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A piezoelectric quartz crystal impedance study on Cu(2+)-induced precipitation of bovine serum albumin in aqueous solution.

Piezoelectric quartz crystal impedance (QCI) technique was used for monitoring the Cu(2+)-induced precipitation of bovine serum albumin onto the gold electrode. The critical precipitate concentration of Cu(2+) reflected by the significant decrease in the resonant frequency was estimated to be 9.98 x 10(-5) mol x l(-1), and the saturated adherence of the precipitate on the electrode occurred when the Cu(2+) concentration was greater than 9.79x10(-3) mol x l(-1). The frequency shift in air was about 85.5% of that in liquid, and the Deltaf(0)/DeltaR(1) ratio found in solution was 82.67 Hz Omega(-1), suggesting that the frequency response was predominated by the mass change due to precipitate adherence to the electrode surface. The response of the resonant frequency was analyzed using an equation Deltaf=a(0) + a(1) e(-t/tau(1)) + a(2) e(-t/tau(2)). The relationship between the total a(0) values and the Cu(2+) concentration was discussed.

Animals↗

Interaction process between ionic surfactant and protein probed by series piezoelectric quartz crystal technique.

A method for probing the interaction process between ionic surfactant and protein was developed with series piezoelectric quartz crystal (SPQC) sensing technique. It was based on the sensitive response of the SPQC sensor to the change in solution conductivity. A new relationship between the sensor response and the properties of ionic species in solution was derived. The method was used to examine the interaction process of two surfactants, cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulfate (SDS), with lysozyme in aqueous solution. The obtained experimental results were in agreement with those of other methods from references. These results had been discussed. It was shown that the new method developed here was a useful and promising tool for probing the ionic surfactant-protein interaction process and might find more applications in similar studies.

Biophysics↗

Monitoring of Lactobacillus fermentation process by using ion chromatography with a series piezoelectric quartz crystal detector.

A new method monitoring Lactobacillus fermentation process, which combines ion chromatography (IC) with a series piezoelectric quartz crystal (SPQC) technique, is presented in this paper. Monitoring of the fermentation process was realized by examining the rate of production of lactic acid. An automatic membrane dialyser was used for the pretreatment of the sample in on-line monitoring. A mixture of p-hydroxybenzoic acid and N,N-diethylethanolamine was adopted as mobile phase and its flow rate was 0.8 ml/min. The effects of some fermentation conditions were also discussed in detail. Accordingly, the optimal fermentation conditions were obtained. This method is simple and convenient while the results obtained are accurate and reliable.

Carboxylic Acids↗

Monitoring for adsorption of human serum albumin and bovine serum albumin onto bare and polystyrene-modified silver electrodes by quartz crystal impedance analysis.

The adsorption of human serum albumin (HSA) and bovine serum albumin (BSA) from PBS (pH 7.4) onto bare and polystyrene (PS)-modified silver electrodes was in situ monitored using quartz crystal impedance analysis. The adsorption characteristics of HSA and BSA were discussed by analyzing piezoelectric parameter simultaneous responses. Experimental results indicated that for both HSA and BSA, the amount adsorbed on bare silver was more than that on PS-modified surface. The BSA amount adsorbed on the two surfaces was more than that of adsorbed HSA. A kinetic model was developed to describe the adsorption process and fitted to the experimental data of frequency shift. It was shown that HSA adsorption could be described by a kinetic equation involving two consecutive reactions. At lower concentration, BSA adsorption only involved the first reaction. At higher concentration, BSA adsorption on PS-modified surface involved two consecutive reactions. All fitted results were well in agreement with the corresponding experimental results. The regression values of reaction rate constants for the HSA and BSA adsorption were obtained. These data exhibited difference in adsorption kinetics under different conditions.

Adsorption↗

Real-time investigation of the interaction between primaquine phosphate and bovine serum albumin (BSA) by piezoelectric quartz crystal impedance analysis.

