Purification of Vero cell derived live replication deficient influenza A and B virus by ion exchange monolith chromatography

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

“…Superiority of monolithic columns in this respect was previously shown for different nanoparticles [6,10,11] A very important part of a nanoparticle production represents the set-up of appropriate analytical methods for downstream and upstream monitoring of the process and for the final product analysis. Reproducible DSP and final products are possible only when appropriate analytical assays are in place.There iscurrently a variety of analytical methods available for these purposes, but due to their relatively high variability and different information that they provide usually a combination of several are needed to get reliable and informative results.…”

“…The second step in the DSP scheme can incorporate a concentration step which is very important especially for low titer virus productions and can be performed with precipitation Some viruses bind to both anion and cation exchange matrixes at neutral pH [11], although their isoelectric point is below 6. This phenomenon can be explained by the fact that viruses are large macromolecular assemblies with non-uniform surface charge distribution.…”

“…New generation chromatography media, CIM monolithic columns were used in DSPs of different viruses like influenza [11,13,37] adenovirus [8,29], adeno-associated virus [9,38], lentivirus [10,39], rubella [40], different phages [26,27,28,41,42] and different VLPs [6,43] with good virus/phage/VLP recoveries while reaching high dynamic binding capacities at the same time.…”

“…Due to the large void volume of membranes compared to monoliths, the resolution, regardless of the flow rate used, is superior with monoliths. This advantage of monoliths is especially important and has already been recognized in different DSP steps of nanoparticles [6,8,9,10,11]. Excellent resolution of monoliths was demonstrated by Lock's group [9] who developed a method for separation of two very similar viral particles (viral capsids with and without genetic material) using CIM monolithic column and a very shallow linear gradient.…”

Methacrylate monolith chromatography as a tool for waterborne virus removal

“…Highly porous material with interconnected flowthrough channels (average diameter of 1.5, 2 or even 6 m) enables convective mass transport of the molecules (in contrast to the diffusive transport of bead based supports), leading to flow independent dynamic binding capacity and separation [18,19]. Anion exchange monoliths (QA and DEAE) have proven to be very effective for fast separation or concentration of different viruses such as tomato mosaic virus [20,21], potato virus Y [22], orthoreovirus [23], rotaviruses [24,25], hepatitis A virus and caliciviruses [26], rubella virus [27], influenza virus [28], adenovirus [29], adeno-associated virus [30], lenti virus [31] bacteriophages [32][33][34] and virus-like particles [35,36]. They have also been successfully applied to the concentration of waterborne viruses [25,26] where fast flow rates and large binding capacity are essential.…”