Abstract: An assay apparatus includes a cell with a working electrode and a sonicating device structurally coupled to the cell for sonication the contents of the cell.

Abstract: Apparatus for acquiring an image of a specimen comprising a cassette having an optical portion holding a specimen array on a TIR surface and being removably matable to a processing portion having a polarized light beam source and a processing polarization-sensitive portion to image the spatially distributed charges in polarization of the specimen array. In one form the array optical portion comprises a transparent slide having a bottom surface with first and second gratings located to direct polarized light to the TIR surface and to direct light reflected by that (TIR) surface to an imager, respectively. The apparatus may include a flow cell integral with the optical portion as well as means for selecting the direction and wavelength of the polarized light.

Abstract: Antibodies and methods are described for the detection and quantitation of cardiac specific troponin I in samples. Cardiac-specific troponin isoforms exist in various forms in the blood, including free and complexed forms. By selecting antibodies that are insensitive and/or sensitive to these various forms, the present invention can provide immunoassays that more accurately reflect the clinical state of an individual. These described antibodies and methods can be used for providing indicators of myocardial infarction and other cardiac pathologies.

Abstract: The invention relates to a microfluidic device with microchannels that have separated regions which have a member of a specific binding pair member such as DNA or RNA bound to porous polymer, beads or structures fabricated into the microchannel. The microchannels of the invention are fabricated from plastic and are operatively associated with a fluid propelling component and detector.

Abstract: The present invention relates to non-isotopic immunoassays for thiopurine methyltransferase (TPMT). The immunoassays of this invention may be homogenous or heterogenous, in which detection of the TPMT-catalyzed reaction product relies upon specific binding of antibody to 6-MMP or other TPMT-catalyzed reaction products. Preferred embodiments of this invention include a Rapid Immunomigration Cassette and an assay carried out in ELISA assay format.

Abstract: The invention relates to a method of evaluating the immunological status of a subject comprising the steps of 1) determining the content of an antibody in a liquid sample from the subject using an immunoassay, wherein the reaction between the antibody of the sample and a ligand in the form of an antigen, an antibody or a hapten, the ligand being directed to the Fab region of the sample antibody, is carried out in the presence of other constituents of the sample to obtain a measurement 1, 2) determining the content of an antibody in the liquid sample using an immunoassay, wherein the reaction between the antibody of the sample and a ligand in the form of an antigen, an antibody or a hapten, the ligand being directed to the Fab region of the sample antibody, is carried out in the absence of other constituents of the sample to obtain a measurement 2, and 3) interrelating measurements 1 and 2 to express the interference and using the interference as a parameter for evaluating the immunological status of the subje

Abstract: Assay systems and specialized antibodies for the detection and quantitation of troponin I and troponin T in body fluids as an indicator of myocardial infarction. Since troponin I and T exist in various conformations in the blood, the ratios of the monomeric troponin I an T and the binary and ternary complexes, as well as which form of troponin present in the blood, may be related to the metabolic state of the heart. Disclosed is a system to determine the presence of a troponin form or a group of troponin forms in a sample of whole blood, serum or plasma. Disclosed is a stabilized composition of troponin; the stabilized composition can comprise a stabilized composition of troponin I, wherein the troponin I is oxidized, the troponin I can be unbound or the troponin I can be in a complex. Disclosed is a method for improving the recovery of troponin I or T from a surface used in immunoassays.

Abstract: In a bio-separation system, incident radiation (e.g., from a laser or LED source) for detection of separated analytes is directed at the detection zone axially along the separation medium, instead of through the boundary walls of the detection zone. In one embodiment, incident radiation at one or more wavelengths is directed via at least one optic fiber that extends axially along the separation medium to the proximity of the detection zone. Emitted radiation from the detection zone passes through the boundary walls about the detection zone for off-column detection, and/or is directed axially along the separation medium for on-column detection. In another aspect of the present invention, the detection zone is located at a widened zone along the separation channel. In a further aspect of the present invention, the optical detection configuration may be scaled up and implemented in a multi-channel CE system that comprises multiple capillary separation channels.

