Cambridge Healthtech Instituteの第11回年次
- 5:00 pm Main Conference Registration1:00 pm
Recommended Pre-Conference Short Course2:00 pm
SC2: Introduction to Lipid Nanoparticle Characterization and Formulation
*Separate registration required. See short courses page for details.
Dessert Break in the Exhibit Hall with Poster Viewing1:40 pm
INCREASING THE THROUGHPUT OF BIOPHYSICAL METHODS
Contemporary Biophysical Methods for More Efficient, Higher Resolution Analysis of Biopharmaceutical Higher Order Structure
Protein higher order structure (HOS) is an important product quality attribute that governs the structure-function characteristics, safety, and efficacy of therapeutic proteins. Biophysics analyses help establish process and product knowledge, understand the impact of upstream and downstream process conditions, monitor product stability, and assess product comparability when process improvements are implemented. In this presentation, we are going to highlight contemporary biophysical methods for more efficient, higher resolution analysis of biopharmaceutical HOS with automated CD, DSC, microfluidic modulation IR, and NMR for protein characterization and comparability/biosimilarity studies for biopharmaceutical process and product development.
High-Throughput Bioanalytical Analyses of Bispecific Antibodies Using Intact Protein Mass Spectrometry Combined with Affinity Capture, Sample Stream, and FAIMS
Here, we present an intact mass spectrometry (MS)-based assay that combines affinity capture with the SampleStream (SS) platform and FAIMS. The captured samples are directly loaded into the SS platform where each sample is injected within 30 seconds. FAIMS further offers improvements in signal-to-noise by separating ions prior to MS analysis. The established methods enable the high-throughput measurement of drug concentration and biotransformation for bispecific antibodies during preclinical studies.
- Commercial Biotherapeutics Stabilized with Trehalose / Sucrose
- Understanding physicochemical properties of Trehalose and Sucrose
- Advantages of Trehalose over Sucrose
- From Liposome to m-RNA vaccines - importance of highly purified characterized Excipients
- Typical components in mRNA-LNP vaccine Excipients
- Examples for utilizations of Sucrose and Trehalose in Covid 19 related formulations
- Amino Acid Buffers in commercial formulations - Importance of highly characterized AAs as Excipients
- Methionine as Biopharmaceutical Stabilizer and Antioxidant
Mass photometry is a revolutionary new way to analyse biomolecules. It enables the accurate mass measurement of single molecules in solution, in their native state and without the need for labels. This approach opens up a wide variety of applicatons in the biophyscial characterization space, including but not limited to: sample characterization, oligomerization studies, interaction studies, monitoring molecular assemblies, and quantifying AAV empty/partial/full ratios.
Refreshment Break in the Exhibit Hall with Poster Viewing3:50 pm
EMERGING METHODS AND INSTRUMENTS
Cryo-Electron Microscopy Revolutionizing Rational Biologics Drug Discovery
Structural biology’s utility in drug discovery lies in its ability to rationalize targeting approaches for both large and small molecules projects, facilitate project execution, and to make these projects both more time- and cost-effective. Cryo-EM has revolutionized structural biology providing atomic resolution data to elucidate MOA, to map epitope/paratope, to modulate affinity, etc., in just a day’s time. A couple of examples on multi-specific drugs and ADCs will be discussed.
Epitope Mapping of Biologics Using Carbene Chemical Footprinting and Mass Spectrometry
Antibody epitope characterization is an important component of therapeutic drug discovery as the binding site directly affects the biological activity. Chemical footprinting with mass spectrometry using carbenes generated from irradiation of diazirine-containing reagents was used to characterize antibody binding sites at the residue level. The structural resolution obtained allows panels of epitope-binned antibodies to be mapped and enables mechanistic understanding of function to support antibody therapeutic lead selection.
Automated Westerns for Product and Impurity Characterization
Gene therapy drugs using adeno-associated viruses (AAV) have been shown to be safe and effective in multiple clinical trials and two commercial products approved in the US. Understanding the physicochemical properties of these gene therapies is critical to patient safety. Here we used a capillary-electrophoresis Western system (CE-Western) to develop assays for characterizing process-related impurities and product attributes.
Close of Day6:00 pm
Dinner Short Course Registration6:00 pm
Recommended Dinner Short Course6:30 pm
SC5: Introduction to Gene Therapy Product Manufacturing and Analytics
*Separate registration required. See short courses page for details.
