Bio Molecule

Integrated NanoparticleBiomolecule Hybrid System: Synthesis, Properties, and Applications

Introduction

• Functional Biomolecule Nanoparticle Structure on Surfaces for Application as sensors (生物奈米粒子材料表面應用在感測器) • Biomolecule Functionalized Magnetic Particles (生物分子應用於磁性粒子) • Biomolecule Based Nanocircuitry (生物分子應用奈米迴路)

Angew. Chem. Int. Ed. 2004, 43, 6042-6108

• Synthesis and Properties of Biomolecule Functionalized Nanoparticles (生物分子改質奈米粒子) • Biomolecule Functionalized Nanoparticles for Controlled Chemical Reactivity (生物分子改質奈米粒子調控化學反應) • The Aggregation of Biomolecule Functionalized Nanoparticles (生物分子聚集改變奈米粒子功能性) • Assembly of Biomolecule Nanoparticle Architectures on Surfaces (生物奈米粒子自組裝材料表面)

Synthesis and Properties of Biomolecule Functionalized Nanoparticles (生物分子改質奈米粒子)

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Biomolecule-nanoparticle (生物分子結合奈米粒子) Formation of the biomoleculenanoparticle (NP) hybrids (生物分子與奈米粒子雜合)

Nanoparticles could be encapsulated by some natural proteins to provide their affinity binding （一些蛋白質可能將奈米粒子封入提 供彼此結合性）

Chaperonin proteins as ATPresponsive barrels for the inclusion of nanoparticles

• a. Top and side view of GroEL and T.th cpn • b. The formation of GroEL-Cds NP complexes by inclusion of Cds NPs into the cylindrical cavity of GroEL, and ATPtriggered release of the guest

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Transmission electron micrographs Properties of NanoparticleBiomolecule Hybrid System (奈米粒子與生物分子雜合系統的 特性)

T.th cpn-Cds NP complexes

Intact T.th cpn

The electrochemistry controlled recognition of flavin by a pyridineduamide-functionalized nanoparticle

Biomolecule Functionalized Nanoparticles for Controlled Chemical Reactivity (生物分子改質奈米粒子調控化學反 應)

flavin

Biomolecule Functionalized Nanoparticles for Controlling DNA Reactivity

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奈米粒子控制DNA反應

奈米粒子顯示DNA反應

Radio-frequency (1-GHz)

奈米粒子控制酵素反應 （CdS NP-capped mesoporous）

奈米粒子控制酵素反應 （CdS NP-capped mesoporous)

Carboxylic acids modified Cds NPs

The convalent coupling of the Cds NPs to the amine function of the matrix Matrix by a siloxane anchoring site

The use of biotin-streptavidin (SAv ) interactions to build nanoparticle networks End-to-end assembly

12: dibiotin

13: biotin disulfide

TEM: SAv-interconnected Au nanorods

14: Ester derivative of biotin

Biotin-functionalized ferritin 15: bifunctional linker

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Nucleic Acid Functionalized NPs for Controlled Aggregation

TEM of gold nanorods that are organized by DNA hybridization

16: 3’-thiol-TACCGTTG-5’ 17: 5’-AGTCGTTT-3’-thiol 18: A DNA linker

B: cuvettes with a mixture of the Au NPs and the added DNA strand responsible for the assembly process C: variations in the absorption spectrum of the DNA-linked NP networks as function of temperature D: the satellite system consists of two different sizes of nanoparticles

DNAzyme system for the analysis of metal ions

DNAzyme system for the analysis of metal ions

active 17E only

17E/17Ec is 1:20

Enzyme strand a: the active 17E DNAzyme-NP sensor 1: absence Pb2+ ions 2: presence Pb2+ ions

Controlled association of Au NPs based on biocatalytic transformation of oligonucleotide

b: an inactive 17Ec DNAzyme- c: Calibration plots for the analysis of Pb2+ ions NP sensor 1: absence Pb2+ ions 2: presence Pb2+ ions

Controlled association of Au NPs based on biocatalytic transformation of oligonucleotide DNA-NP complex before

DNA-NP complex

Treatment with EcoRI and after subsequent treatment with DNA ligase

• TEM analysis of the DNA-NP complexes before a. and after b. treatment with EcoRI and after subsequent with DNA ligase c.

