PEGS Europe Summit Biologics Dev Stream

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.

Final Agenda

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Unpublished Data Icon: Unpublished Data | Case Study Icon: Case Study



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



13:35 Massive de novo Sequencing of IgG Variants in Human Blood by Mass Spectrometry

Roman ZubarevRoman 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.



Case Study Icon
Alison Turner
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 

Goupil_AnneAnne Goupil-Lamy, Principal Field Application Scientist, BIOVIA Science Council Fellow, BIOVIA

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

Case Study and Unpublished Data Icon
Zang Li
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., Senior Scientist, 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.)

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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 KamatVishal 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.

Unpublished Data Icon
Stefanie Wohlrab
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.

Case Study and Unpublished Data Icon
Kelly Loyet
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 HollerEggehard 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 ShatzWhitney 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_PassJesper 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 Luncheon Presentation
(Sponsorship Opportunity Available) or 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 SchlothauerTilman 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 RappErdmann 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 LiHuijuan 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_SharmaAlok 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.


Case Study Icon
James Sutter
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 Icon: Unpublished Data | Case Study Icon: Case Study

Day 1 | Day 2 | Speaker Biographies | Download Brochure