Ultrasensitively sensing acephate using molecular imprinting techniques on a surface plasmon resonance sensor

An ultrathin molecularly imprinted polymer film was anchored on an Au surface for fabricating a surface plasmon resonance sensor sensitive to acephate by a surface-bound photo-radical initiator. The polymerization in the presence of acephate resulted in a molecular-imprinted matrix for the enhanced binding of acephate. Analysis of the SPR wavenumber changes in the presence of different concentrations of acephate gave a calibration curve that included the ultrasensitive detection of acephate by the imprinted sites in the composite, K_ass for the association of acephate to the imprinted sites, 7.7 × 10¹² M⁻¹. The imprinted ultrathin film revealed impressive selectivity. The selectivity efficiencies for acephate and other structurally related analogues were 1.0 and 0.11-0.37, respectively. Based on a signal to noise ratio of 3, the detection limits were 1.14 × 10⁻¹³ M for apple sample and 4.29 × 10⁻¹⁴ M for cole sample. The method showed good recoveries and precision for the apple and cole samples spiked with acephate solution. This suggests that a combination of SPR sensing with MIP film is a promising alternative method for the detection of organophosphate compounds.     

Detection of parathion methyl using a surface plasmon resonance sensor combined with molecularly imprinted films

An ultra-sensitive and highly selective parathion methyl(PM) detection method by surface plasmon resonance(SPR) combined with molecularly imprinted films(MIF) was developed. The PM-imprinted film was prepared by thermo initiated polymerization on the bare Au surface of an SPR sensor chip.Template PM molecules were quickly removed by an organic solution of acetonitrile/acetic acid(9:1,v/v), causing a shift of 0.58 in SPR angle. In the concentrations range of 10à13–10à10mol/L, the refractive index showed a gradual increase with higher concentrations of template PM and the changes of SPR angles were linear with the negative logarithm of PM concentrations. In the experiment, the minimum detectable concentration was 10à13mol/L. The selectivity of the thin PM-imprinted film against diuron,tetrachlorvinphose and fenitrothion was examined, but no observable binding was detected. The results in the experiment suggested that the MIF had the advantages of high sensitivity and selectivity.     

A novel method to prepare SPR sensor chips based on photografting molecularly imprinted polymer

A novel method to prepare surface plasmon resonance(SPR) sensor chips based on grafted imprinted polymer is explored. Benzophenone photografting system is used to grow molecularly imprinted polymer(MIP) films from the modified surface of gold substrate.The surface morphology and thickness of MIP films were investigated by scanning electronic microscope(SEM).The adsorption properties of sensor chip were studied by SPR spectroscopy.The results demonstrate that nano-MIP films can be constructed on the surface of gold substrate with the good adsorption of template molecules.     

Surface plasmon resonance sensor chips for the recognition of bovine serum albumin via electropolymerized molecularly imprinted polymers

In this paper,a surface plasmon resonance(SPR)sensor chip for detection of bovine serum album(BSA)was prepared by electropolymerization of 3-aminophenylboronic acid(3-APBA)based on molecularly imprinted polymer(MIP)technique.The surface morphology of MIP and non-imprinted(NIP)flms were characterized by scanning electroscopy(SEM).SEM images exhibited nanoscale cavities formed on the MIP films surface homogeneously due to the removal of BSA templates.The effects of pH,ion strength of rebinding BSA,the specific binding and selective recognition were studied for MIP films.Results indicated that the BSA-imprinted films exhibited a good adsorption of template protein(0.02–0.8 mg/mL)in0.05 mol/L sodium phosphate buffer at pH 5.0 with the limit of detection(LOD)of 0.02 mg/mL.     