Real-time investigation of the interaction between primaquine phosphate and bovine serum albumin by the piezoelectric quartz crystal impedance (PQCI) analysis was carried out for the first time. Three kinds of electrodes were investigated. Compared with bare gold (Au) electrode, the gold electrode self-assembled of nanogold colloids exhibits maintained biocompatibility, increased capacity and more bioactivity. Additionally, on the basis of the multi-dimensional information provided by the PQCI analysis, the real-time interaction information and the kinetics of the binding process was investigated and a response model was deduced. At 37 degrees C, the binding rate (k1), dissociation rate (k(-1)) and equilibrium constants (Ka) were 4.19x10(2) (mol l(-1))(-1) s(-1), 1.01x10(-3) s(-1) and 4.15x10(5) (mol l(-1))(-1) for the electrode modified by nanogold particles; 3.83x10(2) (mol l(-1))(-1) s(-1), 9.70x10(-4) s(-1) and 3.95x10(5) (mol l(-1))(-1) for the bare gold electrode, respectively.

Animals↗

The in vitro effects of high-Tc-superconducting particles (YBa2Cu3O6-7) and quartz (SiO2) on bovine alveolar macrophages.

The cytotoxic effects of YBa2Cu3O6--7-particles on cultured bovine alveolar macrophages were investigated by monitoring the release of lactate dehydrogenase and N-acetylglucosaminidase into the culture medium and testing the viability of the cells. After 20 hours of incubation the effects of YBa2Cu3O6--7-particles were very similar to those of analogous mass concentrations of DQ 12 quartz.

Acetylglucosaminidase↗

Antibody/antigen affinity behavior in liquid environment with electrical impedance analysis of quartz crystal microbalances.

Electrical impedance analysis has been used to study anti-human immunoglobulin G (anti-h IgG) adsorption and the subsequent human immunoglobulin G (hIgG) or rabbit immunoglobulin G (rIgG) affinity reaction in aqueous liquids on a polystyrene (PS)-modified quartz crystal microbalance (QCM) surface. Time-dependent adsorption data of both the frequency shift and the electrical equivalent parameters (motional resistance, shunt capacitance, quality factor, etc) are monitored. It was found that the motional resistance, R, increases while the resonance frequency, f, decreases during both the anti-h IgG immobilization and the subsequent affinity process. Decreasing f primarily arises from the increased mass loading. Increasing R indicates more power dissipation (increased losses) in the system. The change in motional resistance, delta R, in the affinity reaction is considerably larger than that in anti-h IgG immobilization adsorption process, although the resonant frequency shifts, delta f, are very close in these two processes. Specifically, for a saturated solution, the ratio of delta R/delta f is 9.45 x 10 (-3) Omega/Hz for anti-h IgG adsorption and 28.1 x 10 (-3) omega/Hz for anti-h IgG/hIgG binding respectively, indicating the increased power dissipation with the increasing binding molecules. The shunt capacitance changes little in the hIgG binding process ( approximately 0.01 pF).

Animals↗

Real-time monitoring of enzymatic cleavage of nucleic acids using a quartz crystal microbalance.

The use of quartz crystal microbalance (QCM) for monitoring in situ the enzymatic cleavage of surface-confined nucleic acids by nucleases is described. Such real-time monitoring of mass changes associated with the enzymatic digestion indicates that the activity and specificity of nucleases is preserved at the gold surface, and can be used for manipulating surface-confined DNAs and RNAs. These observations indicate great promise for using QCM for elucidating the interactions of nucleic acids with enzymes, and for enhancing the power of hybridization biosensors.

Crystallization↗

Quantitative measurements of the interaction between monosialoganglioside monolayers and wheat germ agglutinin (WGA) by a quartz-crystal microbalance.

