Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Quartz”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 577 records · Page 32Linked to original sources
PubMed2005

Quartz crystal microbalance detection of glutathione-protected nanoclusters using antibody recognition.

A quartz crystal microbalance (QCM) immunosensor was developed for the quantitative detection of glutathione-protected nanoclusters. Advantages intrinsic to QCM were employed to make it an attractive alternative to other immunosensing techniques. We have addressed challenges in the area of QCM mass sensing through experimental correlation between damping resistance and frequency change for a reliable mass measurement. Electrode functionalization was optimized with the use of protein A to immobilize and present polyclonal IgG for antigen binding. This method was developed for the detection of glutathione (antigen)-protected clusters of nanometer size with high surface area and thiolate valency. Quantitation of glutathione-nanocluster binding to immobilized polyclonal antibody provides equilibrium constants (K(a) = (3.6 +/- 0.2) x 10(5) M(-1)) and kinetic rate constants (k(f) = (5.4 +/- 0.7) x 10(1) M(-1) s(-1) and k(r) = (1.5 +/- 0.4) x10(-4) s(-1)) comparable to literature reports. These observations further imply that immunoreactive nanoparticles have potential in medical diagnostics and materials assembly.

Antibodies↗
PubMed2004

Simultaneous surface plasmon resonance and quartz crystal microbalance with dissipation monitoring measurements of biomolecular adsorption events involving structural transformations and variations in coupled water.

Simultaneous quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) measurements are used to analyze the surface kinetics of two biomacromolecular systems, one lipid and one protein based, undergoing surface-induced conformational changes. First we establish a theoretical platform, which allows quantitative analysis of the combined SPR and QCM-D data. With this theoretical base, new information can be extracted, not obtainable with either technique alone. As an example we demonstrate how time-resolved measurements with these two techniques in combination--yielding three independent measured quantities--add new information about (i) kinetics, i.e. number of adsorbed molecules per unit area versus time, and (ii) temporal variation in the mass fraction of coupled water versus coverage. In particular, it is demonstrated for the first time, how the kinetics of the process during which adsorbed vesicles are spontaneously transformed into a supported phospholipid bilayer (SPB) on SiO(2) can be quantitatively separated into its two dominating states: adsorbed vesicles and supported planar bilayer patches. In addition, the relevance of dynamically coupled water for interpretation and modeling of the QCM-D response during bilayer formation is discussed and further illustrated with a second model system: streptavidin adsorption on a biotin-modified SPB. A very strong coverage dependence in the number of water molecules per protein sensed by the QCM is demonstrated, with strong implications for the use of QCM as a tool for quantitative determination of protein mass uptake kinetics.

Adsorption↗
PubMed2004

Electrochemical quartz crystal nanobalance to detect solvent displacement by pH-induced conformational changes of proteins at Pt.

The electrochemical quartz crystal nanobalance (EQCN) techniques of simultaneous measurements of frequency and cyclic voltammetry (CV) were used to investigate protein adsorption behavior resulting from pH-induced conformational changes at the Pt electrode at 298 K. The adsorption behavior of holo- and apo-alpha-lactalbumin was studied in electrolyte solutions of pH < 2, 7.4, and 11. The EQCN frequency measurements did not directly monitor the mass of the adsorbed protein at anodic potentials, but instead, at a potential characteristic of the double layer for platinum, gave a measure of the extent of solvent displacement by the adsorbed protein (i.e., a "footprint"), which correlated well with known pH-induced conformational changes of the protein. Simultaneous CV charge transfer measurements provided information on the number of layers of protein adsorbed to the surface. This ability of the EQCN to detect solvent displacement by protein adsorption is potentially useful for biosensors to detect and to monitor protein conformational changes in the bulk and during the adsorption process. The Langmuir adsorption isotherm provided the Gibbs energy of adsorption, DeltaG(ADS), and showed excellent agreement between the CV and EQCN frequency measurements.

