Expression of an RSV-gag virus-like particle in insect cell lines and silkworm larvae

J Virol Methods. 2011 Nov;177(2):147-52. doi: 10.1016/j.jviromet.2011.07.012. Epub 2011 Jul 27.

Abstract

Rous sarcoma virus group antigen protein-based virus-like particles (VLPs) are well known for their structural integrity and ease of handling. VLPs play an important role in drug delivery systems because they can be manipulated with ease. In this study, a new method was established for expressing Rous sarcoma virus group antigen protein based VLPs in silkworm larvae and establishing stably expressing insect cell lines. These VLPs have been isolated by ultracentrifugation using a sucrose step gradient of 10-60% (v/v), and their spherical structure has been confirmed using transmission electron microscopy (TEM). The spherical morphology is similar in both the silkworm larvae and in stably expressing cell lines. Silkworm larvae are better suited for producing Rous sarcoma virus group antigen protein-based VLPs on a large scale; yields from silkworm larvae were approximately 8.2-fold higher than yields from stable cell lines. These VLPs provide a new method for large-scale application in vaccine development and drug delivery systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Viral / metabolism
  • Blotting, Western
  • Bombyx / genetics
  • Bombyx / metabolism*
  • Cell Line
  • Gene Products, gag / metabolism*
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Genome, Insect*
  • Hemolymph / metabolism
  • Larva / genetics
  • Larva / metabolism
  • Microscopy, Electron, Transmission
  • Rous sarcoma virus / genetics
  • Rous sarcoma virus / metabolism*
  • Ultracentrifugation
  • Viral Proteins / genetics
  • Viral Proteins / metabolism

Substances

  • Antigens, Viral
  • Gene Products, gag
  • Viral Proteins