Downstream processing of virus, virus-like particles and nanoparticulate inclusion bodies to be used as gene delivery vehicles for human gene therapy applications

Design and analyze future affinity ligands such as antibodies, aptamers, metal affinity, peptides, triazine dyes, and red blood cell carriers to increase monolithic chromatography selectivity and two-phase affinity-aqueous systems. Monolithic chromatography systems may be constructed and optimized by examining viruses, virus-like particles, and nanoparticulate inclusion bodies from difficult culture media. Different chromatographic characteristics such as monolithic supports, active monolith groups, binding and elution buffers, elution gradients, column equilibrium volumes, and regeneration should be investigated with the aim of optimizing monolithic chromatography separation and recovery. To develop and optimize aqueous two-phase systems, several different parameters need to be investigated and compared, such as polymer type and concentration, salt solution, surfactants and ions, polymer molecular weight, ion nature and ionic strength, present affinity ligands, volumetric ratio, temperature, pH, and sample load, to name a few.Several relevant biological models, such as Newcastle disease (NDV), herpes (HPr), or viral vectors bluetongue (BTV), can be employed to establish different patterns and predict the generalized mechanistic process of enhanced purification systems. Because optimal systems are intended to be deployed at the industrial level, a comparison of typical downstream processing and alternatives in terms of recovery, technological advantages, economic feasibility, and environmental sustainability is needed. Cell separation, viral concentration and viral purification may all be reduced to one step utilizing integrated chromatography platforms on monolith supports and aqueous two-phase systems. In addition, the efficiency of separation and purification yields achieved after these integrated systems are predicted to be substantially higher than those produced in conventional procedures. As a result, the benefits of these alternative and cutting-edge technologies are predicted to contribute to more efficient, long-term, and cost-effective downstream processing of virus-like particles.