Animals↗
PubMed2005

Selection of ligands for affinity chromatography using quartz crystal biosensor.

This paper described a new strategy for rapid selecting ligands for application in affinity chromatography using a quartz crystal microbalance (QCM) biosensor. An aminoglycoside antibiotic drug, kanamycin (KM), was immobilized on the gold electrodes of the QCM sensor chip. The binding interactions of the immobilized KM with various proteins in solution were monitored as the variations of the resonant frequency of the modified sensor. Such a rapid screen analysis of interactions indicated clearly that KM-immobilized sensor showed strong specific interaction only with lysozyme (LZM). The resultant sensorgrams were rapidly analyzed by using a kinetic analysis software based on a genetic algorithm to derive both the kinetic rate constants (k(ass) and k(diss)) and equilibrium dissociation constants (K(D)) for LZM-KM interactions. The immobilized KM showed higher affinity to LZM with a dissociation constant on the order of 10(-5) M, which is within the range of 10(-4)-10(-8) M and suitable for an affinity ligand. Therefore, KM was demonstrated for the first time as a novel affinity ligand for purification of LZM and immobilized onto the epoxy-activated silica in the presence of a high potassium phosphate concentration. The KM immobilized affinity column has proved useful for a very convenient purification of LZM from chicken egg white. The purity of LZM obtained was higher than 90%, as determined by densitometric scanning of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified fraction. These results confirmed that the selected KM ligand is indeed a valuable affinity ligand for purification of LZM. The new screening strategy based on a QCM biosensor is expected to be a promising way for rapid selecting specific ligands for purifying other valuable proteins.

Algorithms↗
PubMed2006

Immunophenotyping of acute leukemias using a quartz crystal microbalance and monoclonal antibody-coated magnetic microspheres.

Immunophenotyping, which utilizes panels of lineage-associated monoclonal antibodies to recognize clusters of differentiation (CD) antigens expressed on various leukocytes, plays a key role in the clinical diagnosis of acute leukemias. In this paper, a rapid, simple, and automatic immunophenotyping technique for acute leukemias has been initially proposed by incorporating immunomagnetic separation and quartz crystal microbalance (QCM) measurement. Core-shell paramagnetic microspheres of silica-coated ZnFe2O4 were newly fabricated following a W/O microemulsion route and further functionalized for anchoring separately CD antibodies of lineage-different leukocytes. The resultant immunomagnetic microspheres were introduced, in turn, to capture target leukocytes from the samples and were magnetically accumulated onto the protein A-modified QCM to be analyzed. The response characteristics of the developed QCM system for immunophenotyping various lineage-defined leukocytes were investigated in detail. Results indicate that this new technique can allow for easy and clear identification of acute leukocytes of lymphoid and myeloid origins as well as their subsets. It may also permit the quantitative determination of acute leukocytes with cell concentration down to approximately 10(3) cells mL(-1). Moreover, the applicable feasibility of the QCM immunophenotyping method was validated through assessing a number of clinical specimens, which phenotypes are in acceptable agreement with those obtained by the immunoenzyme assay clinically used for this purpose.

Acute Disease↗
PubMed2006

Quartz crystal microbalance studies of polymer gels and solutions in liquid environments.

Spreading of liquids and soft solids on a rigid surface in a surrounding liquid medium is investigated by utilizing the lateral sensitivity of the quartz crystal microbalance (QCM). While the QCM has been used extensively to study systems with spatial variations in the direction normal to the crystal's electrodes, few studies have exploited the QCM's ability to sense changes in loading in the plane of the electrodes. We propose equations to describe the predicted response of the QCM to a generalized viscoelastic material spreading at the QCM surface at the expense of the surrounding liquid medium. Several experimental examples are given in order to support the validity of the proposed equations, including situations where the spreading material is a Newtonian liquid, a viscoelastic liquid, or one of two viscoelastic solids. The first viscoelastic solid is a physically cross-linked gel based on a styrene/ethylene-butene/styrene triblock copolymer in mineral oil, and the second is a cross-linked poly(ethylene glycol) hydrogel.