Monosialogangliosides (GM1, GM2, GM3 and GM4) were reconstituted in lipid monolayers at the air-water interface. The binding amounts and the initial binding rates of wheat germ agglutinin (WGA) to the monosialoganglioside monolayers were quantitatively studied by use of a quartz-crystal microbalance (QCM). A QCM was horizontally attached to the monolayer from the air phase, and the binding behavior (mass increase) was followed by the frequency decrease of the QCM. WGA binding affinities for the ganglioside monolayers were influenced by hydrophilic head groups of lipid matrices, densities of gangliosides, and sequences of oligosaccharide in gangliosides. Binding of WGA to the gangliosides reconstituted in a phosphatidylcholine (sphingomyelin and distearoylphosphatidylcholine) matrix was strongly suppressed, but not in a neutral glycolipids (GlcCer, GalCer, and LacCer), dipalmitoylphosphatidylethanolamine, and dipalmitoylphosphatidylethanolamine matrix. WGA showed high affinity for monolayers containing 20 mol% gangliosides, but only low affinity for 100% ganglioside monolayers. WGA preferably binds to gangliosides in the following sequence: GM3 > GM4 >> GM2 = GM1. No affinities of WGA for GM2 and GM1 were observed. The combined techniques of monolayer and QCM have the advantages of investigating recognition properties of gangliosides.

Air↗

Aerodynamic particle size of metered-dose inhalers determined by the quartz crystal microbalance and the Andersen cascade impactor.

Given the rapid sizing capability and high sensitivity, the quartz crystal microbalance (QCM) cascade impactor has been evaluated for the size determination of metered-dose inhaler (MDI) aerosols. The effects of surfactants present in MDI formulations, crystal coating, particle bounce and crystal overloading on the QCM cascade impactor are investigated. To reduce particle bounce, it is necessary to coat the crystals and use new coated surfaces for each measurement. Mass median aerodynamic diameters (MMADs) obtained from the QCM cascade impactor are compared to those from the commonly used Andersen cascade impactor. For MDI formulations containing little or no surfactants, MMADs obtained from the QCM and Andersen cascade impactors are comparable. For MDI formulations containing a significant amount of surfactant (or any non-volatile excipients), the QCM cascade impactor measures the combined size distribution of the drug and non-volatile excipients. A technique is devised in this study to deduce the drug-only size distribution from the QCM impactor for surfactant-containing MDI formulations and show comparable results to the Andersen cascade impactor except for high drug load Intal. The QCM impactor has proved to be a useful tool for rapid size measurement of MDI formulations.

Nebulizers and Vaporizers↗

Determination of divalent carboxylic acids in traditional Chinese herbal and patent medicines using HPLC with double cell quartz crystal detector.

A rapid, simple and sensitive high performance liquid chromatography (HPLC) method combined with a novel double cell quartz crystal (DCQC) detector is described for the determination of divalent carboxylic acids in traditional Chinese herbal and patent medicines. The chromatographic system involves the use of a muBondapak-NH2 column with 20 mM KH2PO4 (pH 3.4, adjusted by H3PO4)/acetonitrile (95:5%) as a mobile phase. The DCQC detector has a low temperature coefficient and high conductance sensitivity independent of the background from 7.2 to 2500 microS. Analysis of sample is completed without the use of an ion suppression device.

Calibration↗

Voltammetric and electrochemical quartz crystal microbalance studies of sulforaphane and its Zn(II) complexes.

Cyclic voltammetry (CV) and electrochemical quartz crystal microbalance (EQCM) methods were used to study the behavior of sulforaphane (SFR). To established electrochemical properties of SFR methyl-, ethyl-, 2-chloroethyl-, propyl-, butyl- and tert-butyl-isothiocyanates were examined at gold electrode. Optimal measurement parameters were established and stock solutions developed. The electrode activity of the studied compounds (cathodic peak E(p)=-0.6 V) was found to be primarily due to the oxidation of the isothiocyanate group. By the EQCM method it was found that during the reduction of SFR the mass of the electrode systematically increased in successive scans by ca. 120 ng per scan and in the mixture of SFR and Zn(2+) ions ca. 350 ng. Formation of SFR complexes with Zn(2+) ions was confirmed by EQCM and UV methods. Cyclic voltammetric method has been elaborated for SFR determination and compared with UV method.

