Purification and concentration of non-infectious West Nile virus-like particles and infectious virions using a pseudo-affinity Cellufine Sulfate column

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

“…), XRX-001 has been tested in phase I clinical trial [115] and is now developed by Pnuvax (Montreal, Canada). In Table 2 are quoted two studies that detail the performance of cellufine sulfate column chromatography in purifying West Nile (VLPs and virions) [40] and dengue [44] viruses, two other members of the Flaviviridae family. Ohtaki et al [40] captured infectious West Nile virions from a (Vero cell) culture supernatant with high recovery yields (up to 93%).…”

“…In Table 2 are quoted two studies that detail the performance of cellufine sulfate column chromatography in purifying West Nile (VLPs and virions) [40] and dengue [44] viruses, two other members of the Flaviviridae family. Ohtaki et al [40] captured infectious West Nile virions from a (Vero cell) culture supernatant with high recovery yields (up to 93%). The concentration of contaminant proteins in the purified dengue virus suspension obtained by Kanlaya and Thongboonkerd [44] was 6 times lower than in the virus stock but infectivity (PFU number/mg protein) was only doubled due to poor virus recovery.…”

Improving dengue viral antigens detection in dengue patient serum specimens using a low pH glycine buffer treatment

“…Media containing VLPs were concentrated by polyethylene glycol precipitation (10% PEG), and media containing NS1 were concentrated by ultrafiltration using a 10-kD pore size centricon (Millipore, Billerica, MA, USA); finally, concentrated VLPs and NS1 antigens were further purified using Cellufine Sulfate columns (Chisso, Tokyo, Japan) as previously suggested. 19 Columns were washed with 10 mL adsorption buffer (0.01M sodium phosphate, 0.1M NaCl, pH 7.5) and then 0.4M and 0.6M NaCl in adsorption buffer were used to elute NS1 and VLPs, respectively. The purity of DENV-2 VLPs and NS1 was determined using western blot and ELISA using MHIAF.…”

Downstream processing and chromatography based analytical methods for production of vaccines, gene therapy vectors, and bacteriophages

“…In some cases the concentration step is placed also after the chromatography based purification step[10].Chromatography part of the DSP can consist of one or several steps, using different chromatography modes/chemistries depending on purity requirements of the final product and the type of nanoparticle applied. Purification with chromatography methods can be either based on size (size exclusion chromatography; SEC)-usually used as a polishing step in the virus DSP, hydrophobicity (hydrophobic interaction chromatography, HIC), (pseudo) affinity(30,31,32,33] or charge (anion/cation exchange chromatography).The most widely used chromatography mode for the capture step of industrial scale virus DSP is charge specific separation where viruses are separated from impurities on the basis of the difference in their surface charge distribution. Since majority of viruses have isoelectric points below 6, they efficiently bind to anion exchange matrixes at neutral pH.…”