The Analytical Characterisation of Biotherapeutics at PEGS Europe brings analytical scientists together to share their knowledge and experiences in developing new approaches to characterize molecules and residual impurities; while expanding their toolbox
to include exciting technologies such as ARMES, aptamers, native MS and MAM, etc.
Final Agenda
WEDNESDAY 20 NOVEMBER
07:45 Registration (Foyer A) and Morning Coffee (Foyer D)
08:30 Chairperson’s Opening Remarks
Jonathan Bones, PhD, Principal Investigator, CCL, National Institute for Bioprocessing Research and Training
08:35 KEYNOTE PRESENTATION: An International Collaboration: Towards the Standardisation of Gene Therapy
Yuan Zhao, PhD, Principal Scientist, Leader, Gene Therapy Section, Advanced Therapy Division, NIBSC, Medicines & Healthcare Products Regulatory
Agency
Potential safety risks, limited efficacy, or ethical conflicts may present challenges in the success of developing GTMP. Manufacturing hurdles, including changes in production sites and manufacturing processes, pose challenges to developers regarding
reproducibility and comparability of results for gene therapy. Introduction of an International Standard for gene therapy is especially important, given the usually orphan nature of the diseases to be treated with gene therapy, hampering the comparison
of cross-trial and cross-manufacturing results. This presentation will discuss challenges and regulatory perspectives in quality control and standardization of gene therapy and an international effort in developing the 1st WHO International Standard
for gene therapy products.
09:05 USP Standards and Best Practices for Advanced Therapies
Fouad Atouf, PhD, Vice President, Global Biologics, U.S. Pharmacopeia
The development of advanced therapy medicinal products offers great opportunities for therapeutic innovation; some challenges remain to be resolved for successful development and entry of these products to the healthcare market. Some of the challenges
relate to the lack of consistency in quality of raw materials and the lack of harmonized analytical methods across the industry. The United States Pharmacopeia (USP) is committed to working with regulators and developers of advanced therapies on the
standardization of analytical methods to assess the quality of these products throughout their lifecycle. This presentation will provide an overview of best practices and standardized procedures and associated physical reference materials in support
of this important segment of the industry.
09:35 Mapping Analytical Methods for Quality Assessment of Biotherapeutics and Biosimilars; Quality Attributes and Regulatory Considerations Perspectives
Maha Hegazy, PhD, Professor,
Analytical Chemistry, Cairo University
The FDA, EMA, and ICH have drawn attention to a number of structural features that have to be assessed to confirm consistent batch production and ensuring control of the manufacturing process for regulatory acceptance. Significant differences between
batches need to be investigated, thus integrated advanced analytical methods with new tools of design of experiments (DOE) and data analysis are needed to generate maximum information for quality assessment of biotherapeutics and comparability of
biosimilars to the reference product. Mapping analytical methods for multiple quality attributes is also required to ensure the method’s ability to detect relevant differences between samples.
10:05 Bispecific Binding Kinetics Analysed with a Two-Colour switchSENSE® Biosensor
Ulrich Rant, PhD,
CEO, Dynamic Biosensors GmbH
- Simultaneous detection of interactions of bispecific binders with two antigens
- Analysis of on- and off-rates
- Avidity vs. affinity analysis
- Engineering of binding selectivity
10:35 Coffee Break in the Exhibit Hall with Poster Viewing (Rio Pavilion)
11:15 Anisotropy Resolved Multi-Dimensional Emission Spectroscopy (ARMES): A Multivariate Approach to Intrinsic Protein Emission Analysis
Alan G. Ryder, PhD, Professor,
Nanoscale Biophotonics Laboratory, National University of Ireland Galway
Fluorescence anisotropy can be related to protein structure, size, and aggregation profile. When implemented using multi-dimensional measurements of intrinsic protein emission and combined with multivariate analysis, one can extract potentially very useful
diagnostic information. Here we show how these 4D measurements can be applied to the study of protein structure changes, PEGylation reactions, and for bioreactor monitoring.
