Direct Electrochemistry Of Proteins Research Articles

Direct electrochemistry of proteins and nucleic acids: The focus on 3D structure

The current knowledge regarding direct electrochemistry of proteins and nucleic acids on solid electrodes is reviewed and discussed from the point of view of their 3D structure. The accent on the spatial organization of these biopolymers allows for deeper understanding of their redox transformations on electrode surface and for making some parallels with redox processes in living systems. The 3D concept of biopolymer electrochemistry is presented as having the evident practical potential, enabling to register such molecular events as complex formation, aggregation, and degradation, as well as post-translational or post-replicative modifications of proteins and nucleic acids, which are closely connected to various biochemical processes in health and disease. This mini review aims at drawing attention of bioelectrochemical community to problems related to the interpretation of oxidation and reduction signals from biomolecules which are polymeric and possess particular 3D structures. Also, a present lack of systematic studies in terms of ‘structure-properties’ in the field of direct biopolymer electrochemistry, coupled with a profound investigation of reaction mechanisms is emphasized.

Electrochemiluminescence-Repurposed Abiological Catalysts in Full Protein Tag for Ultrasensitive Immunoassay.

Being announced as one of the "2019 Top Ten Emerging Technologies in Chemistry" by IUPAC, the directed evolution of artificial metalloenzymes has led to a broad scope of abiotic processes. Here, inspired by those key proteins in bioluminescence, a rudimentary expression of bio-electrochemiluminescent (ECL) macromolecules was achieved via the complexation of zinc proto-porphyrin IX (ZnPPIX) within apo-hemoglobin (apo-Hb). A high-yield monochromic irradiation at 644 nm could be provoked potentiostatically from the reconstituted holo-HbZnPPIX in solutions. Its secondary structure integrity was elucidated by UV and circular dichroism spectrometry, while voltammetry-hyphenated surface plasmon resonance authenticated its ligation conservativeness in electrical fields. Further conjugation with streptavidin rendered a homogeneous Janus fusion of both receptor and reporter domains, enabling a new abiological catalyst-linked ECL bioassay. On the other hand, singular ZnPPIX inside each tetrameric subunit of Hb accomplished an overall signal amplification without the bother of luminogenic heterojunctions. This pH-tolerant and non-photobleaching optics was essentialized to be the unique configuration interaction between Zn and O2, by which the direct electrochemistry of proteins catalyzed the transient progression of O2 → O2·- → O2* + hυ selectively. Such principle was implemented as a signal-on strategy for the determination of a characteristic cancer biomarker, the vascular endothelial growth factor, resulting in competent performance at a low detection limit of 0.6 pg·mL-1 and a wide calibration range along with good stability and reliability in real practices. This simple mutation repurposed the O2-transport Hb in the erythrocytes of almost all vertebrates into a cluster of oxidoreductases with intrinsic ECL activity, which would enrich the chromophore library. More importantly, its genetically engineered variants may come in handy in biomedical diagnosis and visual electrophysiology.

Bacterial Respiratory Chain Diversity Reveals a Cytochrome c Oxidase Reducing O2 at Low Overpotentials.

Cytochrome c oxidases (CcOs) are the terminal enzymes in energy-converting chains of microorganisms, where they reduce oxygen into water. Their affinity for O2 makes them attractive biocatalysts for technological devices in which O2 concentration is limited, but the high overpotentials they display on electrodes severely limit their applicative use. Here, the CcO of the acidophilic bacterium Acidithiobacillus ferrooxidans is studied on various carbon materials by direct protein electrochemistry and mediated one with redox mediators either diffusing or co-immobilized at the electrode surface. The entrapment of the CcO in a network of hydrophobic carbon nanofibers permits a direct electrochemical communication between the enzyme and the electrode. We demonstrate that the CcO displays a μM affinity for O2 and reduces O2 at exceptionally high electrode potentials in the range of +700 to +540 mV vs NHE over a pH range of 4-6. The kinetics of interactions between the enzyme and its physiological partners are fully quantified. Based on these results, an electron transfer pathway allowing O2 reduction in the acidic metabolic chain is proposed.

