Evaluation of viral contamination in a baculovirus expression system

Microbiol Immunol. 2018 Mar;62(3):200-204. doi: 10.1111/1348-0421.12572. Epub 2018 Feb 19.

Abstract

Insect expression systems based on baculovirus are widely used for generating recombinant proteins. Here, the infectivity of baculoviruses under the physiological stresses of 'freeze-thaw' and sonication and the baculoviral contamination of recombinant proteins after protein purification were evaluated. Our findings suggest that Nonidet P-40 (NP-40) treatment of baculoviruses completely abolishes their infectivity and that recombinant proteins purified with affinity beads do not include infectious baculoviruses. We therefore suggest that baculovirus is completely inactivated by NP-40 treatment and that recombinant proteins are unlikely to be contaminated with infectious baculoviruses after their affinity purification.

Keywords: baculovirus; insect expression system.

MeSH terms

  • Baculoviridae / drug effects
  • Baculoviridae / genetics*
  • Baculoviridae / radiation effects*
  • Chromatography, Affinity
  • DNA Contamination*
  • Freezing / adverse effects*
  • Genetic Vectors / genetics
  • Octoxynol
  • Polyethylene Glycols / pharmacology
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / isolation & purification
  • Sonication / adverse effects*
  • Stress, Physiological*
  • Virus Inactivation / drug effects

Substances

  • Recombinant Proteins
  • Polyethylene Glycols
  • Octoxynol
  • Nonidet P-40