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M E Meyerhoff

Publications and source records attributed to M E Meyerhoff.

At least 37 records · Page 2Linked to original sources
PubMed1997

Photo-cross-linked decyl methacrylate films for electrochemical and optical polyion probes.

Potentiometric and optical polyion probes based on photo-cross-linked thin films of decyl methacrylate (DMA) are described, and the effects of film composition on the response toward heparin are examined in detail. In accordance with existing theory governing potentiometric polyion response, lowering the amounts of plasticizer and tridodecylmethylammonium chloride ion exchanger within the film enhances its sensitivity toward heparin. Varying the cross-linker content of a DMA-based film, however, provides an additional mechanism to regulate its physical structure and, hence, the observed potentiometric polyion response. Films with low hexanedioldimethacrylate cross-linker content yield optimal potentiometric heparin detection limits (0.04 microM), suggesting a lower diffusion coefficient within such films, apparently due to interactions between adjacent pendant decyl groups. Increasing crosslinker content interrupts these interactions and facilitates diffusion. This knowledge is applied to optimize optical heparin sensing via DMA films covalently attached to glass substrates. When used in a limited volume/fixed exposure time measurement mode, such optically sensitive films can detect clinically relevant levels of heparin (0.5-5 units/mL) in undiluted human plasma.

Cross-Linking Reagents↗
PubMed1997

Electrochemical assay of proteinase inhibitors using polycation-sensitive membrane electrode detection.

A simple and sensitive electrochemical method suitable for real time detection of trypsin-like proteinase inhibitors is described. The method is based on utilizing a protamine (a polycationic substrate for trypsin-like proteinases)-sensitive membrane electrode to monitor, potentiometrically, the initial rate of protamine decomposition upon the addition of a proteinase-antiproteinase test solution. In the presence of proteinase inhibitors, the initial rate of change in electromotive force is dependent on the concentration of proteinase inhibitor in the sample solution. The feasibility of this new assay method is demonstrated by detecting four inhibitors of trypsin-like proteinases: alpha1-antiproteinase inhibitor, alpha2-macroglobulin, aprotinin, and soybean inhibitor, using trypsin as the indicator proteinase. The efficacy of inhibition by each species, as expressed by I50 values (concentration of the inhibitor that induces 50% of the maximum proteinase inhibition), is shown to correlate well with literature values for the association constant of the proteinase-antiproteinase complex (k[assoc]). The proposed electrochemical assay for aprotinin is examined further using trypsin, plasmin, and kallikrein as the proteinase indicator reagents. It is found that the trypsin-aprotinin system offers the highest sensitivity and lowest detection limit for aprotinin detection. Application of the proposed method for measuring aprotinin in pretreated plasma samples is also reported.

Aprotinin↗
PubMed1996

Retention behavior of amino acids and peptides on protoporphyrin-silica stationary phases with varying metal ion centers.

Various metalloprotoporphyrins (MProP) covalently linked to silica supports are examined as novel immobilized metal ion affinity chromatography (IMAC) stationary phases for separations of amino acids/peptides. Under reversed-phase HPLC conditions, the MProP-silicas exhibit high affinity toward L-histidine via metal-nitrogen axial ligation interactions, with an increasing degree of histidine retention highly dependent on the specific metal ion (M) in the center of the protoporphyrin (ProP) structure: Fe(III) > Ni(II) > Cu(II) > Zn (II) approximately Cd(II). Aromatic amino acids (i.e., L-trytophan and L-phenylalanine) are also retained on MProP columns through pi-pi interactions with the immobilized porphyrins, with the greatest affinity for L-trytophan observed on CuProP-silica columns. Peptides rich in L-histidine and L-tryptophan residues are selectively retained on most of the MProP-silica phases examined; however, the addition of an organic modifier and/or lowering the pH of the mobile phase can be used independently to attenuate the pi-pi and metal ion-nitrogen ligation interactions, respectively. Reproducible separations of His-Phe and trytophan releasing hormone are achieved on a FeProP-silica column even after extensive washing with 50 mM EDTA, demonstrating a fundamental advantage of the new MProP-silica over existing IMAC stationary phases, in which the metal ion is anchored weakly to the support via immobilized iminodiacetate and related ligands.

Amino Acid Sequence↗
PubMed1996

Mixed potential response mechanism of cobalt electrodes toward inorganic phosphate.

