We do this for both gs system cell lines and cell lines brought to us by our customers.
Cell line construction.
Stable cell line construction service.
We have titrated the amount required for our hela cell lines and this can be a starting point for other cell lines.
Thaw a new vial of cells after 20 30 passages.
The health of the the target cell line is critical for obtaining accurate results.
According to the upstream regulator of the reporter gene reporter cell lines can be further divided into two groups.
1 constitutive reporter cell lines in which reporters are constitutively expressed under the control of a constitutive strong promoter such as cmv promoter.
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.
The first step in the process is the isolation of single viable cells.
Lenti x 293t cells or alternative cell lines target cell line any human cancer cell line titered lentivirus containing your sequence of interest.
The development of a stable cell line is time consuming and complicated.
Delivery of the target gene into the host cell screening of stable expression clones and stability test.
Methods to generate stable cell lines.
Stable cell line construction for protein antibody production.
The amount of g418 required to kill cells not expressing the construct will vary from cell line to cell line.
We use a modified frt flp in recombinase system to produce human proteins in cho cells and 293 cells for stable expression.
Check the cells for mycoplasma regularly.
Do not over or under grow your cells.
Construction of a stable cell line usually takes 3 steps.
And 2 inducible reporter cell lines in which reporters are inducibly.
Infection efficiency will vary between cell lines.
To create stable cell lines is a key method in reducing the production costs of biopharmaceuticals.
Depending on the scope of the experiment several options are used for the generation of a stable cell line.
Over time this will select for cells that have stably incorporated the gfp plasmid into their genomic dna.
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.
Cell line development requires the discovery of single cell derived clones that produce high and consistent levels of the target therapeutic protein.
Selection antibiotic e g puromycin microcentrifuge tubes.
Limiting dilution is a technique that relies on statistical probability but is time consuming.