High-yield CHO Stable Cell Line Development

Biotechnology companies, pharmaceutical companies, and academic laboratories frequently require gram quantities of antibodies or proteins for research, diagnostic, or clinical purposes. The production of such large quantities of antibodies or proteins recombinantly in mammalian cells is a challenging obstacle. While cell culture media and culturing conditions play a role in protein production levels, the essential driver for high production yield are the cells used to produce the recombinant protein. While several large biotechnology and pharmaceutical companies (e.g. Genentech/Roche) have been able to successfully develop and utilize high-yield stable cell lines, their cell line development technology is either secret or very expensive to contract. Recognizing the market need in this area, LakePharma has set out to develop a process that can make high-yield mammalian stable cell lines relatively inexpensively and with the utmost efficiency, making the development of high-yield stable cell lines accessible to all.

LakePharma has also developed proprietary bicistronic vectors for recombinant antibody production. These vectors contain uniquely positioned restriction enzyme sites that enable efficient cloning of antibody genes. Unlike the popular bicistronic vectors that employ two copies of identical CMV promoters and are susceptible to instability, LakePharma vectors do not use CMV promoters and do not have stability issues.

LakePharma uses a variety of gene amplification-compatible selectable markers, including a new patented and proprietary selectable marker system developed completely in-house. This new selectable marker has radically improved the stable cell line generation process and has significantly increased the success rate of stable cell line development for difficult targets. Utilizing this selection process, the expression levels and stability of target genes can now be tightly controlled.

By combining LakePharma's proprietary expression vectors and high-yield stable cell line development systems, LakePharma has engineered CHO stable lines that produce therapeutic proteins and antibodies at levels exceeding 50 PCD. Using this cell line development process, the path to 1 gram per liter production yield is as short as 2 months. With additional gene amplification and clone selection, the production yield can be further improved to 5 gram per liter, which will reduce the amount of media consumed and the number of bioreactors required. Such technology milestones enable efficient and inexpensive large-scale mammalian protein and antibody production.