Hewlett-packard Company

Agilent Technologies, Inc.

Kay Lichtenwalter

Gary B Gordon

William S Hancock

Scott A Conradson

James A Apffel

John A Chakel

Philip John Kuekes

Calvin B Ward

Hewlett E Melton, Jr

Karen M Hahnenberger

Gargi Choudhary

An assay plate for detecting the presence of a mobile reactant that binds to a immobilized reactant and the methods of making and using the same. An assay plate according to the present invention includes a substrate and at least one dried aliquot of the immobilized reactant, the immobilized reactant being bound to the surface of the substrate. The immobilized reactant binds the mobile reactant when a solution containing the mobile reactant is brought into contact with the immobilized reactant. The mobile and immobilized reactants may be any pair of biological compounds that have a specific affinity for one another. For example the reactants may be nucleic acids or antibody-antigen pairs. The preferred embodiment of an assay plate according to the present invention includes a plurality of assay spots, each spot having a different immobilized reactant or concentration thereof. The preferred method for fabricating an assay plate according to the present invention includes the steps of binding the immobilized reactant to the substrate, washing the substrate to remove any immobilized reactant that is not bound to the substrate and then drying the substrate. The dried assay plates are preferably stored in a water-proof container until used. An assay utilizing an assay plate according to the present invention is carried out by bringing a solution containing the mobile reactant into contact with the dried aliquot or aliquots on the assay plate. The assay plate is then washed to removed unbound material and the amount of mobile reactant bound to the washed assay plate determined. In the preferred embodiment of the present invention, the washed assay plate is dried prior to measuring the amount of mobile reactant bound to the washed assay plate.

Dry biochemical assay plate and method for making the same

An assay plate for detecting the presence of a mobile reactant that binds to a immobilized reactant and the methods of making and using the same. An assay plate according to the present invention includes a substrate and at least one dried aliquot of the immobilized reactant, the immobilized reactant being bound to the surface of the substrate. The immobilized reactant binds the mobile reactant when a solution containing the mobile reactant is brought into contact with the immobilized reactant. The mobile and immobilized reactants may be any pair of biological compounds that have a specific affinity for one another For example the reactants may be nucleic acids or antibody-antigen pairs. The preferred embodiment of an assay plate according to the present invention includes a plurality of assay spots, each spot having a different immobilized reactant or concentration thereof The preferred method for fabricating an assay plate according to the present invention includes the is steps of binding the immobilized reactant to the substrate, washing the substrate to remove any immobilized reactant that is not bound to the substrate and then drying the substrate. The dried assay plates are preferably stored in a water-proof container until used. An assay utilizing an assay plate according to the present invention is carried out by bringing a solution containing the mobile reactant into contact with the dried aliquot or aliquots on the assay plate. The assay plate is then washed to removed unbound material and the amount of mobile reactant bound to the washed assay plate determined. In the preferred embodiment of the present invention, the washed assay plate is dried prior to measuring the amount of mobile reactant bound to the washed assay plate.

An electrophoretic technique is provided for moving a plurality of chemicals into distinct zones for immobilization on a solid surface. The technique includes introducing a first electrolyte and a second electrolyte into a channel, and interposing between the first and second electrolytes at least one solution containing a plurality of chemicals. Under a given electric field, the first electrolyte has anions with higher effective mobility than the chemicals and the second electrolyte has anions with lower effective mobility than the chemicals. When an electrical potential is applied across the length of the channel the plurality of chemicals in the solution are moved into spatial zones. The chemicals in the zones can then be bound to the interior surface of the channel. Chemicals so bound to the wall surface can be used to build very small structures such as arrays and electrical conducting structures.

Method of binding a plurality of chemicals on a substrate by

A method for detecting the presence of a target nucleic acid species having first, second and third target sequences. In the present invention, a solution to be tested is brought into contact with a solid support having a probe nucleic acid species attached thereto, the probe nucleic acid sequence comprising a sequence that is complementary to the first target sequence. A dye solution is then brought into contact with the solid support. The dye solution includes a first dye attachment nucleic acid sequence coupled to a first dye and a second dye attachment nucleic acid sequence coupled to a second dye. The first dye attachment nucleic acid sequence is complementary to the second target sequence and the second dye attachment nucleic acid sequence is complementary to the third target sequence. The ratio of concentrations of the first and second dye attachment nucleic acid sequences in the dye solution is different from the ratio of the first and second dye attachment sequences specifically bound by the target nucleic acid species. By measuring the ratio of the first and second dyes, non-specific binding of the dye attachment sequences can be detected. In one embodiment of the invention, the target nucleic acid sequence comprises a fourth target sequence, and the dye solution includes a third dye attachment nucleic acid sequence complementary to the fourth target sequence and coupled to a third dye.

