Electrochemical biosensor modified with dsDNA monolayer for restriction enzyme activity determination
Joanna Zajda , Łukasz Górski , Elżbieta Malinowska
AbstractA simple and cost effective method for the determination of restriction endonuclease activity is presented. dsDNA immobilized at a gold electrode surface is used as the enzymatic substrate, and an external cationic redox probe is employed in voltammetric measurements for analytical signal generation. The assessment of enzyme activity is based on a decrease of a current signal derived from reduction of methylene blue which is present in the sample solution. For this reason, the covalent attachment of the label molecule is not required which significantly reduces costs of the analysis and simplifies the entire determination procedure. The influence of buffer components on utilized dsDNA/MCH monolayer stability and integrity is also verified. Electrochemical impedance spectroscopy measurements reveal that due to pinhole formation during enzyme activity measurement the presence of any surfactants should be avoided. Additionally, it is shown that the sensitivity of the electrochemical biosensor can be tuned by changing the restriction site location along the DNA length. Under optimal conditions the proposed biosensor exhibits a linear response toward PvuII activity within a range from 0.25 to 1.50 U/μL.
|Journal series||Bioelectrochemistry, ISSN 1567-5394|
|Publication size in sheets||0.5|
|Keywords in English||DNA-based biosensor; Electrochemical sensor; Restriction endonuclease activity determination|
|ASJC Classification||; ; ;|
|Score|| = 35.0, 31-01-2020, ArticleFromJournal|
= 35.0, 31-01-2020, ArticleFromJournal
|Publication indicators||= 2; = 2; : 2016 = 1.014; : 2016 = 3.346 (2) - 2016=3.413 (5)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.