Crystallization↗
PubMed2006

Combinational application of surface plasmon resonance spectroscopy and quartz crystal microbalance for studying nuclear hormone receptor-response element interactions.

Conventional methodologies for studying protein-DNA complexes, such as electrophoretic mobility shift assays (EMSAs), lack the real-time sensitivity and precision to accurately characterize the complex dynamics of interactions between transcription factors and their binding sites. To better understand the interactions between estrogen receptor (ER) subtypes and the estrogen response elements (EREs), we employed surface plasmon resonance (SPR) spectroscopy and quartz crystal microbalance with dissipation measurement (QCM-D) and made the following observations: (1) base substitutions in ERE half-sites reduced binding affinity for both ERalpha and ERbeta, (2) ERalpha has a higher sequence specificity than ERbeta or there were more nonspecific interactions between ERbeta and control DNA, and (3) ERalpha bound ERE as dimers and ERbeta bound as tetramers. These findings highlight intrinsic differences in DNA-binding properties between receptor subtypes, which are not apparent based on the high degree of conservation (96% identity) in their DNA-binding domains and results from EMSA studies. With this study, we demonstrate the potential of utilizing SPR and QCM in combination for a comprehensive characterization of ER-DNA interactions, including sequence-dependent binding mechanisms and structural differences in ERalpha-DNA and ERbeta-DNA complexes.

DNA↗
PubMed1998

Use of a quartz crystal microbalance to monitor immunoliposome--antigen interaction.

We have used quartz crystal microbalance (QCM)-based real-time biospecific interaction measurement to analyze the binding of immunoliposomes to antigen and examined the use of liposomes as signal-enhancing reagents in competitive QCM immunoassay. For the preparation of immunoliposomes, various amounts of bacterially produced lipid-tagged single-chain antibody against 2-phenyloxazolone were incorporated in phosphatidylcholine liposomes. The immunoliposomes bound specifically to immobilized hapten, and this binding was inhibited by soluble hapten in a concentration-dependent manner. In this competitive assay, antigen could be measured in the concentration range from 10(-5) to 10(-8) M.

Antibodies↗
PubMed1998

Kinetic measurements of DNA hybridization on an oligonucleotide-immobilized 27-MHz quartz crystal microbalance.

A highly sensitive 27-MHz quartz-crystal microbalance, on which a 10-30-mer oligonucleotide was immobilized as a probe molecule, was employed to detect hybridization of complementary oligonucleotides in aqueous solution. From frequency decreases (mass increases due to the hybridization) with passage of time, kinetic parameters such as association constants (K(a)) and binding and dissociation rate constants (k(1) and k(-1)) could be obtained, as well as binding (hybridization) amount at the nanogram level (delta m). Kinetic studies were carried out by changing various parameters: (i) the immobilization method of a probe oligonucleotide on Au electrode, (ii) number of mismatching bases in sequences of target oligonucleotides, (iii) length of both probe and target oligonucleotides, (iv) hybridization temperature, and (v) ionic strength in solution. The obtained results were compared with those obtained by a surface plasmon resonance method using a BIAcore system.

Avidin↗
PubMed1999

Precipitation of an insoluble product on enzyme monolayer electrodes for biosensor applications: characterization by Faradaic impedance spectroscopy, cyclic voltammetry, and microgravimetric quartz crystal microbalance analyses.

Precipitation of an insoluble, insulating product on monolayer-functionalized electrodes enables the development of new electrochemical biosensors. Faradaic impedance spectroscopy and cyclic voltammetry are used to probe the electron-transfer resistance at the conductive support upon the accumulation of the insoluble product on the electrode surface. Similarly, microgravimetric quartz crystal microbalance, QCM, analyses were used to assay the formation of the precipitate on the electrode. A horseradish peroxidase, HRP, monolayer electrode is used to analyze H2O2 via the biocatalyzed oxidation of 4-chloro-1-naphthol (1) and the precipitation of the insoluble product (2). A bienzyme-layered electrode consisting of HRP and glucose oxidase, GOx, is used to sense glucose. Biocatalyzed oxidation of glucose by O2, in the presence of GOx, yields H2O2, and the generated hydrogen peroxide effects the formation of the insoluble product (2) in the presence of HRP. The insoluble product accumulated on the electrode, and the extent of the resulting electron-transfer resistance, correlated with the amounts of H2O2 or glucose, and appropriate calibration curves are extracted.