Abstract: In a bio-separation system, emitted radiation signals representative of sample analytes are collected from the detection zone axially along the separation medium, instead of through the boundary walls of the detection zone or the separation column. In one embodiment, emitted signals are collected via an optic fiber that extends from the proximity of the detection zone along the detection collar. According to another embodiment, a single dual purpose (excitation and emission) fiber or dual fibers (one for excitation radiation and the other for emitted radiation detection) are incorporated into detection collar. In another aspect of the present invention, the detection zone is located at a widened zone along the separation channel. In a further aspect of the present invention, the optical detection configuration may be scaled up and implemented in a multi-channel CE system that comprises multiple capillary separation channels.

Abstract: This invention relates to an improved method and system for sensing of one or more analytes. A host molecule, which serves as an adapter/carrier, is used to facilitate interaction between the analyte and the sensor element. A detectable signal is produced reflecting the identity and concentration of analyte present.

Abstract: An assay method and kit for detecting the presence of a predesignated, target IgG antibody in a sample selected from one or more patient bodily fluids. The method comprises the following steps: (a) contacting the sample of one or more patient bodily fluids with a membrane-bound recombinant protective antigen to bind to the target IgG antibody in the sample; (b) previously, simultaneously or subsequently to step (a), binding the protective antigen (PA) with a conjugated label producing a detectable signal; and (c) detecting the signal whereby the presence of the target IgG antibody is determined in the sample by the intensity of the signal. The method can further comprise the step of evaluating immunization status of the patient from whom the sample came by comparing the signal or lack thereof with immunizations previously received by the patient. In a preferred embodiment, the recombinant protective antigen (PA) specifically binds to anthrax protective antigen-specific IgG antibodies.

Abstract: The amount of platelet surface proteins in a sample may be measured by collecting a sample containing platelets into a collection tube containing a platelet stabilizing composition, labeling the platelet surface protein and detecting by cytometry.

Abstract: A method for the assay of N samples each containing a compound to be tested, comprises providing N reaction vessels each containing a population of carrier beads and other reagents for performing the assay, where N is at least 2 e.g. 80-4000. Each population of carrier beads is distinguishable from every other population. After adding the samples to the reaction vessels and performing the assays, the contents of all the reaction vessels are mixed and subjected to analysis by flow cytometry. By means of flow cytometry, each carrier bead is rapidly analysed to identify its population and also to determine the presence or concentration or biological activity of the compound to be tested.

Abstract: Apparatus for acquiring an image of a specimen comprising a cassette having an optical portion holding a specimen array on a TIR surface and being removably matable to a processing portion having a polarized light beam source and a processing polarization-sensitive portion to image the spatially distributed charges in polarization of the specimen array. In one form the array optical portion comprises a transparent slide having a bottom surface with first and second gratings located to direct polarized light to the TIR surface and to direct light reflected by that (TIR) surface to an imager, respectively. The apparatus may include a flow cell integral with the optical portion as well as means for selecting the direction and wavelength of the polarized light.

Abstract: A method for conveniently detecting binding between the von Willebrand factor and glycoprotein Ib and a means to be used therein. The von Willebrand factor fixed in a reactor immobilized in a reaction vessel in the presence of bottrocetin is bound to a chimeric protein constructed by fusing the carboxyl terminal of a partial protein containing the von Willebrand factor-binding site of glycoprotein Ib with the amino terminal of the Fc region of an immunoglobulin molecule. Then the Fc region of the above immunoglobulin molecule is detected to thereby detect the binding between the von Willebrand factor and the glycoprotein Ib or inhibition of this binding.

Abstract: Imaging apparatus and method which uses change of polarization state of a light beam passed through a total internal reflection structure by a single reflection at a TIR surface in which a specimen is placed in the evanescent field associated with the total internal reflection of the light beam, the specimen being the subject of biological, chemical or genetic investigation.

