Cell Line Construction

1 constitutive reporter cell lines in which reporters are constitutively expressed under the control of a constitutive strong promoter such as cmv promoter.

Cell line construction.

Thaw a new vial of cells after 20 30 passages. We use a modified frt flp in recombinase system to produce human proteins in cho cells and 293 cells for stable expression. Stable cell line construction service. The development of a stable cell line is time consuming and complicated.

Do not over or under grow your cells. We do this for both gs system cell lines and cell lines brought to us by our customers. 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.

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. Construction of a stable cell line usually takes 3 steps. Stable cell line construction for protein antibody production.

Depending on the scope of the experiment several options are used for the generation of a stable cell line. The first step in the process is the isolation of single viable cells. Methods to generate stable cell lines. Check the cells for mycoplasma regularly.

Infection efficiency will vary between cell lines. Lenti x 293t cells or alternative cell lines target cell line any human cancer cell line titered lentivirus containing your sequence of interest. 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. Cell line development requires the discovery of single cell derived clones that produce high and consistent levels of the target therapeutic protein.

According to the upstream regulator of the reporter gene reporter cell lines can be further divided into two groups. Over time this will select for cells that have stably incorporated the gfp plasmid into their genomic dna. 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.

Limiting dilution is a technique that relies on statistical probability but is time consuming. Delivery of the target gene into the host cell screening of stable expression clones and stability test. To create stable cell lines is a key method in reducing the production costs of biopharmaceuticals.