2015 Archived Content
Cambridge Healthtech Institute’s Second Annual
Analytical Characterisation of Biotherapeutics
Advances in Analytical Methods for Better Developability Assessment of Molecules
5-6 November 2015
Analytics play an important role throughout a product’s lifecycle, from candidate lead selection, to preparation for IND and characterizing for lot release. At Analytical Characterization of Biotherapeutics, we will take a closer
look at advances in analytical technologies today that offer more accurate and faster methods to characterize structure-function relationships; examine developments in high throughput testing and bioassays for binding and stability; analyze post-translational
modifications and glycosylation effects on quality and stability of biologics as well as characterize complex molecules and novel formats such as bispecifics, ADCs and fusion proteins.
Thursday, 5 November
12:30 Registration
13:00 Dessert Break in the Exhibit Hall with Poster Viewing
13:30 Chairperson’s Opening Remarks
Roman A. Zubarev, Ph.D., Professor, Division of Chemistry I; Head, Department of Medical Biochemistry & Biophysics, Karolinska Institutet
KEYNOTE PRESENTATION
13:35 Massive de novo Sequencing of IgG Variants in Human Blood by Mass Spectrometry
Roman A. Zubarev, Ph.D., Professor, Division of Chemistry I; Head, Department of Medical Biochemistry &
Biophysics, Karolinska Institutet
The traditional proteomics assay is complemented by profiling both IgGs and co-extracted proteins using de novo sequencing by HCD and ETD MS/ MS. Each of these two additional domains (IgGs and co-extracted proteins) adds at least as much information
to patient stratification as the direct proteomics assay. New IgG peptides discovered by de novo sequencing contribute significantly to the predictive power of IgG-omics, and are potentially indicative of the disease etiology.
14:20
Biophysical Characterisation for Selection of Robust Humanised Therapeutic Antibody Candidates
Alison Turner, Group Leader, Biophysics, Biology, UCB Celltech
Panels of humanised antibodies from UCB’s new Core Discovery Platform are transiently expressed and purified, then screened using a number of assays and biophysical techniques to select therapeutic candidates with optimal chemical and physical stability.
This process provides early information on ‘manufacturability’ by reducing the aggregation risk during different purification steps (shear stress, buffer effects) and confidence in the stability of the drug product during storage and administration
(high concentration effects).
14:50 Computational Approaches to Optimise Antibody Efficacy & Pharmaceutical Developability as Therapeutic Agents
Hugues-Olivier Bertrand, Ph.D., Senior Director, Biologics Presales, Senior Fellow, BIOVIA Science Council
We will present how the convergence of BIOVIA capabilities supported by a common platform can be designed to help with the discovery and optimization of biotherapeutic candidates, in particular the management and analysis of all scientific and quality data generated throughout the process. We will highlight predictive analysis in Discovery Studio for early candidate selection and optimization. This includes high throughput antibody annotation and structure prediction, developability assessment to help improve stability, solubility and viscosity.
15:20 Refreshment Break in the Exhibit Hall with Poster Viewing
16:05
Characterisation of Acidic Species in Monoclonal Antibody
Li Zang, Ph.D., Senior Scientist, Analytical Development, Biogen
Acidic species in monoclonal antibody are highly heterogeneous and challenging for detailed characterization. They may post impacts on the function and stability of the monoclonal antibody. A detailed characterization of acidic species in a monoclonal
antibody biopharmaceutical will be presented in this talk. The potential impact of acidic species on function and stability of the antibody will be discussed.
16:35 Late-Stage Characterisation of a Therapeutic Enzyme
Peter Bernhardt, Ph.D., SeniorScientist, Analytical Development, Shire
An approach for late-stage characterization will be discussed that focuses on critical quality attributes. This approach includes developing a comprehensive understanding of product-derived substances and impurities formed during manufacturing and
under relevant storage conditions, as well as structure elucidation of product variants and understanding of structure/function relationships. A complex glycoprotein used for enzyme replacement therapy will be used as an example.
17:05 End of Day
17:00 – 17:30 Dinner Short Course Registration*
SC8: The Challenge of Protein Aggregation and Formation of Sub Visible Particles in the Development of Biopharmaceuticals
SC9: Advanced Techniques for Characterisation of Protein Aggregates, Particulates and Contaminants
(*Separate registration required.)
