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Biomedical subjects

F V Bright

Publications and source records attributed to F V Bright.

12 recordsLinked to original sources
PubMed1995

Dynamics surrounding Cys-34 in native, chemically denatured, and silica-adsorbed bovine serum albumin.

We report the steady-state and time-resolved fluorescence of 6-acryloyl(dimethylamino)naphthalene (acrylodan) covalently attached to Cys-34 in bovine serum albumin (BSA). For this conceptually simple system, complicated fluorescence intensity and anisotropy decay kinetics are observed. The steady-state and time-resolved results demonstrate the presence of an excited-state reaction for the BSA-acrylodan system. Additional analysis shows that dipolar relaxation of the environment surrounding acrylodan within BSA is responsible for most of the observed time-dependent evolution of the emission spectrum. The effects of temperature, chemical denaturation, and protein adsorption to a bare silica substrate are also investigated. These results demonstrate the complexity of the changes within a protein/biorecognition element that affect the signal from a single fluorescent reporter group.

2-Naphthylamine↗
PubMed1995

Synthesis and characterization of fluoropolymeric substrata with immobilized minimal peptide sequences for cell adhesion studies. I.

In this work, poly(tetrafluoroethylene-co-hexafluoropropylene) (also known as fluorinated ethylene propylene; FEP) was functionalized at the surface using a radio frequency glow discharge plasma. This particular surface modification produced controlled densities of hydroxyl functionality on the FEP surface. These surface hydroxyl groups provided sites for the covalent attachment of minimal peptide sequences, that are specific for neuronal attachment. FSCA, ATR-FTIR, ToF-SIMS, and fluorescence spectroscopy were used to evaluate peptide reaction efficiencies and to verify that intact peptide sequences were covalently attached to the FEP surfaces. These modified substrata were then used to study the cell attachment and response to covalently bound minimal peptide sequences. Cell attachment and differentiation results using NG108-15 and PC12 neuronal cell lines are presented in the adjoining paper by Ranieri et al.

Amino Acid Sequence↗
PubMed1990

Regenerable fiber-optic-based immunosensor.

An immunosensor is described that is based on fluorescently labeled F(ab') anti-human serum albumin antibody fragments covalently immobilized to the distal end of a fiber-optic probe. When human serum albumin is present, it is bound to the sensor and shields the fluorescent label from the solvent water, and a significant increase in the label fluorescence results. The sensor can be regenerated by simply immersing the sensing tip in chaotropic media. Under these conditions the antigen-antibody complex is selectively disrupted without adversely affecting the sensor. In the present configuration, the same sensor can be recycled over 50 times before the immunosurface inactivates significantly. With proper storage the sensor can last for up to 4 months.

Electrodes↗
PubMed1990

Effects of inaccurate reference lifetimes on interpreting frequency-domain fluorescence data.

The effects of assigning inaccurate reference lifetimes in lifetime determinations are predicted theoretically by using standard equations. This theory leads to a method to remove reference error effects using common least-squares software. This method cannot, however, be used to deconvolute data collected with isochronal references. Uncorrected data can always be exactly solved with models containing one more degree of freedom than the true model. Monoexponential decays are fit by double-exponential decays or excited-state processes. Unimodal distributed decays often appear as discrete, double-exponential decays.

Kinetics↗
PubMed1989

Multicomponent fluorometric analysis using a fiber-optic probe.

Single-frequency phase-resolved fluorometry through single and bifurcated fiber-optic probes is used to quantify mixtures of spectrally similar fluorophores. Correlation coefficients from correlation plots are greater than 0.98, and standard errors of estimate are less than 0.13 microM for 80 binary mixtures. Quantification of fluorophores at up to 10:1 molar ratios with lifetime separations of less than 200 ps is possible. The simultaneous quantification of the individual components of ternary and quaternary synthetic mixtures is demonstrated. Application of phase resolution to ambient light rejection is reported also. In this case, the emission spectrum of 10 nM Rhodamine 6G is easily obtained in the presence of a fluctuating, unmodulated background.

Anthracenes↗
PubMed1989

Thermodynamic study on the effects of beta-cyclodextrin inclusion with anilinonaphthalenesulfonates.

Thermodynamic parameters and stoichiometries for the binding of anilinonaphthalenesulfonates to beta-cyclodextrin are obtained from steady-state fluorescence intensity and anisotropy measurements. Specifically, formation constant, enthalpy, and entropy values are obtained for complexes of beta-cyclodextrin with eight different substrate molecules at five different temperatures and six different pH values, and their associated errors are given. We propose an explanation of the relative magnitudes of the values obtained with regard to the geometry of the substrate and the importance of the various noncovalent interactions responsible for the complexation.

Anilino Naphthalenesulfonates↗
PubMed1989

Multifrequency phase fluorescence study of hapten-antibody complexation.

The rotational dynamics of a phenytoin fluoroimmunoassay system is investigated by using multifrequency phase and modulation fluorescence spectroscopy. Results indicated that the fluorescein label on this relatively small hapten undergoes significant (greater than 45%) local motion even when bound to the phenytoin antibody. We attribute this local motion to unhindered rotation of the fluorescein moiety around the bond attaching it to phenytoin. This local motion affects the sensitivity of a steady-state polarization immunoassay dramatically.

Antibodies↗
PubMedDate not supplied

In situ studies of protein conformation in supercritical fluids: trypsin in carbon dioxide.

The conformation of the monomeric enzyme trypsin has been studied in supercritical carbon dioxide. Steady-state fluorescence spectroscopy is used to follow the conformation of trypsin in situ as a function of CO2 density. Our results show for the first time that protein denaturation can occur during the fluid compression step and that the native trypsin is only slightly more stable (1.2 kcal/mol) than the unfolded form. These results demonstrate the power of fluorescence spectroscopy as a tool for studying protein conformation and dynamics in supercritical fluids.

Carbon Dioxide↗