Synthesis, characterization, and electrochemical applications of multifunctional Fe3O4@C–Au nanocomposites

Fabrication of multifunctional nanocomposites that are simultaneously magnetic, stable, and biocompatible is critical for their application in bioanalysis but remains a challenge. In this study, Fe3O4@C–Au nanocomposites were prepared by a rapid microwave-hydrothermal procedure followed by the layer-by-layer assembly for this intention. The morphologies, composition, phase structures, and magnetic properties of the nanocomposites were well-characterized by field emission scanning electron microscopy, transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectra, and superconducting quantum interference device. The nanocomposites have the sizes of about 200 nm in diameter. They integrate the advantages of the satisfactory stability of carbon shell, the good biocompatibility and conductivity of Au nanoparticles as well as the excellent magnetic properties of Fe3O4 core. Furthermore, a novel biosensor based on the multifunctional nanocomposites was fabricated after the immobilization of hemoglobin and the direct electrochemistry was studied. The proposed biosensor exhibits wide linear range, high sensitivity, good reproducibility, and stability. The presented multifunctional nanocomposites provide an alternative and effective matrix for the direct electrochemistry of proteins and might have potential applications in biosensing and drug delivery.

A novel mesoporous silica nanosphere matrix for the immobilization of proteins and their applications as electrochemical biosensor

A mesoporous silica nanoshpere (MSN) was proposed to modify glassy carbon electrode (GCE) for the immobilization of protein. Using glucose oxidase (GOD) as a model, direct electrochemistry of protein and biosensing at the MSN modified GCE was studied for the first time. The MNS had large surface area and offered a favorable microenvironment for facilitating the direct electron transfer between enzyme and electrode surface. Scanning electron microscopy, transmission electron microscopy, UV–vis spectroscopy and cyclic voltammetry were used to examine the interaction between GOD and the MSN matrix. The results demonstrated that the immobilized enzyme on the MSN retained its native structure and bioactivity. In addition, the electrochemical reaction showed a surface controlled, reversible two-proton and two-electron transfer process with the apparent electron transfer rate constant of 3.96s−1. The MNS-based glucose biosensor exhibited the two linear ranges of 0.04–2.0mM and 2.0–4.8mM, a high sensitivity of 14.5mAM−1cm−2 and a low detection limit of 0.02mM at signal-to-noise of 3. The proposed biosensor showed excellent selectivity, good reproducibility, acceptable stability and could be successfully applied in the reagentless detection of glucose in real samples at −0.45V. The work displayed that mesoporous silica nanosphere provided a promising approach for immobilizing proteins and fabrication of excellent biosensors.

Electroanalysis in micro- and nano-scales

The tremendous advances in nanoscience and nanoengineering have sparked the accelerated development of electroanalysis towards highly sensitive and specific detections. In particular, novel nanomaterials with unique physical and chemical properties have been synthesized for electroanalysis in micro- and nano-scales to qualitatively and/or quantitatively examine the changes of compositions and structures for scientific insights and practical applications. In this review, we mainly survey our recent works on electroanalysis in micro- and nano-scales, which covers micro- and nano-fabrication techniques, microelectrodes and powder microelectrodes, direct electrochemistry of proteins, electrochemical analysis of living cells, and electrochemical immunoassay, with emphasis on the performance improvement by engineered functional nanomaterials. These electroanalytical methods have demonstrated their ability and reliability to study fundamental science and provide practical applications. It is our intention that this review could attract increasing interest to explore new electroanalytical methods in micro- and nano-scales and further develop broad important applications for critical chemical/biological sciences.

Novel urchin-like In2O3–chitosan modified electrode for direct electrochemistry of glucose oxidase and biosensing

A novel urchin-like In2O3–chitosan modified glassy carbon electrode (GCE) is for the first time prepared. The direct electrochemistry of glucose oxidase (GOD) immobilized on the surface of modified GCE and biosensing are studied. The urchin-like In2O3 nanostructure-based matrix has large specific surface area and provides a favorable and biocompatible microenvironment for promoting the direct electron transfer between proteins and electrode surface. The properties of different modified electrode are characterized by scanning electron microscopy (SEM), electrochemical impedance spectra (EIS), UV–vis spectroscopy, Fourier transform infrared spectroscopy (FTIR) and cyclic voltammetry, respectively. The constructed glucose biosensor shows wide linear range (5.0 × 10−6 to 1.3 × 10−3 M), low detection limit (1.9 × 10−6 M), a Michaelis–Menten constant of 0.37 mM. The proposed biosensor has good sensitivity, excellent selectivity, good reproducibility and stability. This urchin-like In2O3–chitosan matrix provides a new approach and efficient matrix for the direct electrochemistry of proteins and developing novel types of biosensors.

