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Biomedical subjects

M E Meyerhoff

Publications and source records attributed to M E Meyerhoff.

At least 19 recordsLinked to original sources
PubMed1992

Reversible potentiometric oxygen sensors based on polymeric and metallic film electrodes.

Various materials and sensor configurations that exhibit reversible potentiometric responses to the partial pressure of oxygen at room temperature in neutral pH solution are examined. In one arrangement, platinum electrodes are coated with plasticized poly(vinyl chloride) films doped with a cobalt(II) tetraethylene pentamine complex. For such sensors, potentiometric oxygen response is attributed to a mixed potential originating from the underlying platinum electrode surface as well as a change in redox potential of the Co(II)-tetren-doped film as the complex binds oxygen reversibly. The response due to the platinum surface is prolonged by the presence of the Co(II)-tetren/PVC film. Alternately, thin films of metallic copper, electrochemically deposited on platinum and/or sputtered or vapor deposited on a single crystal silicon substrate, may be used for reversible oxygen sensing. The long-term reversibility and potentiometric stability of such copper film-based sensors is enhanced (up to 1 month) by preventing the formation of cuprous oxide on the surfaces via the application of an external nonpolarizing cathodic current through the working electrode or by specifically using sputtered copper films that have [100] preferred crystal structures as determined by X-ray diffraction. The implications of these findings in relation to fabricating analytically useful potentiometric oxygen sensors are discussed.

Cobalt↗
PubMed1992

Lectin-based homogeneous enzyme-linked binding assay for estimating the type and relative amount of carbohydrate within intact glycoproteins.

The feasibility of using a new lectin-based homogeneous enzyme-linked binding assay for estimating the type and relative amount of specific carbohydrate structures within intact glycoproteins is examined. Malate dehydrogenase-galactose, -mannose, and -N-acetylglucosamine conjugates are utilized in conjunction with Jacalin, concanavalin A, and wheat germ agglutinin, respectively. The catalytic activity of the glyco-enzyme conjugates is inhibited significantly (greater than 60%) in solution in the presence of the respective lectins. The observed inhibition for each reagent set is reversed in proportion to the type and relative amount of specific carbohydrates present within test glycoproteins added to the assay mixture. Competitive binding ED50 values for a number of synthetic and native model glycoproteins correlate well with the known carbohydrate content of these species. The proposed method is much faster than previous solid-phase lectin-based enzyme-linked methods used to probe carbohydrate content/structure (less than 15 min) and has the potential to be fully automated.

Acetylglucosamine↗
PubMed1991

Operation of ion-selective electrode detectors in the sub-Nernstian/linear response range: application to flow-injection/enzymatic determination of L-glutamine in bioreactor media.

A novel approach for eliminating positive errors from endogenous ionic interferences when using ion-selective electrodes as detectors in flow-injection enzyme-based blosensing configurations is described. The method involves using a high background level of interfering ions in the sample diluent/carrier stream to convert the normally logarithmic potentiometric sensor into a linear detector over a given concentration range of primary ions. A split-stream single-detector arrangement provides a convenient means to compensate for varying levels of background interferent ions in the injected samples. One portion of the split stream passes directly to the ion-electrode detector, yielding a signal linearly related to the concentration of endogenous primary ions in the sample. The second portion of the split sample is delayed while passing through an immobilized enzyme that generates electrode detectable primary ions in proportion to the concentration of the substrate analyte in the sample. Two linear equations with two unknowns describe the twin potentiometric responses observed. The concept is demonstrated by the accurate determination of L-glutamine in hybridoma bioreactor media via the use of an ammonium-ion-selective membrane electrode detector and immobilized glutaminase enzyme.

Biosensing Techniques↗
PubMed1991

Electrochemical performance, biocompatibility, and adhesion of new polymer matrices for solid-state ion sensors.

