Highly Sensitive Method for Titration of Adenovirus Vectors
Clinical development of vaccines based on adenovirus (Ad) vectors requires accurate techniques to determine vector doses including contents of infectious particles. For vectors derived from Ad virus of human serotype 5 content of infectious particles can readily be determined by plaque assays. Vaccine vectors based on alternative Ad serotypes such as those derived from chimpanzees or so-called rare serotype plaque poorly and titration by plaque assays underestimates the content of infectious particle by 50-100 fold. Here we describe a simple technique that was initially developed for titration of HAdV-5 vectors and that we modified for titration of Ad vectors from alternative serotypes.
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This is a list of supplementary files associated with this preprint. Click to download.
Table Primers Primers All primers used in this protocol are shown in below table.
Component DNA Table Template RNA Table and Concentration
Component RNA Table Template RNA Table and Concentration
Adenovirus Table Ad vector and PCR reaction Table
Table 1 Troubleshooting
Table 3 Virus titers of Ad-EGFP determined by hexon staining and EGFP visualization
Table 2 Virus titers determined by hexon staining and nested PCR
Posted 13 Oct, 2011
Highly Sensitive Method for Titration of Adenovirus Vectors
Posted 13 Oct, 2011
Clinical development of vaccines based on adenovirus (Ad) vectors requires accurate techniques to determine vector doses including contents of infectious particles. For vectors derived from Ad virus of human serotype 5 content of infectious particles can readily be determined by plaque assays. Vaccine vectors based on alternative Ad serotypes such as those derived from chimpanzees or so-called rare serotype plaque poorly and titration by plaque assays underestimates the content of infectious particle by 50-100 fold. Here we describe a simple technique that was initially developed for titration of HAdV-5 vectors and that we modified for titration of Ad vectors from alternative serotypes.
Figure 1
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