Cambridge Healthtech Institute’s 13th Annual

Characterizing Protein Aggregates and Impurities

Strategies and Tools for Detection and Characterization of Aggregates and Impurities in Biologics

January 18 - 19, 2022 ALL TIMES PST

The popular Cambridge Healthtech Institute’s 13th Annual Characterizing Protein Aggregates and Impurities conference covers the latest trends, challenges, and solutions in understanding characterization and mitigation of problems generated by protein aggregation, impurities, and contaminants in biopharmaceuticals. This conference will feature case studies, new and unpublished data, and interactive discussions on how to carry out a risk assessment and mitigation for aggregates and impurities arising from products, excipients, processes, and packaging, immunogenicity concerns, prediction and modeling, and new tools for detection and quantitation. This conference is preceded by a sister conference, the 8th Annual Characterization of Biotherapeutics.

Tuesday, January 18

1:00 pm Registration (Sapphire West Foyer)
1:30 pm Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)


Session Room: Sapphire H

2:00 pm Organizer's Welcome Remarks

Nandini Kashyap, MPharm, Conference Director, Cambridge Healthtech Institute

2:05 pm

Chairperson's Opening Remarks

Tobias Werk, PhD, CEO, Bionter AG, Switzerland

Requirements for Particulates in the European Pharmacopeia

Hanns-Christian Mahler, PhD, CEO, ten23 health

This talk aims to discuss current requirements in the European Pharmacopeia (Ph.Eur.) related to particulates in parenteral preparations, specifically for biopharmaceuticals. Specific focus will be given on recent changes in the monographs and requirements.


2:40 pm

Qualitative Analysis of Protein Aggregates in Biotherapeutics by Backgrounded Membrane Imaging (BMI)

Markela Murphy, Dosage Form Design & Development, Biopharmaceuticals Development, R&D, AstraZeneca, Gaithersburg, US

Protein degradation via aggregation route is a common source of SVPs. Well-established methods for SVP analysis, such as light obscuration and microflow imaging, are not high-throughput and require significant amounts of sample volume. The focus of this work was to evaluate the backgrounded membrane imaging (BMI) for SVP analysis and identify critical experimental parameters. In conclusion, BMI can be used as a high-throughput method for qualitative particle analysis.

3:10 pm Sponsored Presentation (Opportunity Available)
David Sloan, PhD, Director, Applications, RedShiftBio

The AQS3pro system, built upon microfluidic modulation spectroscopy, is an automated infrared platform optimized to determine a biomolecule’s secondary structure in a complex buffer and at formulation concentrations.  The AQS3pro combines a 24 or 96-well plate, a quantum cascade laser, and a microfluidic flow cell to produce highly precise, higher order structure measurements for the determination of stability, aggregation, lot-to-lot similarity, and formulation optimization of biomolecules.

3:40 pm Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)


4:30 pm

Reliable Counting of Protein Aggregate Particles

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

Quantitative, robust counting of protein aggregates in biotherapeutics continues to be an important priority as a critical quality attribute. It is well known that due to the small index of refraction difference between protein aggregates and the buffer solution, that aggregate particles can be wrongly sized or missed altogether by methods that use imaging or light scattering as detection methods.  We will show a case study of how the new NIST ETFE standard reference material can be used to help calibrate measurements. We will also show how aggregates made from fluorescently labeled NISTmAb can be used in simultaneous fluorescence/brightfield imaging and flow cytometry measurements data to estimate how many particles are being in the brightfield imaging or light scattering measurements. The effects become increasingly important when the particle diameter is < 2 mm.

5:00 pm

How to Bring Sub-Visible Particle Analysis by Light Obscuration to the Next Level

Tobias Werk, PhD, CEO, Bionter AG, Switzerland

Particles remain a number one issue and reason for product recalls because they are critical for patient safety. Thus, sub-visible particle testing is mandatory for any bio-pharmaceutical preparation. Until today the commonly used technology comprises many manual steps and destroys the sample during testing. We show how little modifications can tailor the mature light obscuration technology to the biotech needs, allowing scientists only requiring small sample volumes while staying compliant to the regulations. It has the potential to save millions worth of precious sample material every year.

