Purification of poliovirus obtained from human diploid cells by means of DEAE-Sepharose.
Mahoney, MEF and Saukett poliovirus strains were grown on human diploid cells (MRC 5) and concentrated on Amicon filter. Concentrated viral suspensions containing 3H-uridine labelled virus were mixed with 14C-amino acid labelled cell extracts containing calf serum and passed through a DEAE-Sepharose column. Fractions eluted at various ionic strengths were analyzed for infectivity, radio-activity and serum content by counter immunoelectrophoresis in the presence of a rabbit anti-calf serum. A peak containing 40--60% of the input infectivity was easily obtained by elution with 0.01 M phosphate buffer at pH 7. The virus peak did not contain 14C or calf serum and its purity was controlled by PAGE-SDS electrophoresis and electron microscopy. This technique may be useful in the large-scale purification of viruses used in the preparation of inactivated vaccines.
D002460:Cell Line; D002845:Chromatography / Q000379:methods*; D004171:Diploidy; D006801:Humans; D017955:Poliovirus / Q000302:isolation & purification* / Q000472:pathogenicity / Q000648:ultrastructure; D012685:Sepharose
D012685:Sepharose