Stable Cell Line Construction Service
To create stable cell lines is a key method in reducing the production costs of biopharmaceuticals. We use a modified Flp-FRT recombination system to produce human proteins in CHO cells and 293 cells for stable expression. We have pre-selected CHO cell lines [and 293 cell lines] that contain a single stably integrated FRT site at a transcriptionally active genomic locus and we have proven that they can produce high level expressers of recombinant proteins.
We have also developed a unique series of high-yield vector system, in which two GC-rich regions flanking the gene expression cassette makes the promoter and enhancer continuously exposed to transcriptional machinery by preventing chromatin condensing. This revolutionary ground-breaking technology allows the available highest yields in all common mammalian cell protein production host cell lines, such as 293, CHO and NSO. Antibody yields of up to 2.0- 5.5 grams/liter [80-120pg/cell/day] have been achieved using this system in chemically defined, animal component-free medium. The selection strategy has been designed to identify cell lines that are highly productive and have the ability to grow well in suspension culture in a state-of-the-art, fed-batch fermentation process, in which cell density can reach 1.5×107/mL.
Fig.1 Schematic representation of the development of stable cell lines for the production of therapeutic recombinant proteins.1
In addition, Creative Biolabs is also specialized in Engineered Bacterial construction and large scale fermentation. As an example, our specific bacterial expression vector pCD-Bac is a temperature-inducible prokaryotic expression vector, which contains a PRPL promoter, cIts857 gene [temperature-sensitive CI repressor], multiple cloning sites and two strong transcription terminators. With genetic codons optimized into that bacterial preference, this vector enables high-yield protein expression of most proteins in the host bacterial strain Escherichia coli DH5α when our propriety fermentation protocol is employed. Our fermentation process is a batch process, not a continuous process.
Reference
All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.
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