title: Robust nanoplasmonic substrates for aptamer macroarrays with single-step detection of PDGF-BB.
abstract: An aptamer macroarray on a robust nanoplasmonic substrate with fluorescence enhancement is developed for a single-step sensitive detection of human platelet-derived growth factor-BB (PDGF-BB), a predominant cancer biomarker in cancer angiogenesis. A hybrid Au-nanoparticles-poly (dimethylsiloxane) (PDMS) as nanoplasmonic substrate is prepared via the in-situ reduction of AuCl4(-) ions in PDMS matrixes onto 96 or 384 well plates. In the absence of target molecules, unfolded PDGF-BB aptamer conjugated with dye TAMRA is electrostatically bound to a positively charged poly-L-lysine (PLL)-coated Au nanocomposites film surface, and the fluorescence enhancement effects can be optimized by varying the distance between TAMRA and the Au nanocomposites film, which is easily adjusted by varying the thickness of the biocompatible poly-(acrylic acid) (PAA/PLL) multilayers, and thus metal-enhanced fluorescence of dye TAMRA conjugated with the aptamer is generated up to 15.2-fold. The interaction of the aptamer to its target induces the reversible conformation change of the aptamer, and consequently, the electrostatic potential is overcome by binding force. As a result, the target-binding interaction of the aptamer causes the irreversible detachment of the aptamer from the nanostructured Au film surface to decrease fluorescence of TAMRA. The aptamer macroarray provides not only the appropriate sensitivity for clinical diagnostics with a wide range of linear detection from 10pg/mL to 10μg/mL, high specificity for PDGF-BB against VEGF-165, VEGF-121, NaCl and IgG, and temporal biological stability, but also a single-step detection. We envision that the efficient and robust aptamer macroarray can be extended to the detection of other biomarkers.
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China. Electronic address: dongzhunjutcm@hotmail.com.
authors.forename: Dong
authors.initials: D
authors.lastname: Zhu
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Rui Xiang
authors.initials: RX
authors.lastname: Yang
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China. Electronic address: yupingtang@njucm.edu.cn.
authors.forename: Yu-Ping
authors.initials: YP
authors.lastname: Tang
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Zhao Yi
authors.initials: ZY
authors.lastname: Miao
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Hu
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Yu
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, PR China.
authors.forename: Chen Yang
authors.initials: CY
authors.lastname: Xu
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D000077214:Becaplermin; D015374:Biosensing Techniques / Q000295:instrumentation*; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D004867:Equipment Design; D006046:Gold / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D046228:Microarray Analysis / Q000295:instrumentation*; D053761:Nanocomposites / Q000737:chemistry / Q000648:ultrastructure; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D011107:Polylysine / Q000737:chemistry; D020574:Proto-Oncogene Proteins c-sis / Q000097:blood*
chemical_list: D052157:Aptamers, Nucleotide; D004129:Dimethylpolysiloxanes; D020574:Proto-Oncogene Proteins c-sis; D000077214:Becaplermin; D011107:Polylysine; D006046:Gold
keywords: Aptamer macroarrays; Human platelet-derived growth factor-BB (PDGF-BB); Nanoplasmonic; Robust; Single-step detection