Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase
Figure 4
Expression of secreted GCR from transfected CHO-DXB11 cells selected at 700 nM MTX and cultivated for 45 days in MTX-free medium, by western blotting analysis. (a) Conditioned media of the high-producer clone (C 12) collected on days 15, 30, and 45. (b) Conditioned media of 11 subclones (SC) collected on day 45. Nontransfected CHO-DXB11 cells were used as negative control. Clone 12 cultivated at 700 nM MTX was a positive control, as well as the nonglycosylated recombinant GCR of 56 kDa purified from E. coli. Glycosylated recombinant GCR is indicated by arrows. The volume of culture medium applied to the gel corresponded to the supernatant of 15,000 cells.
