Role of environmental conditions on the expression levels, glycoform pattern and levels of sialyltransferase for hFSH produced by recombinant CHO cells

A recombinant CHO cell line in which the expresison of human follicle stimulating hormone (hFSH) was under the control of the β actin promoter was maintained in steady state perfusion cultures on a protein free medium. The level of expression of the hFSH was controlled by varying the steady state level of dissolved oxygen (10–90% of air saturation) and of sodium butyrate (0–1.5mM). Under these conditions, the specific productivity of hFSH (qFSH) varied from 0.7 to 4.8 ng hFSH/106 cells/h. As the specific productivity of hFSH increased, there was a shift in the FSH isoforms to the lower pI fractions, corresponding to increased sialic acid content. As the specific productivity of hFSH increased, shifting the isoform distribution towards the lower pI isoforms, that the sialyltransferase enzymic activity also increased.

[1]  A. Ulloa-Aguirre,et al.  Immunological and biological potencies of the different molecular species of gonadotrophins. , 1988, Human reproduction.

[2]  P. Louisot,et al.  Comparative study of the N-glycoprotein synthesis through dolichol intermediates in mitochondria, Golgi apparatus-rich fraction and endoplasmic reticulum-rich fraction. , 1990, The International journal of biochemistry.

[3]  B. Shah,et al.  Comparison of N‐Linked Oligosaccharides of Recombinant Human Tissue Kallikrein Produced by Chinese Hamster Ovary Cells on Microcarrier Beads and in Serum‐Free Suspension Culture , 1994, Biotechnology progress.

[4]  Charles F. Goochee,et al.  Environmental Effects on Protein Glycosylation , 1990, Bio/Technology.

[5]  P. Gray,et al.  Expression of FSH in CHO cells I. Comparison of promoter types and effects of their respective inducers , 2004, Cytotechnology.

[6]  J. Paulson,et al.  Sialylation of glycoprotein oligosaccharides N-linked to asparagine. Enzymatic characterization of a Gal beta 1 to 3(4)GlcNAc alpha 2 to 3 sialyltransferase and a Gal beta 1 to 4GlcNAc alpha 2 to 6 sialyltransferase from rat liver. , 1982, The Journal of biological chemistry.

[7]  C. Coutifaris,et al.  Biosynthesis and secretion of follicle-stimulating hormone. , 1983, Endocrine reviews.

[8]  W. Walker,et al.  Developmental changes in the activities of sialyl- and fucosyltransferases in rat small intestine. , 1986, Biochimica et biophysica acta.

[9]  F. Naider,et al.  Partial characterization and purification of the glycosylation site recognition component of oligosaccharyltransferase. , 1988, Journal of Biological Chemistry.