Electrochemistry↗

Comparison of surface plasmon resonance and quartz crystal microbalance in the study of whole blood and plasma coagulation.

The coagulation of blood plasma and whole blood was studied with a surface plasmon resonance (SPR) based device and a quartz crystal microbalance instrument with energy dissipation detection (QCM-D). The SPR and QCM-D response signals were similar in shape but differing in time scales, reflecting differences in detection mechanisms. The QCM-D response time was longer than SPR, as a physical coupling of the sample to the substrate is required for molecules to be detected by the QCM-method. Change of sample properties within the evanescent field is sufficient for detection with SPR. Both the SPR signals and the QCM-D frequency and dissipation shifts showed dependency on concentrations of coagulation activator and sensitivity to heparin additions. The ratio of dissipation to frequency shifts, commonly considered to reflect viscoelastic properties of the sample, varied with the concentration of activator in blood plasma but not in whole blood. Additions of heparin to the thromboplastin activated whole blood sample, however, made the ratio variation reoccur. Implications of these observations for the understanding of the blood coagulation processes as well as the potential of the two methods in the clinic and in research are discussed.

Animals↗

Microgravimetric DNA sensor based on quartz crystal microbalance: comparison of oligonucleotide immobilization methods and the application in genetic diagnosis.

We report on the study of immobilization DNA probes onto quartz crystal oscillators by self-assembly technique to form variety types of mono- and multi-layered sensing films towards the realization of DNA diagnostic devices. A 18-mer DNA probe complementary to the site of genetic beta-thalassaemia mutations was immobilized on the electrodes of QCM by covalent bonding or electrostatic adsorption on polyelectrolyte films to form mono- or multi-layered sensing films by self-assembled process. Hybridization was induced by exposure of the QCMs immobilized with DNA probe to a test solution containing the target nucleic acid sequences. The kinetics of DNA probe immobilization and hybridization with the fabricated DNA sensors were studied via in-situ frequency changes. The characteristics of QCM sensors containing mono- or multi-layered DNA probe constructed by direct chemical bonding, avidin-biotin interaction or electrostatic adsorption on polyelectrolyte films were compared. Results indicated that the DNA sensing films fabricated by immobilization of biotinylated DNA probe to avidin provide fast sensor response and high hybridization efficiencies. The effects of ionic strength of the buffer solution and the concentration of target nucleic acid used in hybridization were also studied. The fabricated DNA biosensor was used to detect a set of real samples. We conclude that the microgravimetric DNA sensor with its direct detection of amplified products provide a rapid, low cost and convenient diagnostic method for genetic disease.

Biosensing Techniques↗

A quartz crystal microbalance cell biosensor: detection of microtubule alterations in living cells at nM nocodazole concentrations.

The quartz crystal microbalance (QCM) was used to create a piezoelectric biosensor utilizing living endothelial cells (ECs) as the biological signal transduction element. ECs adhere to the hydrophilically treated gold QCM surface under growth media containing serum. At 24 h following cell addition, calibration curves were constructed relating the steady state Deltaf and DeltaR shift values observed to the numbers of electronically counted cells requiring trypsinization to be removed from the surface. We then utilized this EC QCM biosensor for the detection of the effect of [nocodazole] on the steady state Deltaf and DeltaR shift values. Nocodazole, a known microtubule binding drug, alters the cytoskeletal properties of living cells. At the doses used in these studies (0.11-15 microM), nocodazole, in a dose dependent fashion, causes the depolymerization of microtubules in living cells. This leads a monolayer of well spread ECs to gradually occupy a smaller area, lose cell to cell contact, exhibit actin stress fibers at the cell periphery and acquire a rounded cell shape. We observed the negative Deltaf shift values and the positive DeltaR shift values to increase significantly in magnitude over a 4-h incubation period following nocodazole addition, in a dose dependent fashion, with a transition midpoint of 900 nM. Fluorescence microscopy of the ECs, fixed on the gold QCM surface and stained for actin, demonstrated that the shape and cytoskeleton of ECs were affected by as little as 330 nM nocodazole. These results indicate that the EC QCM biosensor can be used for the study of EC attachment and to detect EC cytoskeletal alterations. We suggest the potential of this cellular biosensor for the real time identification or screening of all classes of biologically active drugs or biological macromolecules that affect cellular attachment, regardless of their molecular mechanism of action.

