Cambridge Healthtech Institute’s 6th Annual

Detection and Characterization of Particulates and Impurities

Hot Topics, Emerging Contaminants and Impurities, Case Studies, and New Technologies

January 21-22, 2020

Part of the Analytics & Impurities pipeline

Cambridge Healthtech Institute’s 6th Annual Detection and Characterization of Particulates and Impurities conference will bring together leading researchers to discuss hot topics, emerging contaminants and impurities and new characterization tools for impurities that may come from various sources and stages of product development. Through new presentations, high-level poster presentations, and interactive discussions, top scientists will share new insights into characterization and control of various impurities. Some of the hot topics for this year will be new and novel technologies for aggregates and impurities in gene therapies, AAVs, virus and pathogen detection, host cell proteins, lipases and enzymatic degradation, other particles and aggregations, and chemistry and manufacturing controls (CMC).

Final Agenda

TUESDAY, JANUARY 21

1:00 pm Registration (Sapphire West Foyer)

1:30 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

Particles, Aggregates, and Stability
Aqua aB

2:00 Chairperson’s Opening Remarks

Boxu Yan, PhD, Senior Director, Analytical Development and Quality Control, Acceleron Pharma

KEYNOTE PRESENTATION

2:05 Microfluidic Approaches for the Characterization of Biopharmaceuticals

Paolo Arosio, PhD, Professor, Biochemical Engineering, Department of Chemistry and Applied Biosciences, ETH Zurich

We highlight the emerging possibilities offered by advances in microfluidic technology for the analysis of therapeutic proteins during manufacturing and formulation. We discuss a diffusion microfluidic platform for the characterization of: a) polydisperse size distributions in the sub-micron range; b) sizes and interactions in solutions at high protein concentration; and c) specific interactions in complex mixtures.

2:45 Cell-Based FcgR Binding Assay for Sensitive Detection of Biologics Aggregates

Joel F. Cohen-Solal, PhD, Senior Scientist, Global Protein Sciences, AbbVie Bioresearch Center, Inc.

Low-affinity Fc gamma receptors of different species have the ability to specifically bind to immune complexes or IgG aggregates, thus acting as either physiological sensors or uptake receptors. We will review the literature and internal data showing that a cell-based FcgR binding assay is, therefore, the relevant assay to monitor the presence of physiologically active aggregates in biologics solution or in serum.

3:15 Fast, Low Volume Subvisible Particle Analysis with HORIZON

Gabriel Mercado, PhD, Senior Marketing Applications Scientist, Halo Labs

The HORIZON is the industry’s first analytical system to address the need for rapid, comprehensive subvisible particle analysis even when limited sample material is available. Based on the USP <788> method of Membrane Microscopy, image analysis on HORIZON is fully automated, uses a simple 96-well plate based approach, and requires only 25µL per sample. HORIZON enables incorporation of subvisible particle analysis into biologics workflows as early as developability assessment through late stage formulation and QC.

3:45 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

4:30 Moving towards Harmonization in Subvisible Particle Measurements: Advanced Data Analytics and New Reference Standards

Richard Cavicchi, PhD, Research Physicist, Biomolecular Measurement Division, Material Measurement Laboratory, National Institute of Standards and Technology

Improved quantitative measurements of subvisible particles will require advanced analytic methods to relate measured quantities to particle attributes (e.g., an equivalent diameter). New NIST reference materials can help with calibration and validation of these measurements.

5:00 Insight into Process Development of Oligonucleotide/Polymeric Carrier Formulation

Rui Fang, PhD, Senior Scientist, Sterile Formulation Sciences, Merck & Co., Inc.

This presentation highlights investigations into formulation and process variables to understand the root cause of particle formation in an oligonucleotide/polymeric carrier formulation during manufacturing. Process variables that potentially lead to the failure were identified. Innovative approaches were applied to study pump pulsation. Predictive tools including CFD modeling and the 4th Bourne reaction were used to study mixing efficiency of the mixing chamber that potentially influences formulation composition and stability.

