Electrochemical studies of glucose oxidase immobilized on glutathione coated gold nanoparticles

Glutathione (L-gamma-glutamyl-L-cysteinyl-L-glycine; GSH) forms a surface monolayer on gold nanoparticles by tethering via sulfur bonds (Au:GSH). In the present study, glucose oxidase (GOx; EC 1.1.3.4) was immobilized by covalent chemical coupling reactions on to Au:GSH nanoparticles and the enzyme coupled nanoparticles formed a stable colloid (stable for several weeks) in water. The immobilized enzyme was investigated for electrochemical characteristics to monitor the FAD (prosthetic group of the GOx) redox potentials. Various concentrations of substrate (glucose) were added to check the oxidation characteristics. It was observed that with increase in substrate concentrations, the oxidation rate increased proportionally with the current. The present study demonstrated that GOx was effectively coupled to the gold nanoparticle (Au:GSH). The coupled nanoparticle system could be used in a potential biosensor application. Similarly, other enzymes (e.g., horseradish peroxidase) could be immobilized to the Au:GSH nanoparticles via the peptide arm (GSH) to achieve the desired characteristics needed for a specific application in biosensor.     

Structure and activity of apoferritin-stabilized gold nanoparticles

A simple method for synthesizing gold nanoparticles stabilized by horse spleen apoferritin (HSAF) is reported using NaBH(4) or 3-(N-morpholino)propanesulfonic acid (MOPS) as the reducing agent. AuCl(4)(-) reduction by NaBH(4) was complete within a few seconds, whereas reduction by MOPS was much slower; in all cases, protein was required during reduction to keep the gold particles in aqueous solution. Transmission electron microscopy (TEM) showed that the gold nanoparticles were associated with the outer surface of the protein. The average particle diameters were 3.6 and 15.4 nm for NaBH(4)-reduced and MOPS-reduced Au-HSAF, respectively. A 5-nm difference in the UV-Vis absorption maximum was observed for NaBH(4)-reduced (530 nm) and MOPS-reduced Au-HSAF (535 nm), which was attributed to the greater size and aggregation of the MOPS-reduced gold sample. NaBH(4)-reduced Au-HSAF was much more effective than MOPS-reduced Au-HSAF in catalyzing the reduction of 4-nitrophenol by NaBH(4), based on the greater accessibility of the NaBH(4)-reduced gold particle to the substrate. Rapid reduction of AuCl(4)(-) by NaBH(4) was determined to result in less surface passivation by the protein. Methods for studying ferritin-gold nanoparticle assemblies may be readily applied to other protein-metal colloid systems.     

Investigation of Gold and Silver Nanoparticles on Absorption Heating and Scattering Imaging

Efficient conversion of absorbed light to heat energy and strong scattering by gold and silver nanoparticles suggest these nanoparticles as the agents of heating and imaging. Absorption efficiency and scattering efficiency of gold and silver nanoparticles were studied through numerical simulation using the discrete dipole approximation method. This study shows that the size of gold and silver nanoparticles can effect gold and silver nanoparticles’ absorption efficiency and scattering efficiency. The gold nanoparticle is found to possess the maximum absorption efficiency when the size of gold nanoparticle is 50 nm and the incident wavelength is 540 nm, and the increasing scattering efficiency with the increasing size of gold nanoparticle in the medium, and refractive index of the medium is around 1.33. However, the silver nanoparticle owns the maximum absorption efficiency when the size of silver nanoparticle is 20 nm and the incident wavelength is 396 nm, and the maximum scattering efficiency when the size of silver nanoparticle is 30 nm and the incident wavelength is 410 nm in the same medium. The conditions for achieving the maximum adsorption efficiency and scattering efficiency of gold and silver nanoparticle can be used for heating and imaging using visible and near-infrared light.     