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Enzyme-controlled distance between DNA-bridged Au NPs (酵素控制奈米金粒子DNA橋)

Composite Assemblies of Nucleic Acids, Proteins, and Nanoparticles (奈米立子與核酸,蛋白質自組裝)

EcoRI SAv Biotinylated antibodies

The hybrid with 59° bend Following the binding of EcoRI

The assembly of controlled multiparticle composites upon Hybridization on DNA template

The use of conjugate for the sensing of antigens (偵測抗原)

The construction of four-nanoparticle clusters (金奈米粒子四聚體)

Cowpea mosaic virus (CPMV) labeled with 1.4nm Au clusters

Assembly of Layered NanoparticleProtein Arrays on Surface Assembly of Biomolecule-Nanoparticle Architectures on Surface (生物分子與奈米立子結構表面自組裝)

Construction of TiO2/cty c multilayers on a QCM

Optical absorbance at lamda=409 nm of assemblies of different thicknesses

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Nucleic Acid-Nanoparticle Architectures on Surfaces (核酸與奈米立子結構表面自組裝)

Electrostatic deposition of CdS NPs on a DNA chain at the air/water interface

Fig. B: AFM image of a DNA strand With Au NPs that specifically bound to the template through biotin-SAv interaction

38. Cationic surfactant molecules 39. Positively charged CdS NPs(3 nm) capped with thiocholine

The 2D assembly of Au NPs by using a DNA-based method

The use of dip-pen lithography and DNA to produce a predesigned multinanoparticle pattern 43. TCTCAACTCGTAA10 44. A10CGCATTCAGGAT 45. TACGAGTTGA -GAATCCTGAATGCG

AFM image of the resulting 2D Au 40. A gold NP is attached to oligonucleotide NP-DNA network 41. Hybridized with oligonucleotide 42. DNA network that consists of oligonucleotide

AFM image of the assembly , which consists of large and small nanoparticles Scale bar=1 um

The use of DNA as linker to construct nanoparticle multilayer on surface (利用DNA結合奈米粒子在表面形成多層連結)

46. Monolayer of an oligonucleotide 47. Composed of two domains (46 and 48) 48. Au NPs (13nm) that were functionalized with oligonucleotide

The absorbance and the fluorescence spectrum: respectively, of the CdS NP multulayer assemblies: a to d =1 to 4 layers

Functional Biomolecule Nanoparticle Structure on Surfaces for Application as sensors (生物奈米粒子材料表面應用在感 測器)

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Bioelectronic Systems Based on nanoparticle-Enzyme Hybrids as System (奈米粒子結合酵素生物電子感測系統)

FAD cofactor

Assembly of the CdS NP-AChE hybrid system for the photoelectrochemical detection of enzyme activity

Apo-glucose oxidase

acetylthiocholine as the substrate The reconstituted GOx electrode in the presence of different concentration of glucose

The enzyme generated thiocholine

Photocurrent action spectra observed in the presence of acetylthiocholine • • • • • •

Calibration curve of the photocurrent at lamda=380nm at variable concentration

a: 0mM b: 6mM c: 10mM d: 12mM e: 16mM f: 30mM 53: acetylthiocholine

Photocurrent spectra for the CdSAChE system Electrochemical detection of DNA by the deposition of catalytic silver clusters on the DNA strand 55: dibromibe (inhibits the photocurrent formation) a: in the absence of inhibitor 55 b: upon addition of the inhibitor 55 (1X10-6 M) c: after rinsing the system, exclusion of the inhibitor, and addition of 53

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Hybridization of the complementary target DNA with the DNA probe

Loading of the Ag+ ions onto immobilized DNA

56: DNA probe

57: target DNA Reduction of Ag+ ions by hydroquinone to form silver aggregates on the DNA backbone

Dissolution of the silver aggregates in acidic solution Immunosensing at microsized Au electrodes by the change of conductivity

PSA: potentiometric stripping analysis

Immunosensing at microsized Au electrodes

The use of a DNA-NP conjugate

stabilized by an anionic protective

58: A probe nucleic acid 59: The target 27-mer nucleotide 60: Au NPs were functionalized with a nucleic acid

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Stripping potentiograms measured upon the sensing of different concentrations of DNA

Different concentrations of DNA that are bound to magnetic particles and labeled with CdS NPs a: 0.2 mgL-1 b: 0.4mgL-1 c: 0.6mgL-1 d: control

Multitarget electrochemical DNA detection with different nanocrystal labels

a: probe-modified magnetic beads b: hybridization with the DNA targets c: second hybridization with the NP-labeled probe d: dissolution of the NPs and the electrochemical detection

The amplified detection of DNA by using nucleic acid-Au NP-functionalized beads

Stripping potentiogram measured upon the sensing of different concentration

The DNA molecular are labeled with ZnS NPs (T1), CdS NPs (T2), PbSNPs (T3)

