Quantification of recombinant core-like particles of bluetongue virus using immunosorbent electron microscopy

J Virol Methods. 1999 Jun;80(1):1-9. doi: 10.1016/s0166-0934(98)00170-0.

Abstract

Immunosorbent electron microscopy was used to quantify recombinant baculovirus-generated bluetongue virus (BTV) core-like particles (CLP) in either purified preparations or lysates of recombinant baculovirus-infected cells. The capture antibody was an anti-BTV VP7 monoclonal antibody. The CLP concentration in purified preparations was determined to be 6.6 x 10(15) particles/l. CLP concentration in lysates of recombinant baculovirus-infected cells was determined at various times post-infection and shown to reach a value of 3 x 10(15) particles/l of culture medium at 96 h post-infection. The results indicated that immunosorbent electron microscopy, aided by an improved particle counting method, is a simple, rapid and accurate technique for the quantification of virus and virus-like particles produced in large scale in vitro systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bluetongue virus / genetics
  • Bluetongue virus / ultrastructure*
  • Cell Line
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Spodoptera / cytology
  • Viral Core Proteins / analysis*
  • Viral Core Proteins / genetics

Substances

  • Recombinant Fusion Proteins
  • VP3 protein, Bluetongue virus
  • Viral Core Proteins
  • VP7 protein, orbivirus