Anionic Conjugated Polymer with Aptamer-Functionalized Silica Nanoparticle for Label-Free Naked-Eye Detection of Lysozyme in Protein Mixtures

被引:51
作者
Wang, Yanyan [1 ]
Pu, Kan-Yi [1 ]
Liu, Bin [1 ]
机构
[1] Natl Univ Singapore, Dept Chem & Biomol Engn, Singapore 117576, Singapore
关键词
EGG-WHITE LYSOZYME; SENSITIVE ELISA; DNA; FLUORESCENT; DENSITY; PROBE; POLYELECTROLYTES; SENSORS; ASSAY;
D O I
10.1021/la100139p
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
An assay triggered by recognition-induced charge switching is developed for protein detection and quantification. Aptamer-functionalized silica nanoparticles (NPs) have been synthesized to capture lysozyme, resulting in an alternation of the surface charge from negative to partially positive. The binding event is then translated and monitored by the fluorescence signal of a highly fluorescent anionic poly(fluorene-alt-vinylene)(PFVSO3), which "stains" on protein/aptamcr-NP complexes via electrostatic interaction. Blue-greenish fluorescence of PFVSO3 is observed in the presence of lysozyme by the naked eye, while no fluorescence is obtained for NPs upon treatment with a mixture of foreign proteins. A linear relationship between NP fluorescence and lysozyme is observed in the concentration range of 0-22.5 mu g/mL, which gives a limit of detection as similar to 0.36 mu g/mL. This work demonstrates a convenient label-free conjugated polyelectrolyte (CPE)-based protein detection with high specificity and sensitivity, which has potential applications in medical diagnosis.
引用
收藏
页码:10025 / 10030
页数:6
相关论文
共 48 条
[1]   Protein detecting arrays based on cationic polythiophene-DNA-aptamer complexes [J].
Aberem, Maite Bera ;
Najari, Ahmed ;
Ho, Hoang-Anh ;
Gravel, Jean-Francois ;
Nobert, Philippe ;
Boudreau, Denis ;
Leclerc, Mario .
ADVANCED MATERIALS, 2006, 18 (20) :2703-+
[2]   Fluorescent polyelectrolytes as protein sensors [J].
Ambade, Ashootosh Vasant ;
Sandanaraj, Britto Selvaraj ;
Klaikherd, Akamol ;
Thayumanavan, S. .
POLYMER INTERNATIONAL, 2007, 56 (04) :474-481
[3]   STRUCTURE OF HEN EGG-WHITE LYSOZYME - A 3-DIMENSIONAL FOURIER SYNTHESIS AT 2A RESOLUTION [J].
BLAKE, CCF ;
KOENIG, DF ;
MAIR, GA ;
NORTH, ACT ;
PHILLIPS, DC ;
SARMA, VR .
NATURE, 1965, 206 (4986) :757-&
[4]   Protein carbonyl measurement by a sensitive ELISA method [J].
Buss, H ;
Chan, TP ;
Sluis, KB ;
Domigan, NM ;
Winterbourn, CC .
FREE RADICAL BIOLOGY AND MEDICINE, 1997, 23 (03) :361-366
[5]   Aptamer-based biosensors for label-free voltammetric detection of lysozyme [J].
Cheng, Alan K. H. ;
Ge, Bixia ;
Yu, Hua-Zhong .
ANALYTICAL CHEMISTRY, 2007, 79 (14) :5158-5164
[6]   Automated selection of anti-protein aptamers [J].
Cox, JC ;
Ellington, AD .
BIOORGANIC & MEDICINAL CHEMISTRY, 2001, 9 (10) :2525-2531
[7]   ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) QUANTITATIVE ASSAY OF IMMUNOGLOBULIN-G [J].
ENGVALL, E ;
PERLMANN, P .
IMMUNOCHEMISTRY, 1971, 8 (09) :871-&
[8]   High-efficiency fluorescence quenching of conjugated polymers by proteins [J].
Fan, CH ;
Plaxco, KW ;
Heeger, AJ .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (20) :5642-5643
[9]   Reduced nonspecific adsorption on covalently immobilized protein surfaces using poly(ethylene oxide) containing blocking agents [J].
Frederix, F ;
Bonroy, K ;
Reekmans, G ;
Laureyn, W ;
Campitelli, A ;
Abramov, MA ;
Dehaen, W ;
Maes, G .
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 2004, 58 (01) :67-74
[10]   DNA surface hybridization regimes [J].
Gong, Ping ;
Levicky, Rastislav .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (14) :5301-5306