Development of a lipase-based optical assay for detection of DNA.

• Published in: Organic & Biomolecular Chemistry
• Main Authors: Pinijsuwan, Suttiporn, Shipovskov, Stepan, Surareungchai, Werasak, Ferapontova, Elena, Gothelf, Kurt Vesterager
• Format: Article in Journal/Newspaper
• Language: English
• Published: 2011
• Subjects: Antarc*; Antarctica
• Online Access: https://pure.au.dk/portal/da/publications/development-of-a-lipasebased-optical-assay-for-detection-of-dna(0f425783-9535-4449-8550-59220f54a6f7).html; https://doi.org/10.1039/C0OB01165G

A lipase-based assay for detection of specific DNA sequences has been developed. Lipase from Candida antarctica was conjugated to DNA and captured on magnetic beads in a sandwich assay, in which the binding was dependent on the presence of a specific target DNA. For amplification and to generate a detectable readout the captured lipase was applied to an optical assay that takes advantage of the enzymatic activity of lipase. The assay applies p-nitrophenol octanoate (NPO) as the substrate and in the presence of lipase the ester is hydrolyzed to p-nitrophenolate which has a strong absorbance at 405 nm. The method provides detection a detection limit of 200 fmol target DNA and it was able to distinguish single base mismatches from the fully complementary target.