytical strategy that can be applied to
control and characterize the individual mAb products in a combination I-O product. Several separation methods will be discussed for the control and characterization of key quality attributes of the combination mAb product including size and
Anthony Mire-Sluis, PhD, Head, Global Quality, AstraZeneca
Novel modalities can challenge existing paradigms because they often require science and risk-based thinking that does not ‘fit’ familiar development pathways. Understanding the quality attributes of a new molecular entity can challenge
standard analytical technology both physicochemical, and in particular, with biological assays. Creating robust manufacturing processes and control strategies can be challenging with even mature molecular platforms, so ‘out of the box’
thinking is necessary for novel modalities.
12:20 pm Comprehensive Workflow by Capillary Electrophoresis – Mass Spectrometry (CE-MS) for Biotherapeutics Drug Development
Mei Han, Senior Scientist, Pharmacokinetics & Drug Metabolism, Amgen, Inc.
Characterization of protein therapeutics by mass spectrometry is an essential part of all stages of drug discovery and development. Herein we discuss our CE-MS workflow for analysis protein therapeutics from pre-dose and post-dose samples.
The discussion will include measuring intact mass, reduced samples, charge variants, and characterization of in vivo biotransformation.
12:50 Luncheon Presentation I: Next-Generation Capillary Electrophoresis Technology for Protein Analysis
Karyssa Edwards, Assistant Scientist, Analytical Development, Celldex Therapeutics
Biotherapeutic proteins are complex molecules and challenging to analyze. Capillary electrophoresis (CE) is a highly efficient separation method which affords superior resolution, short separation time, and small sample volumes. The Maurice system innovates
the conventional CE technology by combining both cIEF and CE-SDS detection schemes into a fully automated instrument, allowing your protein profiling either by size or charge. In this presentation, we demonstrate the application of Maurice system
for analysis of biotherapeutic proteins.
1:20 Automated Data Processing and Analysis for Quality Monitoring of Biotherapeutics by Multi-Attribute Method (MAM)
Joe Shambaugh, Head Genedata Expressionist, Genedata
Mass spectrometry (MS) enables simultaneously measurement of multiple biotherapeutic critical quality attributes (CQAs) at the molecular level. Applying a multi-attribute method (MAM) can increase product quality while reducing development and manufacturing
costs. We present a MAM implementation using a single software platform for processing, analysis, and management of MS data. Dedicated workflows for a given biomolecule were tailored to measure CQAs, test for impurities, and check system suitability.
1:50 Session Break
2:20 Problem-Solving Breakout Discussions (Commonwealth Hall)
Analytical Issues in the Development of Gene Therapy Products
Zhenhong Li, Ph.D., Senior Director, Analytics, RegenxBio
- How to design and execute In vitro potency assay for gene therapy applications?
- How to mitigate the limitation on sample requirements for gene and cell therapy quality control testing?
- How to carry out stability studies leverage data by using platform approaches?
Implementing MAM in GMP Environment
Da Ren, PhD, Principal Scientist, Amgen
Rich Rogers, PhD, Scientist 4, Just Biotherapeutics
- What is the MAM?
- Instrumentation and software
- Implementation timeline
- Challenges and concerns
- Communication with regulatory agencies
High Throughput Predictive Methods and Technologies for Better Candidate Selection
Deniz B. Temel, PhD, Senior Research Investigator, Bristol-Myers Squibb
- Emerging technologies predicting aggregation, viscosity and long-term stability
- How predictive are the predictive techniques?
- Challenges in automating analytical assays
- How to improve our developability assessment by using automation?
- What kind of strategies we might implement for some of the techniques to make them more robust and automated?
3:20 Networking Refreshment Break (Harbor & Mezzanine Level)
4:00 Chairperson’s Remarks
Peter Fung, PhD, Senior Manager Product Marketing, NanoTemper Technologies
4:10 Challenges and Opportunities in Engineering Protein Biopharmaceuticals
K. Dane Wittrup, PhD, C.P. Dubbs Professor, Chemical Engineering and Biological Engineering; Associate Director, Koch Institute
for Integrative Cancer Research, Massachusetts Institute of Technology (MIT)
Arthur C. Clarke’s First Law posits that “When a distinguished but elderly scientist states that something is possible, he is almost certainly right. When he states that something is impossible, he is very probably wrong.” Bearing
this in mind, in this talk, I will highlight areas of protein drug development that appear poised for breakthroughs in the coming decade or so.
