cGMP compliant Stable Cell Line Options
Fut8 Knockout CHO Cell Lines
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.
GFT Knockout CHO Cell Lines
GDP-fucose transporter GFT (GDP-fucose-transporter) is an important protein required for fucosylation of N-oligosaccharide core in mammalian cells. Creative Biolabs uses genetic engineering technology to modify the expression system and knock out the key protein GFT in the fucose modification pathway in the antibody expression host cell CHO-K1, which can effectively reduce the level of antibody fucose modification. This technology has two advantages:
1. Block the classic fucosylation pathway and the compensatory pathway, thereby achieving the goal of completely removing fucosylation (the fucose knockout rate is as high as over 98%).
2. At the same time, construct a stable GFT-/- CHO-K1 cell line for further ADCC research.
Fucoanalog™ CHO Cell Lines
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.1 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).1,2
cGMP compliant Cell Banking
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.
Scale-Up
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
cGMP compliant manufacturing facilities.
Cell culture optimization—conduct media optimization including
both animal or animal-free compositions; Determine cell viability, behavior, productivity and stability.
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.
ADCC / CDC⁺ Antibody CQA Optimization
To optimize the critical quality attributes of ADCC / CDC⁺ products, we have established a comprehensive
approach, including but not limited to:
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Aggregate control
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Charging variant control
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Glycosylation optimization
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Sialic acid content optimization
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ADCC / CDC activity optimization
For more details about our stable cell line service, please do not hesitate to contact us.
Reference
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Rosenlöcher, Julia, et al. "Applying acylated fucose analogues to metabolic glycoengineering." Bioengineering 2.4 (2015): 213-234.
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under Open Access License CC BY 4.0, without modification.
