Cambridge Healthtech Instituteの第13回年次
Characterization for Novel Biotherapeutics
- 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.
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
WORKFLOWS AND ASSAYS FOR ASSESSING AND CHARACTERIZING NOVEL MODALITIES
Characterizing the in vivo Stability of Atypical Large Molecule Modalities Using Complementary Bioanalytical Tools
Novel protein modalities are emerging to deliver sophisticated mechanisms of action canonical antibodies cannot. However, little is known about the in vivo stability liabilities (i.e.; biotransformation) with these new modalities. A multi-pronged approach was established using LC-MS and capillary electrophoresis-based methods to characterize and quantify biotransformation liabilities and the in vitro/ex vivo vs. in vivo translatability including but not limited to chemical linker deconjugation, clipping, and amino acid level modifications.
KEYNOTE PRESENTATION: What’s in Your Toolbox? Analytical Strategies for Agile Viral Vector PD
Viral vectors are used in both gene and cell therapy applications. However, analytical methods for viral vectors are far from plug-and-play. What do you do when you want to launch a new vector program, but you lack basic tools to measure quantity or quality? We’ll discuss a virologist’s approach to deciding where to save, spend, and splurge when it comes to building analytical tools to enable early vector development.
CE is a powerful analytical technique for the characterization of next generation medicines. Fast, high resolution separations with UV or Laser Induced Fluorescence (LIF) detection can enable quantitative purity and heterogeneity analysis of complex biologics. Learn how validated CE workflows for proteins, nucleic acids, and viral vectors can be applied with high precision in validated laboratory environments.
Coffee Break in the Exhibit Hall with Poster Viewing10:00 am
Characterization of Antibody-Drug Conjugates Created by New Site-Specific Conjugation Methods
With 11 FDA-approved and marketed entries, antibody-drug conjugates (ADCs) have become clinically and commercially successful cancer treatments that maximize therapeutic potency and limit systemic toxicity through the selective delivery of highly potent drugs. Among many conjugation strategies, site-specific bioconjugation methods including our dual variable domain (DVD)-based technology have been developed to improve the homogeneity and stability of ADCs, aiming to advance clinical outcomes. With the growing interest in the innovation of site-specific and orthogonal bioconjugation methods for single and dual payloads, analytical characterization and functional evaluation are important to provide a baseline for their safety and efficacy profiles.
Bioanalytical Strategy and Streamlined Methods to Characterize Novel Drug Conjugates to Understand Efficacy/Toxicity
ADCs are amongst the fastest-growing drug classes in oncology evidenced by the boom of recent new approvals. In vivo PK, biotransformation, and payload tumor delivery are key information to guide linker drug design and selection. This presentation will share our bioanalytical strategy and methods to characterize novel drug conjugates from in vivo samples, and how the data is used to understand efficacy/toxicity to select molecules with optimized therapeutic index.
Investigation of a Novel Site-Specific Antibody-Drug Conjugate
Antibody-drug conjugates (ADCs) have become a promising class of antitumor agents for treating cancer patients. For the ADC conjugations, traditional and site-specific approaches are being considered. To overcome the heterogeneity observed by traditional conjugations, site-specific conjugations are becoming more and more prevalent. They have been shown to eliminate heterogeneity and improve conjugate stability. This presentation will focus on the various analytical techniques used for analyzing a site-specific ADC molecule.
Luncheon in the Exhibit Hall and Last Chance for Poster Viewing12:10 pm
RNA THERAPEUTICS, LNPS, AND VIRAL VECTORS
Characterization of mRNA Fragments to Evaluate Risk of Truncated or Off-Target Antigen Expression
mRNA vaccines are a newly established class of safe and effective biotherapeutics. The mRNA is manufactured using an in vitro transcription process followed by purification, and the drug product process involves formation of mRNA-containing lipid nanoparticles. A comprehensive control strategy ensures consistent quality, and characterization studies strengthen process and product knowledge. This presentation introduces novel approaches to mRNA fragment characterization to evaluate the risk of off-target or truncated antigen translation.
Analytical Characterization of Therapeutic siRNAs
RNA interference (RNAi) offers a promising therapeutic approach for the treatment of genetic diseases. Triggered by small interfering RNAs (siRNAs), RNAi silences genes by inhibiting translation of problematic transcripts. Fundamentally distinct from antibodies and small molecules, siRNAs require different strategies to evaluate purity and stability as well as support developmental and regulatory processes. Here, we show a series of analytical methods characterizing the purity and biophysical properties of therapeutic siRNAs.
Revolutionary cell and gene therapies offer significant promise to treat life-threatening diseases. However, getting these therapies to market quickly and efficiently is challenging. Rapid and accurate testing of critical quality attributes of viral vectors is necessary but can be impeded by old-school analytics like SDS-PAGE. Come learn how next-generation analytical solutions from Bio-Techne are designed to remove these analytical bottlenecks to get therapeutics to patients sooner.
Networking Refreshment Break2:50 pm
Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC) is a Must-Have, First-Principles Tool in Gene Therapy Characterization
While SV-AUC is revered as a gold-standard technique, it is also maligned as an outdated and cumbersome technique. This talk seeks to introduce the audience to the modern AUC platform and to highlight the unique insights offered by SV-AUC with case studies covering the characterization of AAVs, LNPs, and nucleic acid. Along the way, several myths regarding SV-AUC will be dispelled showing SV-AUC can be a medium-throughput, must-have technique!
