Description
• More accurate adenoviral titer than traditional plaque-forming unit assays
• Faster results: 2.5 days vs. 10 days
• No agar overlay steps
Accurate measurement of adenovirus titer is critical for gene delivery. Traditional plaque-forming unit (PFU) assays are long and suffer from high inter-assay variability. The QuickTiter Adenovirus Titer ELISA Kit provides a quick, complete system to functionally titer virus infectivity. The assay recognizes all 41 serotypes of adenovirus, and can be used with any adenovirus system that can amplify in HEK 293 cells.
Introduction: 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 (1010 VP (viral particles)/mL, which can be concentrated up to 1013 VP/mL) and purified by MyBioSource's 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. These methods are time-consuming (10 days), require a long infection period, and suffer from a high degree of inter-assay variability and are affected by virus-cell interactions. MyBioSource's QuickTiter Adenovirus Titer ELISA Kit utilizes an antibody against adenovirus hexon proteins to quantitate infected cells. The hexon proteins are the largest and most abundant of the structural proteins in the adenovirus capsid, and they are distributed symmetrically to form capsid facets.
MyBioSource's QuickTiter Adenovirus Titer ELISA Kit provides a quick and complete system to functionally titer virus infectivity; it provides sufficient reagents for up to 192 tests in 96-well plates. In contrast to the 10-day infection of a classical plaque assay, the kit only requires a 2-day infection. Detection sensitivity is 104 ifu/mL (106-107 VP/mL), which is sufficient for most adenoviral samples. The kit recognizes all 41 serotypes of adenovirus by immunocytochemistry and can be used with any adenovirus system as long as the virus is able to amplify in HEK 293 cells