Direct electron transfer of cytochrome C and its electrocatalytic properties on multiwalled carbon nanotubes/ciprofloxacin films

In this study, stable and homogenous thin films of multiwalled carbon nanotubes (MWCNTs) were obtained on conducting surface using ciprofloxacin (CF, fluoroquinolone antibiotic) as an effective-dispersing agent. Further, MWCNTs/CF film modified electrodes (glassy carbon and indium tin oxide-coated glass electrode) are used successfully to study the direct electrochemistry of proteins. Here, cytochrome C (Cyt-C) was used as a model protein for investigation. A MWCNTs/CF film modified electrode was used as a biocompatible material for immobilization of Cyt-C from a neutral buffer solution (pH 7.2) using cyclic voltammetry (CV). Interestingly, Cyt-C retained its native state on the MWCNTs/CF film. The Cyt-C adsorbed MWCNTs/CF film was characterized by scanning electron microscopy (SEM), UV–visible spectrophotometry (UV-vis) and CV. SEM images showed the evidence for the adsorption of Cyt-C on the MWCNTs/CF film, and UV–vis spectrum confirmed that Cyt-C was in its native state on MWCNTs/CF film. Using CV, it was found that the electrochemical signal of Cyt-C was highly stable in the neutral buffer solution and its redox peak potential was pH dependent. The formal potential (−0.27 V) and electron transfer rate constant (13 ± 1 s−1) were calculated for Cyt-C on MWCNTs/CF film modified electrode. A potential application of the Cyt-C/MWCNTs/CF electrode as a biosensor to monitor H2O2 has been investigated. The steady-state current response increases linearly with H2O2 concentration from 2 × 10−6 to 7.8 × 10−5 M. The detection limit for determination of H2O2 has been found to be 1.0 × 10−6 M (S/N = 3). Thus, Cyt-C/MWCNTs/CF film modified electrode can be used as a biosensing material for sensor applications.

Electrodeposition of Co Nanoparticles on the Carbon Ionic Liquid Electrode as a Platform for Myoglobin Electrochemical Biosensor

The electrochemical deposition of Co nanoparticles on carbon ionic liquid electrode (CILE) was described and further used as the platform to construct a myoglobin (Mb) electrochemical biosensor. CILE was prepared by mixing a certain ratio of carbon powder, 1-ethyl-3-methylimidazolium tetrafluoroborate (EMIMBF4), and liquid paraffin together. The presence of ionic liquid on the electrode surface facilitated the formation of Co nanoparticles, and a layer of Co nanoparticles was deposited on the surface of CILE with the average diameter of 300 nm after the cyclic voltammetic scan in the CoCl2 solution. The formed Co/CILE was used as a new basal electrode for the investigation on the direct electrochemistry of protein. Mb molecules were further cast on the surface of Co/CILE and immobilized with Nafion film. The fabricated Nafion/Mb/Co/CILE showed good electrochemical behaviors with a pair of well-defined quasi-reversible redox peaks of Mb obtained, which was attributed to the electrochemical reaction of heme Fe(III)/Fe(II) redox couples. The results indicated that the presence of Co nanoparticles exhibited great promotion to the direct electron transfer of Mb. The Mb electrochemical biosensor showed good electrocatalytic activity to the reduction of hydrogen peroxide and trichloroacetic acid (TCA). The modified electrodes showed good stability and reproducibility, which had potential application in third generation biosensor.

Evaporative deposition of lipophilic quantum dots for an enzyme modified electrode

Lipophilic quantum dots were placed on the surface of a horseradish peroxidase (HRP) modified glassy carbon (GC) electrode by evaporative deposition. Direct electron transfer of immobilized HRP was found to occur. The deposition of quantum dots (QDs) was characterized by various ways, and the results indicated that the direct electron transfer of HRP was promoted preferably when QDs were deposited for 7 h at 35 °C. Under these conditions, the average coverages of QDs and HRP on the electrode surface is 56 nmol cm−2 and 65 pmol cm−2, respectively, and the resulting heterogeneous electron transfer rate constant k of HRP is 6.0 s−1. This work provides a new clue for research on the direct electrochemistry of proteins, and on biosensors based on it.

