Biomedical subjects

Gabriel P Lopez

Publications and source records attributed to Gabriel P Lopez.

6 recordsLinked to original sources

PubMed2004

Near-simultaneous and real-time detection of multiple analytes in affinity microcolumns.

A miniaturized immunoassay system based on beads in poly(dimethylsiloxane) microchannels for analyzing multiple analytes has been developed. The method involves real-time detection of soluble molecules binding to receptor-bearing microspheres, sequestered in affinity column format inside a microfluidic channel. Identification and quantitation of analytes occurs via direct fluorescence measurements or fluorescence resonance energy transfer. A preliminary account of this work based on single-analyte format has been published in this journal (Buranda, T.; Huang, J.; Perez-Luna, V. H.; Schreyer, B.; Sklar, L. A.; Lopez, G. P. Anal. Chem. 2002, 74, 1149-1156). We have extended the work to a multianalyte model system composed of discrete segments of beads that bear distinct receptors. Near-simultaneous and real-time detection of diverse analytes is demonstrated. The importance of this work is established in the exploration of important factors related to the design, assessment, and utility of affinity microcolumn sensors. First, beads derivatized with surface chemistry suitable for the attachment of fluorescently labeled biomolecules of interest are prepared and characterized in terms of functionality and receptor site densities by flow cytometry. Second, calibrated beads are incorporated in microfluidic channels. The analytical device that emerges replicates the basic elements of affinity chromatography with the advantages of microscale and real-time direct measurement of bound analyte on beads rather than the indirect determination from eluted sample typical of affinity chromatography. In addition, the two-compartment analysis of the assay data as demonstrated in single-analyte columns provides a template upon which the dynamics of multiple-analyte assays can be characterized using existing theoretical models and be tested experimentally. The assay can potentially detect subfemtomole quantities of protein with high signal-to-noise ratio and a large dynamic range spanning nearly 4 orders of magnitude in analyte concentration in microliter to submicroliter volumes of analyte fluid. The approach has the potential to be generalized to a host of bioaffinity assay methods including analysis of protein complexes (e.g., biomolecular indicators of diseases). Proof-of-principle analytes include FLAG peptide and carcinoembryonic antigen detected at physiologically relevant concentration levels.

Carcinoembryonic Antigen↗

PubMed2004

Effect of substratum surface chemistry and surface energy on attachment of marine bacteria and algal spores.

Two series of self-assembled monolayers (SAMs) of omega-substituted alkanethiolates on gold were used to systematically examine the effects of varying substratum surface chemistry and energy on the attachment of two model organisms of interest to the study of marine biofouling, the bacterium Cobetia marina (formerly Halomonas marina) and zoospores of the alga Ulva linza (formerly Enteromorpha linza). SAMs were formed on gold-coated glass slides from solutions containing mixtures of methyl- and carboxylic acid-terminated alkanethiols and mixtures of methyl- and hydroxyl-terminated alkanethiols. C. marina attached in increasing numbers to SAMs with decreasing advancing water contact angles (theta(AW)), in accordance with equation-of-state models of colloidal attachment. Previous studies of Ulva zoospore attachment to a series of mixed methyl- and hydroxyl-terminated SAMs showed a similar correlation between substratum theta(AW) and zoospore attachment. When the hydrophilic component of the SAMs was changed to carboxylate, however, the profile of attachment of Ulva was significantly different, suggesting that a more complex model of interfacial energetics is required.

Bacterial Adhesion↗

PubMed2003

Microchip countercurrent electroseparation.

We report a microchip-based method for separating charged molecules according to electrophoretic mobility. The method is based on opposed electroosmotic, electrophoretic and convective forces. Similar to isoelectric focusing, solute can be accumulated into stationary zones, but without use of ampholytes. The method of "microchip countercurrent electroseparation" described here has potential application in the design of microfluidic separation chips.

Journal Article↗

PubMed2003

Functional nanocomposites prepared by self-assembly and polymerization of diacetylene surfactants and silicic acid.