Enhanced oligonucleotide-nanoparticle conjugate stability using thioctic acid modified oligonucleotides

Metallic nanoparticles of gold functionalized with oligonucleotides conventionally use a terminal thiol modification and have been used in a wide range of applications. Although readily available, the oligonucleotide–nanoparticle conjugates prepared in this way suffer from a lack of stability when exposed to a variety of small molecules or elevated temperatures. If silver is used in place of gold then this lack of stability is even more pronounced. In this study we report the synthesis of highly stabilized oligonucleotide–nanoparticle conjugates using a simple oligonucleotide modification. A modified solid support was used to generate 3′-thioctic acid modified oligonucleotides by treatment with an N-hydroxysuccimidyl ester of thioctic acid. Unusually, both gold and silver nanoparticles have been investigated in this study and show that these disulphide-modified oligonucleotide probes offer significant improvements in nanoparticle stability when treated with dithiothreitol (DTT) compared with monothiol analogues. This is a significant advance in oligonucleotide–nanoparticle conjugate stability and for the first time allows silver nanoparticles to be prepared that are more stable than standard gold-thiol functionalized nanoparticles. This opens up the possibility of using silver nanoparticles functionalized with oligonucleotides as an alternative to gold.     

Preparation,Optimization, and Characterization of SERS Sensor Substrates Based on Two-Dimensional Structures of Gold Colloid

Generally, the immobilization of two-dimensional nanoparticles in immersion procedures is time-consuming (over 24 h). In this paper, we report a very effective and simple method to fabricate two-dimensional gold nanoparticle patterns over large areas with high regularity for surface-enhanced Raman scattering (SERS). We achieved a highly sensitive SERS colloid surface by optimizing temperature and immersion time. The surfaces were characterized by X-ray photoelectron spectroscopy, UV–Vis, atomic force microscopy, and scanning electron microscopy. The SERS activity of surfaces was compared by using two techniques: “dip” and “dip and dry” in an aqueous solution of 10⁻⁶ M crystal violet. The influence of the morphology of the surface was investigated with both the dip and dip and dry techniques.     

Preparation of gold nanopowders and nanoparticles using chitosan suspensions

Simple methods for preparation of gold nanopowders and nanoparticles are reported. Gold/chitosan nanoparticles were prepared by using basic chitosan suspension as a dispersant and as a reductant. The resulting nanoparticles were processed by pyrolysis and thus obtain black gold nanopowder. The FESEM images indicate that most diameters of the nanopowder prepared were in the range of 50 and 200 nm. Hydrolysis is another quick decomposition method for chitosan. Acetic acid was adopted to implement the hydrolysis. The AEM images of the auberginic suspension show that the average gold nanoparticle diameter was less than 40 nm with good dispersion. Use of chitosan suspensions can produce gold nanopowder as well as gold nanoparticle without using toxic organic chemicals.     

On prediction of optical properties of two- and multiphase nanocomposites for nanomedicine

The theory of optical properties of nanoparticles is considered with the aid of dispersion relations, which are based on the Kramers-Kronig analysis. It is shown that one can utilize rather general dispersion relations, which hold for liquid matrices that contain nanoparticles. Wiener bounds incorporating the Kramers-Kronig analysis are utilized in assessment of the complex permittivity of a nanoparticle.     

Fluorescence Quenching of CdTe Nanocrystals by Bound Gold Nanoparticles in Aqueous Solution

Water-soluble gold nanoparticles with an average diameter of 5 nm were prepared with carboxylic acid terminated thiol ligands. These ligands contain zero to eight methylene moieties. CdTe nanocrystals with an average diameter of 5 nm were synthesized with aminoethanethiol capping. These nanocrystals displayed characteristic absorption and emission spectra of quantum dots. The amine terminated CdTe nanocrystals and carboxylic-acid-terminated gold nanoparticles were conjugated in aqueous solution at pH 5.0 by electrostatic interaction, and the conjugation was monitored with fluorescence spectroscopy. The CdTe nanocrystals were significantly quenched upon binding with gold nanoparticles. The quenching efficiency was affected by both the concentration of gold nanoparticles in the complex and the length of spacer between the CdTe nanocrystal and Au nanoparticle. The observed quenching was explained using Förster resonance energy transfer (FRET) mechanism, and the Förster distance was estimated to be 3.8 nm between the donor–acceptor pair.     

