Cell Line Construction

Do not over or under grow your cells.

Cell line construction.

We use a modified frt flp in recombinase system to produce human proteins in cho cells and 293 cells for stable expression. Thaw a new vial of cells after 20 30 passages. We have titrated the amount required for our hela cell lines and this can be a starting point for other cell lines. 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. Infection efficiency will vary between cell lines. Stable cell line construction for protein antibody production. According to the upstream regulator of the reporter gene reporter cell lines can be further divided into two groups.

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. To create stable cell lines is a key method in reducing the production costs of biopharmaceuticals. 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. Methods to generate stable cell lines. Do not add penicillin streptomycin to the media. Selection antibiotic e g puromycin microcentrifuge tubes.

The amount of g418 required to kill cells not expressing the construct will vary from cell line to cell line. 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. Lenti x 293t cells or alternative cell lines target cell line any human cancer cell line titered lentivirus containing your sequence of interest. 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. The development of a stable cell line is time consuming and complicated. Cell line development requires the discovery of single cell derived clones that produce high and consistent levels of the target therapeutic protein.

Check the cells for mycoplasma regularly. When large amount of recombinant protein is required stable expression line construction is recommended for reproducible and cost effective protein production. The health of the the target cell line is critical for obtaining accurate results.