Biosensing Techniques↗
PubMed2004

A piezoelectric quartz crystal biosensor: the use of two single cysteine mutants of the periplasmic Escherichia coli glucose/galactose receptor as target proteins for the detection of glucose.

We have examined the potential utility of a glucose biosensor that employs the glucose/galactose receptor of Escherichia coli with a quartz crystal microbalance (QCM). Two different genetically engineered mutant proteins were utilized, each involving the incorporation of a single cysteine into the amino acid sequence of the protein. The proteins were immobilized on the surface of a piezoelectric crystal by a direct sulfur-gold linkage. Since the cysteines were located at different positions in the sequence, the receptors attach to the surface with different orientations. Considering only mass effects, the target sugars for this receptor are predicted to be too small to be detectable with a QCM. However, our sensors indicated measurable and reproducible frequency responses when immobilized receptor was exposed to sugar. This unexpectedly large frequency response occurs because the protein film is transformed from a viscous layer to a more rigid nondissipative film. The QCM can detect these changes because of the direct linkage of the proteins to the surface. Calculations of the frequency response expected for a viscoelastic film with different rheological characteristics support this hypothesis. This study is significant because it illustrates a widened applicability for the QCM methodology to protein systems that bind small molecules and undergo ligand-induced conformational changes.

Alanine↗
PubMed2005

Kinetic studies of DNA cleavage reactions catalyzed by an ATP-dependent deoxyribonuclease on a 27-MHz quartz-crystal microbalance.

Catalytic DNA cleavage reactions by an ATP-dependent deoxyribonuclease (DNase) from Micrococcus luteus were monitored directly with a DNA-immobilized 27-MHz quartz-crystal microbalance (QCM). The 27-MHz QCM is a very sensitive mass-measuring device in aqueous solution, as the frequency decreases linearly with increasing mass on the electrode at a nanogram level. Three steps in ATP-dependent DNA hydrolysis reactions, including (1) binding of DNase to the end of double-stranded DNA (dsDNA) on the QCM electrode (mass increase), (2) degradation of one strand of dsDNA in the 3' --> 5' direction depending on ATP (mass decrease), and (3) release of the enzyme from the nonhydrolyzed 5'-free-ssDNA (mass decrease), could be monitored stepwise from the time dependencies of QCM frequency changes. Kinetic parameters for each step were obtained as follows. The binding constant (K(a)) of DNase to the dsDNA was determined as (28 +/- 2) x 10(6) M(-)(1) (k(on) = (8.0 +/- 0.3) x 10(3) M (-)(1) s(-)(1) and k(off) = (0.29 +/-0.01) x 10(-)(3) s(-)(1)), and it decreased to (0.79 +/- 0.16) x 10(6) M(-)(1) (k'(on) = (2.3 +/- 0.2) x 10(3) M (-)(1) s(-)(1) and k'(off) = (2.9 +/- 0.1) x 10(-)(3) s(-)(1)) for the completely nonhydrolyzed 5'-free ssDNA. This is the reason the DNase bound to the dsDNA substrate can easily release from the nonhydrolyzed 5'-free-ssDNA after the complete hydrolysis of the 3' --> 5' direction of the complementary ssDNA. K(a) values depended on the DNA structures on the QCM, and the order of these values was as follows: the dsDNA having a 4-base-mismatched base-pair end (3) > the dsDNA having a 5' 15-base overhanging end (2) > the dsDNA having a blunt end (1) > the ssDNA having a 3'-free end (4) >> the ssDNA having a 5'-free end (5). Thus, DNase hardly recognized the free 5' end of ssDNA. Michaelis-Menten parameters (K(m) for ATP and k(cat)) of the hydrolysis process also could be obtained, and the order of k(cat)/K(m) was as follows: the dsDNA having a blunt end (1) approximately the dsDNA having a 4-base-mismatched base-pair end (3) > the ssDNA having a free 3' end (4) >> the ssDNA having a free 5' end (5). Thus, DNase could not recognize and not hydrolyze the free 5' end of ssDNA. The DNA hydrolysis reaction could be driven by dATP and GTP (purine base) as well as ATP, whereas the cleavage efficiency was very low driven with UTP, CTP (pyrimidine base), ADP, and AMP.

