Analysis of Recombinant Adenoviruses Using an Integrated Microfluidic Chip-Based System

✍ Scribed by Michael T. McCaman; Pete Murakami; Erno Pungor Jr.; Karen M. Hahnenberger; William S. Hancock

• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 139 KB
• Volume: 291
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2001.5043

✦ Synopsis

The use of recombinant virus for gene therapy requires rigorous quality control methods to ensure that the viral vector preparations are functional and safe. A viral identity test is performed in which the viral payload, or transgene, is PCR amplified, followed by digestion with restriction enzymes that yields a characteristic "fingerprint." These DNA fragments are typically analyzed by agarose gel electrophoresis. The ethidium bromide-stained gels are photographed or scanned and the results are sufficient for a qualitative or semiquantitative identity confirmation of the viral product. We have investigated the use of an integrated microfluidic chip-based system as a new tool in the quality control testing of a recombinant, adenoviral, gene therapy product. The chip-based method was found to be very sensitive, requiring 100-fold less sample and only one-third the time compared to the agarose gel method. The automated data analysis sizes and quantitates the DNA fragments, thus yielding a more thorough, reproducible, sensitive, and rapid analysis.