Ammonium and potassium ion-selective membranes formulated with PVC/hydroxylated PVC, polyurethane/hydroxylated PVC, and moisture-curable silicone rubber matrices are studied in an effort to extend the lifetime of solid-state ion sensors through improved membrane adhesion. The PVC/membranes exhibit electrochemical performance equivalent to that of conventional PVC membranes in terms of slope, detection limit, and selectivity. The polyurethane- and silicone-rubber-based membranes have better adhesion to silicon nitride than do PVC or hydroxylated PVC matrices. Incorporating a silanizing reagent (silicon tetrachloride) significantly improves the adhesion of the polyurethane matrix. The use of silicon tetrachloride in membrane matrices also enhances the electrochemical stability of the interfacial potential between ion-selective polymer-matrix membranes and silver epoxy inner reference electrodes of solid-state sensors. The biocompatibility of the polymer matrices is examined via radiotracer protein adsorption studies and whole blood clotting time measurements. The polyurethane- and silicone-rubber-based membranes exhibit less overall nonspecific protein adsorption than the PVC or hydroxylated PVC matrices.

Adhesiveness↗
PubMed1991

Potentiometric enzyme channeling immunosensor for proteins.

A potentiometric immunosensor for the detection of human IgG has been developed using an asymmetric, ion-selective membrane with immobilized adenosine deaminase and IgG. A protein A-alkaline phosphatase conjugate binds to the immobilized IgG, creating a bienzymatic catalytic layer. In the presence of sample IgG, the conjugate does not bind to the membrane. Instead, the intermediate in the two-step reaction (adenosine) must diffuse to the membrane surface, reducing the rate of product (ammonium) formation within the diffusion layer detected by the membrane. The immunosensor demonstrated is for the determination of IgG. A simplified model is described to predict the maximum rate enhancement for the 'channeled' versus 'unchanneled' reaction mechanisms.

Biosensing Techniques↗
PubMed1990

Homogeneous enzyme-linked binding assay for studying the interaction of lectins with carbohydrates and glycoproteins.

A simple and rapid homogeneous enzyme-linked binding assay method for studying lectin-carbohydrate interactions is described. The method is based on the homogeneous inhibition of appropriate enzyme-saccharide conjugates by specific carbohydrate-binding lectins. In the presence of carbohydrate structures recognized by the lectins, enzyme activity is regained in an amount of proportional to the concentration of carbohydrate. The new method can be used to rapidly assess the relative carbohydrate specificity of the various lectins and for the selective analytical detection of simple saccharides and complex glycoproteins. Indeed, when Jacalin lectin is used in conjunction with a malate dehydrogenase-galactose conjugate, selective measurement of human IgA (immunoglobulin A) at microgram per milliliter levels in less than 10 min is possible. The potential for using this analytical methodology for determining changes in the carbohydrate structure of intact recombinant glycoproteins is also discussed.

Carbohydrates↗
PubMed1990

Use of ionomer membranes to enhance the selectivity of electrode-based biosensors in flow-injection analysis.

The use of ionomer membranes to enhance the selectivity of potentiometric enzyme electrodes in flow-injection measurement arrangements is examined. The ionomer membranes employed are permeable to analyte substrates but relatively impermeable to detectable ions that would normally interfere with the measurement of the substrates if the enzyme electrodes were in direct contact with the sample. As a model system, the selectivity of enzyme electrodes prepared with nonactin-based ammonium-sensitive polymeric membranes is evaluated. In the preferred configuration, a thin hydrophilic anion-exchange membrane is incorporated within a flow-through dialysis unit upstream from the enzyme-electrode detector. As the sample passes through the dialysis unit, neutral or anionic analyte molecules (urea or glutamine) move through the membrane while the permeation of endogenous ammonium ions and other cations in the sample is retarded. A flowing recipient buffer on the other side of the membrane carries the analyte substrate to the enzyme-electrode detector. Enhancements in selectivity for analyte substrates over endogenous ammonium and potassium ions are greater than or equal to 9-fold when compared to enzyme-electrode flow-injection analysis (FIA) systems assembled without the ionomer membrane unit. The analytical utility of the proposed system is demonstrated by the accurate measurements of urea in blood serum and L-glutamine in hybridoma bioreactor media.

Biosensing Techniques↗
PubMed1990

Current and future directions in the technology relating to bedside testing of critically ill patients.

Significant progress has been made recently in the measurement methods and instrumental approaches applicable to bedside testing of critically ill patients. While the "ideal" technology would involve the ability to obtain accurate stat profile values on a continuous basis via noninvasive methods, given the present state of noninvasive sensing technologies, this capability is unlikely to be achieved in the foreseeable future. In principle, invasive and on-line techniques offer more hope for future success in continuous bedside monitoring of all the key critical care analyses. However, success in these directions will come only when issues regarding sensor stability and sampling device/sensor biocompatibility are completely solved. Until then, it appears that the user-friendly point of care type stat analyzers that can provide accurate values for all the key analytes, used in conjunction with existing noninvasive trend monitors (eg, pulse oximetry), will offer the most attractive approach for the effective treatment of critically ill patients.