Association temperature governs structure and apparent thermodynamics of DNA-gold nanoparticles

Apparent thermodynamics of association of DNA-modified gold nanoparticles has been characterized by UV spectroscopy and dynamic light scattering (DLS). Extinction coefficients of unlabelled and DNA-labelled gold nanoparticles have been determined to permit quantitative analysis of the absorption measurements. In contrast to previous studies the associating gold nanoparticles were furnished with complementary oligonucleotide DNA single strands. This resulted in direct complex formation between the nanoparticles on mixing without the requirement of a DNA linker sequence for initiation of cluster formation. Melting curves of the nanoparticle assemblies formed at different temperatures were subjected to two-state analysis. A comparison of the apparent thermodynamic parameters obtained for the dissociation of these aggregates suggests that both thermodynamically and structurally different nanoparticle clusters are obtained depending on the temperature at which assembly proceeds. The van't Hoff enthalpies permit an estimate of the DNA duplexes: gold nanoparticle ratio involved in network formation.     

金溶胶基底的 SERS 增强效果的实验研究

本文以结晶紫作为探针分子,研究了以金溶胶膜、pH ＝6以及 pH ＝13的金溶胶溶液为活性基底的表面增强拉曼光谱的增强效果。采用化学还原法制备金溶胶,加入氢氧化钠改变其 pH 值,并以自组装法制备金溶胶膜。通过比较金溶胶膜、pH ＝6及 pH ＝13时金溶胶溶液的增强因子以及在这三种金溶胶基底上结晶紫的检测限,分析不同活性基底增强效果的差异。三种活性基底的增强因子分别可达到5．9×103、1．5×105、2．3×107,pH ＝13的金溶胶溶液有最佳的增强效果。以这三种金溶胶为基底对结晶紫进行表面增强拉曼光谱探测,可得到检测限为70．7 nmol／L、9．6 nmol／L、1．8 nmol／L。结果表明,金溶胶溶液的增强效果明显优于金溶胶膜,而通过改变金溶胶体系的 pH 值可以改变金纳米颗粒的聚合程度及对探测物的吸附特性从而获得更高灵敏度的活性基底。     

Enhancing the efficiency of a PCR using gold nanoparticles

We found that the PCR could be dramatically enhanced by Au nanoparticles. With the addition of 0.7 nM of 13 nm Au nanoparticles into the PCR reagent, the PCR efficiency was increased. Especially when maintaining the same or higher amplification yields, the reaction time could be shortened, and the heating/cooling rates could be increased. The excellent heat transfer property of the nanoparticles should be the major factor in improving the PCR efficiency. Different PCR systems, DNA polymerases, DNA sizes and complex samples were compared in this study. Our results demonstrated that Au nanoparticles increase the sensitivity of PCR detection 5- to 10-fold in a slower PCR system (i.e. conventional PCR) and at least 10⁴-fold in a quicker PCR system (i.e. real-time PCR). After the PCR time was shortened by half, the 100 copies/µl DNA were detectable in real-time PCR with gold colloid added, however, at least 10⁶ copies/µl of DNA were needed to reach a detectable signal level using the PCR reagent without gold colloid. This innovation could improve the PCR efficiency using non-expensive polymerases, and general PCR reagent. It is a new viewpoint in PCR, that nanoparticles can be used to enhance PCR efficiency and shorten reaction times.     

A new amplification strategy for ultrasensitive electrochemical aptasensor with network-like thiocyanuric acid/gold nanoparticles

An ultrasensitive and highly specific electrochemical aptasensor for detection of thrombin based on gold nanoparticles and thiocyanuric acid is presented. For this proposed aptasensor, aptamerI was immobilized on the magnetic nanoparticles, aptamerII was labeled with gold nanoparticles. The magnetic nanoparticle was used for separation and collection, and gold nanoparticle offered excellent electrochemical signal transduction. Through the specific recognition for thrombin, a sandwich format of magnetic nanoparticle/thrombin/gold nanoparticle was fabricated, and the signal amplification was further implemented by forming network-like thiocyanuric acid/gold nanoparticles. A significant sensitivity enhancement had been obtained, and the detection limit was down to 7.82 aM. The presence of other proteins such as BSA and lysozyme did not affect the detection of thrombin, which indicates a high specificity of thrombin detection could be achieved. This electrochemical aptasensor is expected to have wide applications in protein monitoring and disease diagnosis.     