11:45 Aptamers and Enzyme Cascades as New Tools for Analytical Characterization of Biopharmaceuticals
Urs Lohrig, PhD, Lab
Head, Physico-Chemical Characterisation, Novartis
Probing higher order structure of biopharmaceuticals is the domain of instrumental analytics like CD, FT-IR, NMR and X-ray crystallography. Here, we present two simple approaches to supplement the analytical toolbox: Aptamer technology and an Analytical
Cascade of Enzymes (ACE) – both probing molecular structures. Aptamers offer an adoptable, not immunogenic-driven selection process and long-term supply of critical reagent in contrast to polyclonal antibodies. ACE detects structural differences
in mAbs at a 1% level – a range inaccessible by most instrumental methods.
12:15 NEW: Characterization from Developability to BLA
Jean-Michel Menet, PhD, Head, Characterization, Biologics Development/BioAnalytics, Sanofi
Characterization toolbox is evolving along the development phase of therapeutic proteins (i.e., from developability studies to the filing of the BLA) and to suit protein modality (e.g., IgG1 to multispecific). Examples applied to monospecific and multispecific
antibodies will be given showing toolbox used for early phase projects and for late phase projects. Approaches under development for CQA-driven CMC development will also be presented: high order structure technologies such as HDX-MS and NMR, native
MS, MAM.
12:45 How
Merck is Going to Simplify the Paradigm of Mass Spectrometry: from Characterization
Tool to Routine QC Method
Angelo
Palmese, PhD, Head, Characterization & Innovative Analytics, Analytical
Development, Biotech, Merck
13:15 Luncheon Presentation I: Automated Multi Attribute Method Analyses for Process Development and Characterization of mAbs
Martin Hoffmann, Senior Scientist, Research & Development – Bioanalytics Frankfurt – Bioprocess Analytics, Sanofi-Aventis Deutschland GmbH
We present a mass spectrometry based approach to simultaneously monitor critical quality attributes (CQA) of therapeutic monoclonal antibodies (mAbs) - which has been termed multi-attribute method (MAM). With MAM we are supporting the process development
of mAbs. For CQA quantification, mAbs samples were digested on an automated liquid handling robot and analyzed by HPLC separation in combination with high resolution mass spectrometric detection.
13:45 Luncheon Presentation II (Sponsorship Opportunity Available)
14:15 Session Break
14:30 Chairperson’s Remarks
Fouad Atouf, PhD, Vice President, Global Biologics, U.S. Pharmacopeia
14:35 Quantitation of HCP by Mass Spectrometry as a Method to Control the Quality of Biopharmaceuticals
Annick Gervais, PhD, Director, Analytical Development, Biologicals, UCB Pharma SA
15:05 Monitoring of Clearance of Lipase Host Cell Proteins in mAb Manufacturing Using a LC-MRM Quantitation Method
Rachel Chen, PhD, Scientist II, Analytical Development, Biogen
Successful removal of host cell proteins (HCPs) is very important for biopharmaceutical product development to ensure product quality and safety. Recently, it has been demonstrated that certain lipases may be the cause for enzymatic degradation of polysorbate
20 and 80, which are common surfactants used in protein formulations. An LC-MS/MS method was developed to achieve sub ppm quantitation level of three lipases. The method has been applied to monitor the clearance of lipases for various mAbs under different
downstream processes.
15:35 Refreshment Break in the Exhibit Hall with Poster Viewing (Rio Pavilion)
16:15 The Transition to Native MS in a Biopharmaceutical Development Lab – Lessons Learned and the Road Ahead
Dan Bach Kristensen, PhD, Principal Scientist, Symphogen
In recent years, native MS has gained significant popularity as a tool for intact mass analysis of large biomolecules. Key strengths include the ability to interface a variety of chromatographic techniques with the MS, and the excellent quality of the
spectral data. At Symphogen, native MS is established as the method of choice for intact mass analysis, and here learnings from the transition to native MS and thoughts on future applications will be presented.
