University of Utah Research Foundation

Biocentrex, LLC

Plc Diagnostics, Inc.

Ivd Systems, LLC

Duke University

James N. Herron

W. Monty Reichert

Douglas A. Christensen

Hsu-kun Wang

James N Herron

Douglas A Christensen

Hsu-Kun Wang

Karin D Caldwell

Shao-Chie Huang

Ai-Ping Wei

W Monty Reichert

Eric M Simon

Richard D McEachern

Vera Janatova

Victor A Pollak

Jacob D Durtschi

Vera Janatov&aacute;

Vera Janatová

Scott D Miles

Steven M Blair

Reuven Duer

Joseph D Andrade

Farhad Mahdavi

Samuel Tolley

Yongdong Liu

Victor A Pollack

Ajay Nahata

A biomolecular assay includes a substrate with a metallic layer on at least one surface thereof. The metallic film includes nanocavities. The nanocavities are configured to enhance signals that are representative of the presence or amount of one or more analytes in a sample or sample solution, and may be configured to enhance the signal by a factor of about two or more or by a factor of about three or more. Such signal enhancement may be achieved with nanocavities that are organized in an array, randomly positioned nanocavities, or nanocavities that are surrounded by increased surface area features, such as corrugation or patterning, or nanocavities that have quadrilateral or triangular shapes with tailored edge lengths, or with a plurality of nanoparticles. Methods for fabricating biomolecular substrates and assay techniques in which such biomolecular substrates are used are also disclosed.

Biosensors including metallic nanocavities

A system and methods of sequencing a nucleic acid by detecting the identity of a fluorescent nucleotide analogue incorporated at the 3' end of a growing nucleic acid strand are provided. The system may include a substrate comprising a plurality of substantially parallel excitation waveguides, and a plurality of substantially parallel collection waveguides, the excitation waveguides and collection waveguides crossing to form a two-dimensional array of intersection regions, a plurality of optical sensing sites in optical communication with the intersection regions, one or more switchable light sources and a detector coupled to the light dispersive module. Methods of using these systems are also described.

System and method for nucleic acids sequencing by phased synthesis

Methods and apparatus for detecting single nucleotide polymorphisms in genes of interest are disclosed. A plurality of probes is immobilized on a planar waveguide. The probes comprise sequences complementary to a wildtype sequence of the gene of interest and complementary to a sequence of a known SNP in the gene of interest. A fluorescently-labeled analyte is flowed over the planar waveguide. The binding between the labeled analyte and each of the probes causes a change in the fluorescence signal. The SNP is detected by comparing the hybridization kinetics of the analyte with each of the probes. A method of detecting single nucleotide polymorphisms in a gene of interest by sequencing by hybridization is also disclosed.

Detection of single nucleotide polymorphisms using planar waveguides

A composite waveguide for evanescent sensing in fluorescent binding assays comprising a substrate layer having one or more thin-film waveguide channels deposited thereon. Binding molecules having the property of binding with specificity to an analyte are immobilized on the surface of the thin-film channels. In preferred embodiments, the composite waveguide includes integral light input coupling means adapted to the thin-film channels. Light coupling means may include a grating etched into the substrate prior to deposition of the thin film, or a waveguide coupler affixed to the upper surface of the thin film. The waveguide coupler has an input waveguide of high refractive index which receives the laser light through one end, propagating it by total internal reflection. Propagated light is coupled evanescently into the thin film across a spacer layer of precise thickness with a lower index of refraction than that of the input waveguide or the thin-film waveguide.

Integrated optic waveguide immunosensor

Improvements in a biosensor of the type having reservoirs or wells for analyzing a biological liquid are disclosed. A biosensor () includes a waveguide () placed between a plurality of members such as plates (), at least one of the members () being formed to define the walls () of the reservoirs where the liquid is biologically analyzed. The walls of the reservoirs are made of an inert, opaque material such as a metal. Although the biosensor may include a gasket (), the gasket is associated with the members and waveguide in such a way (e.g. by recessing the gasket into a channel formed into a metal plate) so that the gasket does not form any significant portion of the reservoir wall. Waveguides of varying composition (e.g. plastic, quartz or glass) may be associated with the members to form the biosensor. The metal plate of the biosensor has input and output ports for infusing, draining, or oscillating the liquid to be analyzed in the reaction reservoir. Also disclosed is a sled-shaped waveguide associated with a rear lens to couple light out of the waveguide to serve as a quality control measure thus insuring that the biosensor is properly placed and that the light is working.

Lens and associatable flow cell

Fluorescent energy transfer dyes capable of moving between a more stacked configuration to exhibit fluorescent quenching and a more spaced configuration to exhibit fluorescence can be conjugated to a peptide epitope or nucleic acid for use in the detection of an unknown antibody in bulk solution. The resulting labeled peptide reagent can be used in an immunoassay procedure by placing it in bulk solution along with the unknown antibody to be detected. When the antibody binds to the peptide epitope, the pair of dyes carried by the peptide epitope will have their configuration altered from a stacked to an unstacked configuration and will exhibit a fluorescent increase in response thereto.

Assay procedure using fluorogenic tracers

Methods and apparatus for evanescent light fluoroimmunoassays are disclosed. The apparatus employs a planar waveguide and optionally has multi-well features and improved evanescent field intensity. The preferred biosensor and assay method have the capture molecules immobilized to the waveguide surface by site-specific coupling chemistry. Additionally, the coatings used to immobilize the capture molecules provide reduced non-specific protein adsorption.

