Research Article

Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase

Figure 2

Western blotting analysis of the expression of recombinant human GCR in stably transfected CHO-DXB11 cells, after the third round of amplification with 150 nM MTX. (a) Conditioned media of 5 selected clones (C 12, C 20, C 22, C 37, and C 45). (b) Cellular extracts from the selected clones and the corresponding membrane stained with Ponceau S after proteins transfer. The total protein from conditioned media was previously concentrated by precipitation with 10% TCA. Nonglycosylated recombinant human GCR of 56 kDa purified from E. coli was used as positive control. Conditioned medium and cellular extract of nontransfected CHO-DXB11 cells were the negative control. Glycosylated recombinant GCR bands of 59–69 kDa are indicated by the arrows. The volume of sample applied to the gel corresponded to the supernatant (a) or extract (b) of 150,000 cells.
875383.fig.002a
(a)
875383.fig.002b
(b)