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Method Article
Hildegund Ertl
Ertl Lab, Wistar Institute
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
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.
Keywords
adenoviral vectors
Figure 1
Figure 2
Figure 3
The authors have no conflicts of interest to disclose.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.doc
Table Primers Primers All primers used in this protocol are shown in below table.
- supplement0.doc
Component DNA Table Template RNA Table and Concentration
- supplement0.doc
Component RNA Table Template RNA Table and Concentration
- supplement0.doc
Adenovirus Table Ad vector and PCR reaction Table
- supplement0.doc
Table 1 Troubleshooting
- supplement0.doc
Table 3 Virus titers of Ad-EGFP determined by hexon staining and EGFP visualization
- supplement0.doc
Table 2 Virus titers determined by hexon staining and nested PCR
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Version History
CURRENT STATUS: Posted
Version 1
Posted 13 Oct, 2011
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Subject Areas
Microbiology
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Hildegund Ertl
Ertl Lab, Wistar Institute
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 13 Oct, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Microbiology
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
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
Figure 2
Figure 3
The authors have no conflicts of interest to disclose.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.doc
Table Primers Primers All primers used in this protocol are shown in below table.
- supplement0.doc
Component DNA Table Template RNA Table and Concentration
- supplement0.doc
Component RNA Table Template RNA Table and Concentration
- supplement0.doc
Adenovirus Table Ad vector and PCR reaction Table
- supplement0.doc
Table 1 Troubleshooting
- supplement0.doc
Table 3 Virus titers of Ad-EGFP determined by hexon staining and EGFP visualization
- supplement0.doc
Table 2 Virus titers determined by hexon staining and nested PCR
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