Actins↗

Immunosensor for the differentiation and detection of Salmonella species based on a quartz crystal microbalance.

Immunosensors based on the microgravimetric quartz crystal microbalance (QCM) technique have been developed for the detection of Salmonella species from serogroups A, B and D. Salmonella serogroup-specific murine monoclonal antibodies, respectively, raised against these serogroups were immobilized onto the silver electrodes of piezoelectric (PZ) crystals by cross-linkage via glutaraldehyde (GA) to the electrode surfaces pre-coated with thin polyethyleneimine (PEI) layer. The specific immunosensors developed gave responses in linear ranges from 10(5) to 5x10(8) cells per ml with no significant interference from other strains of Salmonella and Escherichia coli up to 10(8) cells per ml. They showed good repeatability and excellent linear range, achieving detection limits down to 10(4) cells per ml with ability to distinguish different strains of Salmonella. These biosensors exhibited an exquisite specificity evidenced by their ability to discriminate antigens, the structures of which differ only by the isomeric form of di-deoxyhexose. The antibody-modified crystals showed no loss in activity over 4 days under storage at 4 degrees C.

Animals↗

Improved procedures for immobilisation of oligonucleotides on gold-coated piezoelectric quartz crystals.

The high sensitivity and specificity of DNA hybridisation techniques makes them powerful tools for environmental or clinical analysis. This work describes the development of a DNA piezoelectric biosensor for the detection of the hybridisation reaction. Attention was focused on the choice of the coating chemistry that could be used for the immobilisation of oligonucleotides onto the gold surface of the quartz crystal. Four immobilisation procedures were tested and compared considering the amount of immobilised probe, the extent of the hybridisation reaction, the possibility of regeneration and the absence of non-specific adsorption. All the experiments were performed with oligonucleotides of 25 bases (probe, target and non-complementary oligonucleotide). The four coating methods were all based on the use of self-assembled monolayers (SAM). Three of them employed the interaction between streptavidin and biotin for the immobilisation of a biotinylated probe. Results indicated that immobilisation of a biotinylated probe on streptavidin linked to a layer of carboxylated dextran provides higher sensitivity for the detection of the hybridisation reaction, absence of non-specific adsorption and a higher stability with respect to the regeneration step.

Apolipoproteins E↗

Quartz crystal microbalance (QCM) affinity biosensor for genetically modified organisms (GMOs) detection.

A DNA piezoelectric sensor has been developed for the detection of genetically modified organisms (GMOs). Single stranded DNA (ssDNA) probes were immobilised on the sensor surface of a quartz crystal microbalance (QCM) device and the hybridisation between the immobilised probe and the target complementary sequence in solution was monitored. The probe sequences were internal to the sequence of the 35S promoter (P) and Nos terminator (T), which are inserted sequences in the genome of GMOs regulating the transgene expression. Two different probe immobilisation procedures were applied: (a) a thiol-dextran procedure and (b) a thiol-derivatised probe and blocking thiol procedure. The system has been optimised using synthetic oligonucleotides, which were then applied to samples of plasmidic and genomic DNA isolated from the pBI121 plasmid, certified reference materials (CRM), and real samples amplified by the polymerase chain reaction (PCR). The analytical parameters of the sensor have been investigated (sensitivity, reproducibility, lifetime etc.). The results obtained showed that both immobilisation procedures enabled sensitive and specific detection of GMOs, providing a useful tool for screening analysis in food samples.

Affinity Labels↗