Preparation of purified, sterilized, and stable adenovirus vectors using albumin

J Virol Methods. 2002 May 16;103(2):191-9. doi: 10.1016/s0166-0934(02)00028-9.

Abstract

Cesium chloride ultracentrifugation combined with dialysis is a standard method for preparing a high titer of purified recombinant adenovirus. However, it was found that sterilization of the filter membrane after dialysis led to a complete loss of adenovirus activity. A virus pellet was visualized after centrifugation shortly and electron microscopy revealed an aggregation of recombinant virus within the filter membrane following dialysis with 10% (V/V) glycerol in phosphate buffered saline. The entrapment of aggregated adenovirus by the filter membrane explains why adenovirus activity is lost following sterilization. The addition of 1% albumin prevented viral aggregation and allowed the purified virus to retain its activity after filter sterilization. Furthermore, viral activity was retained within the 1% albumin solution for at least 1 week at 37 degrees C and for 2 weeks at 4 degrees C, whereas viral activity within the albumin-free solution was quickly lost.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Adenoviridae / isolation & purification*
  • Adenoviridae / physiology
  • Animals
  • Cell Line
  • Genetic Vectors / isolation & purification*
  • Genetic Vectors / physiology
  • Humans
  • Recombination, Genetic
  • Serum Albumin, Bovine
  • Virus Cultivation / methods

Substances

  • Serum Albumin, Bovine