Cell line development is a critical first step in the manufacturing of recombinant proteins.
At this stage, you are setting the foundation for all of your future work. Fermentation, expression and purification problems can be circumvented by cell line selection and modification, vector construction, and clone selection.
Through the development of our Lotus system, we have gained extensive experience in cell line development and engineering by modifying the genome of bacterial cell lines. Our toolkit involves various methods of recombineering such as gene knockouts with homologous recombination via lambda-Red and the FLP/FRT method, and the CRISPR Type I system.
Production of recombinant proteins is a 3 step process.
Cell Line Development Services
- Gene knockouts
- Gene knockdown /suppression
- Knock-in of genetic segments
- Point mutations
- Addition of tags to genes on the chromosome
- Host cell protein analysis
- Promoter modification/replacement
- Clone selection
- Expression vector construction
Lotus Cell Line Optimization Platform
A System for Reduced Host Cell Protein Contamination
Boston Mountain Biotech has developed the Lotus platform, a patented method of optimizing cell lines that offer improved downstream purification of recombinant protein products without the reliance on affinity tags or costly resins. Currently, Lotus has been optimized for use with E. coli based proteins purified with ion exchange (IEX) based chromatography. However, we are continuing to expand the suite into other resins and hosts common to the industry. To support the use of these modified cell lines, BMB offers cell line development services to further provide integration with the client’s existing processes.
Comparative utilization studies
- BMB will express and purify the client’s desired recombinant protein in both the Lotus modified cell line as well as a cell line of their choice
- Data will be provided comparing the two cell lines in the areas of growth, target protein expression, host cell protein expression, initial capture efficiency, overall purification efficiency and target protein activity.
Additional cell line development
- BMB will provide additional modifications to design a fully customized cell line based upon the client’s particular needs, this can include gene knockouts, gene suppression or other modifications.
- Data analytic services can be supplied to the client to guide rational choices that support efficient growth, expression and purification methodology