Cell Line Construction

Cell line development requires the discovery of single cell derived clones that produce high and consistent levels of the target therapeutic protein.

Cell line construction.

Methods to generate stable cell lines. Thaw a new vial of cells after 20 30 passages. And 2 inducible reporter cell lines in which reporters are inducibly. To create stable cell lines is a key method in reducing the production costs of biopharmaceuticals.

We use a modified frt flp in recombinase system to produce human proteins in cho cells and 293 cells for stable expression. Selection antibiotic e g puromycin microcentrifuge tubes. When large amount of recombinant protein is required stable expression line construction is recommended for reproducible and cost effective protein production. We have titrated the amount required for our hela cell lines and this can be a starting point for other cell lines.

Check the cells for mycoplasma regularly. The development of a stable cell line is time consuming and complicated. Stable cell line construction for protein antibody production. 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.

The first step in the process is the isolation of single viable cells. 1 constitutive reporter cell lines in which reporters are constitutively expressed under the control of a constitutive strong promoter such as cmv promoter. Stable cell line construction service. Over time this will select for cells that have stably incorporated the gfp plasmid into their genomic dna.

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. Depending on the scope of the experiment several options are used for the generation of a stable cell line. Delivery of the target gene into the host cell screening of stable expression clones and stability test. Infection efficiency will vary between cell lines.

Do not add penicillin streptomycin to the media. Construction of a stable cell line usually takes 3 steps. We do this for both gs system cell lines and cell lines brought to us by our customers. Limiting dilution is a technique that relies on statistical probability but is time consuming.

Lenti x 293t cells or alternative cell lines target cell line any human cancer cell line titered lentivirus containing your sequence of interest. 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. The amount of g418 required to kill cells not expressing the construct will vary from cell line to cell line.