The use of Stable Producer HEK293 Cell lines for Viral Vector Manufacturing
Gene and cell therapy workflows frequently utilize stable producer HEK293/293T cells in the production of viral vectors and are an example of an advanced therapy medicinal products (ATMPs). Compared to transient transfection methodologies, stable producer lines can provide a more scalable, higher titer production method, provide improved batch-to-batch consistency and also be a source for “the material for sequencing” as described in FDA draft guidelines from 2018.
Seeding and Evidence of Clonality with HEK293 Cells
The 2020 FDA guidance on CMC and INDs for cell and gene therapy spoke to both regulators and manufacturers looking to standardize best practices around viral vector production workflows. This includes avoiding the use of animal-derived reagents and using assurance of clonality as part of production quality control was highlighted.
Incorporate the Solentim VIPS® for high efficiency single cell seeding and in-well verification by imaging and Cell Metric® for Day 0 single cell assurance and outgrowth imaging into your workflow.
Increase clonal outgrowth and protect the cells during freezing and thawing with our range of animal-free reagents, InstiGRO, InstiSHAKE and InstiTHAW for HEK Cells.
Improving Yields with Osmolality
Consistency is critical throughout gene therapy vector production for therapeutics, including viral vector delivery and gene editing. Small deviations in the composition of cell growth media can have unwanted effects on cell density, cell composition, and virus yield.
Osmolality can help you meet these goals as well as optimize the physiological range for transfection complexes, identify media needs, and determine the effects of osmotic pressure on the stability of a complex.
Delivering Assurance For Viral Vector Production
The 2020 FDA guidance on CMC and INDs for cell and gene therapy spoke to both regulators and manufacturers looking to standardize best practices around viral vector production workflows. This includes avoiding the use of animal-derived reagents and using assurance of clonality as part of production quality control was highlighted.
With the OsmoTECH XT, we are able to test 20 microliter samples. Other osmometers have a sample volume of 100 microliters, which doesn’t seem like a lot. But when you are in the cell and gene therapy field, where you are working on therapies that will go into patients right away, or you are doing patient samples, you want to use the smallest sample possible.
Kayla Pasake, Research Associate for Analytical Development at The Center for Breakthrough Medicines
Understanding the Impact of Osmolality on Upstream AAV Manufacturing
Osmolality, a measure of solute concentration, is a critical attribute in the bioprocess workflow, whether one is manufacturing therapeutic antibodies or in the production of viral vectors for use in Cell & Gene Therapy applications. Understanding the osmolality of solutions used in each step provides critical information on culture conditions, material quality at each step of purification, and final formulation composition.
Popular Educational Resources
Learn more about how our technologies can be utilized to help accelerate your gene therapy vector production by checking out our educational resources below
Application Note
Clonality and improved outgrowth for HEK 293 cells when developing stable producer cell lines for viral vector production in gene therapy.
Associated Products
OsmoTECH® XT Single-Sample Micro-Osmometer
Best-in-class osmolality performance, designed for the toughest samples
OsmoTECH® PRO Multi-Sample Micro-Osmometer
Combines ease-of-use with accurate and precise measurements for reliable results
Solentim VIPS®
High efficiency, single cell seeding with image-based proof of clonality for GMP-compatible workflows
Next Generation Cell Culture Growth Supplements
Animal component-free cell culture supplements for CHO and HEK cells
Bioprocessing Services
Get the best performance from your osmometers and cell line development instruments from the people who designed and built them. We provide a range of services and service plans to meet the different needs of our customers.
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