4:55 The Next Generation of Cancer Immunotherapy: Targeting Myeloid Immune Checkpoints
Kipp Weiskopf, MD, PhD, Resident Physician, Internal Medicine, Brigham and Women’s Hospital
Immune cells of the myeloid lineage hold tremendous potential as effectors of cancer immunotherapy. The CD47/SIRPα axis is a key molecular pathway that governs the interaction between myeloid cells and tumors. Therapies that target the interaction
are effective across multiple preclinical models of cancer and are now under investigation in clinical trials. Further studies have revealed additional regulators of myeloid cell activation that can be exploited as myeloid immune checkpoints.
5:40 Welcome Reception in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
7:15 End of Day
TUESDAY, MAY 1
8:00 am Registration (Commonwealth Hall) and Morning Coffee(Harbor Level)
8:25 Chairperson’s Remarks
Zhimei Du, PhD, Director, Bioprocess, Merck & Company, Inc.
8:30 The Impact of Antibody Variants on the Quality Attributes of Immunoconjugates
Alex Lazar, PhD, Director, Analytical and Pharmaceutical Sciences, ImmunoGen, Inc.
One of the main components of immunoconjugates is the targeting antibody to which the payload molecules are attached. The antibody variants can have substantial impact on the consistency of the conjugation process and/or on the quality of the
conjugated material. For monitoring the impact of antibody variants (e.g. tri-sulfides, incomplete masking) on the attributes of immunoconjugates, appropriate analytical methods were developed and results will be presented.
9:00 In vitro Biological Characterization of Kadcyla (Trastuzumab Emtansine)
Shan Chung, PhD, Principal Scientist and Group Leader, Genentech
Kadcyla (a.k.a. trastuzumab emtansine/ado-trastuzumab emtansine) is an antibody-drug conjugate composed of a humanized anti-HER2 antibody (trastuzumab) covalently conjugated to a cytotoxic drug (DM1). Kadcyla is indicated for the treatment of
patients with HER2-positive metastatic breast cancer. This presentation will describe in vitro characterization of Kadcyla for biological activities pertinent to its mechanisms of actions, including DM1-induced
apoptosis and trastuzumab-mediated inhibition of cell proliferation and induction of effector functions.
9:30 Size Exclusion Chromatography Mass Spectrometry (SEC-MS) – A Universal Approach for Quantitating DAR and Drug-Distribution of ADCs in a Research and Development Environment
Jay Jones, Senior Research Associate, Seattle Genetics
Native-intact mass analysis by SEC-MS can be used to quantitatively determine the drug-to-antibody ratio (DAR) of ADCs. To demonstrate suitability for use with a variety of ADC modalities, we successfully qualified this high-throughput DAR determination
method using a variety of cysteinyl-linked ADC modalities. The SEC-MS method could be a universal approach for DAR determination to support process development in early phase, prior to the development of chromatographic assays used for release.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
10:50 Linking Analytical Findings to Potency, Safety and Immunogenicity Risk Assessment
Vibha Jawa, PhD, Director, Biologics and Vaccine Development, Merck
Even with stringent control specifications, risk factors are often based on bioanalytical characterizations not necessarily related to biological thresholds and safety risks. It is difficult to evaluate clinical findings with lot to lot changes
in product quality attributes. This talk reviews existing risk assessment tools and their applications as well as gaps in being able to perform a risk assessment related to changes in product quality attributes during different stages
of biotherapeutic development.
11:20 Proteomics Identifies a CHO Host Cell Protein that May Impact Polysorbate Degradation
Kelvin Lee, PhD, Professor, Chemical & Biomolecular Engineering, University of Delaware
There is great interest in a better understanding of difficult to remove host cell proteins. We used proteomics approaches to identify different classes of difficult to remove CHO host cell proteins. One of the proteins that was identified
may play a role in polysorbate degradation. We evaluated cell line development strategies to mitigate the expression of this protein and any resulting impact on polysorbate degradation.