Dual tag binding assay

An integrated nucleic acid sample analysis system for matrix-assisted laser-desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS) is described. The integrated system comprises a miniaturized sample preparation compartment interfaced directly with a MALDI-TOF ionization surface for amplifying and/or otherwise chemically manipulating an oligonucleotide analyte and presenting the analyte to a MALDI ionization surface for mass spectrometry analysis. The miniaturized integrated sample handling system disclosed herein finds application in the amplification and analysis of DNA samples for genetic diagnosis and other uses.

Integrated nucleic acid analysis system for MALDI-TOF MS

An integrated liquid sample handling system for matrix-assisted laser-desorption/ionization time-of-flight mass spectroscopy (MALDI-TF MS) is described. The integrated system comprises a miniaturized sample handling compartment containing a MALDI ionization surface for separating, chemically manipulating and detecting analytes by MALDI -TOF MS. The miniaturized integrated sample handling system disclosed herein is applicable to a wide range of analytical problems that require separation and/or chemical manipulation of a sample prior to mass analysis as part of the analytical procedure.

Integrated planar liquid handling system for maldi-TOF MS

A method is provided for detecting a target nucleic acid analyte in a sample. The method involves preparing an immobilized capture oligonucleotide that is complementary to the target analyte using a magnetic cycling method, incubating the sample with the immobilized capture oligonucleotide to capture the target analyte thereby forming a capture oligonucleotide-analyte complex, and detecting the presence of the capture oligonucleotide-analyte complex.

Method for detecting a target nucleic acid analyte in a sample

An assay system for detecting the binding of a mobile reactant to an immobilized reactant on an assay plate. The assay plate includes a substrate having an assay spot deposited thereon. The assay spot includes the immobilized reactant. In the present invention, a carrier dye is included in the assay spot, the amount of the carrier dye indicating the amount of the immobilized reactant that is present in the assay spot. In the preferred embodiment of the present invention, the assay spot is generated by depositing liquid on the assay plate at a location corresponding to the assay spot. The carrier dye is dissolved in the liquid and remains after the liquid evaporates. The amount of carrier dye in each spot may be measured spectroscopically and provides a means for identifying defective assay plates. The amount of carrier dye in each spot may also be recorded on the assay plate and used in conjunction with measurements of the amount of material bound to the assay spot to determine the concentration of a reactant in a solution being tested for the reactant.

Biochemical assay plate and method for making the same

A method and apparatus is provided for spotting a biological probe onto an array. A micropipette containing a quantity of the biological probe in solution is manipulated to a position above a selected location within the array. The micropipette is pressurized sufficiently to produce a droplet of the biological probe at an open tip of the micropipette. Formation of the droplet is simultaneously visually monitored during the pressurization of the micropipette in order to estimate a volume measurement of the droplet. Upon reaching a predetermined volume for the droplet, the pressurizing of the micropipette is discontinued. The droplet of the predetermined volume is then dispensed onto the selected location.

Manufacturing method and apparatus for biological probe arrays using

A method and apparatus for dispensing immobilized reactants onto substrates for use in generating a biological assay plate for detecting the presence of mobile reactants that bind to the immobilized reactants. The apparatus includes a dispensing bed that holds a plurality of substrates. Each potential immobilized reactant is held in a cartridge. The apparatus includes a holder that allows any of these cartridges to be used in dispensing an immobilized reactant. The apparatus includes a positioning mechanism for positioning the cartridges over each of the substrates. The positioning mechanism also allows a cartridge so positioned to be brought into contact with each of the substrates at a predetermined location on each of the substrates so as to dispense a known quantity of the immobilized reactant onto the substrate at that location. In the preferred embodiment of the present invention, the cartridges include a chamber for holding the immobilized reactant and a channel having a first end communicating with the chamber. The second end of the channel provides the exit path through which the immobilized reactant exits when the cartridge is brought into contact with the substrates.

Micro-dispenser for preparing assay plates

An apparatus detects a target reactant that binds to an immobilized reactant. The apparatus generates a holographic image at a predetermined location when the reactants are present and bound to one another. The immobilized reactant is bound to a support surface at selected locations. The locations are chosen such that a holographic plate is generated when the target reactant binds to the immobilized reactant. An associated method may be used to detect antibody-antigen reactions, the binding of two strands of nucleic acid, the binding of an enzyme to one of its substrates, and so on.

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San Jose, CA, United States of America

Kay Lichtenwalter