Abstract: Apparatus for acquiring an image of a specimen comprising a cassette having an optical portion holding a specimen array on a TIR surface and being removably matable to a processing portion having a polarized light beam source and a processing polarization-sensitive portion to image the spatially distributed charges in polarization of the specimen array. In one form the array optical portion comprises a transparent slide having a bottom surface with first and second gratings located to direct polarized light to the TIR surface and to direct light reflected by that (TIR) surface to an imager, respectively. The apparatus may include a flow cell integral with the optical portion as well as means for selecting the direction and wavelength of the polarized light.

Abstract: The invention relates to novel fluorescence-based assays for protein kinases and phosphatases which can be used in high throughput screening. The methods of the invention utilize a competitive immunoassay to determine the amount of substrate that is phosphorylated or dephosphorylated during the course of a kinase or phosphatase reaction to yield a product, as well as the phosphorylating or dephosphorylating activity of a kinase or phosphatase.

Abstract: Probe sets for the multiplexed detection of the binding of, or interaction between, one or more ligands and target antiligands are provided. Detection involves the release of identifying tags as a consequence of target recognition. The probe sets include electrophoretic tag probes or e-tag probes, comprising a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. Target antiligands are contacted with a set of e-tag probes and the contacted antiligands are treated with a selected cleaving agent resulting in a mixture of e-tag reporters and uncleaved and/or partially cleaved e-tag probes. The mixture is exposed to a capture agent effective to bind to uncleaved or partially cleaved e-tag probes, followed by electrophoretic separation. In a multiplexed assay, different released e-tag reporters may be separated and detected providing for target identification.

Abstract: According to the present invention, the biological or pharmacological activity of a test material like a plant or herbal material, an extract of a plant or herbal material, a natural or synthetic compound or some combination thereof, can be quantified by observing the pattern of structural changes induced in a eukaryotic cell's proteins. These structural changes may be evidenced by protein phosphorylation, by protein-protein interactions and the like. The amount and nature of protein phosphorylation is qualitatively and quantitatively related to the in vitro concentration of biologically/pharmacologically active components to which the mammalian cells are exposed. Additionally, formation or loss of protein-protein complexes may be determined in whole cell homogenates through the use of non-denaturing electrophoresis and staining for proteins or protein phosphorylation.

Abstract: The present invention relates to an immunoassay and an apparatus for the same. A method of the present invention comprises following processes: (1) an analyte is labeled to detect antigen-antibody reaction, (2) the magnetic material label is magnetized by a magnetic field, (3) the magnetized magnetic material label detected by a SQUID which detect a magnetic field having right angle to the magnetic field. At the same time, the present invention contains an apparatus to execute the method provided by the present invention. The apparatus comprises a magnetic field generation means that generates a magnetic field to magnetize the labels. The apparatus comprises a SQUID that measures magnetic field.

Abstract: The present invention relates to systems, apparatuses and methods for multidimensional protein separation. In particular, the present invention relates to 3-dimension protein separation and characterization systems and methods.

Abstract: Disclosed is a method for determining the fertility of mammals, in particular of humans, wherein a body or organ fluid is taken from a mammal, the afamin content is determined in this body or organ fluid, and the determined afamin content is compared with a reference value so as to determine the fertility, the use of afamin for determining the fertility of mammals, as well as a kit for carrying out this method.

Abstract: A method for the qualitative and quantitative determination of multimers of fibrinogen and of von Willebrand factor by gel electrophoresis, in which a sample containing von Willebrand factor (vWF) or fibrinogen is fractionated by submarine electrophoresis using a continuous, homogeneous agarose gel free of lumps, and the multimer bands are visualized immunochemically after a western blot analysis by a specific antibody-enzyme conjugate on the blotting membrane or by a suitable dye, preferably with a blue stain, in the gel, is described.

Abstract: Proteins can be rapidly separated to a high degree of resolution by electrophoresis on polymeric membranes that have high protein binding capacity. The electrophoretic separation is carried out in a low conductivity, water-miscible organic solvent buffer. The low conductivity of the organic solvent buffer minimizes heat generation, and the water-miscible nature of the organic solvent buffer permits the analysis of hydrophobic and low molecular weight proteins as well as hydrophilic proteins. When electrophoresis is conducted under non-denaturing conditions, it allows the detection of enzymatic activities, protein-protein interactions and protein-ligand interactions.

