Monitoring of the tissue distribution of fibroblast growth factor containing a high mannose-type sugar chain produced in mutant yeast

Glycoconj J. 2004;20(6):385-97. doi: 10.1023/B:GLYC.0000033995.95412.1f.

Abstract

Most therapeutic glycoproteins have been produced in mammalian cell lines. However, the mammalian cell culture system has various disadvantages, i.e., a high culture cost, difficulty in performing a large scale-up because of complicated handling requirements, and the risk of contamination by prion or other unknown pathogenic components through cultivation in the presence of bovine serum. There is thus a growing need for other host cells in which the recombinant glycoproteins can be produced. Recently, we successfully developed a mutant yeast strain engineered in a glycosylation system. The sugar chain produced in the mutant yeast is not immunogenic to the human immuno-surveillance system. In the present study, we selected fibroblast growth factor (FGF) as a model glycoprotein and assessed the bioactivity of FGF produced in yeast in terms of its proliferating activity and tissue distribution in mammalian cells and in the whole body. Structural changes in the sugar chains of FGFs derived from mutant yeast, as compared with those from mammalian cells, did not affect the proliferating activity remarkably. However, the tissue distribution in the mouse differed significantly; a high-mannose type sugar chain was the major determinant of the specific distribution of FGF to the kidney. The mechanism of this phenomenon is still unclear, but our observations suggest that recombinant glycoproteins derived from mutant yeasts producing high-mannose type sugar chains would be applicable for tissue-targeting therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • CHO Cells
  • Cells, Cultured
  • Cricetinae
  • Electrophoresis, Polyacrylamide Gel
  • Endothelium, Vascular / cytology
  • Escherichia coli / metabolism
  • Fibroblast Growth Factors / biosynthesis*
  • Fibroblast Growth Factors / chemistry*
  • Fungal Proteins / chemistry
  • Genetic Vectors
  • Genotype
  • Glycoproteins / chemistry
  • Glycoproteins / metabolism
  • Glycosylation
  • Male
  • Mannose / chemistry*
  • Mannosidases / biosynthesis
  • Mannosyltransferases / genetics*
  • Membrane Glycoproteins / genetics*
  • Mice
  • Models, Genetic
  • Mutation
  • Oligosaccharides / chemistry
  • Prions
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / chemistry
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins / genetics*
  • Time Factors
  • Tissue Distribution

Substances

  • Fungal Proteins
  • Glycoproteins
  • Membrane Glycoproteins
  • Oligosaccharides
  • Prions
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • Fibroblast Growth Factors
  • MNN1 protein, S cerevisiae
  • Mannosyltransferases
  • Mannosidases
  • mannosyl-oligosaccharide 1,2-alpha-mannosidase
  • Mannose