Development and modeling of two-dimensional fast protein liquid chromatography for producing nonstructural protein-free food-and-mouth diseases virus vaccine.

摘要: Abstract Concerns for the use of non-purified or incompletely purified inactivated foot-and-mouth disease (FMD) vaccine, like difficulties differentiation vaccinated from infected animals, can be a motivation in order to develop methods based on size exclusion chromatography (SEC). In this study, two dimensional (2D-SEC) system was successfully constructed using different SEC column media achieve high-throughput purification cell culture-derived foot and mouth diseases virus (FMDV). A mathematical model also utilized predict get better insight into separation process. Column packing particles characteristics such as void volume, total particle porosity accessible acquired experimentally. Retention times elution profile molecules, blue dextran bovine serum albumin, were used evaluating capability separating critical impurities (residual DNA (rDNA) non-structural protein (NSP)) active ingredient vaccine (FMDV particle). Experiments carried out with commercial columns (XK 26/60) 16/100) four superdex 200 prep grade, sephacryl S-500 HR, Sephacryl S-400 HR S-300HR. The first validated by experimental chromatographic data then propose best 2D-SEC downstream processing FMDV vaccine. loading capacity sample increased 12.5% volume purity final product more than 90%. entire process performed 77% recovery 79.1% yield. Based high-performance (HPSEC), NSP-free about 90% over 94.6% host removed. Analyses HPSEC, transmission electron microscopy (TEM) dynamic light scattering (DLS) indicated that had spherical shape mean 30 nm their structure remained intact.