Adenosine Diphosphate↗
PubMed2005

Kinetic studies of site-directed mutational isomalto-dextranase-catalyzed hydrolytic reactions on a 27 MHz quartz-crystal microbalance.

A quartz-crystal microbalance (QCM) technique was applied to analyze effects of site-directed mutagenesis of a glycosidase (isomalto-dextranase) on the hydrolysis mechanism of the substrate binding (k(on), k(off), and K(d)) and the catalytic process (k(cat)), separately, by using a dextran-immobilized QCM in buffer solution. D266N, D198N, and D313N mutants, which are predicted as critical residues of the isomalto-dextranase hydrolytic activity, dramatically decreased the apparent enzyme activity. The D266N mutant, however, did not change the substrate binding ability (K(d)), and the D198N and D313N mutants largely increased K(d) values due to the increase of k(off) and/or the decrease of k(on) values, as well as the negatively small k(cat) values. From these results, we estimate the reaction mechanism, in which Asp266 acts as only a general acid in the catalytic process, Asp198 acts as both nucleophile in the catalytic process and binding the substrate, and Asp313 acts as only the substrate binding.

Amino Acid Sequence↗
PubMed1998

Kinetic studies of sequence-specific binding of GCN4-bZIP peptides to DNA strands immobilized on a 27-MHz quartz-crystal microbalance.

Specific protein-DNA interaction was studied quantitatively by using a highly sensitive 27-MHz quartz-crystal microbalance (QCM). Biotinylated DNA double strands (21 bp, having a CRE site of 5'ATGACGTCAT3') were immobilized on an avidin-bound QCM surface, and sequence-specific binding of bZIP 56-mer peptides (having both the basic region for binding and the leucine zipper region for dimerization) to the DNA strand on the QCM was observed. The binding amount (Deltam) at the nanogram level and kinetic parameters such as association constants (Ka) and binding and dissociation rate constants (k1 and k-1) could be obtained from time courses of QCM frequency decreases. A bZIP peptide as a dimer was observed to bind sequence-specifically to one DNA strand having a CRE site. Ka values of ss-bZIP, in which the leucine-zipper region of bZIP was substituted by a Cys-Cys linkage, were largely decreased, and the sequence selectivity also disappeared. Ka values obtained by the QCM method showed good agreement with those obtained from the conventional gel mobility shift assay or from circular dichroism spectrum changes. When the specific sequence of the CRE site of DNA strands was partly changed, Ka values decreased by about a half due to the increase of the dissociation rate constant (k-1) independent of the binding rate constant (k1).

Amino Acid Sequence↗
PubMed2001

Analysis of adsorption function of polyhydroxybutyrate depolymerase from Alcaligenes faecalis T1 by using a quartz crystal microbalance.

Enzymatic adsorption and degradation of three types of aliphatic polyester films and two types of polyolefin films by the extracellular PHB depolymerase from Alcaligenes faecalis T1 have been studied by using a quartz crystal microbalance (QCM) technique. Hydrolysis of poly[(R)-3-hydroxybutyrate] was quantitatively followed by the QCM technique. Adsorption of the enzymes to films was also quantitatively detected by the QCM. Kinetic study on enzymatic adsorption suggests that the PHB depolymerase binds to the substrates not only by hydrophobic interaction but also by specific interaction between the ester bonds of polyesters and the binding domain of the enzyme. The results show that the QCM technique is a sensitive tool to study enzymatic degradation kinetics of biodegradable polyesters.

