Development of serum-free media for the production of recombinant proteins from rCHO cells using statistical design of hydrolysates mixture

Abstract

To achieve enhanced cell growth and increased product concentration in serum free media, three non-animal derived hydrolysates were used as additive to serum free basal medium (SFM-G; SFM-YH2) developed by information of the previous work in our laboratory. According to the simplex lattice design, 10 media that have different proportions of yeastolate, soy hydrolysate, and wheat gluten hydrolysate were prepared. After Batch cultures of 3 rCHO cell lines producing anti-4-lbb antibody (H-1-5), hTPO (R-2-3-2), and ABC antibody (ABC 11-08) in 10 media prepared before, the effect of each medium was analyzed by statistical program. For H-1-5 and ABC 11-08 cells, yestolate showed strong effect on enhancing specific production rate (q) and negative effect on growth of cells. Wheat gluten hydrolysate showed strong effect on growth of cells when mixed with other hydrolysate. Soy hydrolysate showed positive (not highest) effect on growth and production. For R-2-3-2 cells, yeastolate was most suitable additives that do not need to mix with other hydrolysate. Then, batch cultures of 3 rCHO cell line using optimized proportions of 3 hydrolysates obtained by program were carried out to confirm the result predicted by the simulation. Further, for ABC 11-08 the effect of hydrolysates mixture on the growth of cells was maintained through over 6 passeges in SFM.