title: Novel sensory surface for creatine kinase electrochemical detection.
abstract: This work describes a novel concept of biosensor for quantifying enzymes, where the substrate is immobilized directly over the working area of a screen printed electrode (Au-SPE). This concept is applied here to creatine kinase isoenzyme (CK-MB), a cardiac biomarker in ischemic conditions. It acts as a phospho-transferase on creatine (Crea), requiring the presence of phosphate. So, the phosphorylated form of creatine (Pcrea) was immobilized on the Au/SPE previously aminated with cysteamine (Cys) by self-assembling monolayer technique. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) studies were used to follow the chemical modifications in the Au-SPE. Since Pcrea is an electroactive species at low potential, its consumption over the platform by the enzyme changed the electrical response of the biosensor. So, CK-MB determination has been achieved in mediator free-conditions due the redox proprieties of the Pcrea. The analytical features of the resulting biosensor were studied by square wave voltammetry (SWV). The limit of detection was 0.11 µg/mL and the slope was -0.029(± 0.0035) µA × mL/µg. The interference effect of troponin T (TnT), bovine serum albumin (BSA) and myoglobin (Myo) in the performance of the sensor was tested and good selectivity was observed. The biosensor was successfully applied to biological fluids, showing good stability at room temperature and excellent sensitivity and selectivity. This new concept of biosensor is especially useful for point of care (POC) applications, due to the low cost and small size of the final device.
authors: BioMark/ISEP, Instituto Superior de Engenharia do Porto, Porto, Portugal; UOSE, INESC-Porto, Faculdade Ciências/Universidade Porto, Porto, Portugal; REQUIMTE/FCT-UNL, Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa, Portugal.
authors: Felismina T C
authors: FT
authors: Moreira
authors: PROCAPE, LAPED, Universidade Federal de Pernambuco, Recife, Brazil.
authors: Rosa A F
authors: RA
authors: Dutra
authors: REQUIMTE/FCT-UNL, Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa, Portugal.
authors: João P
authors: JP
authors: Noronha
authors: BioMark/ISEP, Instituto Superior de Engenharia do Porto, Porto, Portugal. Electronic address: goreti.sales@gmail.com.
authors: M Goreti F
authors: MG
authors: Sales
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D002417:Cattle; D003401:Creatine / Q000378:metabolism; D052279:Creatine Kinase, MB Form / Q000378:metabolism* / Q000652:urine*; D058266:Dielectric Spectroscopy; D055664:Electrochemical Techniques / Q000295:instrumentation*; D004566:Electrodes; D057075:Enzyme Assays / Q000295:instrumentation; D004867:Equipment Design; D006801:Humans; D057230:Limit of Detection; D009206:Myocardium / Q000201:enzymology; D010725:Phosphocreatine / Q000378:metabolism
chemical_list: D010725:Phosphocreatine; D052279:Creatine Kinase, MB Form; D003401:Creatine
keywords: CK-MB; Creatine kinase isoenzyme; Screen-printed electrodes; Self-assembled monolayer; Senso