5:30 Close of Day

5:30 - 5:45 Short Course Registration (Sapphire West Foyer)

5:45 - 8:45 Recommended Dinner Short Course*

SC5: Protein Aggregation: Mechanism, Characterization, and Consequences - Detailed Agenda

Instructors:

Thomas Laue, PhD, Professor Emeritus, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire

Kevin Mattison, PhD, Principal Scientist, Malvern Pananalytical, Inc.

*Separate registration required

WEDNESDAY, JANUARY 22

7:45 am Registration (Sapphire West Foyer) and Morning Coffee (Sapphire West & Aqua West Foyer)

Surfactant-Related and Product Impurities
AQUA AB

8:15 Chairperson’s Remarks

Paolo Arosio, PhD, Professor, Biochemical Engineering, Department of Chemistry and Applied Biosciences, ETH Zurich

FEATURED PRESENTATION

8:20 Human IgG1 Hinge Fragmentation as the Result of Radical Mediated Cleavage

Boxu Yan, PhD, Senior Director, Analytical Development and Quality Control, Acceleron Pharma

Hinge cleavage of a recombinant human IgG1 antibody is commonly observed from purified drug substance and in stability samples. The hinge cleavage was found initiated by radical-induced breakage of the disulfide bond between the two hinge cysteines. As hydroxyl radicals exist in healthy cells’ tissues and most buffer solutions, they could therefore be the source for the radical-induced fragmentation of human IgG1 antibodies in vivo.

8:50 Impact of Buffer Species and Stainless-Steel Contact on Polysorbate and Protein Stability

Hieu (Vinnie) La, Associate Scientist, Formulation and Process Development, Pharmaceutical R&D, BioTherapeutics Pharmaceutical Sciences, Pfizer, Inc.

We explore the impact of buffer species, Fe spike and stainless-steel contact on polysorbate 80 (PS80) stability to enable a better understanding of histidine/EDTA-based formulations. We monitored PS80 degradation and protein methionine oxidation (MetOx) in formulations of a monoclonal antibody (mAb) containing several different buffers. It was found that polysorbate stability was sensitive to buffer species and that formulations containing histidine had much faster PS80 and MetOx rates. The metal exposure study suggested that PS80 was extremely sensitive to metal leachables. Stainless steel exposure in histidine buffer resulted in more rapid degradation of PS80 compared to what was observed for the other buffering systems examined. Addition of EDTA, however, was able to inhibit both PS80 and methionine oxidation in the histidine-containing formulation.

9:20 Selected Poster Presentation: Optimization of an HTP Two-Step Antibody Purification Process and Robust Analytical Characterizations

Richard Yuan, MS, Consultant Biologist, Eli Lilly & Co.

In the early stages of antibody discovery and development, generating 10-100 mgs of purified materials quickly is crucial for functional screening assays and develop-ability assessment.  We optimized an HTP two-step purification process (affinity capture and preparative SEC) that completes 24 mAbs purification in < 20 hours.  A stringent CIP process for system and columns was established to minimize endotoxin contamination.  The plate-based analytical characterization and QC of the purified materials is reported

9:50 Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

10:35  Selected Poster Presentation: Lipase Induced Degradation of Polysorbates in Solution

Stephen Rumbelow, Research and Technology Manager, Health Care R&D, Croda Inc.

Polysorbates are widely used to stabilize biological formulations, in order to reduce aggregation of the API’s and to minimize any adhesion to internal surfaces.  This poster describes the results of a comparative study of lipase induced degradation of various grades of polysorbates in buffered solutions, via the simultaneous use of a combination of analytical methods.

11:05 Opportunities and Pitfalls in the Characterization of Submicron Particles during Biologics Product Development

Danny K. Chou, PharmD, PhD, President, Biopharmaceutical Characterization and Formulation Development, Compassion BioSolution, LLC

The presentation will focus on how one can take advantage of the newly available technologies for submicron and microsized particles while avoiding potential pitfalls.

11:35 Monitoring Clearance of Lipase Host Cell Proteins during Biotherapeutics Process Development Using a LC-MRM Quantitation Method

Rachel Chen, 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 a sub ppm quantitation level of three lipases. The method has been applied to monitor the clearance of lipases for various mAbs and fusion proteins under different downstream processes.

