Production of rAAV at industrial scale typically involves a series of complex midstream steps, such as clarification by depth filtration and concentration/buffer exchange by tangential flow filtration. We developed a filter-less flow capture method for purification of rAAV serotype 5, using a high-gradient magnetic separator and magnetic Mag Sepharose beads coupled to an AVB affinity ligand. In under 2 h, we captured and eluted rAAV5 directly from ∼ 5 L of cell lysate with satisfying recovery, yield, and purity. The developed process simplifies purification of rAAV.

Speaker:

Adam Wegelius, PhD
Innovation Research Fellow
Cytiva

Efficient viral vector production is essential for the cost-effective manufacture of cell and gene therapies. Many of the current production workflows are reliant on basal media in batch-mode, which can require a long and costly optimization of the HEK293 cell culture conditions to increase their efficiency. For this reason, FUJIFILM Irvine Scientific has introduced BalanCD HEK293 Viral Feed, a chemically defined feed medium designed to boost viral vector yields in fed-batch suspension and adherent cell cultures and tested its effect on viral titer. The results of this study demonstrate that addition of the feed medium 24 hours post-transfection increased adeno-associated virus (AAV) titer by 3-fold to 10-fold and an improved lentivirus (LV) genome and functional titer by 3-fold. As the demand for cell and gene therapy continues to rise, BalanCD HEK293 Viral Feed offers a simplified and effective workflow that can achieve increased viral titer production in high cell density HEK293 cultures.

From this presentation, you will learn more about how HEK293 Viral feed can:

• Increase AAV titer 3-fold to 10-fold across various AAV serotypes (AAV2, AAV5, and AAV9) and improve LV genome and functional titer by 3-fold
• Easily convert to fed-batch with minimal disruption to the workflow
• Scale-up viral vector production from shake flasks to bioreactors

Speaker:

Shan Gao, PhD
Senior Scientist 1
FUJIFILM Irvine Scientific

Viral gene therapy needs much improved processes to substantially cut costs and permit enlarging the population of patients with access to gene therapy cures.

Since the late nineties iBET has been working on continuous unit operations to show they permit large improvement in product quality and yields, reduced footprint, and diminished costs. Furthermore, we have shown such continuous operations can be integrated as a process, further improving performance. Examples of both upstream and downstream operations (perfusion, filtration, chromatography) will be presented and explained, and some integrated continuous processes will be debated.

Speaker:

Manuel J T Carrondo, PhD
Professor of Chemical and Biochemical Engineering, FCT-UNL
Vice-President, IBET

Dr. Ricardo Silva
Senior Scientist
iBET

Mammalian cell platforms offer many advantages for viral vector and recombinant protein production.

HEK293 cells (293SF-3F6 cells) from the National Research Council (NRC) can be grown in suspension culture and serum-free medium, making them ideal for large-scale production of bioproducts. However, creating stable producer cell lines is crucial, and traditional methods such as limiting dilution in multi-well plates and cell sorting can result in low cell survival rates, especially during the cloning process.

In this webinar, Sophie Broussau of National Research Council in Canada will demonstrate how the CellCelector platform can accelerate the development of high-producing HEK 293SF-3F6 lentivirus packaging cell lines, supported by experimental data on monoclonality of the clones isolated by this method.

Key learning objectives:

• Learn how to develop a stable lentivirus packaging cell line
• Hear about the experimental data that demonstrates the monoclonality of cell lines produced by a new method

Speaker:

Sophie Broussau
Cell Line Development Specialist
NRC-Montreal Canada

Producing safe and efficient viral vectors based on adeno-associated virus (AAV) and lentivirus (LV) requires characterization of multiple vector attributes throughout the development process, and quantification of critical quality attributes (CQAs) across manufacturing and quality control. In this webinar we present a platform method for in-depth AAV characterization that quantifies multiple AAV CQAs by combining multi-angle light scattering (MALS) with size-exclusion chromatography (SEC) and multiple concentration detectors. The method enables protein and nucleic acid payload characterization, including capsid concentration, capsid content (empty-full ratio), and aggregate content. Field-flow fractionation (FFF-MALS) offers extended AAV aggregate quantification and is also the ideal solution for larger delivery vehicles like LV and lipid nanoparticles, while dynamic light scattering (DLS) provides a rapid screening solution for all of these vectors.

Speakers:

Dr. Martin Kurnik
Application Scientist II
Waters | Wyatt Technology

Join Yu-Pin Su, Associate Principal Scientist 1, in this insightful webinar where he introduces WuXi Advanced Therapies’ pioneering lentiviral vector production platform. Explore the advanced upstream processes employing both suspension and adherent cell culture systems. For the downstream process, discover our optimized chromatography and UF/DF techniques tailored for efficient lentiviral vector capturing and refining. This results in high-quality lentiviral vectors suitable for either ex vivo or in vivo applications. Our integrated services range from plasmid DNA production to cell therapy, ensuring a holistic solution tailored to our clients' needs. Learn how WuXi Advanced Therapies’ optimized processes and IND express platform can deliver expedited manufacturing processes, reducing your project’s time to the clinic.

Speaker:

Yu-Pin Su
Associate Principal Scientist I, Viral Vector
WuXi Advanced Therapies

The presence of empty capsids in AAV samples is one of the main bottlenecks in AAV production. Quantifying the ratio of empty/full AAV capsids is currently challenging – traditional methods are not quick and simple enough to be employed throughout the process.

In this session, you will gain insights into why mass photometry, a single-molecule bioanalytical technology, stands as the optimal method for the characterization of AAV samples in R&D, process development and manufacturing.

You will discover how the SamuxMP mass photometers can precisely and quickly measure the AAV sample empty/full ratio for capsids of any serotype, with minimal sample requirements and without the need for labeling. You will also learn how the SamuxMP can quantify partially filled and overfilled capsids, as well as provide a rough estimate of titer.

Finally, you will receive an exclusive sneak peek into Refeyn's latest innovation – an analytical instrument tailored for the analysis of adenovirus and lentivirus samples.

Speakers:

Sebastian Joecks
Field Applications Specialist
Refeyn

Join Bibek Ghosh, Associate Principal Scientist I, in this comprehensive webinar where he provides an overview of WuXi Advanced Therapies’ platform. Discover how the seamless integration of upstream and downstream development leads to a robust and scalable transient transfection AAV manufacturing process. Learn about our use of high-performing single clonal cell lines and high-quality plasmids to produce superior viral vectors with high yields. Additionally, Bibek will introduce the streamlined processes of our IND express platform, highlighting pathways to swift approval.

Speaker:

Bibek Ghosh
Associate Principal Scientist I, Viral Vector Process Development
WuXi Advanced Therapies