Stable cell lines are suitable, not only for manufacturing therapeutic and diagnostic antibodies, but also for drug screening, and applications on pharmacological and toxicological studies. Creative Biolabs has extensive experience in stable cell lines development and has already successfully established the stable FUT8−/−-CHO cell lines, Fucoanalog™ cell lines, and Glymax™ cell lines for ADCC⁺ antibody productions. Especially, our stable cell lines can be applied to ADCC/CDC assay development, ADCC/CDC functional studies, and so on.
Proprietary CHO expression platform
Efficient plasmid transfection system
High-throughput screening of cell clones
Clone screening based on mass and high titer
Custom cell culture medium
Assess cell line stability
Upstream process development
GMP cell bank including MCB and WCB
Special expression level suitable for GMP production
Stable cell lines are not always "stable", and stability may change with external factors. To tackle this issue, Creative Biolabs has developed a system that is believed to makes cell lines more stable than ever.
Using our optimized protein expression system, our stable cell line has high protein yield and can also meet your needs for expressing intracellular or secreted target proteins.
By addressing these critical quality attributes on process development as early as possible, we can accelerate the transformation of your product from upstream development to downstream manufacturing.
Chinese hamster ovary (CHO) cells are widely used in therapeutic protein production due to its proper folding rate and turnouts of true post-translational modifications. Deletion of α-1,6 fucosyltransferase (FUT8) in CHO cells produces defucosylated monoclonal antibodies and this process involves: i) introducing a gene-editing vector targeting the FUT8 gene sequence to inhibit Fut8 expression and reducing the fucosylation level of the antibody, ii) obtaining a defucosylated stable CHO cell line using LCA method sorting, thereby to express ADCC enhanced antibody.
Fig.1 ADCC Assay with a hIgG1 expressed in WT and Fut8-/- CHO.
Instead of knocking out any genes, Creative Biolabs' proprietary Glymax™ technology redirects fucose synthesis through a heterologous enzyme that depletes the fucose pool inside the cell. For any successfully established stable CHO cell lines, it can produce defucosylated antibodies with up to 100-fold ADCC activity against tumor cells or virus-infected cells.
Fig.2 Antibodies produced by GlymaxX engineered cells show an increased ADCC response.
Our proprietary GlymaxX® technology can be used on your standard host cell line. We can also re-customize existing clones or provide in-house optimal Glymax CHO cell lines to meet your needs.
On the contrary to genetic engineering at the cellular level, Creative Biolabs has made a breakthrough in reducing the fucose content of the metabolic pathway of the cell. A significant increase in the activity of the monoclonal antibody ADCC is, therefore, achieved.
Fig.3 Mass spectra of N-glycans of total CHO cell membrane proteins after treatment with fucose analog (B) under serum-free culture conditions; in comparison to an untreated control (A).
At Creative Biolabs, we provide cGMP-compliant cell banks in strict accordance with industry standards to ensure that our customers have access to fully characterized and phenotypically consistent cells. Together with our antibody production services, the master cell bank (MCB) and working cell bank (WCB) we offer can be used for routinely laboratory use or therapeutic antibody generation.
Master Cell Banks (MCB) is critical to therapeutic product development, not only in clinical research and manufacture, but also in commercial supply phase.
Working Cell Banks (WCB) is prepared from MCB under far more defined cell culture conditions. WCB can support cell-based assays or work as substrates for other assays. In addition, we also offer comprehensive biosafety testing services, including cell bank characterization and genetic stability assessments of the final producer cell lines to our clients.
Fig.4 Cell Banking.
Creative Biolabs is aiming to provide our customers with a complete portfolio of upstream development services. Our team has extensive pharmaceutical experience in developing and optimizing reproducible and scale-up processes. The goal of our service is to provide the most efficient upstream process development to meet your needs to ADCC/CDC⁺ antibody drugs.
Our upstream process development services include but are not limited to the following:
Raw materials and equipment supply—perform raw material testing in FDA approved cGMP manufacturing facilities.
Cell culture optimization—conduct media optimization including both animal or animal-free compositions; Determine cell viability, behavior, productivity and stability.
Fig.5 The Dasgip Parallel Bioreactor System with four parallel bioreactors used for culture of CHO cells.
Fermentation development—Perform bioreactors range from 1L to 2000L; Available in both GMP and non-GMP manufacturing; Use state-of-the-art cell culture analyzers.
Fig 6. A dramatic decrease in the amount of asparagine (red line ASN) during cell culture was observed as a result of consumption and degradation. An increase in the aspartic acid degradation product aspartic acid was also observed.
To optimize the critical quality attributes of ADCC / CDC⁺ products, we have established a comprehensive approach, including but not limited to:
For more details about our stable cell line service, please do not hesitate to contact us.
Creative Biolabs provides luciferase-based ADCC assay. This Jurkat cell based assay is pioneered by Creative Biolabs, and the methodology is very well accepted by the field. See attached ADCC Reporter Assay Protocol for further details.
All products and services are for Research Use Only. Do Not use in humans.
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Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.