UCOE: Gene Expression Technology
Protein Production Solutions
Millipore's UCOE Technology (Ubiquitous Chromatin Opening Element) Winner of 2007 Frost & Sullivan Technology Innovation Award |
UCOE (Ubiquitous Chromatin Opening Element) expression technology gives major improvements in gene expression in stably-transfected mammalian cells through effects on the structure of chromatin. UCOE expression technology prevents transgene silencing and gives consistent, stable and high-level gene expression irrespective of the chromosomal integration site. UCOE expression elements are small DNA elements (isolated from the area around house-keeping genes, which need to be active most of the time) that create a transcriptionally active, open chromatin environment around an integrated transgene, maximizing its potential to be transcribed into protein, irrespective of the position of the transgene in the chromosome.
| Technical Literature | |
| Data Sheet | UCOE Expression Technology |
| Application Note | An Accelerated Method for Production of Recombinant Proteins using UCOE Expression Technology |
| Posters | |
| Press Releases | |
| Millipore Corporation Signs License Agreement with Bayer HealthCare AG | |
| Millipore Receives Prestigious Frost & Sullivan Innovation Award | |
Advantages
- Over 50% of UCOE-derived clones have higher expression than the best non-UCOE-derived clones
- UCOE-derived vectors produce substantially more expressing clones than non-UCOE-derived vectors
- Cell lines stable over 130 generations
- High-yielding cell lines can be derived in less than 60 days, without amplification
- Small (4-8kb) DNA elements, capable of making two protein chains per plasmid, e.g., antibodies
- Express Antibodies, Receptors, Enzymes, Cytokines, etc.
- Integrates with industry standard platforms, e.g, CHO cells, CMV promoters, plasmids, transfection agents, media, etc.
Successful Integration
The successful integration of a transgene into mammalian cells is largely dependent upon the chromatin structure around the integration site. Most mammalian chromatin exists in the closed (heterochromatin) form, which is transcriptionally silent. With conventional vectors, most integrations suffer from position effects and chromatin shutdown, which leads to gene-silencing and unproductive integration events. As a result, pools of stably-transfected cells generally show low productivity and stability, as only a few cells might be good producers, and gene silencing and overgrowth of non-producers rapidly diminish the overall productivity of the pool. This also makes it very difficult and time consuming to find high-expressing stable clones needed for large-scale manufacturing.» back to top
Benefits Of UCOE Expression Technology
The productivity and stability of individual cells allows pools to be a source for rapidly generating gram quantities of proteins in 3 – 4 weeks. This has potential advantages over transient transfection, minimizing the quantities of DNA and costly transfection agent, as well as allowing aliquots of the pool to be stored for future use if the protein is required at a later stage.3x107 CHO-S cells were transfected with 90µg of an antibody expression plasmid either with or without a UCOE, drug selection was applied and once sufficient cell number was generated cells were seeded at a cell density that would allow growth to 10L. On day 16 a small non-feed addition was made to a proportion of the cultures (+opti). A fraction of the cells were grown and the data shown
Because the majority of cells are stable high-expressors, it is much easier and faster to find high-yielding clones with the productivity and stability required for manufacturing, as opposed to the "needle-in-a-haystack" approach using conventional vectors.
Vectors encoding heavy and light chains (separate plasmids) were cotransfected into CHO-S cells followed by drug selection and screening of 96 randomly selected clones.
