Expression and characterization of albumin fusion protein canine IFNγ-CSA in baculovirus-insect cell expression system

Protein Expr Purif. 2019 Oct:162:32-37. doi: 10.1016/j.pep.2019.02.019. Epub 2019 May 14.

Abstract

In this study, canine IFNγ was fused by a flexible linker with canine serum albumin to construct the fusion protein IFNγ-CSA for the purpose to design a long-acting canine IFNγ. The fusion protein was successfully expressed in baculovirus-infected Sf9 insect cells and was purified by salting-out and ion exchange chromatography. The IFNγ-CSA fusion possessed potent anti-viral assay against vesicular stomatitis virus in cultured cells. IFNγ-CSA was also stable at 37 °C up to 72 h compared with 8 h for IFNγ alone. In vivo pharmacokinetics demonstrated a significantly longer half-life for IFNγ-CSA (15.42 h) than for canine reIFNγ (1.51 h) in KM mice. These results indicate that IFNγ-CSA expression in the baculovirus system was successful and provide a promising long-acting cytokine for veterinary clinical applications.

Keywords: Baculovirus expression system; Canine serum albumin; Fusion interferon-γ; Pharmacokinetics; Stability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / metabolism
  • Antiviral Agents / pharmacokinetics
  • Baculoviridae / genetics*
  • Baculoviridae / metabolism
  • Dogs
  • Female
  • Gene Expression
  • Interferon-gamma / genetics*
  • Interferon-gamma / metabolism
  • Interferon-gamma / pharmacokinetics
  • Mice
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacokinetics
  • Serum Albumin / genetics*
  • Serum Albumin / metabolism
  • Serum Albumin / pharmacokinetics
  • Sf9 Cells
  • Spodoptera
  • Vesicular stomatitis Indiana virus / drug effects

Substances

  • Antiviral Agents
  • Recombinant Fusion Proteins
  • Serum Albumin
  • Interferon-gamma