Sequential two-step chromatographic purification of infectious poliovirus using ceramic fluoroapatite and ceramic hydroxyapatite columns.

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ID: 47753
2019
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Abstract
Infectious virus purification techniques are important for vaccine development and gene therapy applications. However, the standardized one-step purification technique using ceramic hydroxyapatite (CHAp) has proven unsuitable for poliovirus. Therefore, we designed a sequential two-step chromatographic technique for purification of the infectious Sabin type 2 vaccine strain of poliovirus from the cell culture supernatant. In the first step, we removed protein contaminants from the Sabin type 2 virus fraction by pH gradient elution on a ceramic fluoroapatite column. In the second step, we removed double-stranded DNA derived from host cells by diluting the virus fraction, directly loading it on a CHAp column, and purifying it using a phosphate gradient with 1 M sodium chloride. This process achieved removal rates of more than 99.95% and 99.99% for proteins and double-stranded DNA, respectively, and was highly reproducible and scalable. Furthermore, it is likely that it will be applicable to other virus species.
Reference Key
kurosawa2019sequentialplos Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Kurosawa, Yae;Sato, Shigehiro;Okuyama, Tsuneo;Taoka, Masato;
Journal PloS one
Year 2019
DOI
10.1371/journal.pone.0222199
URL
Keywords
apoptosis carboxypeptidase b-like scylla paramamosain vibrio alginolyticus white spot syndrome virus

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