【GS系统】GS Gene Expression System.doc

GS Gene Expression System Principles of the GS Gene Expression System? Glutamine synthetase (GS) is the enzyme responsible for the biosynthesis of glutamine from glutamate and ammonium. This enzymatic reaction is the pathway for glutamine formation in a mammalian cell. In the absence of glutamine in the growth medium, the GS enzyme plays an essential role in the survival of mammalian cells in culture. Some mammalian cell lines, such as mouse myeloma lines, do not express sufficient GS to survive without added glutamine. With these cell lines, a transfected GS gene can function as a selectable marker by permitting growth in a glutamine-free medium. Other cell lines, such as Chinese hamster ovary (CHO) cell lines, express sufficient GS to survive without exogenous glutamine. In these cases, the GS inhibitor, methionine sulphoximine, can be used to inhibit endogenous GS activity such that only transfectants with additional GS activity can survive. A Complete Mammalian Expression System Lonza has developed a highly efficient mammalian expression system which makes use of selection, via glutamine metabolism, and powerful viral promoters. The GS System? is well established with over 85 companies using the technology worldwide. Currently there are over 50 products in clinical trials and 5 products in-market that use the GS System?. Productivity The high manufacturing cost of protein products, particularly for therapeutic proteins, means that maximising volumetric productivity is essential if economic processes are to be established. Use of the GS System? reliably results in high-producing cell lines early in cell line construction programmes. Maximum expression levels attainable depend on the product but cell lines producing over 5 g/L of recombinant antibody have been created, with specific production rates in the range 15-65 pg/cell/day. Productivity in chemically defined, animal compotent-free GS-CHO bioreactor process Productivity in chemically