Cambridge Healthtech Institute’s 4th Annual

Gene Therapy Manufacturing

Production Strategies for Viral Vector Gene Therapies

August 15-16, 2019

It is an exciting time for gene therapy – therapies on the market, strong investment and encouraging clinical data – but manufacturing still represents one of the major stumbling blocks to successful commercialization.

Cambridge Healthtech Institute’s Gene Therapy Manufacturing conference takes a practical, case-study driven approach to the process development, scale-up and production of gene therapies, tackling key topics such as AAV, lentivirus and retrovirus process development and scale-up, CMO management from early to late-stage development.

Final Agenda

Thursday, August 15

11:30 am Registration Open

12:15 pm Enjoy Lunch on Your Own

1:15 Refreshment Break in the Exhibit Hall with Last Chance for Poster Viewing 


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1:55 Chairperson’s Remarks

Meisam Bakhshayeshi, PhD, Head, Purification Development, Gene Therapy, Biogen

2:00 KEYNOTE PRESENTATION: Present and Future of AAV Gene Therapy Technical Development and Manufacturing

Maranga_LuisLuis Maranga, PhD, Chief Technical Operations Officer, Voyager Therapeutics, Inc.

Recombinant AAV is coming of age as a gene therapy vector of choice. Nevertheless, development and manufacturing for AAV is still in its infancy, and, similarly to the evolution of protein biologics in the last 3 decades of the past century, multiple expressions systems and production technologies are still being employed. The current state of technologies for AAV production and how emerging trends will force their evolution will be discussed.

2:45 FEATURED PRESENTATION: Development, Optimization and Technology Transfer of the Scalable Pro10TM rAAV Manufacturing Process

Grieger_JoshJosh Grieger, PhD, CTO, Asklepios BioPharmaceutical

Over a decade ago, we successfully adapted an adherent HEK293 cell line from a qualified clinical master cell bank to grow in animal component-free suspension conditions in shaker flasks, WAVE and stir tank bioreactors allowing for rapid and scalable rAAV production at >250L scale. My presentation will focus on the optimization, technology transfer (2 Pharmaceutical companies and Viralgen) and GMP manufacturing of rAAV vectors utilizing the Pro10 manufacturing process.

3:15 Evolution of a Platform Process Across Clinical Manufacturing

Horvath_JustinJustin Horvath, PhD, Director, Manufacturing, Regenxbio

Pall_Biotech 3:45 Evaluation and Optimization of Filter media for Adeno-Associated Virus (AAV) and Lentivirus Clarification Process

Bharath Raghavan, Senior Engineer, Research and Development, Pall Biotech
Michael Collins, Senior Manager, Research and Development, Pall Biotech

As theImpurity separation- host cells, cell debris and aggregates found in typical viral vector cell culture. The clarification step must combine high capacity for impurity removal, high product yield and ease of scale-up to prepare for downstream operations. Depth & membrane filters investigated and optimized for AAV and lentivirus clarification performance.

4:00 Refreshment Break

4:15 rAAV large scale manufacturing using BEVS technology

Lubelski_JacekJacek Lubelski, PhD, Vice President, Global Pharmaceutical Development, uniQure

Recombinant adeno-associated virus is becoming a vector of choice for a variety of human gene therapy applications. The demand of the vector to treat non-orphan and whole-body administration indications at commercial scale significantly exceeds capabilities of many production systems utilized to date. Therefore, there is an increasing requirement for generation of high quantity of potent rAAV vectors based on various serotypes. Baculoviruses and insect cells have a number of features that make them ideal for development of a robust system that can generate high quantities of rAAV. I will present our experience with baculoviruses/insect cell system in stirred tank bioreactor. Furthermore, I will discuss a comparison between the wave and STR bioreactors for rAAV production with BEVS and discuss the vector quality and potency generated by insect and mammalian cell production systems.