Day 1 | Day 2 | Speaker Biographies | Download Brochure
Friday, 6 November
07:30 Morning Coffee
08:30 Chairperson’s Remarks
Vishal Kamat, Ph.D., Scientist, Biomolecular HTS Center, Therapeutic Proteins, Regeneron Pharmaceuticals
08:35 High-Throughput Label-Free SPR Binding Kinetics Assay for the Characterisation of Fully Human Therapeutic
Antibodies
Vishal Kamat, Ph.D., Scientist, Biomolecular HTS Center, Therapeutic Proteins, Regeneron Pharmaceuticals
Most of the biosensors today have limited throughput capacity and are unconducive to perform high-throughput affinity screening of mAbs. In an effort to support Regeneron’s therapeutic antibody drug discovery, we have successfully optimized
the affinity screening methods to expand the throughput capability using Biacore 4000 and MASS-1, beyond originally anticipated from these two instruments. Strategies adopted to increase throughput and comparability of the binding rates constants
measured using this modified techniques will be presented.
09:05
Analysis of Classical and Novel Biotherapeutics with High-Throughput MCE Assays
Stefanie Wohlrab, Ph.D., Post-Doc, Pharma Technical Development, Roche Diagnostics GmbH
Continuous process automation and Design of Experiments (DoE) increase the number of samples that need to be processed. Microchip capillary electrophoresis (MCE) provides a high-throughput platform to monitor product quality. Here, we demonstrate
the use of Perkin Elmer´s LabChip GXII platform to characterize as well as to monitor antibody fragments during bioprocessing. The MCE assay shows a good linear range, high sensitivity and provides a resolution superior to CE-SDS for
some aspects.
09:35
Bioassays to Quantify a Therapeutic Fc-Fusion Protein in Preclinical Studies
Kelly Loyet, Ph.D., Scientist, Biochemical and Cellular Pharmacology, Genentech, Inc.
Pre-clinical studies were performed to evaluate the pharmacokinetics and efficacy of a potential therapeutic Fc-fusion protein. This talk will summarize the development of bioassays to quantify the pharmacokinetics of the therapeutic protein as
well as markers of its pharmacodynamics. Due to a large number of studies performed and low concentration dosing groups, assays were optimized for efficiency and sensitivity. Both mouse and cynomolgus monkey studies will be discussed.
10:05 Trending and Statistical Tools for the Consistent Monitoring of Bioassay Characteristics
Erica Bortolotto, Ph.D., Scientist, Bioassay Development, Analytical Sciences for Biologics, UCB Pharma SA
How to monitor a method’s performance over time? Which parameters should be trended? How to ensure a continuous validation of the method or the assessment of the right concentration for critical reagents or the bridging between reference
standards? This talk will present an overview of the problems via case studies presenting trending and statistical tools.
10:35 Coffee Break with Poster Viewing
11:00 Method for Postsynthetic Characterisation of Multi Antibody Polymalic Acid Conjugates
Eggehard Holler, Ph.D., Professor & Research Scientist III, Neurosurgery, Cedars-Sinai Medical
Center
The method enables us to characterize arrays of multiple different antibodies covalently attached to biopolymalic acid based multifunctional nanoconjugates. The technology applies mild ammonolysis for cleavage which leaves proteins in their
native state. Antibody content and multiple target (antigen) binding activities are validated by antibody structure and target (antigen) binding analysis using quantitative ELISA and size exclusion HPLC before and after specific cleavage
and degradation of the polymalic acid platform.
11:30 Analytical Methods to Characterise Bispecifics
Samadhi Vitharana, Ph.D., Principal Scientist, Core Sciences & Technology, Takeda California
Bispecific antibodies (BsAbs) targeting multiple antigens are one of the most dynamic classes of protein therapeutics in the pharmaceutical industry. Due to the complexities associated with production and purification processes of this therapeutic
modality, it is important to first assess developability based on purity, functionality, homogeneity, thermal stability, and other important biophysical properties. This presentation will discuss analytical approaches to effectively investigate
these developability and manufacturability properties of BsAbs
12:00 Understanding Size Dependence of Rabbit Vitreal Clearance
Whitney Shatz, Ph.D., Senior Research Associate, Protein Chemistry, Genentech, Inc.
Due to the relatively rapid clearance of protein drugs from the eye following intravitreal administration, maximal efficacy requires frequent dosing, typically monthly or bi-monthly. Although there is some data suggesting molecular weight
(MW) may be important to vitreal clearance and ocular tissue distribution, systematic protein studies have not been performed. Using a rabbit ocular model, we tested this hypothesis to further our understanding of ocular pharmacokinetics.
