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.

Cell line construction.

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 amount of g418 required to kill cells not expressing the construct will vary from cell line to cell line. Over time this will select for cells that have stably incorporated the gfp plasmid into their genomic dna. Depending on the scope of the experiment several options are used for the generation of a stable cell line.

We have titrated the amount required for our hela cell lines and this can be a starting point for other cell lines. Do not add penicillin streptomycin to the media. Stable cell line construction for protein antibody production. Thaw a new vial of cells after 20 30 passages.

Do not over or under grow your cells. Selection antibiotic e g puromycin microcentrifuge tubes. Lenti x 293t cells or alternative cell lines target cell line any human cancer cell line titered lentivirus containing your sequence of interest. Infection efficiency will vary between cell lines.

The health of the the target cell line is critical for obtaining accurate results. To create stable cell lines is a key method in reducing the production costs of biopharmaceuticals. Stable cell line construction service. 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.

Check the cells for mycoplasma regularly. The development of a stable cell line is time consuming and complicated. Methods to generate stable cell lines. Delivery of the target gene into the host cell screening of stable expression clones and stability test.

When large amount of recombinant protein is required stable expression line construction is recommended for reproducible and cost effective protein production. 1 constitutive reporter cell lines in which reporters are constitutively expressed under the control of a constitutive strong promoter such as cmv promoter. Construction of a stable cell line usually takes 3 steps. Limiting dilution is a technique that relies on statistical probability but is time consuming.

The first step in the process is the isolation of single viable cells. And 2 inducible reporter cell lines in which reporters are inducibly. 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.