characterizing biotherapeutics

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


07:45 Registration (Foyer A) and Morning Coffee (Foyer D)

Auditorium VIII

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 AgencyYuan 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

Atouf_FouadFouad 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

Hegazy_MahaMaha 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

Rant_Ulrich_RBEUlrich 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)

Auditorium VIII

11:15 Anisotropy Resolved Multi-Dimensional Emission Spectroscopy (ARMES): A Multivariate Approach to Intrinsic Protein Emission Analysis

Ryder_AlanAlan 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

Lohrig_Urs_PPCUrs 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

Menet_Jean-MichelJean-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 

Hoffmann_MartinMartin 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

Auditorium VIII

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

Gervais_AnnickAnnick 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

Chen_RachelRachel 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)

Auditorium VIII

16:15 The Transition to Native MS in a Biopharmaceutical Development Lab – Lessons Learned and the Road Ahead

Kristensen_DanBachDan 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

Bones_JonathanJonathan 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



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 practices
  • What need to be tested and what shall be the quality requirements for the tests
  • Gearing up for upcoming USP<800>


19:45 End of Day


08:00 Registration (Foyer A) and Morning Coffee (Foyer D)

Auditorium VIII

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

Deuel_JuliaJulia 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

Dubey_SachinSachin 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 KolinkoSebastian 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

Sra_KuldipKuldip 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.


Wyatt Technology 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.

Horiba_Scientific 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*

17:3020:30 Dinner Short Courses

Recommended Short Course*

SC8: Advanced Analytical Technologies for Developability and Early Formulation Assessments - LEARN MORE

*Separate registration required.