Quick method of multimeric protein production for biologically active substances such as human GM-CSF (hGM-CSF)

Biochem Biophys Res Commun. 2009 Aug 14;386(1):40-4. doi: 10.1016/j.bbrc.2009.05.125. Epub 2009 Jun 2.


The C-terminal fragment of C4b-binding protein (C4BP)-based multimerizing system was applied to hGM-CSF to induce dendritic cells (DCs) from peripheral blood monocytes (PBMCs), to see whether the C4BP could stimulate immature DCs, since DCs, equipped with pattern recognition receptors such as toll-like receptors (TLRs), are hypersensitive to various immunologically active molecules like LPS. hGM-CSF gene was merged to the 3'-terminal region of the C4BPalpha-chain gene, and the transfected human 293FT cells produced sufficient amount of octameric hGM-CSF, which resulted in iDCs with the same phenotype and the same response to a TRL4 ligand, LPS and a TLR3 ligand, poly I:C, as those induced with authentic monomeric hGM-CSF. These results suggest that the C4BP-based multimerizing system could facilitate the design of self-associating multimeric recombinant proteins without stimulating iDCs, which might be seen with the other multimerizing systems such as that using Fc fragment of IgM.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Complement C4b-Binding Protein / genetics
  • Complement C4b-Binding Protein / metabolism
  • Dendritic Cells / drug effects
  • Dendritic Cells / metabolism
  • Granulocyte-Macrophage Colony-Stimulating Factor / biosynthesis*
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Protein Biosynthesis*
  • Protein Multimerization
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / pharmacology


  • Complement C4b-Binding Protein
  • Recombinant Proteins
  • Granulocyte-Macrophage Colony-Stimulating Factor