4:45 AAV Upstream Production Process Optimization for the Sf9/Baculovirus System

Slade_PeterPeter Slade, PhD, Director, Cell Culture, Voyager Therapeutics

The baculovirus expression vector system has been used for the expression of recombinant proteins and for some commercially available vaccines. More recently it has been used as a platform for gene therapy and adeno-associated virus (AAV) production. This talk will discuss how the baculovirus system is a scalable robust process for manufacture of AAV and future innovations will allow for significant increases in productivity and product quality control.p>

5:15 End of Day

Friday, August 16

7:30 am Registration Open

7:30 Small-Group Breakout Discussions with Continental Breakfast

This session provides the opportunity to discuss a focused topic with peers from around the world in an open, collegial setting. Select from the list of topics available and join the moderated discussion to share ideas, gain insights, establish collaborations or commiserate about persistent challenges.


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8:30 Chairperson’s Remarks

Johannes C.M. van der Loo, PhD, Director, Clinical Vector Core, The Raymond G. Perelman Center for Molecular and Cellular Therapies, Children’s Hospital of Philadelphia

8:35 Optimization of Transfection and Culture Conditions to Maximize AAV Production in Suspension 293-Based Cells

Piras_BryanBryan Piras, PhD, Lead Scientist, Process Development, Therapeutics Production and Quality, St. Jude Children’s Research Hospital

Production of AAV for early-phase clinical use requires manufacture utilizing hundreds of liters of upstream cell culture. Production facilities for early-phase trials have often relied upon adherent cell-based processes, though suspension cells offer greater flexibility and far superior scalability. We have identified a variety of transfection and culture conditions to help maximize production of an AAV8 vector in 293-based suspension cells.

9:05 Viral Vector Bioprocessing at the National Research Council Canada

Mullick_AlakaAlaka Mullick, PhD, Senior Research Officer, National Research Council Canada

In this presentation, we will summarize expertise in viral vector bioprocessing at NRC. We will focus on strategies that we have developed using our proprietary HEK293SF suspension cell line platform for lentiviral and adeno-associated viral vector (LV, AAV) bioprocessing. We will use case studies to illustrate our progress in these areas, such as development of stable inducible packaging and producer cell lines for LV and scalable workflows for AAV manufacturing.

9:35 High Yield Production Platform for Pre-clinical and Clinical rAAV

Clement_NathalieNathalie Clément, PhD, Associate Director and Associate Professor, Powell Gene Therapy Center, Pediatrics, University of Florida

We will present our latest data on AAV production with two scalable platforms using HEK293 in suspension with either co-infection with recombinant HSV or co-transfection with PEI. Data will include overall yields, quality and purity of the AAV stocks. Case studies will be presented from pre-clinical and clinical manufacturing campaigns.

10:05 Networking Coffee Break


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10:30 Implementing Commercial Ready Processes for Clinical Applications

Dobrowsky_TerrenceTerrence Dobrowsky, PhD, Principal Engineer, Gene Therapy, Biogen

Implementing commercial ready processes early in the development of gene therapy products eliminates scalability and comparability issues later in the product lifecycle. Here we will review development to-date as well as strategies for leveraging both transient transfection and producer cell line (PCL) production systems to quickly establish scalable, manufacturing ready processes at Biogen. This development work includes improvements to cell line and vector generation workflows, shifting from adherent to suspension cell culture systems, media & feed development as well as process steps that are linearly scalable.

11:00 Technology Transfer of an Academic Hematopoietic Stem Cell Gene Therapy Manufacture to Industry: The Mustang Bio X-SCID Experience

Greene_MichaelMichael Greene, MD, Executive Director, Technology Transfer Lead, Mustang Bio, Inc.

Mustang Bio, Inc., a Fortress Biotech Company, has partnered with St. Jude Children’s Hospital to manufacture MB-107, a cell and gene therapeutic to treat the devastating orphan disease X-linked severe combined immunodeficiency (X-SCID). We will present our experience during the technology transfer of an academic manufacturing process to industry.