Direct electrochemistry study of glucose oxidase on Pt nanoparticle-modified aligned carbon nanotubes electrode by the assistance of chitosan–CdS and its biosensoring for glucose

Aligned carbon nanotubes (ACNTs) electrode has been developed for the direct protein electrochemistry and enzyme-biosensor study involving two types of nanoparticles. Pt nanoparticles (Pt nano) were electro-modified on the ACNTs’ each tube, greatly increasing the electrode surface area for locating protein and also its electronic transfer ability. Glucose oxidase (GOD) with chitosan (CS) and CdS nanoparticles electrochemically coated on each tube of ACNTs–Pt nano by the electrodeposition reaction of CS when pH value passing its pKa. The CdS nanoparticles between ACNTs electrode and GOD have stimulated the GOD’s direct electron transfer during its redox reaction of FAD/FADH 2. The CS–GOD–CdS/ACNTs–Pt nano electrode also offer sensitive response to the substrate of glucose with detection limit of 46.8 μM ( S/ N = 3) and apparent Michaelis–Menten constant of 11.86 mM.

Carboxyphenyl Covalent Immobilization of Heme Proteins and its Favorable Biocompatible Electrochemical and Electrocatalytic Characteristics

AbstractA novel carboxyphenyl covalent immobilization technique has been successfully developed to realize the effective attachment of two typical heme proteins, hemoglobin (Hb) and cytochrome c (Cyt‐c), onto underlying glassy carbon electrode (GCE). Primarily, the GCE surface is functionalized with aromatic 4‐carboxyphenyl (4‐CP) group by the electrochemical reduction of diazonium cations, producing covalently linked carboxyl‐terminated active GCE surface to work as a ‘bridge’. Then, Hb and Cyt‐c are readily attached to GCE through the ‘bridge’ by functional covalently combination between NH2 terminal groups of proteins and COOH terminal groups of 4‐CP, obtaining Hb/4‐CP/GCE and Cyt‐c/4‐CP/GCE. On both electrodes, well‐defined peaks attributing to the FeIII/FeII couple of heme group of Hb and Cyt‐c are clearly observed with the electron transfer rate constant (ks) evaluated to be 2.48±0.05 s−1 and 2.73±0.05 s−1, respectively. It is attractive that the formal potential (E°') of the immobilized Hb and Cyt‐c are estimated to be 50 and 100 mV (vs. SCE), respectively, which are closer to the standard redox potential of native Hb and Cyt‐c in solution, owing to the good biocompatibility of 4‐CP groups. The electrodes also exhibit fast response, high sensitivity and well stability for the amperometric detection of H2O2 at a fairly mild potential of 0 V without any mediators, obtaining rather small apparent Michaelis‐Menten constant (KMapp) values of 113 μM for Hb/4‐CP/GCE and 101 μM for Cyt‐c/4‐CP/GCE. All the experimental results indicated that the covalent graft 4‐carboxyphenyl group plays an important role in constructing a “biocompatible bridge” to help the direct electron transfer of Hb and Cyt‐c with favorable biocompatibility and good bio‐ electrocatalytic affinity in virtue of the substituted aryl group only consisting of C, H and O elements, which is similar with the constitutes of organics. It makes the system of functionalized covalent immobilization of proteins onto carbon electrode a promising platform for fabricating the third‐generation biosensors. A new approach for realizing direct electrochemistry of proteins, as well as design of novel bioelectronic devices has been accordingly provided.

Direct electrochemistry behavior of Cytochrome c on silicon dioxide nanoparticles-modified electrode

A newfangled direct electrochemistry behavior of Cytochrome c (Cyt c) was found on glassy carbon (GC) electrode modified with the silicon dioxide (SiO2) nanoparticles by physical adsorption. A pair of stable and well-defined redox peaks of Cyt c′ quasi-reversible electrochemical reaction were obtained with a heterogeneous electron transfer rate constant of 1.66×10−3 cm/s and a formal potential of 0.069 V (vs. Ag/AgCl) (0.263 V versus NHE) in 0.1 mol/L pH 6.8 PBS. Both the size and the amount of SiO2 nanoparticles could influence the electron transfer between Cyt c and the electrode. Electrostatic interaction which is between the negative nanoparticle surface and positively charged amino acid residues on the Cyt c surface is of importance for the stability and reproducibility toward the direct electron transfer of Cyt c. It is suggested that the modification of SiO2 nanoparticles proposes a novel approach to realize the direct electrochemistry of proteins.