Recent advances in surface plasmon resonance based techniques for bioanalysis

Surface plasmon resonance (SPR) is a powerful and versatile spectroscopic method for biomolecular interaction analysis (BIA) and has been well reviewed in previous years. This updated 2006 review of SPR, SPR spectroscopy, and SPR imaging explores cutting-edge technology with a focus on material, method, and instrument development. A number of recent SPR developments and interesting applications for bioanalysis are provided. Three focus topics are discussed in more detail to exemplify recent progress. They include surface plasmon fluorescence spectroscopy, nanoscale glassification of SPR substrates, and enzymatic amplification in SPR imaging. Through these examples it is clear to us that the development of SPR-based methods continues to grow, while the applications continue to diversify. Major trends appear to be present in the development of combined techniques, use of new materials, and development of new methodologies. Together, these works constitute a major thrust that could eventually make SPR a common tool for surface interaction analysis and biosensing. The future outlook for SPR and SPR-associated BIA studies, in our opinion, is very bright. Surface plasmon resonance (SPR) is a powerful and versatile spectroscopic method for biomolecular interaction analysis (BIA) and has been well reviewed in previous years. This updated 2006 review of SPR, SPR spectroscopy, and SPR imaging explores cutting-edge technology with a focus on material, method, and instrument development. A number of recent SPR developments and interesting applications for bioanalysis are provided. Three focus topics are discussed in more detail to exemplify recent progress. They include surface plasmon fluorescence spectroscopy, nanoscale glassification of SPR substrates, and enzymatic amplification in SPR imaging. Through these examples it is clear to us that the development of SPR-based methods continues to grow, while the applications continue to diversify. Major trends appear to be present in the development of combined techniques, use of new materials, and development of new methodologies. Together, these works constitute a major thrust that could eventually make SPR a common tool for surface interaction analysis and biosensing. The future outlook for SPR and SPR-associated BIA studies, in our opinion, is very bright.      

Planar 2,2′-bithiophenes with 3,3′- and 3,3′,4,4′-substituents. A computational study

Ab initio calculations have shown that when carbonyl groups are incorporated into the 3- and 3′-positions of bithiophenes, as part of five-membered rings, the bithiophene system is planar. This is due to electrostatic attraction between the carbonyl oxygen and the sulfur atom in the adjacent ring. In the analogous systems containing a CH₂ group in place of the carbonyl, or containing six-membered rather than five-membered rings, the bithiophenes are twisted. This has implications for preparing planar polythiophenes.     

Visual and surface plasmon resonance sensor for zirconium based on zirconium-induced aggregation of adenosine triphosphate-stabilized gold nanoparticles

Owing to its high affinity with phosphate, Zr(IV) can induce the aggregation of adenosine 5′-triphosphate (ATP)-stabilized AuNPs, leading to the change of surface plasmon resonance (SPR) absorption spectra and color of ATP-stabilized AuNP solutions. Based on these phenomena, visual and SPR sensors for Zr(IV) have been developed for the first time. The A_660 nm/A_518 nm values of ATP-stabilized AuNPs in SPR absorption spectra increase linearly with the concentrations of Zr(IV) from 0.5 μM to 100 μM (r = 0.9971) with a detection limit of 95 nM. A visual Zr(IV) detection is achieved with a detection limit of 30 μM. The sensor shows excellent selectivity against other metal ions, such as Cu²⁺, Fe³⁺, Cd²⁺, and Pb²⁺. The recoveries for the detection of 5 μM, 10 μM, 25 μM and 75 μM Zr(IV) in lake water samples are 96.0%, 97.0%, 95.6% and 102.4%, respectively. The recoveries of the proposed SPR method are comparable with those of ICP-OES method.     

Investigation of 3,3′,5,5′-tetramethylbenzidine as colorimetric substrate for a peroxidatic DNAzyme

In this work, the suitability of 3,3′,5,5′-tetramethylbenzidine sulfate (TMB) as the substrate of a DNAzyme catalytic system composed of a guanine-quadruplex DNA molecule and hemin was investigated. In the presence of H₂O₂, the hemin-DNA complex catalyzes the oxidation of TMB to produce two colored products, much like a peroxidase. The color-generating activity of this system could be influenced by several factors such as buffer type, pH value, DNA sequence, reaction time, and concentrations of both the hemin and H₂O₂. To illustrate the utility of this catalytic system, we designed a colorimetric assay, in which a synthetic oligonucleotide with a sequence complementary to the G-quadruplex DNA was used as the target. A detection limit of 1.86 nM was obtained. Our data have shown that TMB was an excellent colorimetric indicator that reported the peoxidase activities of the widely studied hemin-G-quadruplex DNAzyme system.     

