Sensors 2013, 13(9), 11586-11602; doi:10.3390/s130911586
Article

Oriented Immobilization of His-Tagged Protein on a Redox Active Thiol Derivative of DPTA-Cu(II) Layer Deposited on a Gold Electrode—The Base of Electrochemical Biosensors

1 Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, Olsztyn 10-748, Poland 2 Institute of Biochemistry and Biophysics of Polish Academy of Sciences, Pawińskiego 5a, Warsaw 02-106, Poland 3 Chemistry Department, University of Leuven, Celestijnenlaan 200F, Leuven B-3001, Belgium
* Author to whom correspondence should be addressed.
Received: 8 July 2013; in revised form: 19 August 2013 / Accepted: 27 August 2013 / Published: 3 September 2013
(This article belongs to the Section Biosensors)
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Abstract: This paper concerns the development of an electrochemical biosensor for the determination of Aβ16–23' and Aβ1–40 peptides. The His-tagged V and VC1 domains of Receptor for Advanced Glycation end Products (RAGE) immobilized on a gold electrode surface were used as analytically active molecules. The immobilization of His6–RAGE domains consists of: (i) formation of a mixed layer of N-acetylcysteamine (NAC) and the thiol derivative of pentetic acid (DPTA); (ii) complexation of Cu(II) by DPTA; (iii) oriented immobilization of His6–RAGE domains via coordination bonds between Cu(II) sites from DPTA–Cu(II) complex and imidazole nitrogen atoms of a histidine tag. Each modification step was controlled by cyclic voltammetry (CV), Osteryoung square-wave voltammetry (OSWV), and atomic force microscopy (AFM). The applicability of the proposed biosensor was tested in the presence of human plasma, which had no influence on its performance. The detection limits for Aβ1–40 determination were 1.06 nM and 0.80 nM, in the presence of buffer and human plasma, respectively. These values reach the concentration level of Aβ1–40 which is relevant for determination of its soluble form in human plasma, as well as in brain. This indicates the promising future application of biosensor presented for early diagnosis of neurodegenerative diseases.
Keywords: pentetic acid–Cu(II) complex; gold electrodes; His-tagged proteins; Aβ peptides; electrochemical biosensor

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MDPI and ACS Style

Mikuła, E.; Sulima, M.; Marszałek, I.; Wysłouch-Cieszyńska, A.; Verwilst, P.; Dehaen, W.; Radecki, J.; Radecka, H. Oriented Immobilization of His-Tagged Protein on a Redox Active Thiol Derivative of DPTA-Cu(II) Layer Deposited on a Gold Electrode—The Base of Electrochemical Biosensors. Sensors 2013, 13, 11586-11602.

AMA Style

Mikuła E, Sulima M, Marszałek I, Wysłouch-Cieszyńska A, Verwilst P, Dehaen W, Radecki J, Radecka H. Oriented Immobilization of His-Tagged Protein on a Redox Active Thiol Derivative of DPTA-Cu(II) Layer Deposited on a Gold Electrode—The Base of Electrochemical Biosensors. Sensors. 2013; 13(9):11586-11602.

Chicago/Turabian Style

Mikuła, Edyta; Sulima, Magdalena; Marszałek, Ilona; Wysłouch-Cieszyńska, Aleksandra; Verwilst, Peter; Dehaen, Wim; Radecki, Jerzy; Radecka, Hanna. 2013. "Oriented Immobilization of His-Tagged Protein on a Redox Active Thiol Derivative of DPTA-Cu(II) Layer Deposited on a Gold Electrode—The Base of Electrochemical Biosensors." Sensors 13, no. 9: 11586-11602.

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