Production and characterization of long-acting recombinant human albumin-EPO fusion protein expressed in CHO cell

Protein Expr Purif. 2009 Dec;68(2):137-45. doi: 10.1016/j.pep.2009.07.003. Epub 2009 Jul 10.

Abstract

A long-lasting recombinant human albumin-linker-erythropoietin (EPO) is a human albumin gene fused to the N-terminal of EPO with a (GGSGG)(n)-repeated linker inserted between albumin and EPO. Albumin-EPO fusion genes were co-transfected with the dhfr gene. Albumin-EPO fusion protein has three kinds of sub-types (IALE, AD2LE, AD1LE). Albumin-EPO fusion protein was quantified with human EPO ELISA. The in vitro efficacy of albumin-EPO fusion protein was estimated using F-36E cell, and in vivo efficacy of albumin-EPO fusion protein was estimated using normocythemic mice (B6D2F1). We also determined the in vivo half-life in a Sprague-Dawley rat. A PLA program analysis result demonstrated that the albumin-EPO fusion protein IALE is about 7.8-fold more potent than rHuEPO in increasing the hematocrit of normal mice.

MeSH terms

  • Analysis of Variance
  • Animals
  • Blotting, Western
  • CHO Cells
  • Cell Count
  • Cell Line, Tumor
  • Chromatography, Gel
  • Cloning, Molecular / methods*
  • Cricetinae
  • Cricetulus
  • Erythropoietin / chemistry
  • Erythropoietin / genetics
  • Erythropoietin / metabolism*
  • Half-Life
  • Humans
  • Mice
  • Polymerase Chain Reaction
  • Rats
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Proteins
  • Reticulocytes
  • Serum Albumin / biosynthesis*
  • Serum Albumin / chemistry
  • Serum Albumin / genetics

Substances

  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Serum Albumin
  • Erythropoietin