12:05 pm Session Break

12:15 LUNCHEON PRESENTATION: Dig Deep into Particle Identification With Hound

Robin Sweeney, PhD, Product Manager, Marketing, Unchained Labs

Particles muck up drug quality, cause headaches throughout development, and can shut down production. Hound pairs Raman 785 nm, Raman 532 nm, and laser induced breakdown spectroscopy (LIBS) with automated microscopy to count and ID particles. Hound is equipped to identify organic, inorganic, and elemental particles through built-in and customizable reference databases to track down the source of any contaminant. We’ll discuss Hound applications that highlight its role as an indispensable particle identification tool.

1:15 Session Break

Aqua Salon

1:45 PLENARY KEYNOTE PANEL

R&D CHALLENGES IN AN ERA OF INNOVATION

The PepTalk Plenary Keynote Panel convenes a group of leading scientists working across novel therapeutic modalities and R&D technologies to explore the many challenges associated with discovering, developing, and advancing today’s novel biotherapeutics. The Panel, via a highly interactive format, encourages discussion among both the panelists and the audience members. Please come prepared with your questions and ideas for this spirited discussion.

• Advances and challenges in expression and production for novel modalities
• Implementing next-generation informatics: data collection, standardization, analysis, ML/AI, and considerations for IP landscape and protection
• Implementing R&D and production capacity for gene and cell therapies – where are we heading?
• Modality-specific challenges: multi-specifics for cancer, improving the ADC therapeutic window, improved safety and pharmacology, novel delivery/targeting
• Preclinical and clinical development of drug combinations with focus in IO: How do we select the right combination dose so we can accelerate clinical development?

Moderator:

Mohammad Tabrizi, PhD, Senior Director, Pharmacology, Ascendis Pharma A/S

PANELISTS

Edward Kraft, PhD, Senior Scientific Manager, Biomolecular Resources, Genentech

Ilya Shestopalov, PhD, Associate Director, Cell Analytics, bluebird bio

David E. Szymkowski, PhD, Vice President, Cell Biology, Xencor, Inc.

Alayna George Thompson, PhD, Senior Scientist I, Drug Discovery Science & Technology, AbbVie

3:05 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

Aggregates and Impurities in Gene Therapy Products
AQUA D

4:00 Chairperson’s Remarks

Lisa Lundberg, Lead, QC Bioassay & Cell Culture, Spark Therapeutics

4:05 Monitoring and Control of Aggregates and Impurities in Large Scale AAV Production

Lisa Lundberg, Lead, QC Bioassay & Cell Culture, Spark Therapeutics

4:35 Characterization and Quantification of Empty and Full AAV Capsids

Qin Zou, PhD, Associate Research Fellow and Group Leader, Analytical Research and Development, Pfizer, Inc.

This presentation can highlight various analytical methods for AAV empty and full capsids and emphasize the proper use of analytical ultracentrifugation for this purpose. AAV empty and full capsids is a product-related impurity and a critical quality attribute. Careful consideration of using appropriate analytical techniques to control that is important for a successful product.

5:05 Predicting Viral Clearance: DOE, HTS and AAV Case Studies Utilizing a Non-Infectious MVM Surrogate during Downstream Development

David Cetlin, Founder & CEO, MockV Solutions LLC

Viral clearance studies are expensive and logistically challenging. This presentation will highlight data from the use of a non-infectious MVM surrogate in a variety of downstream applications and processes.

5:35 Selected Poster Presentation: Detection of AAV Capsid Proteins by CE-SDS as an Alternative to Silver Stain SDS-PAGE

April Blodgett, MS, LAT, Biotherapeutics Senior Sales Specialist, PerkinElmer, Inc.

SDS-PAGE followed by silver staining has typically been used to visualize the VP1, VP2, VP3 ratio.  This approach is labor and time-intensive but yields only qualitative data with poor reproducibility.  Here, we describe the use of microfluidic CE-SDS for the characterization of capsid proteins from AAV serotype 8 as the rapid, quantitative, reproducible alternative to SDS-PAGE with silver stain. 

6:05 - 7:00 Networking Reception in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

7:00 Close of Detection and Characterization of Particulates and Impurities Conference