2021 BioMAN Summit

Welcoming, Introduction & Framing of the Summit

30 minutes

Samberg Conference Center (Building E52- 7th Floor) & Virtually

Jacqueline M. Wolfrum

Dr. Wolfrum has been at the MIT Center for Biomedical Innovation (MIT CBI), since 2014. She is Director of the Biomanufacturing Program (BioMAN), a pre-competitive biopharmaceutical industry consortium focused on development of new knowledge, science, technologies, and strategies that advance the manufacture and global delivery of high-quality biologic medicines. She manages sponsored projects and activities […]

Stacy L. Springs

Dr. Stacy Springs is the Executive Director at the MIT Center for Biomedical Innovation (CBI). The Center integrates the Institute’s technical, scientific, and management expertise to solve complex biopharmaceutical challenges. CBI leads multi-stakeholder, multidisciplinary research and educational initiatives with real world impact, including MIT’s Biomanufacturing Consortium, (BioMAN), and it’s Consortium on Adventitious Agent Contamination in […]

Anthony J. Sinskey

Anthony J. Sinskey, Sc.D., is a Professor of Microbiology at the Massachusetts Institute of Technology. He also holds positions as Co-Director of the Malaysia-MIT Biotechnology Partnership Program and as Faculty Director of the MIT Center for Biomedical Innovation. He conducts interdisciplinary research in metabolic engineering focusing on the fundamental physiology, biochemistry and molecular genetics of […]

Base Editing and Prime Editing: Engineered Proteins That Precisely Correct Pathogenic Mutations in Cells, Animals, and Patients

In this lecture, I describe the development and therapeutic application of two precision gene editing technologies that install or correct targeted mutations without requiring double-strand DNA breaks, thereby minimizing undesired consequences of chromosomal cleavage. We developed base editors, proteins that directly perform chemistry on individual DNA bases in living cells to install or correct mutations at targeted positions in genomic DNA. We recently engineered CRISPR-free, all-protein base editors that enabled the first purposeful changes in the sequence of mitochondrial DNA in living cells. By integrating base editors with ex vivo and in vivo delivery strategies that deliver therapeutic proteins, we rescued animal models of human genetic diseases including sickle-cell disease, progeria, and spinal muscular atrophy (SMA). Single-AAV base editing systems enhance the safety and practicality of in vivo base editing. Our development of engineered virus-like particles (eVLPs) provide additional in vivo delivery methods for gene editing proteins that minimize off-target editing and the risk of oncogenic DNA integration. Base editors are in at least six clinical trials to treat diseases including familial hypercholesterolemia, sickle-cell disease, beta-thalassemia, and T-cell leukemia. The first clinical outcomes from ex vivo base editing and from in vivo base editing have also been reported, demonstrating benefit to T-cell leukemia patients and to hypercholesterolemia patients, respectively. I will also describe prime editors, engineered proteins that directly write new genetic information into a specified DNA site, replacing the original sequence, without requiring double-strand DNA breaks or donor DNA templates. Prime editing can mediate any base substitutions, deletions, and/or insertions of up to ~200 base pairs in living cells in vitro and in vivo, and has been applied to directly install or correct pathogenic alleles that previously could not be corrected in therapeutically relevant cells. We illuminated the cellular determinants of prime editing outcomes, and used the resulting insights to develop new prime editing systems with substantially higher editing efficiencies and product purities. Most recently, we used phage-assisted continuous evolution (PACE) to evolve a suite of sixth-generation prime editors (PE6a-6g), which each evolved to specialize in different types of prime edits. The combination of prime editing and site-specific recombinases enable programmable gene-sized (>5 kb) integration and inversion at loci of our choosing in human cells. Prime editing has recently been used to rescue animal models of genetic diseases including sickle-cell disease, metabolic liver diseases, and genetic blindness, and is anticipated to be cleared for clinical trials in 2024. Base editing and prime editing enable precise target gene correction, in addition to target gene disruption, in a wide range of organisms with broad implications for the life sciences and therapeutics.