A palm-sized surface plasmon resonance sensor with microchip flow cell

A small-sized surface plasmon resonance (SPR) sensor with a microchip flow cell has been developed for the purpose of enhancing the sensitivity of the SPR detector for low molecular weight compounds. This portable differential SPR detector consisted of an LED, two cylindrical lenses, a round prism, a divided mirror, a CCD, electronics, and a polydimethylsiloxane/gold microchip with two flow paths (10 mm long, 1 mm wide, 20-100 μm deep). 3-Mercaptopropyltrimethoxysilane was used for sealing the microchip. The performance of the on-site orientated SPR detector was estimated using sucrose and IgA. A drastic change in the SPR intensity appeared. The depth of the flow cell was in inverse proportion to the SPR intensity. Compared to a conventional flow cell having the size of 10 mm (L) × 1 mm (W) × 1 mm (D), its sensitivity to 10% sucrose and 0.9 nM IgA increased about 11 and 39 times, respectively. This phenomenon seemed to be due to the increase in the substance on the SPR sensor based on its size effect. These results showed that the application of the microchip sensor for SPR measurement has the possibility for improvement of the SPR intensity for low molecular substances.     

Single-crystal sapphire-fiber optic sensors based on surface plasmon resonance spectroscopy for in situ monitoring

Single-crystal sapphire-fiber optic sensors based on surface plasmon resonance (SPR) for refractive index (RI) measurements of aqueous and hydrothermal water solutions are described. Accurate measurement of RIs is essential to efficient operation and control of broad range of engineering processes. Some of these processes are carried out with harsh environments, such as high-temperature, high pressure, and chemical corrosion. These extreme physical conditions are proving a limiting factor in application of the conventional silica-based optical sensors. Single-crystal sapphire is an ideal material for sensor applications, where reliable performance is required in the extreme environment conditions. With regard to the liquid species detection, most applications of SPR sensors are designed to function near the refractive index of water (1.3330 RI). The RI changes of aqueous solution can be easily monitored by silica-fiber (RI, 1.4601 at 550 nm) based SPR sensor. However, the sapphire waveguide has a prohibitively high RI (1.7708 at 546 nm) for unmodified monitoring of the RI changes of aqueous solutions. For that purpose, a practical SPR probe geometry has been applied to the ability to tune the SPR coupling wavelength/angle pair with sapphire-fiber based SPR probe.     

Surface plasmon resonance sensor for lysozyme based on molecularly imprinted thin films

Molecularly imprinted polymers (MIPs) selective for lysozyme were prepared on SPR sensor chips by radical co-polymerization with acrylic acid and N,N′-methylenebisacrylamide. Gold-coated SPR sensor chips were modified with N,N′-bis(acryloyl)cystamine, on which MIP thin films were covalently conjugated. The presence of NaCl during the polymerization and the re-binding tests affected the selectivity and the optimization of NaCl concentration in the pre-polymerization mixture and the re-binding buffer could enhance the selectivity in the target protein sensing. When the lysozyme-imprinted polymer thin films were prepared in the presence of 40 mM NaCl, the selectivity factor (target protein bound/reference protein bound) of MIP in the re-binding buffer containing 20 mM NaCl was 9.8, meanwhile, that of MIP in the re-binding buffer without NaCl was 1.2. A combination of SPR sensing technology with protein-imprinted thin films is a promising tool for the construction of selective protein sensors.     

Present and future of surface plasmon resonance biosensors

Surface plasmon resonance (SPR) biosensors are optical sensors exploiting special electromagnetic waves—surface plasmon-polaritons—to probe interactions between an analyte in solution and a biomolecular recognition element immobilized on the SPR sensor surface. Major application areas include detection of biological analytes and analysis of biomolecular interactions where SPR biosensors provide benefits of label-free real-time analytical technology. This paper reviews fundamentals of SPR affinity biosensors and discusses recent advances in development and applications of SPR biosensors.     

