QuickTiter™ Adenovirus Quantitation Kit - 20 assays
Recombinant adenoviruses have tremendous potential in both research and therapeutic applications. There are numerous advantages they provide when introducing 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 (10^10 VP (viral particles)/mL, which can be concentrated up to 10^13 VP/mL) and purified by Cell Biolabs ViraBind™ Adenoviral Purification Kit or traditional CsCl ultracentrifugation.
A particular challenge in the delivery of a gene by a viral vector is the accurate measurement of virus titer. Traditionally, infectivity particles are measured in culture by a plaque-forming unit assay (PFU) that scores the number of viral plaques as a function of dilution. Others utilize antibodies that recognize adenovirus hexon proteins by immunohistochemistry staining or FACS analysis. These methods are time-consuming, require a long infection period, and suffer from a high degree of interassay variability and are affected by virus-cell interactions. For highly purified virus samples, an optical absorbance at 260 nm has been used to estimate the total number of virus particles. However this method can not be used in an unpurified viral supernatant, because some of the components it contains can contributes to the optical absorbance of 260 nm.
The QuickTiter™ Adenoviral Quantitation Kit does not involve cell infection; instead it specifically measures the viral nucleic acid content of purified viruses or unpurified viral supernatant sample (See Test Principle). Especially for unpurified viral supernatant, the kit is very useful for determining the supernatant titer before the purification step. The kit has detection sensitivity limit of 1 X 10^9 VP or 1 X 10^10 VP/mL when 100 µL of adenoviral supernatant is used in the assay, which is sufficient for mid or high-titer adenovirus sample. The entire procedure takes about 45 to 60 minutes. Each kit provides sufficient quantities to perform up to 20 tests for viral samples and controls.
QuickTiter™ Adenoviral Quantitation Kit provides an efficient system for rapid quantitation of
adenovirus titer for both viral supernatant and purified virus.