Description
• Achieve the quality of CsCl procedures in <2 hours, without ultracentrifugation
• Recover >90% adenoviral yield
Purification of viruses via cesium chloride ultracentrifugation procedures can be tedious and time-consuming. ViraBind Adenovirus Purification Kits provide a much more efficient system for fast adenoviral purification with high yields. The ViraBind Adenovirus Miniprep Kit uses a special spin column to purify supernatant from a single T75 flask or 10cm dish per prep in about 30 minutes.
Introduction: Recombinant adenoviruses have tremendous potential in both research and therapeutic applications. There are numerous advantages in using an adenovirus to introduce genetic material into host cells. The permissive host cell range is very wide. The virus has been used to infect many mammalian cell types (both replicative and non-replicative) for high expression of the recombinant protein. Recombinant adenoviruses are especially useful for gene transfer and protein expression in cell lines that have low transfection efficiency with liposome. After entering cells, the virus remains epichromosomal (i.e. does not integrate into the host chromosome so does not activate or inactivate host genes). Recently, recombinant adenoviruses have been used to deliver RNAi into cells.
HEK 293 cells or their variants are used as host cells for viral amplification. Recombinant adenoviruses can be grown at high titer (1010 VP (viral particles)/mL, which can be concentrated up to 1013 VP/mL). The concentrated viral supernatant is subjected to CsCl ultracentrifugation to separate the viruses from the cellular proteins and media components. Following ultracentrifugation, CsCl is then removed by dialysis. The CsCl procedure is both tedious and time consuming (16-24 hrs). ViraBind Adenoviral Miniprep Kit does not involve ultracentrifugation, instead viruses are captured on spin column based on the unique properties of adenoviral capsid proteins. The entire procedure takes about 30 minutes. Each column is designed to purify viruses harvested from one T75 flask or 10-cm plate, and has a capacity of up to 1.0 x 1011 VPs.
ViraBind Adenovirus Miniprep Kit provides an efficient system for quick adenoviral purification with high recovery (>95%). The system may be adapted to purification of other viral types, such as retrovirus and lentivirus