5:30 pm

Opportunities and Challenges in Real-Time Monitoring of Protein Aggregation during Biopharmaceutical Development and Manufacturing

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

Real-time analytical techniques provide an opportunity for more effective monitoring and control of protein aggregation in bioprocessing as well as a better understanding of the mechanisms of protein aggregation. The objective of this presentation is to share these opportunities as well as challenges in the implementation of novel technologies that can aid in the detection of protein aggregates in real-time.

6:00 pm Close of Day

Wednesday, January 19

7:30 am Registration (Sapphire West Foyer)
8:00 am BuzZ Sessions with Continental Breakfast (Sapphire Foyer)

PepTalk BuzZ Sessions are focused, stimulating discussions in which delegates discuss important and interesting topics related to upstream protein expression and production through downstream scale-up and manufacturing. This is a moderated discussion with brainstorming and interactive problem-solving between scientists from diverse areas who share a common interest in the discussion topic. Continue to check the event website for detailed discussion topics and moderators.

BuzZ Table 2: Aggregation in Protein Formulations

Richard Cavicchi, PhD, Research Physicist, Biomolecular Measurement Division, Material Measurement Laboratory, National Institute of Standards and Technology
  • What role do formulation components (including polysorbate and silicone oil) play in the aggregate formation and are there strategies to mitigate this process?
  • What are the stress factors such as pH changes, temperature changes, or physical effects (shaking/dropping), that are receiving the most attention in your process as a source of aggregates, and what steps are taken to mitigate the stress? 
  • What measurement instruments are most useful at different stages of product development and maintenance?
  •  What improvements can help instruments be best utilized for better quantitation and harmonization between instruments?

BuzZ Table 3: Novel Excipients for Sterile Drug Product Formulation

Ankit Kanthe, PhD, Analytical Scientist, Sterile Drug Product Development, Bristol Myers Squibb
  • Scope for new excipients in line with FDA’s new initiative
  • Understanding excipients ability to prevent protein-protein aggregation
  • New experimental tools or computational methods to help screen novel excipients


Session Room: Sapphire H

9:00 am

Chairperson's Remarks

Kevin Zen, PhD, Executive Director, Chemistry, Manufacturing and Controls, AnaptysBio Inc
9:05 am

High Molecular Weight Species in Therapeutic Monoclonal Antibody Products: Physicochemical Characterization and Analysis of Impact on Drug Safety and Efficacy

Nathan Joh, PhD, Senior Scientist, Amgen, Inc.

High molecular weight (HMW) species a size-variant purity attribute of therapeutic monoclonal antibodies, is of potential concern due to its possible impact on immune safety and efficacy. We demonstrate no measurable impact by HMW enriched from drug substance from a typical monoclonal antibody. No response was detected in model immune-cell lines or primary human immune cells exposed to the HMW. The HMW did not cause statistical changes in potency. Secondary and tertiary structures were comparable to monomers. These evidences suggests that HMW poses little or no impact on the immune safety or efficacy of the studied monoclonal antibody drug product.

9:35 am

Detect, Characterize and Control the Impurities of Therapeutic Proteins

Kevin Zen, PhD, Executive Director, Chemistry, Manufacturing and Controls, AnaptysBio Inc

The presentation will overview impurity analytical technologies and highlight analytical control strategy in the context of regulatory filing.

Maria Germana Sanna, PhD, Field Application Scientist, Gyros Protein Technologies

Monitoring impurities levels throughout bioprocess workflows is critical to quality. In this talk we will present data from different case studies in which biotherapeutic bioprocess samples and viral vector samples were assessed for process related impurities using the automated Gyrolab immunoassay system. Results demonstrate dynamic range, precision, dilutional linearity, and spike data that fulfills key regulatory bioanalytic method requirements with increased productivity in downstream process development.

10:35 am Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
11:15 am

Extended Characterization and Impact of Visible Fatty Acid Particles – A Case Study with a mAb Product

Anthony Tomlinson, Technical Development Scientist, Late Stage Pharmaceutical Dev, Genentech Inc

Inherent particle formation and detection has become a topic of increased scrutiny in biopharmaceutical formulations. These particles, derived from degradation products of the API and/or the excipients, can be difficult to identify and characterize due to their fragility. In this talk, a case study will be presented on the characterization of these particles under a variety of different conditions with multiple analytical techniques.

