Application of a statistical design to the optimization of culture medium for recombinant interferon-gamma production by Chinese hamster ovary cells

Paula M. L. Castro*, Paul M. Hayter, Andrew P. Ison, Alan T. Bull

*Autor correspondente para este trabalho

Resultado de pesquisarevisão de pares

96 Citações (Scopus)

Resumo

The importance of serum-free medium components on the growth of Chinese hamster ovary (CHO) cells and production of recombinant human interferon(IFN)-gamma was investigated. The complexity of the medium led to the adoption of a statistical optimization approach based on a Plackett-Burman design. From this analysis a set of nutritional components was identified as important for cell growth and recombinant protein production. Glycine was identified as an important determinant of specific growth rate, whereas for cell production bovine serum albumin (BSA), phenylalanine and tyrosine were also identified as important. BSA, sodium pyruvate, glutamate, methionine, proline, histidine, hydroxyproline, tyrosine and phenylalanine were shown to be important for IFN-gamma production. Other medium components, such as insulin, arginine, aspartate and serine produced an inhibitory effect on both cell growth and IFN-gamma production. The effect of the stimulatory nutrients as a whole group was tested by increasing their concentration in the medium. A significant improvement in specific cell growth rate, cell production and IFN-gamma production (up to 45%) was achieved on both shake-flask and fermentor cultures. An increase in the medium concentration of the negative variables had only a small inhibitory effect (approximately 10%) on the same parameters. Analysis of the effects of the group of stimulatory amino acids and BSA on CHO cell growth showed that the effect of the former was independent of BSA.

Idioma originalEnglish
Páginas (de-até)84-90
Número de páginas7
RevistaApplied Microbiology and Biotechnology
Volume38
Número de emissão1
DOIs
Estado da publicaçãoPublicado - out. 1992
Publicado externamenteSim

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