11:50 Glycosylation Profile Characterization
Nathan Brown, PhD, Senior Scientist, AbbVie Bioresearch Center
Next to the amino acid sequence, protein oligosaccharide content can greatly influence function and disposition of a biologic. Complex therapeutic formats, such as fusion proteins, often contain multiple, surface-exposed sites of glycosylation which can significantly impact pharmacokinetics, tissue penetration, distribution and activity. Here, we present our strategy, utilizing multiple, orthogonal approaches, to characterize micro- and macro- glycan heterogeneity and guide early development.
12:20 pm Luncheon Presentation I: Methods for Process Residuals in Biopharmaceutical Production Processes
Ian Parsons, Director, Analytical Laboratory, Charles River
As companies develop manufacturing processes, they need to characterize them and show they consistently and reliably clear process residuals to acceptable levels in the drug substance. This presentation will describe the development, validation
and application of a number of quantitative and limit test analytical methods for residuals, including complex analytes such as polyethylenimine, as well as traditional bioprocess residuals such as IPTG, antibiotics, antifoam, etc.
12:50 Luncheon Presentation II (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:20 Ice Cream Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
2:00 Chairperson’s Remarks
Krishnan Sampath, PhD, Senior Director, Analytical and Drug Product
2:05 Advancing Multi-Attribute Method in GMP Environment
Da Ren, PhD, Principal Scientist, Amgen
As an emerging technology, Multi-Attribute Method (MAM) is a powerful assay that can monitor product quality attributes at the amino acid level. Recent technology advancement enables MAM to be implemented in GMP environment, which makes
it possible to continuously monitor site-specific product quality attributes (PQAs) throughout the manufacturing process of biotherapeutics from product design to release testing. Considerations on implementing MAM in GMP environment
will be discussed in this presentation.
2:35 MAM Consortium’s New Peak Detection Round Robin
Rich Rogers, PhD, Scientist, Just Biotherapeutics
New peak detection, the purity component of the MAM, ensures novel modifications on biotherapeutics and process impurities are not overlooked. The MAM consortium has completed a New Peak Detection round robin study. The study had 34 participants
from around the world. The NIST mAb was used for the study. The study included a reference sample, peptide spiked sample, stressed sample, and unknown sample. Multiple types of mass spectrometers and software packages were evaluated.
3:05 Lessons from the Development and Execution of an Automated Purification, Digestion, and Targeted LCMS Analysis (MAM) of Biotherapeutics
Anders Lund, PhD, Scientific Director, Bioanalytics Characterization Group, BioPharmaceutics Development, Sanofi
Multi Attribute Monitoring (MAM) can be used to track critical quality attributes (CQAs) all along the product lifecycle. To achieve aggressive TTC goals, a testing paradigm including automated protein A purification, enzymatic digestion
and MAM analysis of biotherapeutic products was tested at three biopharmaceutical development sites (Framingham, Frankfurt, Vitry). The lessons learned, in addition to the challenges to design a proper testing paradigm across automation
for sample purification digestion and LC-MAM will be presented.
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
4:25 Development of Analytical Control Strategy for Late-Stage Biologics Programs
Krishnan Sampath, PhD, Senior Director, Analytical and Drug Product Sciences, MacroGenics
Monoclonal antibodies and bispecific DART® molecules are being developed for a variety of indications including immune-oncology. Stage-appropriate and risk-based analytical control strategy needs to be developed to ensure product
quality as molecules progress from early to late stages of development. This presentation will discuss analytical control strategies developed based on evolving product understanding and process knowledge using the above molecules
as case studies.
4:55 New Approaches for Characterization of Extractables and Leachables in Drug Product
Stacey Helming, PhD, Scientist, Analytical Sciences, Regeneron Pharmaceuticals
Single use components used in biomanufacturing processes require some degree of extractable and/or leachable (EL) risk assessment prior to use. How this assessment is performed, and what data are required, is largely left to the
end user to determine. Improvements to efficiency in EL programs will be discussed, including what constitutes quality extractable data, guidance on leveraging existing data to risk-assess components, and conducting leachable
studies at critical points during the process to ensure a comprehensive EL assessment.
5:25 End of Characterization of Biotherapeutics
5:30 Registration for Dinner Short Courses (Commonwealth Hall)
SC8: Introduction to Biophysical Analysis for Biotherapeutics: Discovery & Development Applications
*Separate registration required.