title: A novel electrochemiluminescence aptasensor for protein based on a sensitive N-(aminobutyl)-N-ethylisoluminol-functionalized gold nanoprobe.
abstract: A novel electrochemiluminescence (ECL) aptasensor for platelet-derived growth factor B chain (PDGF-BB) assay was developed by assembling N-(aminobutyl)-N-ethylisoluminol functionalized gold nanoparticles (ABEI-AuNPs) with aptamers as nanoprobes. In the protocol, the biotinylated aptamer capture probes were first immobilized on a streptavidin coated gold nanoparticle (AuNPs) modified electrode, afterwards, the target PDGF-BB and the ABEI-AuNPs tagged aptamer signal probe were successively attached to the modified electrode by virtue of the dimer structure of PDGF-BB to fabricate a "sandwich" conjugate modified electrode, i.e. an aptasensor. ECL measurement was carried out with a double-step potential in carbonate buffer solution containing H(2)O(2). The aptasensor showed high sensitivity and selectivity toward PDGF-BB and specificity toward PDGF-BB aptamer. The detection limit was as low as 2.7 × 10(-14) M. In this work, the ABEI-AuNPs synthesized by a simple seed growth method have been successfully used as aptamer labels, which greatly amplified the ECL signal by binding numbers of ABEI molecules on the surface of AuNPs. The ABEI-AuNPs signal amplification is superior to other reported signal amplification strategies based on aptamer-related polymerase chain reaction or functionalized nanoparticles in simplicity, stability, labeling property and practical applicability. And the ABEI-AuNPs based nanoprobe is more sensitive than the luminol functionalized AuNPs based nanoprobe. Moreover, such an ultra-sensitive and low-cost assay can be accomplished with a simple and fast procedure by using a simple ECL instrumentation. The aptasensor was also applied for the detection of PDGF-BB in human serum samples, showing great application potential. Given these advantages, the ECL aptasensor is well suited for the direct, sensitive and rapid detection of protein in complex clinical samples.
authors.affiliation: CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science & Technology of China, Hefei, Anhui 230026, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chai
authors.forename: Dayong
authors.initials: D
authors.lastname: Tian
authors.forename: Jie
authors.initials: J
authors.lastname: Gu
authors.forename: Hua
authors.initials: H
authors.lastname: Cui
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D000077214:Becaplermin; D055664:Electrochemical Techniques / Q000379:methods; D004566:Electrodes; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008165:Luminol / Q000031:analogs & derivatives / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry*; D010982:Platelet-Derived Growth Factor / Q000032:analysis*; D020574:Proto-Oncogene Proteins c-sis; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism
chemical_list: D052157:Aptamers, Nucleotide; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D000077214:Becaplermin; D008165:Luminol; C031912:6-(N-(4-aminobutyl)-N-ethyl)amino-2,3-dihydrophthalazine-1,4-dione; D006046:Gold; D019809:Streptavidin