Conjugated polymer/silica nanocomposites with hexagonal, cubic, or lamellar mesoscopic order were synthesized by self-assembly using polymerizable amphiphilic diacetylene molecules as both structure-directing agents and monomers. The self-assembly procedure is rapid and incorporates the organic monomers uniformly within a highly ordered, inorganic environment. By tailoring the size of the oligo(ethylene glycol) headgroup of the diacetylene-containing surfactant, we varied the resulting self-assembled mesophases of the composite material. The nanostructured inorganic host altered the diacetylene polymerization behavior, and the resulting nanocomposites show unique thermo-, mechano-, and solvatochromic properties. Polymerization of the incorporated surfactants resulted in polydiacetylene (PDA)/silica nanocomposites that were optically transparent and mechanically robust. Molecular modeling and quantum calculations and (13)C spin-lattice relaxation times (T(1)) of the PDA/silica nanocomposites indicated that the surfactant monomers can be uniformly organized into precise spatial arrangements prior to polymerization. Nanoindentation and gas transport experiments showed that these nanocomposite films have increased hardness and reduced permeability as compared to pure PDA. Our work demonstrates polymerizable surfactant/silica self-assembly to be an efficient, general approach to the formation of nanostructured conjugated polymers. The nanostructured inorganic framework serves to protect, stabilize, and orient the polymer, mediate its performance, and provide sufficient mechanical and chemical stability to enable integration of conjugated polymers into devices and microsystems.

Acetylene↗

PubMed2002

Biomolecular recognition on well-characterized beads packed in microfluidic channels.

We describe a new approach for the analysis of biomolecular recognition in microfluidic channels. The method involves real-time detection of soluble molecules binding to receptor-bearing microspheres, sequestered in affinity column format inside a microfluidic channel. Identification and quantitation of analytes occurs via direct fluorescence measurements or fluorescence resonance energy transfer (FRET). We establish a model system that detects the FLAG epitope. The assay can potentially detect subfemtomole quantities of antibody with a high signal-to-noise ratio and a large dynamic range spanning nearly 4 orders of magnitude in analyte concentration in microliter-to-submicroliter volumes of analyte fluid. Kinetic and equilibrium constants for the reaction of this receptor-ligand pair are obtained through modeling of kinetic responses of the affinity microcolumn and are consistent with those obtained by flow cytometry. Because of the correlation between kinetic and equilibrium data obtained for the microcolumns, quantitative analysis can be done prior to the steady-state end point of the recognition reaction. This method has the promise of combining the utility of affinity chromatography with the advantage of direct, quantitative, and real-time analysis and the cost-effectiveness of microanalytical devices. The approach has the potential to be generalized to a host of bioaffinity assay methods including analysis of protein complexes and molecular assembly and microsystem-based multianalyte determinations.

Antibodies, Monoclonal↗

PubMedDate not supplied

Dual closed-loop, optoelectronic, auto-oscillatory detection circuit for monitoring fluorescence lifetime-based chemical sensors and biosensors.

We present a new detection instrument for sensor measurements based on excited-state fluorescence lifetimes. This system consists of a primary optoelectronic loop containing a resonance-type rf amplifier, a modulatable fluorescence-excitation light source, a fiber optic feedback loop (with a gap for a fluorescent sensor), and a photomultiplier tube. A secondary, phase-feedback optoelectronic circuit consists of a long-wavelength-pass optical filter, a second photomultiplier tube, a photodiode, an electronic phase detector, a dc amplifier, and an electronic phase shifter (inserted into the main loop). This phase-feedback circuit is new with respect to our previous work. Under the appropriate conditions, the main loop exhibits self-oscillations, manifesting themselves as sinusoidal rf modulation of light intensity. The phase-feedback circuit detects the modulation phase shift resulting from the finite excited-state lifetimes of a fluorophore. As the excited state lifetime changes, the phase shift from the electronic phase shifter also changes, which results in a shift in self-oscillation frequency. The detection system uses self-oscillation frequency as the detection parameter and has excellent resolution with respect to changes in excited-state lifetime ( approximately 1 ps). (c) 2004 Society of Photo-Optical Instrumentation Engineers.

Biosensing Techniques↗