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Protein Aggregation and Emerging Analytical Tools


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FRIDAY, JANUARY 13

7:30 am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee

 

Protein Aggregation and Immunogenicity

8:15 Chairperson’s Opening Remarks

Ulf Nobbmann, Ph.D., Technical Scientific Advisor, Malvern Instruments

8:20 The Regulatory Challenges of Protein Aggregation

Jack Ragheb, Ph.D., Senior Regulatory Research Officer and Principal Investigator, Division of Therapeutic Proteins, FDA

Large protein aggregates are known to be produced during the pharmaceutical manufacturing of therapeutic protein products. However, our understanding of how protein aggregate attributes such as size contribute to this immunogenicity is very limited. This talk will focus on subvisible protein aggregates, how they may interact with the immune system, and the potential impact these particles could have on a product’s safety and efficacy profile.

8:50 Protein Aggregation and the Immune System: Friend or Enemy? In vivo Models to Investigate Impact of Protein Aggregates on Formation of Anti-Drug Antibodies

Melody Sauerborn, Ph.D., Pharmaceutics, University of Utrecht; CEO, ADA InVivo BV

It has become clear that protein aggregates are very potent in inducing formation of anti-drug antibodies and thus causing immunogenicity. The precise immunological mechanism remains unclear. Mice immune tolerant for the protein drug of interest are powerful tools to investigate the immunological mechanisms underlying immunogenicity and they also provide a useful tool in protein drug development.

9:20 Predictive Tools for Immunogenicty

Wim Jiskoot, Ph.D., Professor, Drug Delivery Technology, Leiden University

9:50 Sponsored Presentation (Opportunity Available)

10:05 Networking Coffee Break in the Exhibit Hall with Poster Awards

 

CHALLENGES AND ADVANCES IN ANALYTICS TO CHARACTERIZE PROTEIN AGGREGATES

11:00 Aggregates and their Analysis: Some Perspectives and Lessons from Working with 100+ Products

John Philo, Ph.D., Vice President, Director of Biophysical Chemistry, Alliance Protein Laboratories

This talk will give examples of aggregate analysis and aggregation issues drawn from experience with well over 100 different therapeutic and vaccine products. A particular focus will be things which our clients often forget to do, common sample selection and assay interference pitfalls, and common misunderstandings about aggregate types, aggregation mechanisms, and what AUC and light scattering methods can (and cannot) measure. The examples will include the use of AUC and/or light scattering methods to help qualify or cross-check SEC methods.

11:30 Addressing New Analytical Challenges in Protein Formulation Development

Tizita Mammo, Associate Scientist, Parenterals & Liquids, Drug Product Development, Centocor R&D, Inc., a Johnson & Johnson Company   

12:00 pm Sub-Visible Particle: How do we Know Which Method is Telling the Truth?

Danny Chou, Ph.D., Individual Consultant; Former Head, Bioprocess Analytical Science, Genzyme

The uncertainties that surround the analysis and control of sub-visible particles and its significance in formulation development and quality control of biopharmaceutical products have been receiving considerable attention recently. Despite the uncertainties in the relationship between sub-visible particulates and immunogenicity and technical limitations of current particle detection technologies, it is generally agreed that the best strategy to ensure product quality and patient safety is to utilize complementary technologies in order to obtain the most complete information while allowing one to verify data obtained by orthogonal methods. The goal of this presentation is to share the latest advances using the complementary/orthogonal approach to sub-visible particle characterization.

12:30 Close of Morning Session

12:45 Luncheon Presentations (Sponsorship Opportunity Available) or Lunch on Your Own

 

CHARACTERIZATION OF SUB-VISIBLE PARTICLES

2:00 Chairperson’s Opening Remarks

John Philo, Ph.D., Vice President, Director of Biophysical Chemistry, Alliance Protein Laboratories

2:05 Characterization of Submicron Particle Distributions in Biologics Formulations

Flaviu Gruia, Ph.D., Scientist, Analytical Biochemistry, MedImmune

Nanoparticle Tracking Analysis is a novel technique which has the potential to enhance the current analytical capabilities for detecting, sizing and counting of particles in the sub-micron range. Method performance has been tested with particle standards and several different monoclonal antibody solutions. Sub-micron particle populations and how they evolve over time following different sample manipulations such as dilution, filtration, fill-finish, pH have been measured and will be presented as case studies. Trends observed in the sub-micron particle concentrations suggest that such manipulations can trigger formation of new particles due to different mechanisms. Correlations with techniques that probe different size ranges will be included.

   Sponsored by
Assay Designs Enzo Life Sciences 
2:35 Fluorescence-Based Detection of Protein and Peptide Aggregates:  From Test Tube to Tissue
Wayne F. Patton, Chief Scientific Officer, Enzo Life Sciences
A novel fluorescence-based aggregation assay is described that is suitable for monitoring protein /peptide aggregation generated by thermal-, mechanical agitation- and freeze-thaw-induced stresses.  Stabilized subvisible protein particle reference standards facilitate rapid generation of a standard curve by rehydration, facilitating quantification of the aggregated material.  Advantages of monitoring aggregation using the fluorescence microplate-based assay include simple implementation, ability to quantify minute levels of aggregation, ~30 minute analysis time and robust assay performance.  The described technology has been extended to the detection of inclusion bodies containing aggregates present within cultured cells and formalin-fixed paraffin-embedded brain tissue.

3:05 Characterization of Sub-Visible Particles in Therapeutic Protein Formulations

Tobias Frommknecht, Formulation Scientist, pRED, F. Hoffmann-La Roche Ltd

First a small volume light obscuration (LO) method will be introduced and some limitations to the microflow digital imaging (MDI) methodologies with regards to their ability to characterize sub-visible particles (SbVP) are elucidated. Secondly the effect of fluids with specific physical properties typically observed for aqueous protein formulations – such as turbid, viscous and colored fluids – is presented with regards to counting and sizing abilities of LO and MDI methods using particle suspensions of different buoyancy and contrast. Finally some possibilities of shape analysis in more detail, the limitations and relationships of different shape attributes and the potential of particle morphology parameters will be presented.

3:35 Measurement of Protein Unfolding/Refolding Kinetics and Structural Characterization of Hidden Intermediates by NMR Relaxation Dispersion

Derrick Meinhold, Ph.D., Investigator, Analytical Biochemistry & Biophysics, GlaxoSmithKline

Detailed understanding of protein function and malfunction hinges on the ability to characterize transiently populated conformational states. NMR R2 relaxation dispersion experiments were used to investigate spontaneous unfolding and refolding events of native apomyoglobin revealing a three-state unfolding mechanism leading to a molten globule. NMR chemical shifts of the transient states highlight regions of energetic frustration that crack during unfolding.

4:05 End of Conference


Day 1 | Day 2 | Download Brochure


Links to Companion Meetings

Pipeline 1 

Protein-Device Combinations 

Lyophilization and Emerging Drying Technologies

Optimizing Biologics Formulation Development

Pipeline 1

Higher Throughput Protein Purification

Protein Aggregation and Emerging Analytical Tools