Registration and Morning Coffee7:30 am
MASS SPECTROMETRY APPLICATIONS
High Throughput Mass Spectrometry Applications in Biopharma: from Multi-Specific Antibody Characterization and Quantitation to Evidence-Guided Hot-spot Remediation
Fully-automated high-throughput analytical reversed-phase liquid chromatographic-mass spectrometry is essential to assess large panels of monoclonal and bispecific antibodies at research-stage. Here we present a modality- and target-agnostic MS method that affords the analysis of a 96-well plate in 41.4 mins., as compared to the traditional rpLC-MS method that would typically take 14.4 hrs. Moreover, we show examples utilizing HT LC-MS to assess Fab glycosylation in discovery panels.
A Competitive Binding Mass Spectrometry Strategy for High-Throughput Evaluation of Potential Critical Quality Attributes of Therapeutic Monoclonal Antibodies
Identification of potential CQAs (pCQAs) that impact mAb target binding is important during the development of therapeutic mAbs. Here, we developed a novel competitive binding-MS strategy that enables high-throughput and multiplexed assessment of pCQAs directly from unfractionated and unstressed mAb drug samples. Specifically, by leveraging the differences in target binding capability under competitive binding conditions, the criticality of multiple mAb attributes can be simultaneously evaluated by quantitative mass spectrometry analysis.
Biophysical Characterization of Difficult-to-Express Proteins
The A2A receptor (A2AR), one of four adenosine G protein-coupled receptor subfamily members, shows exceptional expression and membrane trafficking. We created chimeric A1 and A3 receptors with A2AR C-termini that show improved localization to the plasma membrane and bind to their selective ligands as well as native G proteins. In this talk, we will discuss the binding studies of purified receptors and cognate Gsa that elucidate key protein-protein interactions.
In all biologics, subvisible aggregates are a critical quality attribute and key indicator of product stability. Here we present the Aura, a high-throughput, low-volume system to image, size, count, and identify subvisible aggregates and extrinsic materials across all development stages of cell and gene therapy products.
Coffee Break in the Exhibit Hall with Poster Viewing10:30 am
Transition to Plenary Keynote Session11:10 am
PLENARY KEYNOTE SESSION
Advancing Innovative Biologics Modalities from Research to Clinical Application - Novel Platforms, Automation, and Computation
Addressing disease biology in the clinic with protein therapeutics has become increasingly complex. Turning to innovative and novel scaffolds offers opportunities to tailor therapeutics not previously possible due to advances in host cell engineering and protein design approaches. Designing and developing these modalities requires a next-generation approach as we exploit increased potential design space and also growing data sources to leverage as we invent the next wave of therapeutics.
YOUNG SCIENTIST KEYNOTE
Engineering Prime Editor Proteins for Therapeutic Applications
Precision gene editing technologies have the potential to address a wide range of genetic diseases. Prime Editing is a recently developed “search-and-replace” gene editing approach that can precisely perform a wide variety of DNA sequence edits at programmed target sites in human genomes without requiring double-strand DNA breaks or donor DNA templates. I will describe advances to prime editing technology that improve its efficiency, specificity, and capabilities for therapeutic applications.
Session Break1:00 pm
Developing biologics candidates requires evaluation of many critical quality attributes. In early stages, it is prudent to measure multiple attributes in parallel to conserve resources. Prometheus Panta enables measurement of colloidal and conformational stability. Learn how a single platform for light scattering, nanoDSF, and backreflection measurements in parallel along a thermal ramp provides critical stability attributes of candidates and helps you make decisions about which are worth advancing through your pipeline.
Interactive Discussions are informal, moderated discussions, allowing participants to exchange ideas and experiences and develop future collaborations around a focused topic. Each discussion will be led by a facilitator who keeps the discussion on track and the group engaged. To get the most out of this format, please come prepared to share examples from your work, be a part of a collective, problem-solving session, and participate in active idea sharing. Please visit the Interactive Discussions page on the conference website for a complete listing of topics and descriptions.
TABLE 7: Best Practices in Using Biophysical Methods for More Efficient, Higher Resolution Analysis of Biopharmaceutical Higher Order Structure (HOS) - IN-PERSON ONLY
- Automated CD, DSC, microfluidic modulation IR, and NMR for protein characterization
- Best practices for analyzing and interpreting routine biophysical assays (DLS, DSC, SEC-MALS, and IR)
- What NMR methods are currently employed for product characterization and comparability/similarity assessments?
- What analysis software is used for analysis?
- How extensively are NMR data used in regulatory filings for biopharmaceuticals?
TABLE 8: High Throughput Mass Spectrometry in Biopharma: Challenges and Opportunities - IN-PERSON ONLY
- Fast or thorough, can’t it be both?
- High throughput mass spectrometry-based analytics in biopharma
- Current challenges in instrumentation and analysis software for fully automated mass spectrometry applications in biopharma
- Role of native mass spectrometry in characterization of higher order structures and membrane proteins
- Applying mass spectrometry for high throughput epitope mapping, are we there yet?
