2022 WEBINAR SERIES
October 19, 2022, 6:00am (PDT), 9:00am (EDT), 2:00pm (BST), 3:00pm (CEST)
Beyond affinity: antibody discovery, optimization, and developability screening from mammalian display libraries
Peter Slavny, PhD
Chief Technology Officer, IONTAS
ABSTRACT
Clinical drug candidates must have not only appropriate affinity and target specificity, but also optimal biophysical properties. Using nuclease-directed integration, we constructed large mammalian display libraries where each cell contains a single antibody gene inserted at a single locus. IgG-formatted antibodies are displayed on the surface, permitting selection from the library of clones by FACS, which allows the selection for desirable binding properties in the final format. This system is uniquely sensitive to critical biophysical characteristics, such as aggregation propensity and poly-specificity, thereby enabling the detection and avoidance of problematic antibodies during the initial discovery phase.
Learning Objectives:
• Learn about detecting and avoiding antibodies with problematic developability profiles during the early discovery stage
• Discover IONTAS’ novel technology for displaying antibodies on the surface of mammalian cells for antibody screening
• Understand how MaxCyte’s electroporation technology enabled the creation of large antibody mammalian display libraries
May 24th, 2022 9:00am (PDT), 12:00pm (EDT), 6:00pm (CEST)
A cGMP Compatible, Non-Viral CAR T Cell Manufacturing Process
Andrew Mancini, Ph.D.
Field Application Scientist
ABSTRACT
Recent breakthroughs in autologous cellular therapies have generated tremendous excitement for the industry; however, these advances have been confronted with the expense of manufacturing of viral gene delivery and concerns over random integration and the safety of viral vectors. To combat these challenges, researchers at the University of California San Francisco have developed a novel system that offers lower toxicity with a single-stranded DNA (ssDNA) approach.
UCSF scientists used MaxCyte ExPERT technology as a non-viral alternative that allows for site-specific delivery of transgenes through homology-directed repair (HDR) to circumvent complex and expensive manufacturing. This approach resulted in highly efficient knock-in integration efficiencies (46-62%) and yields of 1.5 x 109 CAR+ T cells, well above current adoptive cellular therapy doses.
The groundbreaking experiments discussed during this event demonstrate the ability of the MaxCyte GTx™ to deliver ssCTS HDR templates and CRISPR-Cas9 RNP to modify T cells in clinically relevant amounts.
Learning Objectives:
• Outline large-scale, non-viral manufacturing for engineered CAR T cells
• Demonstrate 5-fold boosted knock-in and >7-fold increased live cell yields relative to dsDNA approaches
• Reveal MaxCyte GTx™ delivery of ssCTS HDR templates and CRISPR-Cas9 RNP to modify T cells for a future clinical setting
2021 WEBINAR SERIES
November 30th, 2021 11:00 AM EST- 8:00am (PST)
ExPERT Interview: Developing a GMP-Compatible, Large-Scale CAR T Cell Manufacturing with nS/MARt Vectors
Matthias Bozza
Researcher – Technology
Manager, DKFZ
ABSTRACT
The DNA Vector Lab at the German Cancer Research Center (DKFZ) has developed Nano-S/MARt (nS/MARt), a novel DNA Vector platform that combines prolonged CAR/TCR expression with minimal disruption of T cell activity. This antibiotic-free, nanovector technology uses scaffold/matrix attachment regions (S/MARs) for DNA vector maintenance and replication and it can be introduced efficiently into primary human T Cells without toxicity. When combined with GMP-compliant MaxCyte Flow Electroporation® and CliniMACS Prodigy™ automated cell processing, nS/MARt enables the production of clinically relevant recombinant CAR or TCR T cells with enhanced anti-tumor activity in a single week.
June 29th, 2021 11:00 AM EDT
Generating CAR or TCR T Cells to clinical scale in a quick GMP-compatible manufacturing workflow using nS/MARt DNA Vectors with MaxCyte Flow Electroporation
Matthias Bozza
ABSTRACT
The compelling need to provide adoptive cell therapy (ACT) to an increasing number of oncology patients within a meaningful therapeutic window makes the development of an efficient, fast, versatile, and safe genetic tool for creating recombinant T cells indispensable. Despite the extraordinary efficacy shown by recombinant T cells expressing Chimeric Antigen Receptors (CARs) in numerous clinical trials, significant challenges remain, ranging from a long lead time, and expensive manufacturing to complicated vector-engineering, optimized gene expression and delivery, and reduced vector-mediated toxicities The DNA Vector Lab at the DKFZ has developed Nano-S/MARt (nS/MARt), a novel DNA Vector platform that combines prolonged CAR-TCR expression with minimal disruption of T cell activity. This antibiotic-free, nanovector technology uses scaffold/matrix attachment regions (S/MARs) for DNA vector maintenance and replication and it can be introduced efficiently into primary human T Cells without toxicity. When combined with GMP-compliant MaxCyte Flow Electroporation® and CliniMACS Prodigy™ automated cell processing, the nS/MARt platform enables the production of clinically relevant recombinant CAR or TCR T cells which provide enhanced anti-tumor activity in a single week.