Christine Probst, Quantitation of Particle Profiles in Biologics and Cell Therapie, Characterizing Protein Aggregates and Impurities, KBI Biopharma

Characterization of particles for injectable therapeutics is increasingly expected to establish product stability and minimize patient risk. We combined machine learning with imaging methods to develop ‘particle profiles’ customized to individual therapeutics, thereby allowing variations to be monitored. Case studies using closed-system transfer devices (CSTD) and cell-based therapeutics will be presented.

12:45 pm Session Break
12:55 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:25 pm Refreshment Break in the Exhibit Hall and Last Chance for Poster Viewing (Sapphire Ballroom)


2:15 pm

Chairperson's Remarks

Wyatt N. Vreeland, Research Chemical Engineer, Chemical Engineering, NIST
2:20 pm

Ultra-Dilute Solution Measurements of Antibody Self-Association

Charles G. Starr, PhD, Scientist, Developability & Preformulation Sciences, Sanofi Group

Identification of mAbs with low levels of self-association has traditionally required relatively concentrated protein solutions, confounding identification of such molecules during early antibody discovery and engineering. We introduce charge-stabilized self-interaction nanoparticle spectroscopy (CS-SINS), a colormetric assay that measures colloidal self-interaction of antibodies in ultra-dilute solutions (0.01 mg/mL). CS-SINS is predictive of high concentration properties such as viscosity and opalescence, which only emerge at orders of magnitude higher concentrations (100+ mg/mL).

2:50 pm

Application of Unfolding Reversibility Studies to Candidate Selection and Formulation Development

Hristo Svilenov, PhD, Associate Professor, Ghent University, Belgium

In this presentation, I will discuss how the reversibility of protein unfolding is related to protein aggregation. Furthermore, I will introduce you to straightforward analytical approaches for studying unfolding reversibility to select aggregation-resistant antibodies and formulations that impede protein aggregation during storage. The presented concepts and techniques can be used for developability assessment and early-stage formulation development of various proteins.

3:20 pm

Synthesis and Characterization of Nanometer-Scale Protein Aggregates Facilitated by Azide

Wyatt N. Vreeland, Research Chemical Engineer, Chemical Engineering, NIST

NIST reference proteins were thermomechanically stressed in sodium azide solutions. Characterization of aggregates by field flow fractionation with light scattering and UV spectrometry showed time-dependent increase in aggregate amount and molar mass. Reducing gel electrophoresis revealed reversible aggregation paths. Kinetic modeling of data implied aggregation mechanism (with azide present) starts by nucleated growth then aggregate-aggregate condensation. Thus, azide preservatives used in ex vivo experiments have demonstrable effects on protein aggregation. 

3:50 pm

Understanding the Dynamics and Phase Behavior of Mixed Antibody-Excipient Adsorption at the Air/Water Interface

Ankit Kanthe, PhD, Analytical Scientist, Sterile Drug Product Development, Bristol Myers Squibb

The interaction of monoclonal antibodies (mAbs) with air/water interfaces plays a crucial role for the manufacture, formulation and storage of these high molecular weight molecules. This talk will focus on the molecular-scale behavior of mAbs that defines an orientational phase space during different stages of adsorption of mAbs to the air/water interface using experimental tools (pendant bubble tensiometry and X-ray reflectivity measurements) and homology modeling. Additionally, competitive adsorption process between mAbs and surfactant based excipients will be discussed to determine the concentration of surfactant necessary to prevent significant mAb adsorption.


4:20 pm

Apply Machine Learning to Predict High Concentration Antibody Aggregation Rate

Pin-Kuang Lai, PhD, Assistant Professor, Department of Chemical Engineering and Materials Science, Stevens Institute of Technology

Predictive models that evaluate aggregation behaviors during early-stage design can facilitate drug development. Machine learning is a widely used tool to train models that predict data with different attributes. However, most machine learning techniques require more data than is typically available in antibody development. In this work, we describe a rational feature selection framework to develop accurate models with a small number of features.

4:50 pm Close of Conference