Springer Protocols: Abstract: 3. Quantifying the Titer and Quality of Adenovirus Stocks

3. Quantifying the Titer and Quality of Adenovirus Stocks

By: Fubao Wang³, Bill C. Mathis³, Allison Montalvo³, Jayanthi J. Wolf³, John A. Lewis³

Abstract

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The broad application of recombinant adenoviruses to the development of vaccines and gene therapy vectors has encouraged the development of molecular assays for the facile quantitation of adenoviral particles and the assignment of their infectious potency. The Genome Quantitation Assay (GQA) and the QPCR-Based Potency Assay (QPA) developed for adenoviruses offer the attributes of precision, rapidity, and high throughput either performed manually or facilitated by simple automated liquid handling systems. These assay attributes allow for accelerated process development support and product characterization and release. The assays for adenovirus could offer the additional advantage in that their quantitation is based on viral replication independent of cytopathology permitting quantitation of serotypes that cause minimal cytopathic effect (CPE) in 293 cells and specificity that allows the components of multivalent vaccines to be discriminated and quantitated for release.

Affiliation(s): (3) Vaccine and Biologics Research, Merck Research Laboratories, Merck & Co., Inc., West Point, PA

Book Title: Gene Therapy Protocols: Design and Characterization of Gene Transfer Vectors

Series: Methods in Molecular Biology | Volume: 434 | Pub. Date: Mar-07-2008 | Page Range: 25-36 | DOI: 10.1007/978-1-60327-248-3_3

Subject: Genetics/Genomics

Key Words: Adenovirus - GQA - QPA - adenoviral genome - infectious potency - QPCR

References

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