The potentiometric response mechanism of a previously reported phosphate ion-sensitive electrode based on a surface-oxidized cobalt metal is examined. Beyond response to phosphate, the cobalt electrode is found to respond to changes in the partial pressure of oxygen in the sample solution. the potentiometric response toward phosphate ions and molecular oxygen is shown to depend on the sample stirring rate as well as the pH, ionic strength, and nature of the buffer salts present within the test solution. X-ray photoelectron spectroscopy studies of the cobalt electrodes, in conjunction with cyclic voltammetric measurements, suggest that the potentiometric response originates from a mixed potential resulting from the slow oxidation of cobalt and simultaneous reduction of both oxygen and Co2+ at the surface of the electrode. In contrast to an originally proposed host-guest mechanism, the present mixed potential mechanism more accurately explains behavior of oxidized cobalt electrodes in the presence of phosphate and oxygen species.

Cobalt↗
PubMed1996

Influence of nonionic surfactants on the potentiometric response of hydrogen ion-selective polymeric membrane electrodes.

The influence of poly(ethylene oxide)-based nonionic surfactants (i.e., Triton X-100 and Brij 35) in the sample phase on the response properties of hydrogen ion-selective polymeric membrane electrodes containing mobile (lipophilic amines) or covalently bound (aminated-poly-(vinyl chloride)) hydrogen ion carriers is reported. In the presence of these nonionic surfactants, membrane electrode response toward interfering cation activity (e.g., Na+) in the sample phase is increased substantially and the pH measuring range shortened. The degree of cation interference for pH measurements is shown to correlate with the basicity of the hydrogen ion carrier doped within the membrane phase. The observed deterioration in selectivity arises from the partitioning of the surfactant into the membrane and concomitant extraction of metal cations by the surfactants in the organic phase. The effect of nonionic surfactants on pH electrodes prepared with aminated-PVC membranes is shown to be more complex, with additional large shifts in EMF values apparently arising from multidentate interactions between the surfactant molecules and the polymeric amine in the membrane, leading to a change in the apparent pKa values for the amine sites. The effects induced by nonionic surfactants on the EMF response function of hydrogen ion-selective polymeric membrane electrodes are modeled, and experimental results are shown to correlate well with theoretical predictions.

Electrodes↗
PubMed1996

Homogeneous enzyme-based binding assay for studying glycosaminoglycan interactions with macromolecules and peptides.

A simple and rapid homogeneous enzyme-based binding assay is described to study the degree of interaction between glycosaminoglycans and various macromolecules/peptides. The method is based on the homogeneous inhibition of a highly positively charged enzyme, acid deoxyribonuclease II (EC 3.1.22.1), by glycosaminoglycan polyanions, such as heparin, chondroitin 4-sulfate, and dermatan sulfate. Catalytic activity of DNase II is inhibited to nearly 100% by relatively small amounts of these glycosaminoglycan molecules. In the presence of species that bind these polyanions, the activity of the enzyme is regained in an amount proportional to the concentration of the species present. Thus, the relative binding affinities of various species with a given GAG can be assessed rapidly by comparing the concentration of the compound required to reverse the enzyme inhibition to 50% of the maximum value (ED(50) values). The feasibility of this binding assay principle is demonstrated by measuring the ED(50) values of five macromolecules: polylysine, polyarginine, protamine, low-density lipoprotein (LDL), and high-density lipoprotein (HDL), using heparins of different size, as well as chondroitin 4-sulfate and dermatan sulfate as the GAG polyanions. The applicability of the assay method is further extended to study GAG-peptide interactions. A variety of small synthetic peptides (8-13 amino acid residues) derived from the heparin-binding domains of protamine and type IV collagen are used as model peptide species. Relative GAG-binding affinities of these macromolecules/peptides are compared to previous literature values and data obtained via a new electrode-based titration method.

Binding, Competitive↗
PubMed1996

A new method of measuring heparin levels in whole blood by protamine titration using a heparin-responsive electrochemical sensor.

OBJECTIVE: To determine the ability of a new electrochemical sensor to determine heparin levels in whole blood and to correlate the heparin levels as determined by this heparin-responsive sensor (HRS) with heparin levels as determined by the Hepcon assay system (Medtronic Hemotec, Parker, CO). DESIGN: Methods comparison study. SETTING: A large academic medical center. INTERVENTIONS: The heparin levels of 162 samples from 24 patients undergoing cardiopulmonary bypass were determined by the HRS system and by the Hepcon system. In 21 samples, heparin levels as measured by anti-Xa activity were determined as well. MEASUREMENTS AND MAIN RESULTS: HRS-determined values correlated highly with Hepcon-determined values (r = 0.942) and with anti-Xa determined values (r = 0.905). Bias +/- precision comparing the HRS and Hepcon methods was 0.211 +/- 0.478 U/mL. CONCLUSIONS: HRS determined that whole blood heparin levels correlate well with Hepcon-determined levels. These limited results indicate that further development and testing of this new technology are warranted.