Clinical Laboratory Techniques↗
PubMed1990

A simple method for estimating association constants between monoclonal antibodies and derivatized or native antigens.

We describe a simple method for estimating the association constant between an antibody and an antigen, based on a theoretical treatment of experimental dose-response data. We used this method to calculate the association constants of an anti-progesterone monoclonal antibody for native progesterone and for progesterone 11 alpha-hemisuccinate. These association constants provide a quantitative measure of the bridging-group recognition by the antibody.

Antibodies, Monoclonal↗
PubMed1990

New in vitro analytical approaches for clinical chemistry measurements in critical care.

The effective management of patients in intensive care units, operating rooms, and emergency rooms requires frequent measurement of a select group of analytes, preferably at or near the patient's bedside. Tests recognized as being essential for such management include blood gases and related variables (pH, pO2, pCO2, HCO3-, hematocrit/hemoglobin, O2 saturation), electrolytes (Na+, K+, Ca2+, Cl-), and in some cases, certain metabolites (glucose, lactate, urea, creatinine). This report describes the measurement principles, practical instrumental designs, analytical performance, and limitations of several newer electrochemical sensor-based approaches useful for in vitro determination of these species in undiluted whole-blood samples. Considerable attention is given to the most recent advances in ion-selective electrode technology as they relate to blood gas and electrolyte determinations. Similar attention is given to modern enzyme-electrode techniques, which are useful for direct measurements of metabolites in whole blood. The challenges of integrating these new analytical methods into convenient, multi-analyte, user-friendly, bedside or stat-lab instruments are also discussed.

Autoanalysis↗
PubMed1989

Potentiometric combination ion/carbon dioxide sensors for in vitro and in vivo blood measurements.

The development and analytical performance of a novel potentiometric combination ion/pCO2 sensor design for in vitro and in vivo measurements are reported. The design is based on incorporating an appropriate ionophore within the outer silicone gas permeable membranes of both conventional macro and new catheter-type pCO2 sensors. Simultaneous measurement of the potentials across the ion-selective/gas permeable membrane and the inner glass or polymer pH sensitive membrane provides the basis for continuous monitoring of both ionic and pCO2 levels with the same device. A macro-sized K+/pCO2 embodiment of the sensor is constructed from a commercial Severinghaus CO2 sensor and is used to demonstrate the principles and capabilities of the proposed design. A flexible, miniaturized (outer diameter = 1.2 mm) combination K+/pCO2 catheter sensor is also described. The catheter-type sensor is fabricated by inserting a tubular polymer membrane pH electrode into an outer silicone rubber tube doped with valinomycin. Continuous measurements of K+ and pCO2 during 6-h blood pump studies using both the macro and catheter-type combination sensors correlate well with those of conventional bench-top analyzers. In addition, continuous (4 h) intravascular measurements with the combination catheter sensor in dogs show good agreement with those of commercial blood analyzers (R = 0.984 and 0.962 for pCO2 and K+, respectively.

Biosensing Techniques↗
PubMed1989

Salicylate-selective membrane electrode based on tin(IV) tetraphenylporphyrin.

The response properties of a new solvent/polymeric membrane electrode with unique selectivity toward anionic salicylate are reported. The electrode is prepared by incorporating 5, 10, 15, 20-tetraphenyl(porphyrinato)tin(IV) dichloride (Sn[TPP]Cl2) into a plasticized poly(vinyl chloride) membrane. The resulting sensor exhibits an anti-Hofmeister selectivity pattern, with high specificity for salicylate over lipophilic inorganic anions (perchlorate, periodate, thiocyanate, iodide, etc.) and biological organic anions (citrate, lactate, acetate). Moderate selectivity over structural analogues of salicylate (3- and 4-hydroxybenzoate, benzoate) is also observed. Radiotracer uptake experiments using [14C]salicylate clearly show that the metal center of the metalloporphyrin is critical for selective salicylate transport in the membrane phase. Minimal response to chloride ions makes the new electrode potentially useful for estimating salicylate levels in biological samples.

Electrodes↗