Cambridge Healthtech Institute’s Inaugural

Protein Aggregates and Particles
Prevent. Detect. Minimize. Eradicate.
5-6 November 2014


Aggregates are under increasing regulatory scrutiny due to their potential to enhance immune responses, thereby compromising the safety and efficacy of the protein drug product. These impurities can be introduced anywhere throughout the development process, from upstream cell culture to downstream purification, from formulation to fill/finish. A thorough understanding of the source and mechanisms of aggregates and particles will help companies better predict the advent of aggregation, mitigate the impact of aggregates and develop safer and more stable drug products.

The Protein Aggregates & Particles conference brings together scientists, engineers and biochemists from analytical development, immunology, formulation, quality, process development and regulation to discuss methods to prevent, monitor, minimize and even eradicate these product-related impurities.


Speaker BiographiesDownload Brochure | Register 

Recommended Short Course*

SC5: Aggregation and Immunogenicity: How Do Formulation, Process and Delivery Influence Immunogenicity of Therapeutic Proteins?

*Separate Registration Required.


Wednesday, 5 November

07:45 Registration and Morning Coffee


MECHANISM OF AGGREGATES AND ENGINEERING TO IMPROVE DEVELOPABILITY

08:30 Chairperson’s Opening Remarks

Murali BilikallahalliMurali Bilikallahalli, Ph.D., Associate Director, Biopharmaceutical Development: Vaccines & Non-mAb Biologics, MedImmune LLC



KEYNOTE PRESENTATION

08:35 Cell and Process Engineering Strategies to Cure Cellular Aggregation of a Difficult to Express Fusion Protein

David JamesDavid C. James, Ph.D., Professor, Chemical & Biological Engineering, University of Sheffield

Based on kinetic modeling of the cellular synthetic process, we compared different strategies (cell engineering, modulation of culture environment) to alleviate cellular aggregation and the associated poor expression of a recombinant Fc-fusion protein by CHO cells. Our analysis serves as a paradigm for multivariate optimisation of DTE protein production.


09:05 Kinetic and Thermodynamic Correlations of Protein Folding & Stability to Aggregate & Particle Formation

Murali BilikallahalliMurali Bilikallahalli, Ph.D., Associate Director, Vaccines & Biologics, Biopharmaceutical Development, MedImmune LLC

Cofactors often have the ability to interact specifically with the unfolded polypeptides. Developing cofactor bound/free protein/protein conjugates is challenging since cofactors in most cases play direct role in structure, stability and activity. Fundamentals of protein folding pathways, cofactors assisted protein folding & stability, and some of the stability challenges in developing metal bound protein as a therapeutic drug will be addressed in this presentation.

 

09:35 A3D: Towards the Design of Aggregation-Resistant Biotherapeutics

SalvatorVenturaSalvador Ventura, Ph.D., Prof Biochemistry & Molecular Biology, Institute of Biotechnology & Biomedicine, University Autonoma of Barcelona

The molecular complexity of protein therapeutics makes them sensitive to stress during production and delivery. A major problem is the formation of aggregates in protein solutions, which impact the product properties and result in adverse immunogenic reactions. Therefore, tools to design aggregation-resistant proteins are receiving increasing interest. Here we introduce A3D which is able to predict and assist the redesign of the aggregation propensity of protein 3D-structures.

10:05 Determining the Stability and Structure of Therapeutic Proteins

Lewis_NeilE. Neil Lewis, CTO, Malvern Instruments & Head, Malvern Bioscience Development Initiative

Aggregation and native-state unfolding of therapeutic proteins is of critical importance to manufacturers and patients alike. The use of hybrid techniques lends itself to more thorough characterization of particulates in therapeutics. An approach that combines dynamic light scattering with Raman spectroscopy can measure the high-order structure of proteins while concurrently monitoring aggregation. Another hybrid technique, the combination of optical microscopy with Raman spectroscopy, allows for both the detection and identification of particulates in protein suspensions.