Abstract: An array (100) of macromolecules (the primary array), typically proteins is generated by 2D electrophoresis, for example, and subsequently transferred to a support membrane (102) by electroblotting or the like. An image of the primary array is captured (202) and the coordinates of the various macromolecular spots in the primary array are determined (402). The next step of the process is to print a secondary (or micro) array of one or more reagents or chemicals onto one or more spots/coordinates of the primary array with a pico-litre (pl) dispenser (702). If the macromolecules are proteins, the reagents may be enzymes such as Trypsin or GluC. Use of two different enzymes deposited onto different coordinates on the same spot will cleave the protein at different amino acid sites and, when the spot is analysed in a MALDI-TOP mass spectrometer, will provide increased coverage or matching of peptides in the protein.

Abstract: Diagnostic methods comprise measuring specifically the level of at least one iso-eosinophilic cationic protein (iso-ECP) in a sample from an individual to be diagnosed, and comparing the measured level of the iso-ECP with a predetermined level of the iso-ECP. The iso-ECP is cytotoxic and the cytotoxicity of the iso-ECP is not capable of neutralization by the monoclonal antibodies EG1 and EG2. Anti-iso-ECP antibody which may be used in the diagnostic methods specifically binds to a cytotoxic isoform of ECP having an epitope which is unique for the native form of the cytotoxic iso-ECP.

Abstract: The present invention provides systems, methods, and screens for an optical system analysis of cells to rapidly determine the distribution, environment, or activity of fluorescently labeled reporter molecules in cells for the purpose of screening large numbers of compounds for those that specifically affect particular biological functions. The invention involves providing cells containing fluorescent reporter molecules in an array of locations and scanning numerous cells in each location with a high magnification fluorescence optical system, converting the optical information into digital data, and utilizing the digital data to determine the distribution, environment or activity of the fluorescently labeled reporter molecules in the cells. The array of locations may be an industry standard 96 well or 384 well microtiter plate or a microplate which is microplate having cells in a micropaterned array of locations.

Abstract: A method is provided for continuously monitoring for the presence or quantity of an analyte in a flowing liquid stream. The method involves binding an analyte-specific receptor species to the surface of a piezoelectric substrate, contacting the surface bound receptor species with the flowing liquid stream and quantitating the presence of the analyte. A novel apparatus for detecting the presence of an analyte in a liquid chromatography eluant is provided as well.

Abstract: The present invention relates to a method for the determination of non-, anti-, or pro-apoptotic and necrotic conditions of cells, newly designed vectors coding for marker proteins, cell lines transfected with such vector, and a method to assay the non-, pro- or anti-apoptotic or necrotic activity of test compounds.

Abstract: The invention concerns the stabilization and amplification of electrochemiluminescence signals in detection methods.

Abstract: The present invention relates to the substantial elimination of errors attributable to carryover microspheres, doublets, or misclassification of microsphere subsets. The present invention is based upon passing a sufficient minimum number microspheres through the flow analyzer during an assay run.

Abstract: Specific genetic deletions are identified in mycobacteria isolates, including variations in the M. tuberculosis genome sequence between isolates, and numerous deletion present in BCG as compared to M. tb. These deletions are used as markers to distinguish between pathogenic and avirulent strains, and as a marker for particular M. tb isolates. Deletions specific to vaccine strains of BCG are useful in determining whether a positive tuberculin skin test is indicative of actual tuberculosis infection. The deleted sequences may be re-introduced into BCG to improve the efficacy of vaccination. Alternatively, the genetic sequence that corresponds to the deletion(s) are deleted from M. bovis or M. tuberculosis to attenuate the pathogenic bacteria.