Effect of gold nanoparticle on the microscopic morphology of white blood cell

Background:  In medicine, there is limited knowledge on the toxicity of nanoparticles. In medicine, there has been limited knowledge on the effect of nanoparticles on the white blood cell.
Objective:  To evaluate the effect of gold nanoparticle on the microscopic morphology of white blood cell.
Setting:  Chulalongkorn Univesity, Bangkok, Thailand.
Method:  This study was performed as an experimental study. Mixture of gold nanoparticle solution and blood sample was prepared and analysed.
Result:  This work revealed that after mixing the blood sample with gold nanoparticle solution, accumulation of gold nanoparticle in the white blood cell was observed.
Conclusion:  The effect of gold nanoparticle on the white blood cell can be detected and this knowledge can be used in cytotoxic drug treatment.     

One-step preparation of antibody and oligonucleotide dual-labeled gold nanoparticle bio-probes and their properties

A novel method of one-step preparation of dual-labeled gold nanoparticle bio-probes was established by the electrostatic adsorption and the covalent bonding of gold nanoparticles with antibodies and thiol-modified oligonucleotides, respectively. Characterization of probes, the coverage and activity of antibodies and oligonucleotides on probe surfaces were detected. The results indicated that the gold nanoparticles labeled with antibodies and oligonucleotides possess good bioactivity and the coverage of oligonucleotide and antibody on a dual-labeled gold nanoparticle bio-probe was (92 ± 20) and (8 ± 3), respectively. The preparative method is simple and stable. The dual-labeled gold nanoparticle bio-probes have an application value in detection of ultramicro protein.     

Biogenic synthesis of floral-shaped gold nanoparticles using a novel strain,Talaromyces flavus

A biogenic route was adopted towards the synthesis of gold nanoparticles using the extract of a novel strain, Talaromyces flavus. Reduction of chloroauric acid by the fungal extract resulted in the production of gold nanoparticle, which was further confirmed by the concordant results obtained from UV–visible spectroscopy, energy dispersive spectroscopy (EDS), and dynamic light scattering (DLS) analysis. Morphology and the crystal nature of the synthesized nanoparticles were characterized using transmission electron microscopy (TEM), X-ray diffraction (XRD) and selected area electron diffraction (SAED). A direct correlation was observed between nanoparticle formation and the concentration of reducing agent present in the fungal extract. The time-dependent kinetic study revealed that the bioreduction process follows an autocatalytic reaction. Crystalline, irregular, and mostly flower-shaped gold nanoparticles with a mean hydrodynamic radius of 38.54?±?10.34 nm were obtained. pH played a significant role on production of mono-dispersed nanoparticle. FTIR analysis partially deciphered the involvement of –NH₂, ?SH, and –CO groups as the probable molecules in the bio-reduction and stabilization process. Compared to the conventional methods, a time-resolved, green, and economically viable method for floral-shaped nanoparticle synthesis was developed.     

The Effect of Ag Nanoparticles on Surface-Enhanced Luminescence from Au Nanovoid Arrays

Studies comparing the effect of adding two different nanoparticle compositions on the plasmonic properties of Au nanovoid arrays were undertaken. Surface-enhanced resonance luminescence and surface-enhanced resonance Raman studies comparing dispersed Ag nanoparticles and Ag nanoparticle aggregates on gold nanovoid arrays were undertaken. These studies showed that using Ag nanoparticle aggregates increased both luminescence and Raman efficiency relative to when dispersed nanoparticles were used; in addition, these studies also showed that adding dispersed Ag nanoparticles supported a more reproducible enhancement in luminescence and Raman across the substrate compared to using Ag nanoparticle aggregates. Finite element analysis simulations indicated that surface plasmon polariton distribution in the sample was affected by the presence of the Ag nanoparticles on the Au nanovoid array.     