Assembly of myoglobin layer-by-layer films with poly(propyleneimine) dendrimer-stabilized gold nanoparticles and its application in electrochemical biosensing

The small-sized Au nanoparticles (3nm) were prepared by reduction of HAuCl4 in the presence of poly(propyleneimine) (PPI) dendrimers, forming the stable PPI–Au nanoclusters in aqueous medium. The PPI–Au nanoclusters might take a kind of “core–shell” structure, in which several PPI molecules were attached on the surface of one gold nanoparticle. The PPI–Au nanoclusters in aqueous dispersions and myoglobin (Mb) in its buffers at pH 5.0 were then alternately adsorbed on the surface of pyrolytic graphite (PG) electrodes and other solid substrates, forming {PPI–Au/Mb}n layer-by-layer films, which was confirmed by cyclic voltammetry (CV) and quartz crystal microbalance (QCM). {PPI–Au/Mb}n films on PG electrodes demonstrated a pair of well-defined and quasi-reversible CV reduction-oxidation peaks for Mb heme FeIII/FeII couple and good electrocatalytic properties toward reduction of oxygen and hydrogen peroxide. Compared with {Au/Mb}n multilayer films containing no dendrimers and {PAMAM/Mb}n films assembled by polyamidoamine (PAMAM) dendrimers and Mb but in the absence of Au nanoparticles, {PPI–Au/Mb}n films showed better electrochemical behaviors and catalytic performances, which may be attributed to the unique structure of PPI–Au nanoclusters and good conductivity of gold nanoparticles. This novel kind of protein multilayer films assembled with dendrimer-stabilized gold nanoparticles may provide a new and general approach to fabricate the biosensors and bioreactors based on the direct electrochemistry of proteins or enzymes.

Free Energy of Transition for the Individual Alkaline Conformers of Yeast Iso-1-cytochrome c,

Direct protein electrochemistry was used to obtain the thermodynamic parameters of transition from the native (state III) to the alkaline (state IV) conformer for untrimethylated Saccharomyces cerevisiae iso-1-cytochrome c expressed in E. coli and its single and multiple lysine-depleted variants. In these variants, one or more of the lysine residues involved in axial Met substitution (Lys72, Lys73, and Lys79) was mutated to alanine. The aim of this work is to determine the thermodynamic affinity of each of the substituting lysines for the heme iron and evaluate the interplay of enthalpic and entropic factors. The equilibrium constants for the deprotonation reaction of Lys72, 73, and 79 were computed for the minimized MD average structures of the wild-type and mutated proteins, applying a modified Tanford-Kirkwood calculation. Solvent accessibility calculations for the substituting lysines in all variants were also performed. The transition enthalpy and entropy values within the protein series show a compensatory behavior, typical of a process involving extensive solvent reorganization effects. The experimental and theoretical data indicate that Lys72 most readily deprotonates and replaces M80 as the axial heme iron ligand, whereas Lys73 and Lys79 show comparably higher pKa values and larger transition free energies. A good correlation is found within the series between the lowest calculated Lys pKa value and the corresponding experimental pKa value, which can be interpreted as indicative of the deprotonating lysine itself acting as the triggering group for the conformational transition. The triple Lys to Ala mutant, in which no lysine residues are available for heme iron binding, features transition thermodynamics consistent with a hydroxide ion replacing the axial methionine ligand.

A Study of Protein Electrochemistry on a Supported Membrane Electrode

Protein electrochemistry offers a direct method to identify and characterize biological electron transfer processes, potentially leading to commercial applications such as biosensors and diagnostic tools. However, establishing a biocompatible electrode interface that maintains the native state of the redox protein involves several challenges. In general, membrane proteins require the presence of a phospholipid bilayer to maintain their biological activity. Synthetic `biomimetic’ membranes are widely used to characterize membrane proteins, however they have seldom been applied to measurements of protein redox activity in electrochemical cells due to their inherent insulating property. In this study we demonstrate the use of the phospholipids: PC, PC/PG and PC/PG/cholesterol membrane mixtures on chemically modified (supported) gold electrode surfaces for direct protein electrochemistry. We compare the electrochemical activity of a relatively small, redox active “test protein”, cytochrome c, in the presence and absence of phospholipid on a gold electrode modified with thiol self assembled monolayers, to explore the effect of chain length and composition of the thiol on the charge coupling. Three thiols were investigated as self assembled monolayers on a gold electrode: octanethiol, mercaptopropionic and mercaptoundecanoic acid. We demonstrate here that the charge transfer efficiency of cytochrome c is better in the presence of the membrane and in addition, a superior redox response is obtained with surfaces modified with a thiol functionalised with a carboxylic acid.