16:45 Probing Biopharmaceutical Microheterogeneity Using Native LC-MS
Jonathan Bones,
PhD, Principal Investigator, CCL, National Institute for Bioprocessing Research and Training
Hyphenation of charge variant analysis using pH gradient cation-exchange chromatography to high-resolution Orbitrap mass spectrometry under native conditions (CVA-MS) has recently been described. Here we demonstrate the power of CVA-MS for profiling microheterogeneity
of biopharmaceutical product quality attributes on both drug substance and drug product. We will also discuss how high-resolution native LC-MS can be applied for automated process monitoring when combined with automation solutions to create a data
generation engine to support Manufacturing 4.0.
17:15 A Reliable and Automated Workflow for LC-MS MAM Analysis of Biopharmaceuticals – From High Throughput Sample Preparation to Data Evaluation
Patrick Sascha Merkle, PhD, Postdoc, Analytical Development & Characterization, Novartis Pharma AG
The LC-MS multi-attribute method (MAM) has emerged as a promising approach for the characterization and relative quantification of critical quality attributes on biopharmaceutical molecules. Here, we present our peptide-level LC-MS MAM workflow that relies
on high-throughput sample preparation, high-resolution MS acquisition, and automated data evaluation in the Genedata Refiner MS software. We envisage that the simplicity and state of automation of the presented LC-MS MAM workflow may allow its routine
use in a non-expert laboratory.
17:45 Networking Reception in the Exhibit Hall with Poster Viewing (Rio Pavilion)
18:45 Problem-Solving Breakout Discussions*
TABLE 16: Native MS in Biopharmaceutical Development
Moderator: Dan Bach Kristensen, PhD, Principal Scientist, Symphogen
- How is native MS applied during biopharmaceutical development?
- What are the advantages/disadvantages relative to conventional intact MS?
- How robust is the native MS platform?
- What kind of non-covalent interactions are people studying with native MS?
- Is native MS currently applied in QC (release testing according to specifications)?
TABLE 17: MAM – A Moving Target
Moderator: Patrick Sascha Merkle, PhD, Postdoc, Analytical Development & Characterization, Novartis Pharma AG
- Peptide mapping versus subunit analysis
- Define the limits of relevance
- High resolution versus low resolution MS instrumentation for MAM
CANCELLED
TABLE 18: In-use Stability for Biologics
Moderator: Sachin Dubey, PhD, Deputy Director/Head, Formulation, Analytical and Drug Product Development, Glenmark Pharmaceuticals
Generally acceptable approaches and good practicesWhat need to be tested and what shall be the quality requirements for the testsGearing up for upcoming USP<800>
19:45 End of Day
THURSDAY 21 NOVEMBER
08:00 Registration (Foyer A) and Morning Coffee (Foyer D)
08:30 Chairperson’s Remarks
Annick Gervais, PhD, Director, Analytical Development, Biologicals, UCB Pharma SA
08:35 Analytical Characterization of a Complex Product: Lentiviral Vector
Julia Deuel, MSc, Senior
Scientist, Analytical Characterization, bluebird Bio
Traditional molecular biology techniques can provide in-depth understanding of lentiviral vector activity and structure, but are often low-throughput and highly variable, contributing disproportionately to COGs, delays in batch release, and potential
batch failures if assays cannot be repeated. Presented here are techniques for characterization of lentiviral vectors with a focus on elucidation of vector structure for evaluation of lot consistency and lentiviral vector comparability following manufacturing
changes. These include modifications to commonly used techniques along with new technologies to provide a broad evaluation of lentiviral vector characteristics and impurities.
09:05 Strategy to Establish Clinically Relevant Specifications at Launch
Monika Meier, PhD, Manager, Development Analytics, Roche Diagnostics GmbH
Specification acceptance criteria are typically based on the understanding of critical quality attributes, clinical experience, and manufacturing capability. With shortened development timelines and few clinical lots, justifications of acceptance criteria
are focused on science- and risk-based assessments of patient impact, providing a balance between appropriate control over high-risk attributes to ensure product quality for the patient, and flexibility for low-risk attributes, as appropriate, for
a robust supply chain.
