title: A conjugated aptamer-gold nanoparticle fluorescent probe for highly sensitive detection of rHuEPO-α.
abstract: We present here a novel conjugated aptamer-gold nanoparticle (Apt-AuNPs) fluorescent probe and its application for specific detection of recombinant human erythropoietin-α (rHuEPO-α). In this nanobiosensor, 12 nm AuNPs function as both a nano-scaffold and a nano-quencher (fluorescent energy acceptor), on the surface of which the complementary sequences are linked (as cODN-AuNPs) and pre-hybridized with carboxymethylfluorescein (FAM)-labeled anti-rHuEPO-α aptamers. Upon target protein binding, the aptamers can be released from the AuNP surface and the fluorescence signal is restored. Key variables such as the length of linker, the hybridization site and length have been designed and optimized. Full performance evaluation including sensitivity, linear range and interference substances are also described. This nanobiosensor provides a promising approach for a simple and direct quantification of rHuEPO-α concentrations as low as 0.92 nM within a few hours.
authors.affiliation: Beijing Institute of Pharmacology and Toxicology, No. 27, Taiping Road, Beijing 100850, China. sunjf2001@163.com
authors.forename: Jiefang
authors.initials: J
authors.lastname: Sun
authors.forename: Aitao
authors.initials: A
authors.lastname: Guo
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Zhang
authors.forename: Lei
authors.initials: L
authors.lastname: Guo
authors.forename: Jianwei
authors.initials: J
authors.lastname: Xie
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D001667:Binding, Competitive; D015374:Biosensing Techniques / Q000379:methods*; D018076:DNA, Complementary / Q000737:chemistry; D004921:Erythropoietin / Q000032:analysis* / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010316:Particle Size; D011061:Poly A / Q000737:chemistry; D011994:Recombinant Proteins / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity
chemical_list: D052157:Aptamers, Nucleotide; D018076:DNA, Complementary; C554270:EPO protein, human; D005452:Fluoresceins; D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D004921:Erythropoietin; D011061:Poly A; C015465:poly(dA); D006046:Gold