30 minutes

Samberg Conference Center

David Liu

David R. Liu is the Richard Merkin Professor and director of theMerkinInstitute of Transformative Technologies in Healthcare, vice-chair of the faculty at the Broad Institute of MIT andHarvard, the Thomas Dudley Cabot Professor of the Natural Sciences at Harvard University, and a Howard Hughes Medical Institute (HHMI)investigator. Liu’s research integrates chemistry and evolution to illuminate […]

Evolving CAR Cell Therapy Engineering: Challenges & Opportunities

Chimeric antigen receptors (CARs) have achieved promising results in treating B-cell malignancies and multiple myeloma, and led to the approval by the FDA of multiple autologous CAR T cell products. CARs allow T-cells to target any antigen of choice, independent of MHC expression. However, there are challenges to extending the use of CAR T cells to solid tumors and other pathologies. Progress in CAR design, cell manufacturing, and genome editing hold the promise of generating safer and more effective genetically instructed immunity. Novel engineered cell types, including innate T-cell types, natural killer cells, macrophages, and induced pluripotent stem cell-derived immune cells, are on the horizon, as are applications of CAR T cells to treat autoimmunity and other pathologies. 

45 minutes

Samberg Conference Center (Building E52- 7th Floor) & Virtually

Isabelle Rivière

Isabelle Rivière received her Ph.D. in Cellular and Molecular Biology from the University ofParis, France. She conducted her graduate studies at the Institut Curie (Paris) and at the Whitehead Institute (Cambridge, MA). After completing her postdoctoral work at NYU, she joined the faculty of Memorial Sloan-Kettering Cancer Center where she developed novel strategies for cell […]

The Central Role of Manufacturing for Complex Biologic Products

Cell and gene therapies are complex biologic products that continue to illustrate the longstanding adage that for biologics the process defines the product. Advancing manufacturing technologies for rapid scale up when large quantities of product are needed and for efficient throughput when small batches are required will be critical moving forward. Use of process automation, real time multiparametric in-line monitoring, and artificial intelligence, among other innovative technologies, may all combine to result in higher quality products produced with notably greater efficiency and at a lower cost. These advances will be particularly important in gene therapy for rare diseases, where the automation of small batch production of high-quality products is likely to be transformational. FDA will continue to advance the research and development of innovative manufacturing technologies while working to make a difference in public health. 

30 minutes

Samberg Conference Center

Peter Marks

Peter Marks, MD, Ph.D Director Center for Biologics Evaluation and Research, FDA Peter Marks received his graduate degree in cell and molecular biology and his medical degree at New York University and completed Internal Medicine residency and Hematology/Medical Oncology training at Brigham and Women’s Hospital in Boston. He has worked in academic settings teaching and […]

The Central Role of Manufacturing for Complex Biologic Products

Cell and gene therapies are complex biologic products that continue to illustrate the longstanding adage that for biologics the process defines the product. Advancing manufacturing technologies for rapid scale up when large quantities of product are needed and for efficient throughput when small batches are required will be critical moving forward. Use of process automation, real time multiparametric in-line monitoring, and artificial intelligence, among other innovative technologies, may all combine to result in higher quality products produced with notably greater efficiency and at a lower cost. These advances will be particularly important in gene therapy for rare diseases, where the automation of small batch production of high-quality products is likely to be transformational. FDA will continue to advance the research and development of innovative manufacturing technologies while working to make a difference in public health.

30 minutes

Samberg Conference Center (Blg E52) 7th floor & Virtually

Refreshment Break

15 minutes

Samberg Conference Center

A Digital Twin for Continuous mRNA Manufacturing

Mechanistic models are being constructed for all unit operations for the end-to-end continuous manufacturing of mRNA biotherapeutics. The dynamic models are integrated with models for constraints, uncertainties, and disturbances to form a digital twin for automated, integrated manufacturing. The digital twin is suitable for (1) evaluation and validation of mechanistic hypotheses to gain mechanistic understanding, (2) comparison of multiple process flowsheet options, (3) optimization of individual unit operations and their control systems, (4) the design of end-to-end operations, and (5) the real-time operation alongside plant operations. Experimentally validated results are presented for multiple unit operations. 

30 minutes

Samberg Conference Center

Richard D. Braatz

Dr. Richard D. Braatzis the Edwin R. Gilliland Professor of Chemical Engineering at MIT, where he conducts research into advanced biomanufacturing systems. He leads process analytics, mechanistic modeling, and control systems design activities within the Center of Biomedical Innovation, and is the Director of the Center for Continuous mRNA Manufacturing.