A novel electroconductive interface based on Fe3O4 magnetic nanoparticle and cysteamine functionalized AuNPs: Preparation and application as signal amplification element to minoring of antigen‐antibody immunocomplex and biosensing of prostate cancer

In this study, a novel electroconductive interface was prepared based on Fe₃O₄ magnetic nanoparticle and cysteamine functionalized gold nanoparticle. The engineered interface was used as signal amplification substrate in the electrochemical analysis of antibody‐antigen binding. For this purpose, biotinilated‐anti‐prostate‐specific antigen (PSA) antibody was bioconjugated with iron oxide magnetic nanoparticles (Fe₃O₄) and drop‐casted on the surface of glassy carbon electrode (GCE). Also, secondary antibody (HRP‐Ab2) encapsulated on gold nanoparticles caped by cysteamine was immobilized on the surface of GCE modified electrode. A transmission electron microscopy images shows that a sandwich immunoreaction was done and binding of Ab1 and Ab2 performed successfully. Various parameters of immunoassay, including the loading of magnetic nanoparticles, the amount of gold nanoparticle conjugate, and the immunoreaction time, were optimized. The detection limit of 0.001 μg. L^?1 of PSA was obtained under optimum experimental conditions. It is found that such magneto‐bioassay could be readily used for simultaneous parallel detection of multiple proteins by using multiple inorganic metal nanoparticle tracers and are expected to open new opportunities for early stage diagnosis of cancer in near future.     

Direct electron transfer between cytochrome P450scc and gold nanoparticles on screen-printed rhodium-graphite electrodes

This paper is concerned with an investigation of electron transfer between cytochrome P450scc (CYP11A1) and gold nanoparticles immobilised on rhodium-graphite electrodes. Thin films of gold nanoparticles were deposited onto the rhodium-graphite electrodes by drop casting. Cytochrome P450scc was deposited onto both gold nanoparticle modified and bare rhodium-graphite electrodes. Cyclic voltammetry indicated enhanced activity of the enzyme at the gold nanoparticle modified surface. The role of the nanoparticles in mediating electron transfer to the cytochrome P450scc was verified using ac impedance spectroscopy. Equivalent circuit analysis of the impedance spectra was performed and the values of the individual components estimated. On addition of aliquots of cholesterol to the electrolyte bioelectrocatalytic reduction currents were obtained. The sensitivity of the nanoparticle modified biosensor to cholesterol was 0.13 microA microM-1 in a detection range between 10 and 70 microM of cholesterol. This confirms that gold nanoparticles enhance electron transfer to the P450scc when present on the rhodium-graphite electrodes.     

Macrophomina phaseolina: microbased biorefinery for gold nanoparticle production

Biofabrication of nanoparticles via the principles of green nanotechnology is a key issue addressed in nanobiotechnology research. There is a growing need for development of a synthesis method for producing biocompatible stable nanoparticles in order to avoid adverse effects in medical applications. We report the use of simple and rapid biosynthesis method for the preparation of gold nanoparticles using Macrophomina phaseolina (Tassi) Goid, a soil-borne pathogen. The effect of pH and temperature on the synthesis of gold nanoparticles by M. phaseolina was also assessed. Different techniques like UV-Visible Spectroscopy, Transmission Electron Microscopy (TEM), Dynamic light scattering (DLS) measurements, Fourier transform infrared (FTIR), and EDX were used to characterize the gold nanoparticles. The movement of these gold nanoparticles inside Escherichia coli (ATCC11103) along with effect on growth and viability was evaluated. The biogenic gold nanoparticle was synthesized at 37 °C temperature and neutral pH. UV-Visible Spectroscopy, TEM, EDX, and DLS measurements confirm the formation of 14 to 16 nm biogenic gold nanoparticles. FTIR substantiates the presence of protein capping on Macrophomina phaseolina-mediated gold nanoparticles. The non-toxicity of gold nanoparticles was confirmed by the growth and viability assay while the TEM images validated the entry of gold nanoparticles without disrupting the structural integrity of E. coli. Biogenic method for the synthesis of nanoparticles using fungi is novel, efficient, without toxic chemicals. These biogenic gold nanoparticles themselves are nontoxic to the microbial cells and offer a better substitute for drug delivery system.