09:35 NEW: Potent Bispecifics, Overcoming Analytical Challenges Enroute Preparation for FIH
Sachin Dubey, PhD, Deputy
Director/Head, Formulation, Analytical and Drug Production Development, Glenmark Pharmaceuticals SA
There are around 130 ongoing clinical trials with different bispecifics formats (including Glenmark’s proprietary BEAT molecules); they are potent and are dosed at extremely low levels (low ng/mL concentration). Preparing for FIH at such low concentration
is a significant challenge from the analytical standpoint – quantification is challenging, release testing has to be adapted, and prevention against surface adsorption has to be ensured. Product characterization, de-risking manufacturing, and
in-use stability for IV infusion are required to be carefully designed and executed with additional controls. Glenmark has three bispecifics in clinical development and experiences gained during their development will be discussed.
10:05 A Platform Approach to Manage Developability and Manufacturability Risks of Biologics Molecules
Sebastian Kolinko, PhD, Scientific Consultant, Biologics, Genedata
We present a workflow system that enables systematic developability and manufacturability assessments, using both in silico and high throughput analytical confirmatory methods, over the entire biologics R&D process
from initial discovery all the way to final candidate selection. We show use cases for mAbs and other complex multi/bispecific formats and discuss building predictive developability models utilizing this system. We also present the underlying molecule
and task management needed for analytical organizations to accomplish this.
10:35 Coffee Break in the Exhibit Hall with Poster Viewing (Rio Pavilion)
11:15 Kinetic Mechanism of Controlled Fab-Arm Exchange for the Formation of Bispecific Immunoglobulin G1 Antibodies
Mark Chiu, PhD, Associate Director, BioTherapeutics Analytical Development, Janssen Research & Development, LLC
A combination of FRET, non-reducing SDS-PAGE, and strategic mutation of the Ab hinge region was employed to characterize the cFAE process. Fluorescence correlation spectroscopy (FCS) was used to determine the affinity of parental (homodimer) and bispecific
(heterodimer) interactions within the CH3 domain to further clarify the thermodynamic basis for bsAb formation. The result is a rate constant mechanism with the dissociation of the K409R parental Ab into half-Ab controlling the overall rate of the
reaction.
11:45 Rapid Release of Autologous Cell Therapy Products to Patients: A Road Less Travelled
Kuldip Sra, PhD, Senior Director,
Technical Operations, CRISPR Therapeutics
For autologous cell therapy products, each patient is a product batch. Manufacturing is a very tedious and manual process. Urgency to release the product quickly to the patient is very high. The presentation will cover the implementation of rapid analytical
methods to release the final product in a desired timeframe to patients.
12:15 Luncheon Presentation I: Beyond Aggregation: a Wealth
of Applications of High-Throughput DLS/SLS for Biologics and NanoDDS
Christian Sieg, PhD, Analytical
Service, Wyatt Technology
The DynaPro® Plate Reader III is used extensively throughout the biopharmaceutical industry to screen protein-based therapeutics for developability and optimal formulation conditions. While detection of aggregation is the most familiar application
of this instrument, it is in fact far more versatile. This seminar will discuss advanced applications to biologics including solubility, aggregation kinetics, conformational and colloidal stability and optimizing crystallization conditions. Further,
native reversible self-association of IgGs associated with high viscosity is addressed.
12:45 Luncheon Presentation II: Quantifying Protein Aggregation
and Subvisible Particle Size and Concentration with Nanoparticle Tracking Analysis
Jeff Bodycomb, Product Line
Manager, HORIBA
Quantifying protein aggregate and subvisible particle size and concentration in formulations remains a continuing challenge. These particles are too small to be imaged by optical microscopy; fractionation techniques change solution conditions and thereby
aggregation; and the size distribution is too polydisperse for classical, single laser NTA. These issues can be addressed with multi-laser NTA, which simultaneously uses three lasers with different wavelengths to collect data on all of the particles
in the suspension.
13:15 Dessert Break in the Exhibit Hall with Poster Viewing (Rio Pavilion)
14:00 End of Analytical Characterisation of Biotherapeutics
17:00 Dinner Short Course Registration*
17:30 – 20:30 Dinner Short Courses
Recommended Short Course*
SC8: Advanced Analytical Technologies for Developability and Early Formulation Assessments - LEARN MORE
*Separate registration required.