The amplified detection of DNA with polystyrene(聚苯乙烯) beads 64: ferrocenecarboxaldehyde (as a redox marker)

a: Hybridization of the NP-functionalized beads with the target DNA b: the enhanced catalytic deposition of gold on the NPs c: Dissolution of the gold clusters d: the detection of the Au3+ ions by stripping voltammetry

65: the probe DNA linked to magnetic particles 66: complementary nucleic acid

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The construction of CdS NP-DNA superstructures

Dendritic amplified DNA sensing by the use of oligonucleotide-functionalized Au NPs

The amplified detection of the 7249base M13mp18 DNA The catalytic deposition of gold on a Au NP conjugate

71: the DNA primer 72: M13mp18 DNA 73: streptavidin-Au

The analysis of a single-base mismatch in DNA

Microgravimetric detection of a single-base mutant b: Arrows indicate (single base mutant)

a: Normal DNA sequence

(1): the attachment of the SAv-Au conjugate (2): the catalytic deposition of gold on the Au NPs

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The construction of mixed-metal “barcode” Nanoparticle-Biomolecule Conjugates for Optical Sensing and Analysis

Au-Ag multistripe “barcode” nanorod

Detection of DNA with “barcodes” 78: a nucleic acid that is labeled with a fluorophore (TAMRA dye) 77: a primer nucleic acid that is linked to Ag 76: analyte DNA

Fluorescence images bound fluorescent DNA

Fluorescence images absence of analyte DNA

Multiple immunoassay by using “barcodes” 81: the antibodies with fluoresceinlabeled anti-human IgG 79: anti-human IgG antibody

82: Texas Red-labeled antirabbit IgG 80: anti-rabbit IgG antibody

Reflectivity images bound fluorescent DNA

Reflectivity images absence of analyte DNA a: Reflectivity images b: Fluorescein (FITC)-labeled anti-human-igG c: Texas Red-labeled anti-rabbit-IgG

Application of Au nanoparticle labels that are encoded with DNA for the amplified immunosensing of prostate-specific antigen(PSA) 83: an antibody which is complementary to PSA was linked to magnetic beads 84: double-strain nucleic acids was linked to the magnetic beads-antibody-PSA 85: triple-component sandwich assay configuration 86: thermal dissociation of the nucleic acid duplex yielded the free nucleic acid 87: single-stranded-oligonucleotide-functionalized Au NPs

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Surface plasmon resonance表面電漿共振(SPR) spectroscopy enhanced by Au NPs for DNA analysis

Localized surface plasmon resonance 局部表面電漿共振(LSPR)

Assembly of Au NP-bound reconstituted glucose oxidase(GOx) on a dithiol monolayer

Assembly of Au NP-bound reconstituted glucose oxidase(GOx) on a dithiol monolayer that is associated with a SPRactive surface

SPR spectra of the Au NP-GOx hybrid system

Calibration plot of the SPR spectra minimum shift as a function of glucose concentration

a: 0 mM b: 0.3 mM c: 1.6 mM d: 8 mM e: 40 mM f: 100 mM

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Assembly of the Au NP-CdS NPacetyl choline hybrid system

Application of Raman dye-functionalized Au NPs for amplified multitarget immunosensing

B: Flatbed scanner images of silverenhanced microarrays upon the immunosensing of different antibodies

C: Typical Raman spectra that correspond to the colored dots in the immunosensing array

Application of Raman dye-functionalized Au NPs for amplified multitarget immunosensing

Replication and telomerization of nucleic acid functionalized CdSecore/ZnSshell NPs

88: primer (complementary M13mp18 DNA ) 89: included Texas Red-functionalized dUTP 90: primer (recognized by telomerase )

Emission spectra upon the time-dependent DNA replication(B) / telomerization(C)

Biomolecule Functionalized Magnetic Particles (生物分子應用於磁性粒子)

a: 0 min b: 10 min c: 30min d: 60 min

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Electrochemical analysis of DNA upon the assembly of DNA molecules at magnetic particles followed by their association with Au NPs

Au NPs are used for the deposition of silver

91: biotin-labeled nucleic acid 92: complementary biotinylated nucleic acid

The Au NPs chemically dissolved

93: the primer biotinylated nucleic acid was linked to magnetic beads through an avidin bridge 94: functionalized with Au NPs a: deposited gold is electrochemically stripped (path a) b: the intermediate enlargement of the Au NPs results in the future amplification of the signal (path b)

The effect of gold enhancement upon the stripping response for the DNA analyte

Application of Redox-functionalized Magnetic Particles for the Triggering and Enhancement of Electrocatalytic and Bioelectrocatalytic Process

PSA signals prior to treatment of the system with gold enhancement solution (a) and after 10 minutes of reaction (b)

