当前位置: X-MOL 学术Biochem. Biophys. Res. Commun. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
qDNase assay: A quantitative method for real-time assessment of DNase activity on coated surfaces
Biochemical and Biophysical Research Communications ( IF 2.5 ) Pub Date : 2020-10-26 , DOI: 10.1016/j.bbrc.2020.10.050
Marie Van der Gucht , Merve Kübra Aktan , Hanne Hendrix , Greetje Vande Velde , Jan Paeshuyse , Annabel Braem , Rob Lavigne

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.



中文翻译:

qDNase分析:实时评估涂层表面DNase活性的定量方法

DNase涂层在医疗植入物,食品和海洋工业的各种应用中显示出防止生物膜形成的巨大潜力。但是,目前尚无直接和定量的方法来表征这些涂层的酶活性。我们在这里介绍qDNase分析,这是一种定量,实时的方法来表征DNase涂层的活性。该测定法结合了(1)使用寡核苷酸探针的作用,该探针在被被包被的DNase裂解后会发出荧光,并且(2)在微孔板荧光计中连续读出荧光信号。这两个属性的组合产生了实时荧光信号,该信号用于直接定量DNase涂层的活性。作为概念证明,-4 U.据我们所知,在qDNase试验提供了第一种方法来报告绝对DNase活性单位DNA酶涂层的活性。该测定法不仅可以更准确地比较现有的DNase涂覆方法,而且还可以在将来合理设计新的DNase涂覆方法。

更新日期:2020-10-30
down
wechat
bug