Proteome analysis of animal cells for development of efficient serum-free culture technology

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Serum-free culture of animal cells is essential for large-scale production of clinically and diagnostically important proteins. With the aim of developing efficient serum-free culture technology, we have undertaken a proteomic analysis of Chinese hamster ovary (CHO) cells, which are widely used as host cells in the production of complex recombinant proteins. CHO cells capable of growing in a serum-free medium were established by stepwise adaptation of cells maintained in a medium supplemented with 10% fetal bovine serum (FBS) to a medium containing lower concentrations of serum and finally to a serum-free medium. Specific growth rate and specific recombinant protein production rate of the cells adapted to growth in serum-free culture were measured and found to he comparable to those of CHO cells maintained in a FBS-supplemented medium. The serum-independent cells were then studied by two-dimensional gel electrophoresis of intracellular proteins with immobilized pH gradient (pH 3-10) in the first dimension and a linear acrylamide gradient (8-18%T) in the second dimension. The resulting protein expression patterns were compared with those in serum-dependent cells. Among over 700 protein spots analyzed, several spots showed marked increase or decrease, indicating that these proteins may play an important role in growth in serum-free culture.