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