Enzyme-linked electrochemical DNA ligation assay using magnetic beads

0431757 - BFÚ 2015 RIV DE eng J - Článek v odborném periodiku
Stejskalová, Eva - Horáková Brázdilová, Petra - Vacek, J. - Bowater, R. P. - Fojta, Miroslav
Enzyme-linked electrochemical DNA ligation assay using magnetic beads.
Analytical and Bioanalytical Chemistry. Roč. 406, č. 17 (2014), s. 4129-4136. ISSN 1618-2642. E-ISSN 1618-2650
Grant CEP: GA ČR(CZ) GPP206/11/P739; GA ČR(CZ) GAP206/11/1638; GA AV ČR(CZ) IAA400040901
Institucionální podpora: RVO:68081707
Klíčová slova: Electrochemistry * Enzyme labeling * DNA ligase
Kód oboru RIV: BO - Biofyzika
Impakt faktor: 3.436, rok: 2014

DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5' biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3' digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode.
Trvalý link: http://hdl.handle.net/11104/0236329