qDNase assay: a quantitative method for real-time assessment of DNase activity on coated surfaces
Van der Gucht, M.; Aktan, M.K.; Hendrix, H.; Vande Velde, G.; Paeshuyse, J.; Braem, A.; Lavigne, R. (2021). qDNase assay: a quantitative method for real-time assessment of DNase activity on coated surfaces. Biochem. Biophys. Res. Commun. 534: 1003-1006. https://hdl.handle.net/10.1016/j.bbrc.2020.10.050 In: Biochemical and Biophysical Research Communications. ACADEMIC PRESS INC ELSEVIER SCIENCE: San Diego etc.. ISSN 0006-291X; e-ISSN 1090-2104, more |  |
| Author keywords | Anti-biofilm; Coatings; Quantitative enzyme assay; DNase; Fluorescence |
| Authors | | Top | - Van der Gucht, M., more
- Aktan, M.K.
- Hendrix, H., more
- Vande Velde, G.
| - Paeshuyse, J.
- Braem, A.
- Lavigne, R., more
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| Abstract | DNase coatings show great potential to prevent biofilm formation in various applications of the medical implant, food and marine industry. However, straightforward and quantitative methods to characterize the enzymatic activity of these coatings are currently not available. We here introduce the qDNase assay, a quantitative, real-time method to characterize the activity of DNase coatings. The assay combines (1) the use of an oligonucleotide probe, which fluoresces upon cleavage by coated DNases, and (2) the continuous read-out of the fluorescent signal within a microplate fluorometer format. The combination of these two properties results in a real-time fluorescent signal that is used to directly quantify the activity of DNase coatings. As a proof of concept, bovine DNase I coatings were immobilized on titanium by means of chemical grafting and their activity was estimated at 3.87 × 10−4 U. To our knowledge, the qDNase assay provides the first approach to report the activity of a DNase coating in absolute DNase activity units. This assay will not only serve to compare existing DNase coating methods more accurately, but will also enable the rational design of new DNase coating methods in the future. |
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