10:35 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 In silico Engineering to Improve Antibody Developability

Bojana PopovicBojana Popovic, Ph.D., Senior Research Scientist, Antibody Discovery and Protein Engineering, MedImmune

Due to mutations introduced during affinity maturation MEDI-1912 displayed an increase propensity to aggregate. Using an in silico spatial aggregation propensity tool we identified surface exposed hydrophobic amino acids responsible for aggregation. These amino acids were reverted to the parental sequence, resulting in a stable antibody without compromising potency or affinity for NGF. By improving the stability of MEDI-1912, we also improved the serum half-life in vivo.

Camera Icon PNDX 11:45 Aggregation of Therapeutic Proteins in Human Plasma and Human Blood: A New Research and Development Field

Tudor ArvinteTudor Arvinte, Ph.D., Chairman & CEO, Therapeomic, Inc.

After injection of a clear, not-aggregated protein solution, aggregation can form at the injection site in tissue or in the blood stream. The talk will present new data on characterisation of the aggregates formed in vitro when different formulations of therapeutic proteins are mixed with human plasma and blood. A model for the aggregation will be presented and some clinical implications of the aggregation induced by biopharmaceuticals will be discussed.

SensiQ 12:15 Identification of High MW Species using diSPR® Pulse Method

Reese_EricEric Reese, Ph.D., Vice President, Marketing, Sales and Marketing, SensiQ Technologies Inc.

The detection and subsequent analysis of protein aggregation is of hallmark importance in the discovery and development of biotherapeutic drugs. Standard methodologies used to detect aggregation can either be time consuming or cumbersome. This presentation describes how the diSPR® technique designated Pulse OneStep® can detect and measure simulated aggregation in proteins or other biomolecules. In addition we demonstrate that by monitoring the diffusion coefficient, aggregation events can be defined in bulk sample. Further research is underway to define the limits of detection. OneStep® marks a quantum innovation in the detection and analysis of sample aggregation by SPR biosensors.

12:45 Enjoy Lunch on Your Own


CHARACTERISING, PREDICTING AND DETECTING AGGREGATES AND PARTICLES

14:00 Chairperson’s Remarks

Pete M. Tessier, Ph.D., Associate Professor, Chemical & Biological Engineering, Rensselaer Polytechnic Institute, USA


KEYNOTE PRESENTATION

14:05 Characterising and Release Testing of Protein Aggregates in Biologics

Ewa MarszalEwa Marszal, Ph.D., CMC Reviewer, Division of Hematology Research and Review, Center for Biologics Evaluation and Research, U.S. Food & Drug Administration

Protein aggregates in biologics present a quality issue and raise safety concerns. Thus, their content should be minimized and controlled. This presentation will cover recent guidelines for particulate matter control and the advancements in particulate matter detection and metrology.


14:35 Using in silico Tools for Prediction of Propensity for Aggregation and for Viscosity

Bernhard HelkBernhard Helk, Ph.D., Head, New Technologies, Novartis Pharma AG

Three in silico tools and their practical application to the prediction and characterisation of protein-protein-interaction are demonstrated: SAP (Spatial Aggregation Propensity) identifies hydrophobic patches and is applied to engineer mAbs and ADCs with increased stability. DI (Developability Index) predicts aggregation propensities based on SAP and net charge. SCM (Spatial Charge Map) ranks mAbs according to viscosity. Case studies for predicting crystallization and viscosity of mAbs are presented.

15:05 Emerging Technologies for Characterising and Detecting Sub-Visible and Visible Particles

Andrea Hawe, Ph.D., CSO, Coriolis Pharma Research

Within the last years emerging technologies such as nanoparticle tracking analysis, resonant mass measurements, flow imaging microscopy or semi-automated visual inspection have been introduced for (sub-) visible particle analysis in biopharmaceutical products. An overview on emerging technologies for sub-visible and visible particle analysis will be given, challenges pointed out and highlighted with relevant examples and case-studies.