Measurement uncertainty in analytical studies based on surface plasmon resonance

The sensitivity of surface plasmon resonance (SPR) transducers depends on the thickness and spatial organization of interfacial structures at their surfaces. This is because the response of the SPR sensor is determined by integrating the distance-dependent refractive index (spatial interfacial architectures), weighted by the square of the electromagnetic field, from zero to infinite distance. The effect of SPR transducer sensitivity variation on the accuracy of SPR analysis is considered. Our quantitative estimation (based on the results of refractometric studies) gave a value for sensitivity variation of about 3% for the formation of a self-assembled thiocyanate layer or a trypsin-soybean trypsin inhibitor surface complex. The estimated accuracy in measured variation (i.e., by 0.01) for the refractive index of the external medium was 3 × 10⁻⁴. This restriction, which follows immediately from the physical mechanism of the SPR phenomenon, should be taken into account when analyzing data obtained with the above technique.     

Organic vapour sensing using localized surface plasmon resonance spectrum of metallic nanoparticles self assemble monolayer

The response of localized surface plasmon resonance (LSPR) spectra of gold and silver nanoparticles, and gold nanoshells to organic vapors was investigated. The surface area of nanomaterials was sufficiently high for quantitative adsorption of volatile organic compounds (VOCs). Surface adsorption and condensation of VOCs caused the environmental refractive index to increase from n = 1.00 in pure air to as high as n = 1.29 in near saturated toluene vapor. The extinction and wavelength shift of the LSPR spectra were very sensitive to changes in the surface refractive index of the nanoparticles. Responses of the LSPR band were measured with a real-time UV-vis spectrometer equipped with a CCD array detector. The response of silver nanoparticles to organic vapors was most sensitive in changes in extinction, while gold nanoshells exhibited red-shifts in wavelength (∼250 nm/RIU) when exposed to organic vapors. The LSPR spectral shifts primarily were determined by the volatility and refractive indices of the organic species. The T₉₀ response time of the VOC-LSPR spectrum was less than 3 s and the response was completely reversible and reproducible.     

Synthesis and crystalline state photochromism of 3,3′-diaryl biindenylidenedione derivatives

A new series of 3,3′-diaryl biindenylidenedione derivatives were synthesized through Grignard reaction. Some of their stereoisomers were obtained by photochemical transformation upon heating and a plausible reaction mechanism was proposed. Most of these compounds exhibited photochromism in crystalline states as well as generation of stable organic radicals. The absolute configurations of the stereoisomers were determined by single crystal X-ray crystallography. The results showed that the position of substituent could dramatically affect molecular structure and photochemical properties of the biindenylidenedione derivatives.     

Biomimetic properties and surface studies of a piezoelectric caffeine sensor based on electrosynthesized polypyrrole

An approach for preparing a chemical sensor for caffeine through the combination of molecularly imprinted polypyrrole and a piezoelectric quartz transducer was proposed. The caffeine-imprinted polymer was synthesized using galvanostatic electropolymerization of pyrrole monomer directly onto one of the gold electrodes of a 9 MHz AT-cut quartz crystal in the presence of caffeine. The optimum conditions for the electrosynthesis of the reagent phase were established. Caffeine molecules were entrapped in the matrix of polymer film, and were removed by subsequent washing with water, leaving behind pores capable of recognizing the target analyte molecule.The caffeine sensor was fixed in a measuring cell and measurement of the resonant frequency of the quartz crystal as it comes in contact with the caffeine solution was carried out in a stopped flow mode. A steady-state response was achieved in about 10 min. The sensor exhibited a linear relationship between the frequency shift and the ln of caffeine concentration in the range of 0.1-10 mg/mL (correlation coefficient, r = 0.9882). The sensitivity of the sensor was about 255 Hz/ln concentration (mg/mL). A good repeatability, R.S.D. = 9 (n = 6) for 0.5 mg/mL caffeine solution was also observed. The use of the sensor can present a potential low-cost option for determining caffeine.Surface analytical techniques such as scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) were performed on the polymer coating in order to elucidate the imprinting process and rebinding of caffeine to the polymer matrix during the sensing process. The SEM micrographs and XPS spectra revealed features and structures that could support the imprinting and recognition of caffeine molecule by the imprinted polymer.     

