Multi-functional crosslinked Au nanoaggregates for the amplified optical DNA detection.

We report a novel sandwich type assay for the optical detection of DNA using multi-component crosslinked Au nanoparticle aggregates. In this design, a DNA bridged multi-functional Au nanoparticle aggregate, which integrates DNA recognition (detection probe), signal amplification (enzyme, horseradish peroxidase) and non-specific blocking (bovine serum albumin, BSA) section, is employed as detection probe. The sandwich type assay includes the capture probe that immobilized on magnetic microparticles, target and the Au nanoparticle aggregates detection probe. In a typical sensing process, the Au aggregates detection probe is brought to the proximity of magnetic particles through the DNA hybridization. Once magnetic field is added, these sandwich complexes are magnetically separated. As a result, HRP that confined at the surface of Au aggregates could catalyze the enzyme substrate and generate an optical signal. This assay is employed for the detection of breast cancer-associated BRCA-1 gene. The detection limit is about 1 fmol, which is significantly improved as compared with the results obtained from individual Au nanoparticle labeled assay. Thus, our results address the possibility of Au nanoaggregates as platform for DNA detection.

[1]  Yi Lu,et al.  A colorimetric lead biosensor using DNAzyme-directed assembly of gold nanoparticles. , 2003, Journal of the American Chemical Society.

[2]  Itamar Willner,et al.  Integrated nanoparticle-biomolecule hybrid systems: synthesis, properties, and applications. , 2004, Angewandte Chemie.

[3]  R. Goody,et al.  Functional Immobilization of the Small GTPase Rab6A on DNA–Gold Nanoparticles by Using a Site‐Specifically Attached Poly(ethylene glycol) Linker and Thiol Place‐Exchange Reaction , 2007, Chembiochem : a European journal of chemical biology.

[4]  J. Zhuang,et al.  Sterically Mediated Two‐Dimensional Architectures in Aggregates of Au Nanoparticles Directed by Phosphorothioate Oligonucleotide‐DNA , 2005 .

[5]  C. Mirkin,et al.  Homogeneous, Nanoparticle-Based Quantitative Colorimetric Detection of Oligonucleotides , 2000 .

[6]  J. Storhoff,et al.  A DNA-based method for rationally assembling nanoparticles into macroscopic materials , 1996, Nature.

[7]  Juewen Liu,et al.  Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes , 2006, Nature Protocols.

[8]  Christof M. Niemeyer Priv.-Doz.,et al.  DNA-Directed Functionalization of Colloidal Gold with Proteins† , 2001 .

[9]  J. Storhoff,et al.  Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. , 1997, Science.

[10]  Chad A Mirkin,et al.  Bio-bar-code-based DNA detection with PCR-like sensitivity. , 2004, Journal of the American Chemical Society.

[11]  Chad A. Mirkin,et al.  One-Pot Colorimetric Differentiation of Polynucleotides with Single Base Imperfections Using Gold Nanoparticle Probes , 1998 .

[12]  W. Smith,et al.  Control of enhanced Raman scattering using a DNA-based assembly process of dye-coded nanoparticles. , 2008, Nature nanotechnology.

[13]  Juewen Liu,et al.  Fast colorimetric sensing of adenosine and cocaine based on a general sensor design involving aptamers and nanoparticles. , 2005, Angewandte Chemie.

[14]  Christof M. Niemeyer,et al.  DNA-Directed Functionalization of Colloidal Gold with Proteins This work was supported by Deutsche Forschungsgemeinschaft and Fonds der Chemischen Industrie. We thank Prof. D. Blohm for helpful discussions and generous support. , 2001, Angewandte Chemie.

[15]  Itamar Willner,et al.  Amplified electrochemical detection of DNA through the aggregation of Au nanoparticles on electrodes and the incorporation of methylene blue into the DNA-crosslinked structure. , 2007, Chemical communications.

[16]  Itamar Willner,et al.  Nucleic acid-functionalized Pt nanoparticles: Catalytic labels for the amplified electrochemical detection of biomolecules. , 2006, Analytical chemistry.

[17]  A Paul Alivisatos,et al.  Enzymatic ligation creates discrete multinanoparticle building blocks for self-assembly. , 2008, Journal of the American Chemical Society.

[18]  Jinghong Li,et al.  In situ amplified chemiluminescent detection of DNA and immunoassay of IgG using special-shaped gold nanoparticles as label. , 2006, Clinical chemistry.

[19]  Itamar Willner,et al.  Dendritic amplification of DNA analysis by oligonucleotide-functionalized Au-nanoparticles , 2000 .

[20]  R. G. Freeman,et al.  Preparation and Characterization of Au Colloid Monolayers , 1995 .

[21]  C. Niemeyer REVIEW Nanoparticles, Proteins, and Nucleic Acids: Biotechnology Meets Materials Science , 2022 .

[22]  C. Niemeyer,et al.  Reversible binding of fluorescent proteins at DNA-gold nanoparticles. , 2006, Angewandte Chemie.

[23]  C. Liu,et al.  One-step homogeneous detection of DNA hybridization with gold nanoparticle probes by using a linear light-scattering technique. , 2006, Angewandte Chemie.

[24]  Chad A Mirkin,et al.  The bio-barcode assay for the detection of protein and nucleic acid targets using DTT-induced ligand exchange , 2006, Nature Protocols.

[25]  Chad A Mirkin,et al.  Nanostructures in biodiagnostics. , 2005, Chemical reviews.

[26]  Chunhai Fan,et al.  Enzyme‐Based Multi‐Component Optical Nanoprobes for Sequence‐ Specific Detection of DNA Hybridization , 2008 .

[27]  Chunhai Fan,et al.  Sequence-specific detection of femtomolar DNA via a chronocoulometric DNA sensor (CDS): effects of nanoparticle-mediated amplification and nanoscale control of DNA assembly at electrodes. , 2006, Journal of the American Chemical Society.

[28]  P. Stenson,et al.  The Human Gene Mutation Database: 2008 update , 2009, Genome Medicine.

[29]  Zhiqiang Gao,et al.  Electrical detection of oligonucleotide using an aggregate of gold nanoparticles as a conductive tag. , 2008, Analytical chemistry.

[30]  Chad A. Mirkin,et al.  DNA-Directed Synthesis of Binary Nanoparticle Network Materials , 1998 .