ADVANCING HIGHER-ORDER STRUCTURE CHARACTERIZATION
Applications of NMR and Statistical Methods in Establishing Analytical Comparability and Process Consistency of HOS in mAbs
During development of mAbs, it is essential to establish comparability of key properties, including higher-order structure, following manufacturing process changes. NMR spectroscopy is a non-destructive and data-rich analytical method that can report on these properties. Recent advances in NMR allow reproducible fingerprint spectra of mAbs to be collected. We show phase-appropriate applications of NMR in characterizing mAb comparability and process consistency, using unbiased statistical approaches to work without residue-specific assignments.
In-Depth Biophysical Characterization of Alpha-Synuclein and Tau Fibrils - Understanding the Critical Properties of Seeding Potent Amyloid Fibrils
Intrinsically disordered proteins aggregation into fibrils is a common pathological feature of neurodegenerative diseases including Alzheimer’s Disease and Parkinson’s Disease. Recombinant preformed fibrils (PFFs) are a widely used tool to mimic disease initiation and progression. However, how monomeric Tau or a-syn is correlated with the corresponding PFF polymorphism and in vitro seeding potency, and how to precisely guide the formation of specific fibril conformers in vitro is currently unknown. Here, we are aiming to establish robust methods to screen PFFs’ seeding potency and understand critical properties that govern PFFs’ seeding potency in biological model systems.
The Fox® Protein Footprinting System is a revolutionary platform for characterizing the HOS of monoclonal antibodies by elucidating changes in peptide and amino acid solvent accessibility. This high-throughput and high-resolution method analyzes epitope/paratope mapping, aggregation-interface identification, formulation effects, and other important mAb discovery and development factors. This presentation will review Fox technology and its successful application to biologics and biotherapeutics.
Ice Cream Break in the Exhibit Hall with Poster Viewing4:35 pm
PEGS BOSTON COMMON: SPEED NETWORKING
How Many New Contacts Can you Make? - IN-PERSON ONLY
Bring yourself, your business cards or e-cards, and be prepared to share and summarize the key elements of your research in a minute. PEGS-Boston will provide a location, timer, and fellow attendees to facilitate the introductions.
METHODS AND MODELS FOR PREDICTION OF AGGREGATION AND STABILITY
Reduction of Therapeutic Antibody Self-Association Using Yeast-Display Selections and Machine Learning
We report a high-throughput protein engineering method for rapidly identifying antibody candidates with both low self-association and high affinity. We conjugate IgGs that strongly self-associate to quantum dots and use these conjugates to enrich yeast-displayed antibody libraries for variants with low levels of immunoconjugate binding. Deep sequencing and machine learning analysis enables identification of extremely rare variants with co-optimized levels of low self-association and high affinity.
Analytical Ultracentrifugation for Aggregate Quantitation - The Dos (and Some Don’ts)
Analytical ultracentrifugation (AUC) is an important technique that is routinely used for aggregate characterization and quantitation. This presentation will review key considerations for design and execution of a successful AUC experiment as well as discuss potential challenges and pitfalls. Emphasis is placed on the instrumentation and analysis methods used in the pharmaceutical industry for assessing aggregates in monoclonal antibody formulations.
In situ Monitoring of Protein Unfolding and Structural States
Biopharma and food proteins are often exposed to high-pressure (HP) and low-temperature conditions during processing and storage. Sub-freezing temperatures, found during lyophilization or freezing, create aggressive environments that pose technical challenges for in situ measurements, both in the presence and absence of ice. Pressure can be used to force degradation in accelerated conditions, as well as to deconvolute cold denaturation from freeze denaturation effects on protein structure. HP-assisted small-angle neutron scattering (SANS) studies of induced unfolding and aggregation of IgG1 monoclonal antibodies will be presented. The advantages of SANS for freeze-thaw studies will be highlighted.
Networking Reception in the Exhibit Hall with Poster Viewing6:40 pm
PEGS BOSTON COMMON: WOMEN IN SCIENCE MEET UP
Women In Science Meet Up - IN-PERSON ONLY
The Women in Science Meet Up celebrates women trailblazers who are setting their own course in science. We invite women and men to come celebrate the successes of these women in breaking down barriers and inspiring future generations of female leaders. Come join fellow scientists and share your personal and professional journey.
- Who or What inspires you to explore a career in science?
- What fuels your imagination and spirit when you’re faced with challenges?
- What is your proudest moment?
- What can each of us do to improve things further?
Close of Biophysical Methods Conference7:40 pm