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
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 molecul...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2013-09-01
|
| Series: | Sensors |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1424-8220/13/9/11586 |
| _version_ | 1797999135951945728 |
|---|---|
| author | Hanna Radecka Jerzy Radecki Wim Dehaen Peter Verwilst Ilona Marszałek Aleksandra Wysłouch-Cieszyńska Magdalena Sulima Edyta Mikuła |
| author_facet | Hanna Radecka Jerzy Radecki Wim Dehaen Peter Verwilst Ilona Marszałek Aleksandra Wysłouch-Cieszyńska Magdalena Sulima Edyta Mikuła |
| author_sort | Hanna Radecka |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T10:59:46Z |
| format | Article |
| id | doaj.art-92e7b57fa99146aea50500cd0c935cc2 |
| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-04-11T10:59:46Z |
| publishDate | 2013-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Sensors |
| spelling | doaj.art-92e7b57fa99146aea50500cd0c935cc22022-12-22T04:28:39ZengMDPI AGSensors1424-82202013-09-01139115861160210.3390/s130911586Oriented 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 BiosensorsHanna RadeckaJerzy RadeckiWim DehaenPeter VerwilstIlona MarszałekAleksandra Wysłouch-CieszyńskaMagdalena SulimaEdyta MikułaThis 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.http://www.mdpi.com/1424-8220/13/9/11586pentetic acid–Cu(II) complexgold electrodesHis-tagged proteinsAβ peptideselectrochemical biosensor |
| spellingShingle | Hanna Radecka Jerzy Radecki Wim Dehaen Peter Verwilst Ilona Marszałek Aleksandra Wysłouch-Cieszyńska Magdalena Sulima Edyta Mikuła 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 pentetic acid–Cu(II) complex gold electrodes His-tagged proteins Aβ peptides electrochemical biosensor |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | pentetic acid–Cu(II) complex gold electrodes His-tagged proteins Aβ peptides electrochemical biosensor |
| url | http://www.mdpi.com/1424-8220/13/9/11586 |
| work_keys_str_mv | AT hannaradecka orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors AT jerzyradecki orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors AT wimdehaen orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors AT peterverwilst orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors AT ilonamarszałek orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors AT aleksandrawysłouchcieszynska orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors AT magdalenasulima orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors AT edytamikuła orientedimmobilizationofhistaggedproteinonaredoxactivethiolderivativeofdptacuiilayerdepositedonagoldelectrodethebaseofelectrochemicalbiosensors |