11:30 Virus Clearance Strategy for AAV Producer Cell Line Process: A Case Study

Morais_JasonJason Morais, DSP Lead, Gene Therapy Development, Sanofi

Wild-type Adenovirus can be used as a ‘helper virus’ to initiate replication of recombinant AAV in producer cells. However, during production, Adenovirus is replicated alongside AAV and is present as a process-related impurity. This Adenovirus must be removed and/or inactivated during purification in order to ensure patient safety. Here we present a virus validation study conducted on an AAV purification process, confirming greater than 20 LRV of Adenovirus reduction.

Fujifilm-Diosynth-Logo 12:00 pm Paving the Way for AAV Late Phase and Commercial Supply for DS and DP
Page_ThomasThomas Page, PhD, Vice President, Engineering and Asset Development, Advanced Therapies, FUJIFILM Diosynth Biotechnologies
Speaking points: Establishing fully closed single use processing for upstream, harvest and downstream for 2000L AAV cell culture process; Using mAb manufacturing processes as a model;  Integrating DP capabilities for vials and syringes to streamline client supply chain; Improvement of quality metrics by removing Freeze-Thaw cycles

12:30 Enjoy Lunch on Your Own


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1:25 Chairperson’s Remarks

Nathalie Clément, PhD, Associate Director and Associate Professor, Powell Gene Therapy Center, Pediatrics, University of Florida

1:30 Development of an Advance AAV Purification Platform Using Integrated Analytics and PAT

Leseigneur_FlorianFlorian Leseigneur, PhD, Viral Vector Development Team, Cell and Gene Therapy Catapult

Traditional AAV manufacturing downstream processes are based upon non-scalable techniques which involve multiple steps and rely on time consuming offline analytical techniques for evaluation. As demand for AAV increases there is a pressing need for simplified viral vector purification trains with real-time titre, quality and purity measurements. Here, we will present an AAV purification platform with integrated analytics and PAT sensor technologies which allows rapid product characterisation

2:00 Chromatography Beads, Monoliths or Membrane Absorbers for Purification of Gene Therapy Vehicles and Bionanoparticles

Jungbauer_AloisAlois Jungbauer, PhD, Professor, Institute of Biotechnology, University of Natural Resources and Life Sciences (BOKU)

Heterogeneous populations of bionanoparticles such as viruses, virus-like particles, or other extra cellular bionanoparticles in gene-therapy or in cancer therapy are released from the cells, carrying different host cell proteins, DNA and RNA fragments. Here we compare the results obtained for the purification particle populations’ separation of bionanoparticles produced in CHO cell culture using different downstream processing approaches. These approaches include polymer grafted anion exchangers, monoliths (anion exchange and hydrophobic interaction), a combination of flow-through and heparin affinity chromatography, and membrane adsorbers.

2:30 Development of a High-Throughput Purification Method to Determine AAV Full/Empty Capsid Ratio

William Kish, PhD., Senior Scientist, Gene Therapy, Purification Process Development, Bioprocess R&D, BioTherapeutics Pharmaceutical Sciences, Pfizer Inc.

This plate-based purification method enables the rapid evaluation of empty/full ratios from up to 96 AMBR® bioreactor cell lysates. AAV crude lysates are purified using Tecan liquid handler and eluted fractions are analyzed using SEC-HPLC. Up to 48 samples can be purified and analyzed within 1.5 work days. Our upstream process development team has increased understanding of relationship between culture conditions and empty/full ratio leading to process improvements. This new tool for gene therapy process development enables us to explore a broader range of bioreactor conditions in less time and with less material.

3:00 Elements of a Robust Downstream Processing Platform for AAV Based Gene Therapy Vectors

Nripen Singh, PhD, Director, Biopharmaceutic Development, Voyager Therapeutics

Adeno associated viruses (AAV) are increasingly becoming the vector of choice for gene therapy applications. Nevertheless, the use of multiple serotypes, with different capsid protein sequences and different biochemical properties, creates challenges in the development of platform purification process. In this presentation, we highlight Voyager’s approach to downstream platform development for AAV vectors expressed with the Sf9/baculovirus system.

3:30 Close of Conference