Millipore Technical Publications |
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A New Vacuum-Based Method for Adenovirus Purification and Concentration |
| Adenovirus vectors have traditionally been purified away from cellular contaminants, expressed recombinant transgenes, and cell culture media serum proteins by cesium chloride or other similar density gradient based methods. However, these traditional methods present several disadvantages to the researcher. Density gradient techniques are lengthy, often requiring several days to complete. The process requires the use of ultracentrifuges which are expensive and not common equipment for the average laboratory. The technique for harvesting adenovirus bands from the density gradient after ultracentrifugation can also be cumbersome for many researchers. We report here an improved method for the rapid purification and concentration of adenovirus serotype 5. This membrane-based method for purifying adenovirus is different from other similar membrane-based methods as it uses a much easier and safer vacuum-based device. Performance in terms of processing time, recovery, purity, and capacity is as good as or better than similar products currently available from other manufactures. The entire procedure including clarification of crude adenovirus, purification of adenovirus, buffer exchange, and concentration can be accomplished in under one hour and results in a concentrated, high titer, pure adenovirus in the buffer of choice. Click the PDF above for the full poster. |
