Analysis of recombinant adenoviruses using an integrated microfluidic chip-based system

The use of recombinant virus for gene therapy requires rigorous quality con trol methods to ensure that the viral vector preparations are functional an d safe. A viral identity test is performed in which the viral payload, or t ransgene, is PCR amplified, followed by digestion with restriction enzymes that yields a characteristic "fingerprint." These DNA fragments are typical ly analyzed by agarose gel electrophoresis. The ethidium bromide-stained ge ls are photographed or scanned and the results are sufficient for a qualita tive or semiquantitative identity confirmation of the viral product. We hav e 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, req uiring 100-fold less sample and only one-third the time compared to the aga rose gel method. The automated data analysis sizes and quantitates the DNA fragments, thus yielding a more thorough, reproducible, sensitive, and rapi d analysis. (C) 2001 Academic Press.