Lunch

1 hour

Bioprocess Strategies to Mitigate the Impact of Proteases on Adeno-Associated Viral (AAV) Vector VP1 Capsid Content

A handful of studies have suggested that cellular and baculovirus proteases may impact the capsid nature of insect cell-produced AAV gene therapy viral vectors. The aim of this investigation was to detect the presence of extracellular protease activity in cell culture, and design bioprocess strategies that may inhibit protease activity, and by extension prevent the reduction of AAV VP1 capsid protein which has been determined to be important for virus infectivity.    We attempted late cell culture process changes (i.e., temperature shifts and pH shifts) to modulate protease activity. We also tried addition of inorganic salts that may have a similar effect. During AAV production, the addition of the cysteine proteinase inhibitor E64 to cell culture led to an increase of normalized AAV VP1 levels by 50-70% compared to the control. A similar outcome was evidenced when the cell culture was exposed to a 23°C shift. Time-course analysis of VP1 levels showed that E64 and temperature shift prevented the continuous reduction of VP1 levels, which was seen in the control condition. Moreover, the late addition of an inorganic salt at micromolar concentration had a similar effect on VP1 content. We confirmed that the salt addition also reduced extracellular protease activity within the last days of the process. Similar trends were seen during the production of a second AAV serotype

30 minutes

Samberg Conference Center

Juan Aponte-Ubillús

Juan Aponte-Ubillus is currently Scientist 2 from the Upstream–Drug Substance Technologies department at Biomarin Pharmaceutical Inc. He has over 7 years of experience characterizing and optimizing cell culture processes to produce complex biologics such as monoclonal antibodies and AAV vectors. Juan obtained his PhD degree from the Keck Graduate Institute of AppliedLife Sciences in California […]

Commercial Production Development of a Complex Plasma Protein Therapeutic; Mitigating the Risks of Revisions to Purification Process Consumables

Therapeutic product characterization throughout the Research, Development and pre-commercialization process is an essential part of ensuring comparability against expected criteria, while further enabling risk mitigation associated with any atypical results representing potential product safety/efficacy concerns. In addition to concerns regarding the impact of process scaling or revision/optimization steps on the active biotherapeutic agent, identifying the source of any atypical result can be complicated by an extensive list of necessary hardware and/or consumables revisions. These revisions may include, but are not limited to, changes to the lots of raw materials, storage media and/or cleaning solutions.  This presentation will outline a recent example of an atypical result that occurred during a tech transfer scale-up phase. The work presented will cover all aspects from the original observation following LC-MS peptide mapping, to the identification of the impurity coupled to planned risk management and mitigative activities. Our data will further summarize the source of this atypical result, impacting multiple pilot scale lots, correlated with a third-party consumable revision. The observation was not only important in mitigating risks ahead of production scale up, but further benefitted other products/modalities using the same third-party consumable, highlighting the importance of communication between both internal and external partners.

30 minutes

Samberg Conference Center

Richard Kerr

Richard graduated with his PhD from University College London (UCL) in 2013, specializing in the higher order structural analysis of proteins by hyphenated mass spectrometry methods. He has held two post-doctoral positions, his first as a research associate at the University of Michigan and the second as an ORISE fellow at the US FDA, in […]

Platform Solutions to Enhance the Safety, Efficacy, and Scalability of Gene Edited Cell Therapies

Since the discovery and description of CRISPR based gene-editing, there has been an explosion in pre-clinical and clinical stage programs focused on the development of gene-edited cell therapies.2023 will likely witness the first ever FDA approval and commercial launch of a CRISPR gene-edited cell therapy. However, there remain many outstanding questions about the design space, manufacturing, and analytical requirements for developing safe and effective CRISPR gene-edited cell therapies. This brief talk will review several novel platform technologies, including high-fidelity Cas9 enzyme, nanoplasmid based HDR donor template, HDR enhancers, and rhAmpSeq for analytical characterization of on-target and off-target edits; this talk will also discuss real-world case studies highlighting the role of these platform technologies in enabling the development of safer and more effective CRISPR gene-edited cell therapies.

30 minutes

Samberg Conference Center (Building E52- 7th Floor) & Virtually

Sadik Kassim

Sadik Kassim, Ph.D. is a scientist and executive with extensive experience in the biotechnology industry with a specific focus on cell and gene therapy bioprocessing and translational research. Currently, he serves as Chief Technology Officer of Genomic Medicines for the Life Sciences companies at Danaher Corporation. Most recently, he was Chief Technology Officer at Vor Bio […]

High-Density Microbioreactor Process Designed for Automated Point-Of-Care Manufacturing of CAR T Cells