An optical immunosensor based on surface plasmon resonance for determination of bFGF

An optical immunosensor based on surface plasmon resonance (SPR) has been developed for immunosensing. The sensor is designed on the basis of fixing incident angle of light and measuring the reflected intensities in the wavelength range of 400-800 nm simultaneously. The SPR spectrum was shown in terms of reflected light intensities versus wavelengths of incident light. The intensity of the reflected light reaches the minimum at the resonant wavelength. Molecular self-assembling in solution is used to form the sensing membrane on gold substrate. The kinetic processes of sensing monolayer formation were studied. The basic fibroblast growth factor, a kind of basic polypeptide, was determined in the concentration range of 0.24-9.6 μg/ml. Under optimum experimental conditions, the sensor has a good repeatability, reversibility and selectivity.     

Flow immunoassay of trinitrophenol based on a surface plasmon resonance sensor using a one-pot immunoreaction with a high molecular weight conjugate

A surface plasmon resonance (SPR) immunosensor based on a competitive immunoreaction for the determination of trinitrophenol (TNP) is described. A goat anti-mouse IgG (1st antibody), which recognizes an Fc moiety of an antibody, was immobilized on a gold film of an SPR sensor chip by physical adsorption. A TNP solution containing a fixed concentration of a mouse anti-TNP monoclonal antibody (2nd antibody) and a TNP-keyhole limpet hemocyanin (KLH) conjugate was incubated in one-pot and introduced into the sensor chip. The TNP-KLH conjugate competes with TNP for binding with the 2nd antibody. The resulting complex of the 2nd antibody with the TNP-KLH conjugate was bound to the 1st antibody, which is immobilized on the sensor chip. The SPR sensor signal based on resonance angle shift is dependent on the concentration of TNP in the incubation solution in the range from 25 ppt to 25 ppb, and the coefficient of variation of the SPR signals for the 25 ppb TNP solution was determined to be 13% (n = 4). The experimental results for the adsorption constant of the 1st antibody on the sensor chip and the binding constant of the 1st antibody complex with the 2nd antibody are discussed, together with theoretical considerations.     

Surface plasmon resonance immunosensor for histamine based on an indirect competitive immunoreaction

The use of a surface plasmon resonance immunosensor for the analysis of histamine (β-imidazole ethylamine) is described. The method is based on an indirect competitive reaction of an anti-histamine antibody in a sample solution with histamine immobilized on a sensor chip and with histamine in the sample solution. A sensor chip immobilized with histamine was prepared using a self-assembly monolayer of 11-mercaptoundecanoic acid (11-MUA) as an anchor membrane, followed by an amino-coupling reaction with histamine after activation of the 11-MUA layer on the sensor chip by treatment with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and N-hydroxysuccinimide. The sensor chip can be reused, after regeneration with a 10 mM HCl solution, which dissociates the anti-histamine antibody complex from histamine on the sensor chip. The affinity constants for the immunocomplex of the anti-histamine antibody with histamine in the solution and for that of the anti-histamine antibody with histamine immobilized on the sensor chip were calculated to be 1.5 × 10⁷ and 7.2 × 10⁵ M⁻¹, respectively, by assuming a Langmuir-type adsorption of the anti-histamine antibody to histamine immobilized on the sensor chip. The detection limit of the method was determined to be 3 ppb.     

Effects of solute-matrix interaction on monitoring the conformational changes of immobilized proteins by surface plasmon resonance sensor

A real-time and labeling-free surface plasmon resonance (SPR) sensor was used to monitor the conformational changes of immobilized globule proteins (RNase A and lysozyme) in chemical unfolding and refolding. The effects of chemical denaturants on the protein structures were investigated. The methodology in protein conformational study on the solid surface is refined through the theoretic calculations and the conformational information of native/denatured proteins in solution. Additionally, our observation illustrates that the ambient buffer solution is merit to influence the refractive index of immobilized protein films and directly be observed from the SPR resonance angle shifts.