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Quantitative electrochemiluminescence detection of proteins: Avidin-based sensor and tris(2,2'-bipyridine) ruthenium(II) label.
Biosens Bioelectron. 2008 Jun 15; 23(11):1645-51.BB

Abstract

Quantitative electrochemiluminescence (ECL) detection of a model protein, bovine serum albumin (BSA) was achieved via biotin-avidin interaction using an avidin-based sensor and a well-developed ECL system of tris(2,2'-bipyridine) ruthenium(II) derivative as label and tri-n-propylamine (TPA) as coreactant. To detect the protein, avidin was linked to the glassy carbon electrode through passive adsorptions and covalent interaction with carboxylate-terminated carbon nanotubes that was used as binder to immobilize avidin onto the electrode. Then, biotinylated BSA tagged with tris(2,2'-bipyridine) ruthenium(II) label was attached to the prepared avidin surface. After binding of BSA labeled with tris(2,2'-bipyridine) ruthenium(II) derivative to the surface-immobilized avidin through biotin, ECL response was generated when the self-assembled modified electrode was immersed in a TPA-containing electrolyte solution. Such double protein labeling protocol with a biotin label for biorecognition and ruthenium label for ECL detection facilitated the detection of protein compared to the classical double antibody sandwich format. The ECL intensity was linearly proportional to the feed concentration of BSA over two orders of magnitude in the range of 15nM to 7.5microM. The detection limit was estimated to be 1.5nM. Further application to the lysozyme analysis was carried out to validate the present approach for an effective and favorable protocol for the quantitative detection of proteins. The dynamic range of lysozyme was from 0.001gL(-1) to 0.1gL(-1) and the detection limit was 0.1mgL(-1). Electrochemical impedance and cyclic voltammetric measurements along with some necessary control experiments were conducted to characterize the successful formation of self-assembled modified electrodes and to grant the whole detection process.

Authors+Show Affiliations

Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Changchun 130022, People's Republic of China.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18337079

Citation

Fang, Lanyun, et al. "Quantitative Electrochemiluminescence Detection of Proteins: Avidin-based Sensor and Tris(2,2'-bipyridine) ruthenium(II) Label." Biosensors & Bioelectronics, vol. 23, no. 11, 2008, pp. 1645-51.
Fang L, Lü Z, Wei H, et al. Quantitative electrochemiluminescence detection of proteins: Avidin-based sensor and tris(2,2'-bipyridine) ruthenium(II) label. Biosens Bioelectron. 2008;23(11):1645-51.
Fang, L., Lü, Z., Wei, H., & Wang, E. (2008). Quantitative electrochemiluminescence detection of proteins: Avidin-based sensor and tris(2,2'-bipyridine) ruthenium(II) label. Biosensors & Bioelectronics, 23(11), 1645-51. https://doi.org/10.1016/j.bios.2008.01.023
Fang L, et al. Quantitative Electrochemiluminescence Detection of Proteins: Avidin-based Sensor and Tris(2,2'-bipyridine) ruthenium(II) Label. Biosens Bioelectron. 2008 Jun 15;23(11):1645-51. PubMed PMID: 18337079.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitative electrochemiluminescence detection of proteins: Avidin-based sensor and tris(2,2'-bipyridine) ruthenium(II) label. AU - Fang,Lanyun, AU - Lü,Zhaozi, AU - Wei,Hui, AU - Wang,Erkang, Y1 - 2008/02/07/ PY - 2007/09/12/received PY - 2007/12/19/revised PY - 2008/01/29/accepted PY - 2008/3/14/pubmed PY - 2008/8/20/medline PY - 2008/3/14/entrez SP - 1645 EP - 51 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 23 IS - 11 N2 - Quantitative electrochemiluminescence (ECL) detection of a model protein, bovine serum albumin (BSA) was achieved via biotin-avidin interaction using an avidin-based sensor and a well-developed ECL system of tris(2,2'-bipyridine) ruthenium(II) derivative as label and tri-n-propylamine (TPA) as coreactant. To detect the protein, avidin was linked to the glassy carbon electrode through passive adsorptions and covalent interaction with carboxylate-terminated carbon nanotubes that was used as binder to immobilize avidin onto the electrode. Then, biotinylated BSA tagged with tris(2,2'-bipyridine) ruthenium(II) label was attached to the prepared avidin surface. After binding of BSA labeled with tris(2,2'-bipyridine) ruthenium(II) derivative to the surface-immobilized avidin through biotin, ECL response was generated when the self-assembled modified electrode was immersed in a TPA-containing electrolyte solution. Such double protein labeling protocol with a biotin label for biorecognition and ruthenium label for ECL detection facilitated the detection of protein compared to the classical double antibody sandwich format. The ECL intensity was linearly proportional to the feed concentration of BSA over two orders of magnitude in the range of 15nM to 7.5microM. The detection limit was estimated to be 1.5nM. Further application to the lysozyme analysis was carried out to validate the present approach for an effective and favorable protocol for the quantitative detection of proteins. The dynamic range of lysozyme was from 0.001gL(-1) to 0.1gL(-1) and the detection limit was 0.1mgL(-1). Electrochemical impedance and cyclic voltammetric measurements along with some necessary control experiments were conducted to characterize the successful formation of self-assembled modified electrodes and to grant the whole detection process. SN - 0956-5663 UR - https://www.unboundmedicine.com/medline/citation/18337079/Quantitative_electrochemiluminescence_detection_of_proteins:_Avidin_based_sensor_and_tris_22'_bipyridine__ruthenium_II__label_ DB - PRIME DP - Unbound Medicine ER -