Gene expression engineering for stable producer cell lines
Improving viral vector and protein production
Producer cell lines are widely used for the manufacturing of medicines including the production of therapeutic proteins, or viral vectors for cell and gene therapies. Commonly used expression systems include CHO, HEK293, PerC6, SF9 and yeast.
Despite improved production yields over the past years, variation in stability and titers are still frontiers for further optimization, specifically for the production of complex and asymmetric molecules. Annogen has leveraged its screening platforms to generate expression vectors for the CHO and the HEK293 platform. Furthermore, Annogen offers its screening platforms for bespoke optimization of any expression system.
Off-the-shelf expression vectors for manufacturing cell lines
Many technologies exist that improve production levels beyond what one would get from a the most typical CMV or EF1a driven constructs, but these technologies are typically costly to access. Annogen has generated expression vectors that can provide a similar titer but are more affordable.
CHO expression vectors
For production of biologics in CHO cells, Annogen has developed an expression vector that improves titers as compared the typical CMV driven constructs by 2-3 fold, particularly for hard-to-produce molecules. This expression vector is available in a standard format for random integration or in a format compatible with PiggyBac mediated integration. Together with the customer Annogen will design constructs depending on the particular need for example for complex and complex and asymmetric molecules.
HEK293 expression vectors
For expression in HEK293, Annogen has developed an expression vector to improves upon levels obtained by CMV by approximately 2 fold in stable integration format. This expression vector is available in a standard format for random integration or in a format compatible with PiggyBac mediated integration. Furthermore, we are developing small molecule inducible promoters for HEK293.
Bespoke promoter identification for manufacturing cell lines
The pharmaceutical industry leverages Annogen’s proprietary SuRE™ platform to identify novel, proprietary promoters for the use in producer cell lines such as CHO and HEK or more exceptional production platforms. SuRE™ allows for the identification of the most optimal (combination) of regulatory elements to drive transgene expression, thereby providing a quick path to high titers, even in a novel expression system. In contrast to other approaches, Annogen exhaustively screens for, and validates, all possible promoter sequences directly in your production system.
Our promoters can subsequently be combined with your preferred expression strategy, e.g. targeted integration, transposase-based integration or transient expression.
Learn more about SuRE™ →
Annogen’s R&D team has extensive experience developing novel expression systems, and has experience for example with inducible systems for AAV production cell lines, toxic transgenes and non-mammalian expression platforms. This experience help us to support diverse biomanufacturers in their development work.
Contact us today to learn how we can develop a bespoke, exclusive expression system that matches your product and requirements.
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Promoter optimization for manufacturing cell lines
In addition to bespoke promoter identification screens, SuRE™ can also be used to optimize existing promoters. As the basis for such a optimization project we perform a saturation mutagenesis (SatMut), coupled to our SuRE readout, which shows the impact of each possible single base mutation.
The results of such a SatMut analysis can form the basis for new promoter designs focused for example on increasing promoter activity, reducing background activity, CpG removal or promoter minimization without the loss of expression.
AIM™ expression locus analysis for manufacturing cell lines
Our AIM platform allows for the identification of gene insertions and their expression levels for 100,000 integrations in parallel.
Transposon mediated integration is now most commonly used for transgene integration, enabling efficient establishment of producer clones with relatively high copy-numbers integrated in a semi-random manner. In some cases researchers prefer an efficient system for single copy, targeted transgene insertion for example through a landing pad. Annogen can help identify stable transgene insertions and asses their expression levels for > 100,000 integrations in parallel by leveraging the proprietary AIM™ platform. The output of this screening for your cell line will give you a comprehensive understanding on the most optimal expression sites giving the highest and most stable titers, helping you determine the most optimal genetic engineering strategy.
To learn more about the added value of AIM™ to your work, check out our poster on the use of AIM™ to identify genomic loci that are induced upon T-cell activation.
Annogen’s AIM™ screen: bespoke expression locus identification for more effective cell therapies
Annogen’s AIM™ screen revolutionizes the level of control researchers have over transgene expression by identifying optimal genomic loci for transgene integration. Choosing the integration site with the right expression profile will enable safer and more effective cell and gene therapies.