Abstract: The present invention provides a method by which UTI concentration can be measured easily with high precision and good reproducibility. The measurement is performed by adding free anti-UTI antibodies to a sample and measuring the degree of the resulting agglutination, for example, from the change in absorbance. As shown in FIG. 3, the UTI concentration and the degree of the agglutination (i.e. the change in absorbance) are correlated. The absorbance can be measured by using a general spectrophotometer, preferably at a wavelength of 300 to 400 nm. Polyethylene glycol is preferably added to the reaction solution as an agglutination accelerator. The polyethylene glycol preferably has an average molecular weight of 2,000 to 20,000, and the concentration of polyethylene glycol in the reaction solution is preferably in the range of 2 to 10 weight %.

Abstract: An analytical test device is described for the immunochromatographic determination of the presence of one or more analytes in fluid samples. The device is configured such that the sample is allowed to enter the detection zone simultaneously from many different directions, eliminating stagnation of the flow of the sample. By selection of the porous substrate, the device also allows for the separation of red blood cells from plasma, providing a rapid test for one or more analytes in a sample of whole blood. The device of the present invention may measure more than one analyte simultaneously from a single sample, either by having multiple immunochromatographic pathways fed by a single sample, or multiple analytes detected in the same pathway by way of multiple capture antibodies.

Abstract: Procedure for the study of the functional activation of leukocytes, platelets and other cells, produced in vivo or induced in vitro, based on the stabilization of cytoplasmic membrane proteins and its detection using quantitative cytometric methods in the absence of any further manipulation of the sample. The procedure includes the sequential incubation of the sample with: 1) either one or a mixture of more than one protease specific inhibitors and, 2) a combination of several fluorochrome-conjugated monoclonal antibodies; to analyze the expression of surface proteins using immunofluorescence methods and quantitative cytometry.

Abstract: The invention provides methods and compositions for detecting binding or unbinding of a molecule to a substrate. The molecule comprises a luminophore and the substrate comprises a semiconductor which acts as a luminescence quencher to provide distance-dependent quenching of the luminophore. Binding or unbinding of the molecule, which may be covalent or noncovalent, is detected as a decrease or increase, respectively, of the detectable luminescence of the luminophore.

Abstract: Analyses of serum samples for the presence and amount of either of the two subunits of human Factor XIII protein are used as a means of eliminating a significant source of error that arises in the testing of serum and plasma. For serum samples, a negative result of an analysis for the presence of subunit a is a means of verifying that a sample is indeed serum, while a negative or positive result for subunit a serves to distinguish serum (negative) from plasma (positive). A positive result for the presence of subunit b is a means of verifying that the sample is either serum or plasma and not any other biological fluid. A quantitative analysis of subunit b is a means of verifying that the sample is of the intended volume rather than having been reduced in volume due to improper sampling. A quantitative analysis of subunit b is also a means of verifying the dilution of a sample of either serum or plasma.

Abstract: The present invention provides internally calibrated competitive assays for use on a solid support. Additionally, the invention provides a method of using such assays.

Abstract: A method for assaying specific binding between a fluorophore-labeled probe and an unlabeled target is provided. The method includes detecting a quenching effect on fluorescence emitted by the fluorophore-labeled probe resulting from binding. The method is conducted without separating complexes of the target and probe from the free target and free probe prior to quenching effect detecting, and without providing a signal quenching agent to quench fluorescent light. Preferably, the probe and target are amino acid-containing compounds, such as proteins. The method can be used for a variety of applications, including screening for drug candidates having optimum binding properties, and quantifying and classifying the binding characteristics between peptide-containing compounds. The method is more sensitive than conventional assays, enabling the analysis of minute samples and low affinity binding interactions between receptors and ligands that are below the detection limits of conventional technology.

Abstract: An assay method for detecting the response or activation of cells, including lymphocytes, and a method for detecting immunological sensitization in a subject, which involves the introduction of cell-activating substance which causes an enzyme of the cells to become available for reaction, and the measurement of the enzymatic reaction using a substrate which generates a detectable product, and a kit for performing such assays.

Abstract: The gene for Streptococcus pyogenes DNase B has been cloned and vectors incorporating the cloned DNA have been used to transform Escherichia coli, allowing efficient and rapid production of the DNase in E. coli without the necessity of growing large quantities of S. pyogenes. The enzyme can be produced with a leader peptide at its amino terminus. An improved method for the purification of naturally occurring S. pyogenes DNase B enzyme is also provided. The DNase B enzyme produced, either by purification of naturally occurring enzyme or by recombinant DNA techniques, can be used to generate antibodies and can also be used in immunochemical assays to detect the presence of anti-DNase B antibodies in serum as a marker of infection by S. pyogenes.