Anticoagulants↗
PubMed1996

Selective monitoring of peptidase activities with synthetic polypeptide substrates and polyion-sensitive membrane electrode detection.

A novel method to monitor specific peptidase activities in biological samples as complex as undiluted plasma/blood is described. The approach is based on the design of synthetic polypeptide substrates in which di- or triarginine sequences are linked to each other via one or more other amino acids recognized specifically by the peptidase to be determined. Detection of chymotrypsin and renin activities using synthetic substrates P4 (F-R-R-R-F-V-R-R-F-NH2) and P5 (R-R-R-L-L-R-R-L-L-R-R-R), respectively, serves to demonstrate the principles of this new assay system. A polyion-sensitive membrane electrode, prepared by doping polymer films with dinonylnaphthalene-sulfonate (DNNS), is shown to exhibit significant nonequilibrium electromotive force (EMF) responses toward these and other polycationic substrates at microgram/milliliter levels under physiological conditions. The same electrode, however, exhibits much smaller total EMF response toward the shorter fragments of the synthetic peptides generated by peptidase activity; hence, the addition of peptidase to a solution containing the synthetic substrate yields a change in electrode EMF response, the rate of which is proportional to the activity of peptidase present. Other synthetic polycationic peptides as well as natural polycationic peptides (e.g., protamine) that lack specific cleavage sites for chymotrypsin and renin, yet are detected by the DNNS-based membrane electrode, do not elicit any significant change in EMF response in the presence of the peptidases, confirming the feasibility and utility of the proposed bioanalytical method.

Amino Acid Sequence↗
PubMed1995

Thrombogenic properties of untreated and poly(ethylene oxide)-modified polymeric matrices useful for preparing intraarterial ion-selective electrodes.

In vitro platelet adhesion studies are used to compare the thrombogenic properties of various polymer matrices useful for preparing implantable ion-selective membrane electrodes. Conventional plasticized poly(vinyl chloride) and alternate polyurethane materials (Tecoflex, Pellethane) doped with proton- (tridodecylamine) and potassium-selective (valinomycin) ionophores are shown to be potentially thrombogenic. Incorporation of high molecular weight block copolymers of poly(ethylene oxide) and poly(propylene oxide) (e.g., Pluronic F108 and Tetronic 1508) within ion-selective membranes reduces platelet adhesion. A more marked decrease in platelet adhesion is, however, observed when the Tecoflex-based membranes are coated with a thin photo-cross-linked layer of poly(ethylene oxide). Such surface-modified membranes are shown to retain potentiometric ion response properties (i.e., selectivity, response times, response slopes, etc.) essentially equivalent to untreated membranes.

Animals↗
PubMed1995

Potentiometric anion selectivity of polymer membranes doped with palladium organophosphine complex.

The potentiometric anion selectivity of polymer membrane-based electrodes formulated with a palladium organophosphine complex (benzylbis(triphenylphosphine)palladium(II) chloride) as the membrane active component is examined. The electrode is shown to exhibit a non-Hofmeister selectivity pattern with a significantly enhanced response toward nitrite over the concentration range of 10 microM-10 mM (log-linear range) and a detection limit 5.0 microM. The effect of lipophilic anionic (tetraphenylborate derivatives) and cationic (tetraalkylammonium) site additives within the membrane on the anion selectivity is examined in detail. Addition of both cationic and anionic sites is shown to improve potentiometric anion selectivity, suggesting that the palladium complex may operate simultaneously as a neutral and charged carrier-type ionophore within the polymer membrane phase. Using optimal membrane formulations (with added 20-30 mol % cationic sites), the sensors prepared with the palladium complex do not exhibit proton/hydroxide response in the range of pH 3.5-12, a potential advantage over previously reported nitrite electrodes prepared with Co(III) corrins and porphyrin complexes.

Electrodes↗
PubMed1995

Optical detection of macromolecular heparin via selective coextraction into thin polymeric films.

