Flow-Through Purification of Viruses- A Novel Approach to Vaccine Purification

Interference chromatography: a novel approach to optimizing chromatographic selectivity and separation performance for virus purification

“…Therefore, the ability of this new technology chromatography membrane to bind virus at resin capacity at membrane flow rates and using interference multivalent ions was evaluated and exploited for a one-step NDV purification and at constant pH of 8.2. The interference chromatography approach creates interference in the sample (for this study, it the interactions were mainly charge shielding and van der Waals forces, similar to methodologies used previously [52,53] and the equilibration buffer in order to influence separation between biomolecules in the matrix, i.e. product-related and process-related molecules.…”

“…This interference method allows for improvements in performance of any chromatographic approach, such as superior resolution, improved binding capacity or augmented flow through productivity. Variations of this technique with anion exchange resins or membranes have been successfully used in the past for virus purification of Influenza H1N1 virus [52] and Influenza A/B virus [53]. One study showed that anions from triprotic kosmostropes, phosphate and citrate, with anion-exchange resins having quaternary amine or primary amine functional groups showed good virus binding, with improved binding by the salt-tolerant primary amine in phosphate buffer at Fig.…”

“…Means ±SD are shown pH 8.0 [52]. The other study used a continuous twocolumn approach with the first column of big bead AEX resin and the second one with a primary amine membrane adsorber with multivalent ions and both chromatography steps run in the negative-binding mode (also known as Flow Through Chromatography) to purify Influenza virus [53]. The study demonstrated the reduction of host DNA on the membrane was very impressive with complete recovery of virus [53].…”

“…Establishment of microenvironments, somewhat like theoretical plates, can effectively separate the target molecule from a complex milieu. Depending on the chromatographic mode, the target molecule can flow through the media while impurities bind (negative mode [53];) or the target molecule is bound to the chromatographic support and the impurities pass through. The latter option, often referred to as bind-and-elute mode ( [52]; and the current study), then requires an elution step to remove the target molecule from the solid phase.…”

Next generation vaccines and vectors: Designing downstream processes for recombinant protein‐based virus‐like particles

“…The same method was used for intermediate purification of HIV gag‐VLPs [51]. In addition, there is also a tendency to use anion exchange membrane adsorbers for flow‐through purification of large biomolecules [117, 118]. Flow‐through chromatography is especially suitable for the purification of lipid‐enveloped and less stable VLPs because the risk of product alterations by binding and elution is reduced.…”