Assembly of layer-by-layer films of heme proteins and single-walled carbon nanotubes: electrochemistry and electrocatalysis

After being treated by mixed acids, single-walled carbon nanotubes (SWNTs) were shortened and had negatively charged groups on the surface. Positively charged hemoglobin or myoglobin at pH 5.0 was successfully assembled with SWNTs into layer-by-layer films on solid surfaces, designated as {SWNT/protein}n. While only those proteins in the first few bilayers closest to the electrode surface exhibited electroactivity, the {SWNT/protein}n films demonstrated a much higher fraction of electroactive proteins and better controllability in film construction compared with cast films of the proteins and carbon nanotubes. The proteins in the {SWNT/protein}n films retained their near-native structure at medium pH. The stable protein film electrode showed good electrocatalytic properties toward reduction of oxygen and hydrogen peroxide, demonstrating the potential application of the {SWNT/protein}n films as a new type of biosensor based on the direct electrochemistry of proteins without using mediators.

Electrochemistry and Electroanalytical Applications of Carbon Nanotubes: A Review

This review addresses recent developments in electrochemistry and electroanalytical chemistry of carbon nanotubes (CNTs). CNTs have been proved to possess unique electronic, chemical and structural features that make them very attractive for electrochemical studies and electrochemical applications. For example, the structural and electronic properties of the CNTs endow them with distinct electrocatalytic activities and capabilities for facilitating direct electrochemistry of proteins and enzymes from other kinds of carbon materials. These striking electrochemical properties of the CNTs pave the way to CNT-based bioelectrochemistry and to bioelectronic nanodevices, such as electrochemical sensors and biosensors. The electrochemistry and bioelectrochemistry of the CNTs are summarized and discussed, along with some common methods for CNT electrode preparation and some recent advances in the rational functionalization of the CNTs for electroanalytical applications.

Thermodynamics of the acid transition in blue copper proteins.

The thermodynamic parameters of the conformational transition occurring at low pH (acid transition, AT) in blue copper proteins, involving protonation and detachment from the Cu(I) ion of one histidine ligand, have been determined electrochemically for spinach and cucumber plastocyanins, Rhus vernicifera stellacyanin, cucumber basic protein (CBP), and Paracoccus versutus amicyanin. These data were obtained from direct protein electrochemistry experiments carried out at varying pH and temperature. For all species but CBP, the overall conformational change turns out to be exothermic. The entropy change is remarkably species-dependent. In particular, we found that (i) the balance of bond breaking/formation favors the acid transition in plastocyanins, which show remarkably negative DeltaH degrees '(AT) values, and (ii) the transition enthalpy turns out to be much less negative (or even positive) for the two phytocyanins (stellacyanin and CBP): for these species, the transition turns out to be observable thanks to the favorable (positive) entropy change. Thus, it is apparent that the thermodynamic "driving force" for this transition is enthalpic for the plastocyanins and entropic for the phytocyanins. Amicyanin is an intermediate case in which both enthalpic and entropic terms favor the transition. Under the assumption that the transition entropy originates from solvent reorganization effects, which are known to involve compensative enthalpy and entropy changes, the free energy change of the transition would also correspond to the enthalpy change due to bond breaking/formation in the first coordination sphere of the metal and in its immediate environment. Indeed, this term turns out to be very similar for the proteins investigated, in line with the conservation of the Cu(I)-His bond strengths in these species, except for amicyanin, for which the greater exothermicity of the transition can be ascribed to peculiar features of the active site.

Direct electron transfer between hemoglobin and a glassy carbon electrode facilitated by lipid-protected gold nanoparticles

We synthesized a kind of gold nanoparticle protected by a synthetic lipid (didodecyldimethylammonium bromide, DDAB). With the help of these gold nanoparticles, hemoglobin can exhibit a direct electron transfer (DET) reaction. The formal potential locates at −169 mV vs. Ag/AgCl. Spectral data indicated the hemoglobin on the electrode was not denatured. The lipid-protected gold nanoparticles were very stable (for at least 8 months). Their average diameter is 6.42 nm. It is the first time to use monolayer-protected nanoparticles to realize the direct electrochemistry of protein.