Abstract
Although ceramic hydroxyapatite (HAp) chromatography has been used as an alternative method ultracentrifugation for the production of vaccines, the mechanism of virus separation is still obscure. In order to begin to understand the mechanisms of virus separation, HAp surfaces were observed by scanning electron microscopy after chromatography with dengue viruses. When these processes were performed without elution and with a 10–207 mM sodium phosphate buffer gradient elution, dengue viruses that were adsorbed to HAp were disproportionately located in the columns. However, when eluted with a 10–600 mM sodium phosphate buffer gradient, few viruses were observed on the HAp surface. After incubating the dengue viruses that were adsorbed on HAp beads at 37°C and 2°C, the sphericity of the dengue viruses were reduced with an increase in incubation temperature. These results suggested that dengue virus was adsorbed to the HAp surface by electronic interactions and could be eluted by high-salt concentration buffers, which are commonly used in protein purification. Furthermore, virus fusion was thought to occur with increasing temperature, which implied that virus-HAp adhesion was similar to virus-cell adhesion.
Highlights
The methodology of density-gradient ultracentrifugation, which has traditionally been applied in the purification of viruses, is able to achieve satisfactory results in vaccine production
HAp Chromatography and HA Test Dengue viruses were adsorbed to HAp particles in Columns A
The consequence of HA units in the fractions and the observation of viruses in the corresponding scanning electron microscope (SEM) images visually confirmed that dengue viruses could be eluted by high-salt concentrations with HAp chromatography, which is consistent with the results reported by other researchers [7,8,9]
Summary
The methodology of density-gradient ultracentrifugation, which has traditionally been applied in the purification of viruses, is able to achieve satisfactory results in vaccine production. Hydroxyapatite (HAp) chromatography is another substitute methodology for ultracentrifugation. HAp chromatography is thought to enable a variety of viruses to be purified. HAp has achieved satisfactory results in large-scale protein purifications. Virus purification by HAp chromatography might be suitable for industry usage. The mechanisms underlying virus adsorption to HAp are still obscure. Reports that have demonstrated favorable isolations of viruses have been published previously, and good results were obtained with high-salt eluting as well as with protein isolation [7,8,9]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
