Depending on the scope of the experiment several options are used for the generation of a stable cell line.
Cell line construction.
To create stable cell lines is a key method in reducing the production costs of biopharmaceuticals.
Check the cells for mycoplasma regularly.
Cell line development requires the discovery of single cell derived clones that produce high and consistent levels of the target therapeutic protein.
With a proven track record in cell culture we offer a complete range of development services to establish a cell line and process that meet your needs and are suitable for manufacturing product for clinical or commercial supply.
Do not add penicillin streptomycin to the media.
Construction of a stable cell line usually takes 3 steps.
1 constitutive reporter cell lines in which reporters are constitutively expressed under the control of a constitutive strong promoter such as cmv promoter.
A mixed population of drug resistant cells can be used directly for experimental analysis with the advantage of generating fast results but also the disadvantage of dealing with an undefined and.
Methods to generate stable cell lines.
Do not over or under grow your cells.
Limiting dilution is a technique that relies on statistical probability but is time consuming.
Over time this will select for cells that have stably incorporated the gfp plasmid into their genomic dna.
We do this for both gs system cell lines and cell lines brought to us by our customers.
The first step in the process is the isolation of single viable cells.
Stable cell line construction for protein antibody production.
Delivery of the target gene into the host cell screening of stable expression clones and stability test.
And 2 inducible reporter cell lines in which reporters are inducibly.
We have titrated the amount required for our hela cell lines and this can be a starting point for other cell lines.
Biologics international corp bic owns highly qualified experts and decades of experience in upstream bio pharmaceutical development providing you with stable cell line development services.
Stable cell line construction service.
The health of the the target cell line is critical for obtaining accurate results.
We use a modified frt flp in recombinase system to produce human proteins in cho cells and 293 cells for stable expression.
Lenti x 293t cells or alternative cell lines target cell line any human cancer cell line titered lentivirus containing your sequence of interest.
Thaw a new vial of cells after 20 30 passages.
Selection antibiotic e g puromycin microcentrifuge tubes.
The development of a stable cell line is time consuming and complicated.
When large amount of recombinant protein is required stable expression line construction is recommended for reproducible and cost effective protein production.