12:30 High-Throughput Characterisation of Monoclonal Antibodies Using the Fortebio Octet HTX
Jesper Pass, Ph.D., Principal Scientist, Novo Nordisk A/S
In order to meet an increasing demand for antibodies
for therapeutic use and as research reagents, a state of the art automated workflow for high throughput generation of monoclonal antibodies (mAbs) has been established. With an
increase in the number of antibodies generated, comprehensive characterization of large panels of antibodies is essential to ensure early selection of mAbs with the desired properties for further development. In order to ensure high
throughput characterization, we utilize label-free Bio-Layer Interferometry (BLI) analysis performed on a Fortebio Octet HTX instrument. The Octet HTX has been implemented in the screening for specific binders, and characterization
of selected mAbs with regards to affinity and epitope binning. The instrument is further used for selection and prioritization of high expressing clones in production cell-line development. Examples of the use of the Octet HTX in the
automated high throughput mAb generation process will be presented.
13:00 Enjoy Lunch on Your Own
13:30 Session Break
14:00 Chairperson’s Remarks
Huijuan Li, Ph.D., Director, Analytical Development, Biologics Bioprocess Development, Merck Research Laboratories
14:05 Multi-Angle Effector Function Analysis of Human Monoclonal IgG Glycovariants
Tilman Schlothauer, Ph.D., Senior Scientist, Biochemical and Analytical Research,
Large Molecule Research, Roche Pharma Research and Early Development (pRED), Roche Innovation Center Penzberg
To assess Fc functionality, many different approaches for characterization exist, more or less reflecting the physiological mechanism of Fc receptor recruitment. Here we combined different ways to measure Fc functionality. In addition
to classical antibody Fc receptor interaction studies of eight different enzymatically engineered IgG1 glycosylation variants, we analyzed also the behavior of these variants as a preformed immune complex on the new developed Fc Receptor
affinity chromatography.
14:35 High-Throughput Glycoprofiling via glyXbox: A High-Performance Glycoanalysis-System Based On xCGE-LIF
Erdmann Rapp, Dr. rer.nat., Head, Bio/Process Analytics, Max Planck Institute for Dynamics of Complex
Technical Systems
We have developed a glycoanalysis approach, based on multiplexed capillary gelelectrophoresis with laser induced fluorescence detection (xCGE-LIF), which shows high potential for high-performance analysis of glycoconjugates. The applicability
of the system is demonstrated for different types of glycosamples (biopharmaceuticals, vaccines, human milk and blood serum). This novel modular high-performance glycoanalysis system allows fully automated, highly sensitive, instrument-,
lab- and operator-independent “real” high-throughput glycoanalysis, a contrast to the prevailing costly and lab-space intensive methods.
15:05 Characterisation of Process Impurities, Product Variants, Post Translational Modifications for Biologics
Huijuan Li, Ph.D., Director, Analytical Development, Biologics Bioprocess Development, Merck Research Laboratories
Multiple case studies will be discussed on characterization of recombinant monoclonal antibody (mAb) drugs and their degraded and/or post-translationally modified counterparts, drug product- related impurities and variants for successful
development of biotherapeutics.
15:35 Biosimilar Development: Comparability and Similarity Analytics
Alok Sharma, Ph.D., Head, Analytical Development, Lupin Pharmaceuticals
Biologics and biosimilars therapeutics have made significant foot-print in pharmaceutical industry. Due to patent exclusivities of early phase biological drugs are reaching to their end, lot of competition is growing to manufacture
these off-patent biologics. As in case of biologics, “Process is the Product”, it becomes critically important to evaluate products manufactured by different routes. On the other, because of the complexity in structures
and the structure–function relationship of biological therapeutics, such changes may lead to changes in molecular structures, which may adversely affect the quality, safety, or efficacy of the drug. Drug manufacturers must
demonstrate the comparability of their products after process and formulation changes to ensure similar quality, safety, and efficacy. Biosimilars also require evaluation of their equivalency to the innovators' products. By complementing
traditional biochemical methodologies, biophysical characterization, using a variety of methodologies, can enhance product knowledge in terms of higher order structure, molecular size distribution, and the properties of aggregates.
The state-of-the-art biophysical and structural techniques in comparability assessments provide critical insight in biosimilarity evaluation.
16:05
Exploring the Generation of Variants in QbD
James Sutter, MSc, MBA, Associate Manager, Biotech Process Sciences, Downstream Process, Merck Serono
This talk will define QbD and its main components such as TPP, CQA, CPP, process characterisation, process & product knowledge, control strategy and design space. In the frame of QbD, optimising quality of biologics by generation,
separation and characterisation of variants is key for product and process understanding. This presentation shows case studies on isolation and characterization of low & high molecular weight and charge antibody variants to
explore various separation techniques for clearance.
16:35 End of Conference
: Unpublished Data | : Case Study
Day 1 | Day 2 | Speaker Biographies | Download Brochure