Abstract: The present invention provides a method for detecting a molecular event, comprising (1) applying an electromagnetic test signal to a sample in which a molecular event is being detected, whereby the sample interacts with and modulates the test signal to produce a modulated test signal, and (2) detecting the modulated test signal, wherein the applying and detecting take place in a temperature-controlled environment, wherein the temperature-controlled environment comprises the sample, a radiating portion of a signal generating circuit, and a receiving portion of a signal detection circuit and wherein the applying and detecting take place in the environment at a temperature controlled to within ±0.5° C.

Abstract: The present invention provides methods of identifying hot-spot residues for one or both members of a receptor-ligand complex of interest. Further provided are methods of using receptor hot-spot residues to identify compounds that functionally bind a receptor in a manner that mimics the binding of a known ligand for the receptor.

Abstract: Imaging apparatus and method which uses change of polarization state of a light beam passed through a total internal reflection structure by a single reflection at a TIR surface in which a specimen is placed in the evanescent field associated with the total internal reflection of the light beam, the specimen being the subject of biological, chemical or genetic investigation.

Abstract: The invention relates to methods, kits and systems for determining an analyte in a liquid sample as well as the use thereof for concentration analysis and screening purposes. In one method, a specific binding partner to the analyte is permitted to compete with a conjugate containing the analyte or an analyte analogue for the binding to free analyte. The conjugate also contains a component that specifically binds to a solid support so that reacted and unreacted conjugate are bound thereto when the reaction solution is contacted with the solid support. The amount of analyte is then determined by measuring by a label-free mass-detection technique, such as surface plasmon resonance, the amount of binding partner immobilized on the solid support via the reacted conjugate. This is made possible by the binding partner having a considerably greater mass than the conjugate. Variants of this method are also disclosed.

Abstract: A mass biosensor uses an intermediate avidin layer to facilitate binding of a biotinylated antibody to a measurement surface of the biosensor. The avidin layer can be added by the manufacturer of the biosensor, while the biotinylated layer can be added by the user. This two-phase method of chemically modifying the measurement surface significantly reduces the user time required to customize the measurement surface to render it capable of binding selected compounds. An organosilane coupling agent attached to the surface provides sites to which avidin is bound. Avidin acts as a universal receptor of biotinylated compounds with specific binding affinities. Biotinylated antibodies or other biotinylated compounds are added and bind to the immobilized avidin. Surface adsorption is reduced by washing the modified surface with biotin to block potential sites of weak bond formation, electrostatic and hydrophobic interactions.

Abstract: Method for specific detection of one or more analytes in a sample. The method includes specifically associating any one or more analytes in the sample with a scattered-light detectable particle, illuminating any particle associated with the analytes with light under conditions which produce scattered light from the particle and in which light scattered from one or more particles can be detected by a human eye with less than 500 times magnification and without electronic amplification. The method also includes detecting the light scattered by any such particles under those conditions as a measure of the presence of the analytes.

Abstract: A method for identifying one or a small number of molecules, especially in a dilution of ≦1 &mgr;M, using laser excited FCS with measuring times ≦500 ms and short diffusion paths of the molecules to be analyzed, wherein the measurement is performed in small volume units of preferably ≦10−14 l, by determining material-specific parameters which are determined by luminescence measurements of molecules to be examined.

Abstract: The present invention provides methods and compositions for screening, diagnosis and prognosis of Cardiac Response, for monitoring the effectiveness of Cardiac Response treatment and for drug development. Cardiac Response-Associated Features (CRFs) detectable by two-dimensional electrophoresis of blood are described. The invention further provides Cardiac Response-Associated Protein Isoforms (CRPIs) detectable in biological sample preparations comprising isolated CRPIs, antibodies specific for CRPIs and kits comprising the aforesaid.