Stable Cell Line Development Services
Transgenic cell lines are suitable for a variety of research applications, such as the production of recombinant antibodies, recombinant proteins, gene editing, functional research, detection and analysis, etc.. Transgenic cell lines serve to improve the productivity of the stable cell lines and improve the efficiency of the experiment.
High quality cell lines start with high quality DNA
Our customers have the option of providing their own DNA for stable cell line generation purposes, however, Biointron recommends gene synthesis to ensure that we use high quality, codon optimized genes for the best expression.
Biointron allows you to choose from either Glutamine Synthetase or DHFR gene expression systems to construct your stable cell line. To start your project, DNA carrying the gene of interest must be introduced into the cell with a stable transfection protocol. Depending on your preference, we will use either plasmid-based stable transfection or lentivirus-based infection as your preferred stable transfection method.
Transient expression evaluation
To ensure that your protein of interest is expressed in the cells, we first characterize your protein expression by ELISA or Western Blot 48 hours after the transfection.
Stable cell line development
Transfected cells are grown in the appropriate selective media to generate stable pools. For packages using a DHFR expression system, we can also subject the stable pools to additional Methotrexate (MTX) amplification and screening, in order to increase expression levels (packages using a Glutamine Synthetase expression system do not require gene amplification). The productivity and quality of the pools is evaluated up to 10 passages for the top 10 clones.
Top 3 clones from each pool are picked and evaluated further, based on their growth rates.
Cell line stability screening
Stability testing is a critical component of characterizing bioproduction cell lines. Stable cell lines in continuous culture suffer from genetic instability as the number of passages increases. Stability test will help you ensure the quality of your production cells over the intended passages.
Final deliverables, QC data and full report
We provide the sequence and construct report (in case of synthesized gene), top 3 stable clones and a comprehensive stable cell line generation report