Concentration and purification of rabies viral and lentiviral vectors

Cold Spring Harb Protoc. 2015 Apr 1;2015(4):386-91. doi: 10.1101/pdb.prot075887.

Abstract

Rabies viral and lentiviral vectors are very useful tools for neuroscientists, but high titer and purity are critical for in vivo applications. Here we present a protocol for concentration and purification of viral stocks by ultracentrifugation on a sucrose step gradient to remove impurities of both higher and lower densities than the virus itself, with sucrose removed by a subsequent pelleting step. The final stocks are concentrated in volume by a factor of up to 1000, with higher expected purity than is obtained following previously published protocols for preparing G-deleted rabies viral vectors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Genetic Vectors / isolation & purification*
  • Lentivirus / isolation & purification*
  • Rabies virus / isolation & purification*
  • Ultracentrifugation / methods*