15:35 Refreshment Break in the Exhibit Hall with Poster Viewing

16:15 Optimising Monoclonal Antibody Selection and Formulation Using Self-Interaction Nanoparticle Spectroscopy

Pete M. TessierPete M. Tessier, Ph.D., Associate Professor, Chemical & Biological Engineering, Rensselaer Polytechnic Institute, USA

Improving selection of highly soluble antibody candidates early in the discovery process requires biophysical methods capable of assaying mAb self-association that are robust for handling hundreds to thousands of samples at extremely dilute antibody concentrations (microgram per mL) and in the presence of contaminants (cell culture supernatants). We report a method (affinity-capture self-interaction nanoparticle spectroscopy) for addressing these challenges and discuss its use in the discovery of new, highly soluble antibodies.

16:45 Stability Challenges for Protein Therapeutics: Structure Alteration and Aggregation Propensity

Joel RichardJoel Richard, Ph.D., Senior Vice President, Peptides, Head, CMC & Engineering Dreux Site, IPSEN

This talk will present biophysical analytical techniques to characterise protein aggregation as well as their aggregation propensities. Appropriate combination of biophysical methods will illustrate structural modifications of the protein in the formulations, display the effect of excipients on these modifications and the subsequent mechanism of formation of sub visible particles. Clinical impact will also be discussed, under the angle of potential safety issues, as identified by the regulatory agencies.


17:15 Problem Solving Roundtable Discussions:

Engineering Aggregate-Resistant Biotherapeutics

SalvatorVenturaModerator: Prof Salvador Ventura, Full Professor, Protein Folding and Conformational Diseases Lab (PFCD), Institute of Biotechnology and Biomedicine, Department of Biochemistry and Molecular Biology, Universitat Autònoma de Barcelona

1. Evolutionary constrains to protein solubility.
2. Can we predict aggregation in protein 3D-structures?
3. Can we predict the aggregation dependence on the solution conditions?
4. Can we engineer the aggregation propensity of proteins without altering other essential properties?
 

Protein Folding in Relation to Stability and Aggregate Formation

Murali BilikallahalliModerator: Murali Bilikallahalli, Ph.D., Associate Director, Vaccines & Biologics, Biopharmaceutical Development, MedImmune LLC





Biophysical Development and Characterisation in Early Drug Discovery

Bojana PopovicModerator: Bojana Popovic, Ph.D., Senior Research Scientist, Antibody Discovery and Protein Engineering, MedImmune  

  • What are some of the key challenges that are very hard to address?
  • Can we improve developability, quality by design approach?
  • Identify symptoms early and prevent rather than treat later once the molecule is in Development.
  • Can early drug discovery truly predict issues that may result at high concentration mAb development?
  • Selecting and screening for desired biophysical characteristics as early in the discovery process as practicable is desirable, to reduce costs and prioritise resources.  

Strategies for Analytical Characterisation of Sub-Visible Particles in Various Stages of Development

Moderator: Andrea Hawe, Ph.D., CSO, Coriolis Pharma Research

  • Which methods are available for sub-visible particle analysis for the nm- and µm-size range?
  • How to select the optimum set of methods for a particular protein/biopharmaceutical drug? 
  • Are emerging methods ready to move to a QC environment?
  • What are the unmet needs in the field of sub-visible particle analysis?
 

18:15 Networking Reception in the Exhibit Hall with Poster Viewing

19:15 End of Day One




Thursday, 6 November

08:00 Morning Coffee


08:30 Chairperson’s Remarks

Tudor ArvinteTudor Arvinte, Ph.D., Chairman & CEO, Therapeomic, Inc.



AGGREGATES AND IMMUNOGENICITY

08:35 Testing the Immunogenic Potential of Aggregates – In silico, in vitro and in vivo

Melody SauerbornMelody Sauerborn, Ph.D., Head of Non-Clinical Development, Mymetics BV and CEO, ADA InVivo

Although the precise characteristics of an aggregate responsible for immunogenic behavior are not known yet, efforts are made to predict the intrinsic probability of a biologic to form aggregates and the impact of aggregates in the clinics. This presentation will give an update on the latest tools and technologies to characterise, predict and test aggregates in silico, in vitro and in vivo.