Automated, High-Throughput Analysis of Multiple RNA Physicochemical Attributes
mRNA-containing lipid nanoparticle (LNP) vaccines allow for a rapid response to seasonal and emerging viral threats given the shortened process/product development timelines. Assay development to measure CQAs like mRNA concentration and average size are rate-limiting steps. We introduce a novel automated analytical technology (commercialized) that assesses mRNA concentration in LNPs and LNP size for 96 samples in ~1 hour, using 2 uL/well and without dyes, which outperforms current methods.
Close of Day4:20 pm
Registration Open7:00 am
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 for In Vitro/In Vivo Biotransformation/PK Analysis of Novel Modalities - IN-PERSON ONLY
- Needs/techniques/workflows to characterize novel biologics PK and biotransformation (multi-specifics, fusions, ADCs, siRNA/mRNA, CAR cells)
- Critical reagent generation to facilitate the bioanalysis
- ADC PK analysis: how many components to monitor, preclinical vs. clinical? Immunoassay vs. LC-MS vs. hybrid?
- ADC DAR analysis: native vs. intact vs. subunits vs. bottom up
- Nucleotide analysis: MS vs. PCR types of analysis
TABLE 8: Characterization Challenges for mRNA Vaccines and Therapeutics - IN-PERSON ONLY
CHARACTERIZATION FOR EMERGING MODALITIES AND FORMATS
Middle-Down and Bottom-Up Analysis of a Tri-Specific Protein Using Electron Activated Dissociation
Mass spectrometry is part of the traditional toolbox for biotherapeutic characterization. For multi-specific proteins, challenges arise when new domains contain complex PTMs or necessary linkers lead to clipping events. Using electron activated dissociation, we are able to characterize clipping events for species annotation and get unambiguous assignment of glycan species using middle-down and bottom-up analysis, respectively, for a more in-depth understanding of our therapeutic candidates.
Genetically-Encoded Degraders: Beyond Small Molecules for Precision Proteome Editing
Ubiquibodies are engineered proteins that direct otherwise stable proteins to the proteasome for degradation. Here, I will discuss the design and engineering of novel ubiquibodies based on AI/ML-derived “guide” peptides and “stand-alone” E3 ligases that together enable selective and customizable target degradation. Overall, our results suggest that engineered ubiquibodies are a modular proteome editing technology with the potential to expand the degradable target space for pharmacological modulation of disease-causing proteins.
Impact of Oxidation, Glycation, and Thermal Stress on the Structure and Function of the Efanesoctocog Alfa (BIVV001) Fusion Protein
Efanesoctocog alfa, BIVV001, is a novel fusion protein consisting of recombinant human B domain deleted FVIII, the D’D3 portion of von Willebrand factor (vWF), half-life extending XTEN polypeptides, and an IgG 1 Fc domain for treatment of hemophilia A. Degradation pathways studies of BIVV001 evaluating oxidation, thermal, and glycation stresses were conducted and employed an array of analytical techniques. The results of these studies provided insight into BIVV001 chemical and physical stability and the functional impact of these routes of degradation.
Heterogeneity of protein therapeutics and the complex nature of protein structure-ligand interaction necessitate detailed characterization and often non-trivial bioassay development skills to support product release testing, stability profiling, or comparability following process changes. This short presentation shows two case studies. The first one focuses on delineating CQAs by characterizing product charge variants and their biological activity to predict the potency of batches following process changes. The second example highlights how the understanding of structure-function requirements of an antibody and its membrane-bound ligand drove development of a bioassay ready for implementation into QC.
Networking Coffee Break10:30 am
NOVEL METHODS AND INSTRUMENTS
Utility of SPR Technology in Biotherapeutic Development: Qualification for Intended Use
Surface plasmon resonance (SPR) has been widely used in biotherapeutic development for decades. However, no systematic study has been performed on how to qualify SPR assays for the various SPR result types need for regulatory documents. This talk will discuss methods and guidelines for SPR assay qualification in various scenarios. We hope our studies can help align SPR applications among the scientific communities involved in drug development.
Leveraging Biotransformation Information to Engineer the Next-Generation of Novel Therapeutic Modalities
In recent years, a great percentage of biotechnology and biopharma companies' portfolios have comprised novel protein and antibody-based therapeutics. Developing and optimizing these molecules requires detailed analytical characterization. In our current work, we utilize liquid chromatography coupled with intact or top-down mass spectrometry in order to understand the biotransformation of novel modalities (trimeric Fab, antibody-drug conjugates, VHH), which facilitates the interpretation of in vivo findings and (re-)engineering of more stable molecules.
Microfluidic Electrophoresis-Based Detection of dsRNA Contaminants in mRNA Vaccines
mRNA vaccines are currently at the forefront of the vaccine industry due to their safety, efficacy, and fast turnaround times between pathogen detection and vaccine development. However, proper analytical methods to assess the purity of these samples are still lagging. Here, we propose a tool that can streamline the characterization of mRNA vaccine purity, with a focus on dsRNA contaminants, through the use of microfluidic electrophoresis and differential labeling. In addition, with its high-throughput compatibility and short analytical time per sample, this method can has great potential for batch-to-batch analysis.
Close of Summit12:30 pm