Apr 7, 2021 09:30 AM EDT
Development of a genome-wide CRISPR screen in CD4+ T cells to identify drug targets for immune-mediated inflammatory diseases
Ricky Trigg
Dr. Trigg is a Senior Scientist in the Department of Functional Genomics at GSK.
ABSTRACT
Immune-mediated inflammatory diseases (IMIDs) are a group of disorders characterized by tissue inflammation as a result of dysregulated immune responses. This webinar will showcase the development of a genome-wide CRISPR screen in primary T cells to identify drug targets for IMIDs, and will cover key areas of workflow optimization, including lentiviral transduction, Cas9 electroporation in bulk cell populations, and fluorescence-activated cell sorting. Preliminary data obtained from a recent ‘mini screen’ will be discussed and the session will close with some key learnings from the workflow development process.
2020 WEBINAR SERIES
October 15th, 2020 11:00 AM EDT
Correction of the Sickle Cell Disease Mutation with CRISPR/Cas9
Dr. Mark DeWitt
Project Scientist at the University of California, Los Angeles
ABSTRACT
Sickle Cell Disease (SCD), one of the world’s most common genetic disorders, causes anemia and progressive multiorgan damage that typically shortens lifespan by decades; currently there is no broadly applicable curative therapy. A universal curative therapy for SCD would address a critical unmet medical need in the United States and worldwide. During this webinar, Dr. Mark DeWitt will discuss the development of a CRISPR/Cas9-based strategy to correct the mutation in CD34+ HSPCs harvested from SCD patients. This technique does not rely on viral vectors that can be challenging to manufacture, instead using synthetic reagents: a Cas9 ribonucleoprotein (RNP) targeting the sickle mutation, and a short single-stranded DNA to program gene correction via homology-directed repair (HDR). These studies will be used to support a 2020 IND filing to initiate a Phase I clinical study in 2021, the first of its kind using CRISPR/Cas9-mediated homology-directed repair in hematopoietic stem/progenitor cells. He will conclude with a critical assessment of the current state of sickle cell disease gene therapy, including progress we have made and challenges that still remain.
September 17, 2020 8:00 AM PT
Scalable Manufacturing and Nanovesicle Delivery of CRISPR-Cas9 Ribonucleoproteins Using a cGMP- Compliant Cell Engineering Platform
Peter Gee
MaxCyte Field Application Scientist
ABSTRACT
CRISPR-Cas9 has tremendous potential as a therapeutic tool for treating human diseases. However, prolonged expression of the nuclease and gRNA from viral vectors in an in vivo context may cause unwanted off-target activity and immunogenicity. To overcome these safety issues, a system was recently developed for transient delivery of CRISPR-Cas9 ribonucleoprotein (RNP), recruiting Cas9 protein by chemically-induced dimerization and sgRNA via a viral RNA packaging signal into extracellular nanovesicles. This system, termed NanoMEDIC (nanomembrane-derived extracellular vesicles for the delivery of macromolecular cargo), demonstrates efficient genome editing in various hard-to-transfect cell types, including human induced pluripotent stem (iPS) cells and myoblasts, and also in vivo in a luciferase reporter mouse model.
August 04, 2020 11:00 AM EDT
Flexible, clinically adaptable, non-viral approaches to CAR TCR methodologies
Rama S Shivakumar
Research Scientist and Customer Application Specialist, MaxCyte Inc.
ABSTRACT
In this webinar for scientists and researchers, Rama Shivakumar, a senior scientist at MaxCyte Inc., will highlight powerful case studies that demonstrate the successful use of MaxCyte’s clinically validated, scalable electroporation system in the pre-clinical and clinical scale engineering of resting and activated T cells using a mesothelin specific CARmRNA; in the enhancement of NK cell cytotoxicities against B cell malignancies using an antiCD19 CAR mRNA; in the transposon (Piggybac and Sleeping Beauty) based gene delivery for manufacture of CAR-T cells ; and finally in the gene editing of T cells for improving the efficacy of a TCR immunotherapy. In particular, during this webinar she will discuss how MaxCyte’s versatile ExPERT platform can enable the next-generation non-viral CAR T therapies including allogeneic, off-the-shelf modalities with the potential for enhanced effectiveness for refractory cancer.