Preparation of functionally active recombinant human interleukin-6

Biochemistry (Mosc). 2007 Apr;72(4):424-9. doi: 10.1134/s0006297907040098.

Abstract

The gene of human interleukin-6 (hIL-6) with an additional 20 amino acids on the N-end, including six histidine residues, was cloned into the expression plasmid pET28b(+). The conditions were elaborated for preparing highly active protein both using denaturing agents and without them. Application of a dialysis cascade allowed us to prepare a functionally active hIL-6 of 90-95% purity with the yield of 3 mg from liter of the cell culture. The highest activity was detected by ELISA in the preparation obtained without denaturing agents. The functional activity of hIL-6 was studied by flow cytofluorimetry. Addition of hIL-6 to the cells of immortal lines of human multiple myeloma resulted in dimerization of the gp130 receptor molecule.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / metabolism
  • Humans
  • Interleukin-6 / biosynthesis*
  • Interleukin-6 / isolation & purification
  • Interleukin-6 / pharmacology
  • Recombinant Proteins / biosynthesis

Substances

  • Interleukin-6
  • Recombinant Proteins