Generation and application of a 293 cell line stably expressing bovine interferon-gamma

Protein Expr Purif. 2014 Jul:99:131-7. doi: 10.1016/j.pep.2014.04.012. Epub 2014 May 2.

Abstract

A stable mammalian cell line expressing highly active bovine interferon-gamma (BoIFN-γ) was generated using Flp recombinase-mediated integration. This recombinant 293 cell line (B1) efficiently secreted FLAG-tagged BoIFN-γ protein into the culture supernatant, as determined by ELISA and Western blot. The recombinant BoIFN-γ exhibited high anti-viral activity, suggesting that the 293 cells expressed BoIFN-γ that structurally and biologically resembled the natural protein. Two monoclonal antibodies (mAbs) with high affinity for the 293 cell-expressed BoIFN-γ were identified using this cell line, and these mAbs can be used for the development of diagnostic kits. Thus, this work demonstrates the successful generation of a 293 cell line that produces large quantities of highly active BoIFN-γ and demonstrates its potential application in the research of bovine infectious diseases.

Keywords: 293 cell line; Antibody; Bovine interferon-gamma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology
  • COS Cells
  • Cattle
  • Cell Line
  • DNA Nucleotidyltransferases
  • HEK293 Cells
  • Humans
  • Interferon-gamma / biosynthesis*
  • Interferon-gamma / immunology
  • Interferon-gamma / pharmacology
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / immunology
  • Recombinant Proteins / pharmacology

Substances

  • Antiviral Agents
  • Recombinant Proteins
  • Interferon-gamma
  • DNA Nucleotidyltransferases
  • FLP recombinase