While adoptive cell therapies have revolutionized cancer immunotherapy, current autologous chimeric antigen receptor (CAR) T cell manufacturing faces challenges in scaling to meet patient demands. CAR T cell production still largely relies on fed-batch, manual, open processes that lack environmental monitoring and control, whereas most perfusion-based, automated, closed-system bioreactors currently suffer from large footprints and working volumes, thus hindering process development and scaling-out. Here, we present a means of conducting anti-CD19 CAR T cell culture-on-a-chip. We show that human primary T cells can be activated, transduced, and expanded to densities exceeding 150 million cells/mL in a 2 mL perfusion-capable microfluidic bioreactor, thus enabling the production of CAR T cells at clinical dose levels in a small footprint. Key functional attributes such as exhaustion phenotype and cytolytic function were comparable to T cells generated in a gas-permeable well. The process intensification and miniaturization, together with the online analytics offered by the microbioreactor, could facilitate high-throughput process optimization studies, as well as enable efficient scale-out of cell therapy manufacturing, while providing insights into the growth and metabolic state of the CAR T cells during ex vivo culture.

30 minutes

Samberg Conference Center

Michael Birnbaum

Michael Birnbaum is an Associate Professor of Biological Engineering at MIT. His research focuses on T cell biology and engineering.During his tenure at the Koch Institute, Birnbaum has received the AACR-TESARO Career Development Award for Immuno-oncology Research, aPackard Fellowship in Science and Engineering, a Pew-Stewart Scholarship for Cancer Research, a V Scholar Grant from the […]

Biophysical Critical Quality Attributes (CQAs) for Cell Therapy Products

One of the critical challenges in cell therapy is the lack of reliable, specific, and non-destructive quality attributes, which are sorely needed for all aspects of the biomanufacturing of these cells, including donor selection, in-process quality monitoring, and release testing. Most biological and biochemical assays are destructive or perturbative, which limits their utility, especially for autologous cell therapy. In this talk, I will showcase some of the emerging ideas of label-free, biophysical critical quality attributes (CQAs) we have been working on, including magnetic, electrical, and mechanical signatures of cells. Once a strong correlation with biochemical cell phenotypes is established, these will serve an important role in improving the overall production of both allogeneic and autologous cell therapy products. 

30 minutes

Samberg Conference Center

Refreshment Break

15 minutes

Advancements in Assessment of Genetic Medicine Quality Attributes

This talk will cover: 

• Setting plasmid DNA starting material specifications 
• Characterization of cell-free DNA starting material 
• Multivalent mRNA drug product identity and ratio determination 

30 minutes

Samberg Conference Center (Building E52- 7th Floor) & Virtually

Lawrence Thompson 

Lawrence (Larry) Thompson is an Associate Research Fellow and Group Leader in Analytical R&D within BioTherapeutic Pharmaceutical Sciences at Pfizer. He is an analytical CMC SME for Pfizer’s adenoviral & plasmid DNA based immunotherapeutics, mRNA drug substances and nucleic acid starting material pipeline (used in rAAV and mRNA production). Prior to joining Pfizer, he worked […]

Safety Strategy for iPSC-derived Allogeneic Cell Therapy Drug Product Release 

Ensuring quality of cell therapy products is required through all stages of development and manufacturing. Quality is confirmed through process design, raw materials testing, in-process testing and release testing. Multiple methods throughout the biomanufacturing process assess the identity, purity, potency and safety of the final product. Here we will explore a strategy case-study for assuring the safety of a final iPSC-derived allogenic drug product while addressing specific regulatory expectations. Topics will include: 

• Meeting regulatory expectations for demonstration of safety 
• Selecting appropriate methods for genetic characterization  
• Identifying informative variant class assessments at each manufacturing stage

30 minutes

Samberg Conference Center

Damien Fink

Damien currently leads the Analytical Development group at Century Therapeutics. He’s a graduate (and avid fan) of Pennsylvania State University and Villanova University. His experience in Analytical Development stems from his tenure at Merck Manufacturing Division (4+ years) and Janssen R&D (16+ years) where he worked extensively on assay development and validation efforts in both […]

Day 1 Closing Remarks

15 minutes

Stacy L. Springs

Dr. Stacy Springs is the Executive Director at the MIT Center for Biomedical Innovation (CBI). The Center integrates the Institute’s technical, scientific, and management expertise to solve complex biopharmaceutical challenges. CBI leads multi-stakeholder, multidisciplinary research and educational initiatives with real world impact, including MIT’s Biomanufacturing Consortium, (BioMAN), and it’s Consortium on Adventitious Agent Contamination in […]

Poster Session and Evening Networking Reception

2 hours

Samberg Conference Center- Salon West