The objective of this study was to examine membrane filtration of a single stranded DNA (ssDNA) with 60 thymine nucleotides, and to elucidate the variables controlling its transmission across track-etched porous membranes. Dead end filtration measurements were performed using different pore size membranes (10, 15, and 30 nm) at different transmembrane pressures in solutions with ionic strength ranging from 0 to 1000 mM NaCl. The diffusivity of the ssDNA was determined using fluorescence recovery after photobleaching, yielding hydrodynamic radii ranging from 1.
View Article and Find Full Text PDFIn this paper, predictions from a theoretical model describing the rejection of a rigid spherical solute from porous membranes are compared to experimental results for a single stranded DNA (ssDNA) with 60 thymine nucleotides. Experiments were conducted with different pore size track-etched membranes at different transmembrane pressures and different NaCl concentrations. The model includes both hydrodynamic and electrostatic solute-pore wall interactions; predictions were made using different size parameters for the ssDNA (radius of gyration, hydrodynamic radius, and root mean square end-to-end distance).
View Article and Find Full Text PDFThe aqueous bulk diffusivities of several near-spherical (icosahedral) and nonspherical (tailed) bacterial viruses were experimentally determined by measuring their flux across large pore membranes and using dynamic light scattering, with excellent agreement between values measured using the two techniques. For the icosahedral viruses, good agreement was also found between measured diffusivity values and values predicted with the Stokes-Einstein equation. However, when the tailed viruses were approximated as spheres, poor agreement was found between measured values and Stokes-Einstein predictions.
View Article and Find Full Text PDFHypotheses: Particle rejection from porous membranes will increase when particle and membrane carry like charges. The influence of charge on particle rejection can be modeled by first solving the Poisson-Boltzmann equation for the electrostatic particle-pore wall interaction energy, enabling one to predict the cross sectional particle concentration in a pore. Rejection coefficients can then be predicted by combining the Boltzmann factor with a hydrodynamic lag coefficient.
View Article and Find Full Text PDFResults from an investigation of the frequency response resulting from ligand binding for a genetically engineered hormone-binding domain of the alpha-estrogen receptor immobilized to a piezoelectric quartz crystal are reported. Two different approaches were used to attach a genetically altered receptor to the gold electrode on the quartz surface: (1) the mutant receptor containing a single solvent-exposed cysteine was directly attached to the crystal via a sulfur to gold covalent bond, forming a self-assembled protein monolayer, and (2) the N-terminal histidine-tagged end was utilized to attach the receptor via a 3,3-dithiobis[N-(5-amino-5-carboxypentyl)propionamide-N',N'-diacetic acid] linker complexed with nickel. Previous studies have shown that these engineered constructs bind 17beta-estradiol and are fully functional.
View Article and Find Full Text PDFThis article describes a biosensor that detects estrogenic substances using a quartz crystal microbalance with a genetically engineered construct of the hormone-binding domain of the alpha-estrogen receptor. The receptor was immobilized to a piezoelectric quartz crystal via a single exposed cysteine, forming a uniform orientation on the crystal surface. Our results illustrate that this sensor responds to a variety of ligands that are known to bind to the estrogen receptor.
View Article and Find Full Text PDFWe 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.
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