09:05 Induction of T Cell Responses by Protein Aggregates: A Key Stage for the Immunogenicity of Biologics

Mark Fogg, Ph.D., Immunology Group Leader, Antitope Limited, an Abzena Company

This presentation will provide case study examples of how preclinical methods measuring drug induced T cell responses can be applied to select drugs with a reduced risk of clinical immunogenicity. Furthermore new data will be presented on how aggregates in some formulations can trigger innate responses that ultimately enhance immunogenicity. The clinical relevance to these findings will be discussed.


ENHANCING STABILITY AND OPTIMISING FORMULATION TO MINIMIZE AGGREGATES & IMMUNOGENICITY

09:35 Investigation of Reversible Self-Association during Early Stage Development of a Low Concentration Antibody-Drug Conjugate

Elizabeth Bartlett, Ph.D., Scientist, Analytical & Pharmaceutical Sciences, Immunogen, Inc.

Reversible self-association is often present in high concentration antibody products, but may also occur in lower concentration preparations. In the case of antibody-drug conjugates (ADC), a novel of molecules for the treatment of cancers, this property can present substantial challenges to successful formulations. In this study, a multi-technique approach was used to identify and investigate the effects of various excipients on reversible self-association in a low concentration ADC.

 Particle Metrix10:05 Reproducible and Fast Exosome Detection - The "Zeta View" System from Particle Metrix GmbH is the Answer!

Muehlbacher_RainerRainer Muehlbacher, DI (FH), MBA, Sales and Marketing Global Manager, Particle Metrix GmbH

Exosomes can potentially be used for prognosis, therapy, and biomarkers for health and disease but the characterization was challenging; Particle Tracking Analysis (NTA/PTA) is a technique where single particles of the sample are visualized and traced. By analysis of the particle traces, the NTA/PTA device (ZetaView®, Particle Metrix, Germany) performs the measurement of particle size, concentration and zeta potential of exosomes in one experiment.

10:35 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 High hydrostatic pressure as a novel method for characterization, identification and mitigation :  Building a better IFN-beta-1b

Amber Fradkin, Ph.D., Director, Analytical Characterization, BaroFold, Inc.

The development of an analytical platform for the detection and characterization of particles in therapeutic protein products has proven to be a challenging task for the pharmaceutical industry.  Many analytical tools have become available over the past 5-10 years to detect and characterize particles ranging in sizes from several nanometers to hundreds of microns.  However, these tools are unable to determine the origination of subvisible particles.  Particles are known to form during purification, process, shipping and handling procedures.  Particles may be stress induced or could be resultant of formulation issues.  In addition, the presence of proteinaceous nanoparticles has potential to lead to subvisible and visible particle formation with storage over shelf life.  We demonstrate how high hydrostatic pressure can be used as a novel tool to purify protein products as well as identify and trouble shoot root cause for particle formation, minimizing immunogenicity.  Case studies based on IFN-beta-1b will be presented.

11:45 NaCl-Induced Aggregation of Monoclonal Antibodies at Low pH: Prevention by Additives

Fabien BickelFabian Bickel, MSc, Ph.D., Student, Institute of Applied Biotechnology, Biberach University of Applied Science

Aggregation is a known phenomenon in downstream processing of monoclonal antibodies. Here we investigate a model system where mAb aggregation is induced by increasing the ionic strength at low pH. Aggregation can be partially reverted by lowering the ionic strength. The effect of protective osmolyte additives on aggregation kinetics and final aggregate concentration is investigated. The system has potential for buffer optimisation and formulation development.

12:15 End of Protein Aggregates and Particles conference



Speaker BiographiesDownload Brochure | Register