Adsorption↗
PubMedDate not supplied

Electropolymerized films formed from the amphiphilic decyl esters of D- and L-tyrosine compared to L-tyrosine using the electrochemical quartz crystal microbalance.

Using the electrochemical quartz crystal microbalance (EQCM), we compared thin films formed on Pt by electropolymerization of l-tyrosine to that of the amphiphilic monomers, decyl esters of d- and l-tyrosine (DEDT and DELT). Mass build-up and film properties were determined as a function of monomer concentration via frequency, f, motional resistance, R, and charge passage, Q, measurements. Films were found to occur by a combination of monomer electropolymerization and adsorption for DEDT and DELT, but only by electropolymerization for l-tyrosine. This difference in film formation process for the monomers is reflected in the net mass build-up for each film, as represented by calculated df/dQ values. For the adsorbing monomers DEDT and DELT, films possessed concentration dependent df/dQ values, more than 100-fold greater than that for l-tyrosine film formation under equivalent electropolymerization conditions. During the entire film growth process, all three films exhibited no significant energy dissipation properties (DeltaR invariant). Concentration dependent adsorption of significant levels of unpolymerized but self-assembled DEDT and DELT monomers account for the subsequent time dependent mass loss observed from the films maintained in buffer in the absence of monomer. Contact angle measurements demonstrated a pH dependent increase in the surface hydrophilicity of films electropolymerized from the DEDT, DELT, and l-tyrosine monomers but not films formed from phenol and 3-nitrophenol monomers. This behavior is consistent with the monomers' known changes in titration/charge state properties with increasing pH. This study provided insight into the film formation, stability, and surface hydrophilicity resulting from electropolymerization of these related tyrosine based monomers. This information is critical to assessing the utility these films may have in the development of new biomaterials and as biological macromolecule or cell immobilization strategies in biosensors.

Electrochemistry↗
PubMedDate not supplied

Enzymatic degradation of poly(L-lactide) film by proteinase K: quartz crystal microbalance and atomic force microscopy study.

Enzymatic degradation of the poly(L-lactide) (PLLA) amorphous film by proteinase K has been investigated by combination of the complementary techniques of quartz crystal microbalance and atomic force microscopy (AFM). The erosion rate increased with increasing enzyme concentrations and attained to be constant under the condition of [proteinase K] > 100 microg/mL. The amount of the enzyme molecules adsorbed to the film was quantitatively evaluated at various concentrations by AFM, and it revealed that the erosion rate is determined by the amount of adsorbed enzyme. Adsorption of proteinase K was irreversible despite lack of the binding domain, so that the enzyme molecules on the film surface could be observed directly by AFM. Transformation of the enzyme molecule caused by packing in high density on the surface was observed at higher enzyme concentrations. The "footprint" of the individual proteinase K molecule on the PLLA film after enzymatic degradation suggests that the enzyme moves on the surface to hydrolyze the film around it.

Adsorption↗
PubMed2006

Effect of ultrasound on DNA polymerase reactions: monitoring on a 27-MHz quartz crystal microbalance.

Effects of ultrasound irradiation on DNA polymerase (Klenow fragment, KF) reactions were studied on the template/primer DNA-immobilized quartz crystal microbalance (QCM). Under ultrasound irradiation, binding of KF to the DNA was suppressed due to the decrease of the binding rate constant (k(1)) and the increase of the dissociation rate constant (k(-)(1)). The catalytic elongation rate (k(cat)) was increased, but the stability of the KF/DNA/monomer ternary complex (K(m)) was decreased by the ultrasound irradiation. Ultrasound effects are discussed in correlation with the conformation changes of domain structures in KF.

Base Sequence↗