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Linkage of 2,3-dichloro-1,4-naphthoquinone (95) to the functionalized particles to yield the aminonaphthoquinone (100)- functionalized magnetic

Carbodiimide coupling of the electron-relay carboxylic derivatives (96-99) to the amino groups of the siloxane layer

particles

Functionalization of magnetic particles with the PQQ-NAD+ Dyed for the electrochemical activation of NAD+-dependent enzymes

Key words

The electrochemical , electrocatalytic and bioelectrocatalytic reaction of functional magnetic particles which are controlled by means of an external magnet

Switching on or off the electrochemical reaction of the redox-relay groups (R)

Covalently bound to the magnetic particles and the electrocatalytic function that is provide by the redox groups

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Differential pulse voltammograms of a Au electrode

Switching on or off

a: 接觸 b: 縮回未接觸

Magnetic activation

Magnetic deactivation 1. Bioelectrocatalytic oxidation of glucose in the presence of glucose oxidase 2. Oxidation of lactase in the presence of lactate dehydrogenase (LDH)

Cyclic voltammograms at a Au electrode

Enhanced bioelectrocatalytic oxidation of glucose in the presence of glucose oxidase (GOx)

upon attraction

retraction

a: 接觸 (on) b: 縮回未接觸(off)

Enhanced bioelectrocatalytic oxidation of glucose in the presence of glucose oxidase (GOx)

Magnetomechanical Detection of Biorecognition Events

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Synthesis of the functionalized magnetic particles for the biorecognition assay Amplified detection of bioaffinity recognition processes by mutilabeled rotating magnetic particles

Amplified biorecognition assay upon the rotation of the functionalized magnetic particles

Labeling of the nucleic acid replica with biotin units by thermal cycles for the amplified detection of viral DNA by multilabeled rotating magnetic particles

Chemiluminescence intensities upon the analysis of M13mp18

Schematic configuration of the instrumental setup and concept for the magnetomechanical analysis of biorecognition processes on functionalizes cantilevers

B: at different rotation speeds a) 0, b) 60, c) 400, d) 2000, e) absence of DNA C: Arrows indicate the time for switching the potential to -0.5V and to 0.0V

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Magnetomechanical deflection or retraction of the cantilever in the analysis of M13mp18

TEM image with Pt NP aggregates

a: the cantilever is subjected to the external magnet b: the external magnet is removed

B: using Taq polymerase and thermal cycles for replication and labeling process C: Dependence of the deflection signal on the concentration of the M13mp18 DNA

Formation of a silver nanowire inside a channel Biomolecule Based Nanocircuitry (生物分子應用奈米迴路)

Formation of a silver nanowire inside a channel (TEM images)

106: β-amyloid

Nanocircuitry produced upon biospecific interaction on surfaces Electron micrograph

Fluoresecence images at different magnifications

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Structure of the Au NP-peptide complex on the histidine-rich polypeptide nanotube template

End –to-end interconnected peptide nanotubes

The assembly of patterned actinbased Au nanowires

AFM images

108: N-hydroxysuccinimidyl ester groups

B: AFM image of the Au wire/actin/Au wire filament C: AFM image of the actin/Au wire/ actin filament

SFM images of nanoparticle networks

AFM image of lamda-DNA AFM image of Lamda-DNA , Which is partially Linked with Au55 clusters

Height profile of both the bare DNA and a decorated part

DNA fragment with Au55 clusters along The phosphate Backbone of the DNA Major grooves

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Assembly of a Au NP wire in the pA/pT template by using Au NPs

AFM image of the Au NP wire in the pA/pT template

pA/pT= polyadenylic acid / polythymidylic acid

The assembly of Au nanowires on a telomer template

The assembly of Au nanowires on a telomer template

The construction of a nanowire that bridges two microelectrodes by deposition of Ag+ ions on a bridging DNA strand

Molecular lithography based on homologous recombination processes carried out by the RacA protein

Current versus voltage (I/V) curves obtained with the structure produced

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AFM image of the patterned DNA template after gold metallization

The fluid-flow-assisted molecular combing of DNA molecules on a surface to yield 1D or 2D arrays

AFM image of a Pd nanowire , a section analysis shows an average Particle height of Approximately 5 nm

AFM image D: a DNA strand cut by an AFM tip

E: and F: AFM tip on the nanometer scale

AFM image of a meshlike 2D array of Pd nanowires

Real-time conductance response from a si nanowire device that is functionalized with a PNA (peptide nucleic acid ) receptor

Si nanowire device with source (S) and drain (D)

The data points shown were obtained from two independent Si nanowire devices

Image of the motility of actin/Au wire/ actin filament on a glass surface The same frame imaged at 5-s time intervals

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