Thin plasticized polymer films, poly(vinyl chloride) doped with a specific ion pairing quaternary ammonium compound, tridodecylmethylammonium chloride, and a lipophilic pH indicator, 3-hydroxy-4-(4-nitrophenylazo)phenyl octadeconate, are shown to exhibit significant and analytically useful optical response toward macromolecular heparin. The response mechanism is based on favorable extraction of heparin into the bulk organic film, owing to the specific ion-pairing complexation reaction between the quaternary ammonium species and the polyanion. A simultaneous coextraction of hydrogen ions results in protonation of the pH chromophore and hence a change in the optical absorbance of the polymeric film. When used in a limited volume/fixed exposure (10 min) detection mode, film absorbances change as a function of the initial heparin concentration in the range of 0.2-3.0 units/mL (1.2-18 micrograms/mL). The practical measurement response time is controlled by heparin diffusion through the stagnant diffusion layer adjacent to the surface of the film as well as within the bulk of the polymer film and is shown to increase with the molecular weight of the heparin species tested. No optical response to heparin is observed when a strong heparin complexing agent (e.g., protamine) is present in the test solution, suggesting that the polymer film can be used to conveniently monitor heparin-protamine (or other antagonist) titrations. The theory relating to the operation of the sensing film in either the equilibrium or the kinetic mode and the selectivity of the optimized film to heparin relative to small anions are presented.

Anions↗
PubMed1995

Protamine-sensitive polymer membrane electrode: characterization and bioanalytical applications.

A polymeric membrane electrode that exhibits significant and analytically useful potentiometric response to submicromolar levels of the heparin antagonist, protamine, is reported. The sensor is prepared by incorporating a lipophilic cation exchanger, potassium tetrakis(4-chlorophenyl)borate (KTpClPB) (at 1 wt%), within a specially formulated polymer membrane composed of 33 wt% 2-nitrophenyl octyl ether (2-NPOE), and 66 wt% poly(vinyl chloride) (PVC). When the polymer film is mounted in an appropriate electrode body, the resulting membrane electrode responds reproducibly to protamine via a nonequilibrium quasi-steady-state change in the phase boundary potential at the membrane/sample interface. Such response can be used to directly monitor, via classical potentiometric titrations, the binding between protamine and a variety of native (porcine and beef) as well as low-molecular-weight heparins. Scatchard analysis of the EMF titration data provides binding constants and stoichiometries for protamine-heparin interactions. The electrode can be further used to follow the enzymatic digestion of protamine by trypsin. In the presence of a given level of protamine, initial rates of potential decrease (-dE/dt) are shown to be linearly related to trypsin activity in solution over the range of 0-130 units/ml. The speed and simplicity of the protamine sensor make it an attractive alternative to classical methods for studying the interaction of protamine with other biologically important macromolecules as well as the proteolytic activity and reaction kinetics of trypsin.

Electrodes↗
PubMed1995

Novel nonseparation sandwich-type electrochemical enzyme immunoassay system for detecting marker proteins in undiluted blood.

A novel nonseparation electrochemical enzyme immunoassay (NEEIA) for detecting marker proteins in undiluted blood is described. The approach is based on preferential electrochemical measurement of surface-bound enzyme-labeled reporter antibody (E-Ab), relative to an excess of this reagent in the sample solution. NEEIAs are carried out on microporous membranes coated with a thin, circular area of gold. The gold serves simultaneously as a working electrode and solid phase for immobilized capture anti-protein antibodies. In the assay, analyte protein is incubated concurrently with the Ab-coated gold surface and excess E-Ab conjugate. Detection of bound E-Ab is achieved by introducing the substrate for the enzyme through the back side of the membrane. The product of bound E-Ab is detected immediately by oxidation or reduction at the gold electrode, and the resulting current is proportional to the concentration of protein in the sample. The feasibility of the NEEIA approach is demonstrated via the detection of prostate-specific antigen in undiluted plasma samples (n = 64), with alkaline phosphatase as the label. Use of multiple gold films deposited on the same porous membrane to perform simultaneous NEEIAs is also described.

Antibodies, Monoclonal↗
PubMed1994

Selectivity of polymer membrane-based ion-selective electrodes: self-consistent model describing the potentiometric response in mixed ion solutions of different charge.

Despite its well-documented limitations, the semiempirical Nicolsky-Eisenman equation is used throughout the existing analytical literature to describe the selectivity of modern polymer membrane-based ion-selective electrodes (ISEs). In this paper, a new quantitative description for the response/selectivity function based on ion-extraction equilibria at the sample/membrane interface is presented. The proposed selectivity formalism clearly illustrates the range of validity for the conventional Nicolsky-Eisenman formalism. Extended equations are derived describing the electrode response in an exact manner, particularly with respect to analyte and interfering ions of different charge. The expression obtained corresponds to the matched potential method proposed previously by Christian and co-workers on the basis of solely empirical observations. Selectivity coefficients required for a given analytical problem with a predefined maximum error can now be predicted more accurately. Such predictions with respect to analyte and interfering ions of varying charges differ by 1-2 orders of magnitude in comparison to the selectivity values required on the basis of the extended Nicolsky-Eisenman formalism.

Electrochemistry↗
PubMed1994

Response mechanism of polymer membrane-based potentiometric polyion sensors.

