Millipore Technical Publications |
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Advanced Method of Cell Line Generation Using UCOE® Technology | ||
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| Year | 2008 | |
IntroductionIsolation of highly-productive stable cell lines is the primary obstacle in the process of therapeutic protein production. Methods such as gene amplification and automated clone selection have been somewhat successful in overcoming this barrier. However, these methods have added additional problems such as increased timelines, instability of expression and increased cost to the process. New technologies that permit isolation of highly-productive cell lines in shorter times are attractive to the industry. UCOE (Ubiquitous Chromatin Opening Element) technology overcomes impediments of cell line development by producing numerous clones from a single transfection that are both highly productive and stable. UCOE technology removes the need for gene amplification or automated clone selection and reduces the time required for cell line development to less than four months. UCOE elements are isolated from regions upstream of constitutively-expressed house-keeping genes and function to prevent cellular transcriptional silencing mechanisms. The UCOE technology is available in a number of vectors containing strong viral promoters, various selection markers and different UCOE elements. These vectors are designed to incorporate either one or two genes. The latter vectors enable expression of to chains of a multimeric protein, such as the immunoglobulin genes, from the same DNA molecule. Alternatively, the utilization of independent single expression vectors gives researchers an opportunity to optimize the ratio of both chains, an approach that in many cases results in an increased productivity. Current UCOE vectors utilize two different elements derived from the murine ribosomal protein S3 (RPS3) and the human HNRPA2B1-CBX3 loci, in a number of different sizes and configurations. Click the PDF above for the full document. | |
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