Waveguide immunosensor with coating chemistry and providing enhanced sensitivity

Methods and apparatus for evanescent light fluoroimmunoassays are disclosed. The apparatus employs a planar waveguide with an integral semicylindrical lens, and has multi-analyte features and calibration features, along with improved evanescent field intensity. A preferred embodiment of the biosensor and assay method has patches of capture molecules, each specific for a different analyte disposed adjacently within a single reservoir. The capture molecules are immobilized to the patches on the waveguide surface by site-specific coupling of thiol groups on the capture molecules to photo-affinity crosslinkers, which in turn are coupled to the waveguide surface or to a nonspecific binding-resistant coating on the surface. The patches of different antibodies are produced by selectively irradiating a portion of the waveguide surface during the process of coupling the photo-affinity crosslinkers, the selective irradiation involving a mask, a laser light source, or the like.

Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays

A step-gradient composite waveguide for evanescent sensing in fluorescent binding assays comprises a thick substrate layer having one or more thin film waveguide channels deposited thereon. In one embodiment, the substrate is silicon dioxide and the thin film is silicon oxynitride. Specific binding molecules having the property of binding with specificity to an analyte are immobilized on the surface of the thin film channels. In preferred embodiments, the composite waveguide further includes light input coupling means integrally adapted to the thin film channels. Such light coupling means can be a grating etched into the substrate prior to deposition of the thin film, or a waveguide coupler affixed to the upper surface of the thin film. The waveguide coupler has a thick input waveguide of high refractive index which receives the laser light through one end and propagates it by total internal reflection. The propagated light is then coupled evanescently into the thin film waveguide across a spacer layer of precise thickness and having an index of refraction lower than either the input waveguide or the thin-film waveguide. The composite waveguide can be constructed by plasma vapor deposition of silicon oxynitride onto the silicon dioxide substrate, masking the channel waveguides with a photoresist, and using reactive ion etching to expose the substrate in the unmasked regions.

Composite waveguide for solid phase binding assays

An improved oligopeptide composition for use in a fluorescent polarization immunoassay for a high molecular weight analyte is disclosed, along with a kit and a method using the composition. The composition comprises an oligopeptide selected by a screening procedure in which a plurality of different oligopeptides having respective amino acid sequences that represent sequential overlapping segments of the analyte amino acid sequence, and a fluorescent label bound thereto. In a preferred embodiment, the oligopeptide has an amino acid sequence which does not form internal disulfide bridges. Such a preferred oligopeptide will generally have no more than one cysteine residue. In a further preferred embodiment, the fluorescent label is tetramethylrhodamine or a cyanine dye. The kit may be packaged with instructions directing a user to prepare an assay solution containing the monoclonal antibody and the oligopeptide in certain respective concentrations. The composition, method and kit are preferably constructed to detect nanomolar concentrations of the analyte.

Compositions and kits for fluorescence polarization assay of large molecules

Improvements in a biosensor are disclosed. A biosensor includes a waveguide, at least a portion of which is substantially planar. One or more reservoirs may be formed adjacent to a chemistry-bearing surface of the waveguide. The biosensor may include a gasket to form a seal between the waveguide and side walls of the reservoir. A sample solution may be introduced into the reservoir or otherwise onto the surface of a waveguide through an input port. Waveguides of varying composition (e.g., plastic, quartz, glass, or other suitable waveguide materials) may be used in the biosensor. Also disclosed is a sled-shaped waveguide, which includes a planar portion and a lens at an end thereof and angled relative thereto for coupling light into the waveguide.

Disclosed are fluorescent energy transfer dyes which are capable of moving between a more stacked configuration to exhibit fluorescent quenching and a more spaced configuration to exhibit fluorescence can be conjugated to a peptide epitope for use in the detection of an unknown antibody in bulk solution. The resulting labeled peptide reagent can be used in an immunoassay procedure by placing it in bulk solution along with the unknown antibody to be detected. When the antibody binds to the peptide epitope, the pair of dyes carried by the peptide epitope will have their configuration altered from a stacked to an unstacked configuration and will exhibit a fluorescent increase in response thereto.

Immunoassay procedure utilizing fluorogenic tracer antigens

Improvements in a biosensor of the type having reservoirs or wells for analyzing a biological liquid are disclosed. A biosensor includes a waveguide placed between a plurality of members such as plates at least one of the members being formed to define the walls of the reservoirs where the liquid is biologically analyzed. The walls of the reservoirs are made of an inert, opaque material such as a metal. Although the biosensor may include a gasket, the gasket is associated with the members and waveguide in such a way (e.g., by recessing the gasket into a channel formed into a metal plate) so that the gasket does not form any significant portion of the reservoir wall. Waveguides of varying composition (e.g., plastic, quartz or glass) may be associated with the members to form the biosensor. The metal plate of the biosensor has input and output ports for infusing, draining, or oscillating the liquid to be analyzed in the reaction reservoir. Also disclosed is a sled-shaped waveguide associated with a rear lens to couple light out of the waveguide to serve as a quality control measure thus insuring that the biosensor is properly placed and that the light is working.

Growing community of inventors

Salt Lake City, UT, United States of America

James N Herron