The potentiometric response mechanism of a previously reported polymer membrane-based electrode sensitive to the polyanion heparin is established. Based on transport and extraction studies, the heparin response is attributed to a nonequilibrium change in the phase boundary potential at the sample/membrane interface. While true equilibrium polyion response, obtained for low heparin concentrations only after very long equilibration times (> 20 h), yields the expected Nernstian response slope of < 1 mV/decade, the observed large and reproducible EMF response to clinically relevant heparin concentrations (approximately 10(-7) M) during typical measurement periods (2-5 min) is ascribed to a steady-state kinetic process defined by the flux of the polyion both to the surface and into the bulk of the polymer membrane. A model describing this nonequilibrium response is presented. With this model, the uniqueness of the polymer membrane composition (e.g., very low plasticizer content, strictly controlled cationic site concentration, etc.) required to achieve analytically useful heparin response becomes clear. Practical working conditions and limitations of the sensor are discussed. To support the generality of the steady-state model proposed, corresponding EMF response data for a newly developed membrane electrode sensitive to a polycationic protein (protamine) are also presented. It is shown that the protamine-responsive membrane electrode appears to operate via the exact same kinetic mechanism as the heparin sensing system.

Biosensing Techniques↗
PubMed1994

Separation-free sandwich enzyme immunoassays using microporous gold electrodes and self-assembled monolayer/immobilized capture antibodies.

A novel separation-free sandwich-type enzyme immunoassay for proteins is performed by designing an electrochemical detection system that enables preferential measurement of surface-bound enzyme-labeled antibody relative to the excess enzyme-labeled reagent in the bulk sample solution. In this initial model system, the assay is carried out using gold-coated microporous nylon membranes (pore size 0.2 micron) which are mounted between two chambers of a diffusion cell. The membrane serves as both a solid phase for the sandwich assay and the working electrode in the three-electrode amperometric detection system. The capture monoclonal antibody is immobilized covalently on the gold side of the membrane via a self-assembled monolayer of thioctic acid. In the separation-free sandwich assay, both model analyte protein (human chorionic gonadotropin; hCG) and alkaline phosphatase labeled anti-hCG (ALP-Ab) are incubated simultaneously with the immobilized capture anti-hCG antibody. Surface-bound ALP-Ab is spatially resolved from the excess conjugate in the bulk sample solution by introducing the enzyme substrate (4-aminophenyl phosphate) through the back side of the porous membrane. The substrate diffuses rapidly through the porous membrane where it first encounters bound ALP-Ab at the gold surface. The enzymatically generated product, aminophenol, is detected immediately by oxidation at the gold electrode (at +0.19 V vs Ag/AgCl), and the magnitude of current is directly proportional to the concentration of hCG in the sample. The response time after substrate addition is less than 1 min, although maximum response toward the analyte protein requires a sample/conjugate preincubation time of 30 min with the porous electrode.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies↗
PubMed1994

Simplified dual-lumen catheter design for simultaneous potentiometric monitoring of carbon dioxide and pH.

A novel dual-lumen catheter electrode design suitable for the simultaneous measurement of PCO2 (partial pressure of carbon dioxide) and pH in flowing blood is described. The probe is fabricated from a single segment of dual-lumen silicone rubber or polyurethane tubing that is impregnated with the proton ionophore tridodecylamine. The impregnation step imparts H+ permselectivity to both inner and outer walls of the tubing. By filling each lumen with a suitable buffer/electrolyte solution and Ag/AgCl reference electrode wire, simultaneous potentiometric detection of both PCO2 and pH is achieved. Careful optimization of incorporated proton carrier (tridodecylamine), plasticizer (o-nitrophenyl octyl ether), and lipophilic counteranion sites (tetrakis[3,5-bis(trifluoromethyl)phenyl]borate) within the tubing walls yields catheter electrodes with resistance values of 10-20 M omega and relatively high stability in flowing blood. Results from continuous measurements of PCO2 and pH during long-term 30-65-h blood loop experiments demonstrate that, after an initial conditioning period, the catheter exhibits low drift rates (PCO2, 4.7 +/- 1.7 mV/30 h; pH, 1.4 +/- 0.5 mV/30 h) and yields continuously measured values in good agreement with those obtained on discrete samples with a commercial blood gas analyzer (PCO2, r2 = 0.997; pH, r2 = 0.915). In vivo evaluation of the catheter sensors, performed by implanting silicone rubber dual-lumen probes in the arteries of anesthetized dogs, indicates that the proposed catheter design can closely follow PCO2/pH changes induced in the animals during 6-13 h of continuous monitoring.

Animals↗