Abstract: The present invention is a method for accurately comparing the levels of cellular components, such as proteins, present in samples which differ in some respect from each other using mass spectroscopy and isotopic labeling. A first sample of biological matter, such as cells, is cultured in a first medium and a second sample of the same biological matter is cultured in a second medium, wherein at least one isotope in the second medium has a different abundance than the abundance of the same isotope in the first medium. One of the samples is modulated, such as by treatment with a bacteria, a virus, a drug, hormone, a chemical or an environmental stimulus. The samples are combined and at least one protein is removed. The removed protein is subjected to mass spectroscopy to develop a mass spectrum. A ratio is computed between the peak intensities of at least one closely spaced pair of peaks to determine the relative abundance of the protein in each sample.

Abstract: This invention relates to methods of assaying one or more analytes simultaneously. The assays of this invention are capable of providing wide dynamic range and rapid processing times. A wide dynamic working range is achieved by simultaneously incubating a sample which may contain the analyte(s) of interest with two or more independently determinable classes of particles coated with an analyte-specific binding partner. The two or more particle classes differ from each other at least in size. The analyte concentration is obtained from readings derived from these two classes by means of a combined standard curve.

Abstract: The present invention relates to the identification of proteins and protein isoforms that are associated with breast cancer and its onset and development, and of genes encoding the same, and to their use for e.g., clinical screening, diagnosis, prognosis, therapy and prophylaxis, as well as for drug screening and development of pharmaceutical products.

Abstract: In a competitive receptor binding assay for detecting TSH-receptor auto-antibodies in a biological sample, the sample is reacted in a reaction mixture which contains (i) a TSH-receptor or TSH-receptor preparation; (ii) a primary competitor, for example labelled TSH; and (iii) an agent for separating a complex composed of the TSH-receptor and the elements bound thereto of the reaction mixture from the liquid phase. According to the invention, the reaction is carried out in the presence of at least one monoclonal or polyclonal antibody specific against a partial peptide sequence of the TSH eceptor. This specific antibody is used to immobilize a complex of TSH-receptor and primary competitor and/or as secondary competitor for another part of the TSH-receptor auto-antibodies expected in a sample. The primary or secondary competitors are or can be selectively labelled.

Abstract: The present invention provides methods and compositions for screening, diagnosis and prognosis of vascular response, for monitoring the effectiveness of vascular response treatment, identifying patients most likely to respond to a particular therapeutic treatment and for drug development. In particular, the screening of drug candidates for their ability to induce a vascular response. Vascular Response-Associated Features (VRFs), detectable by two-dimensional electrophoresis of blood, serum or plasma are described. The invention further provides Vascular Response-Associated Protein Isoforms (VRPIs) detectable in blood, serum or plasma, preparations comprising isolated VRPIs, antibodies immunospecific for VRPIs, and kits comprising the aforesaid.

Abstract: Assay methods are provided for detection or quantitation of any of several proteins which are specifically produced in the endometrium in association with hyperplasia, adenocarcinoma or the proliferative phase of the endometrium. The relevant proteins have been identified by 2D gel electrophoresis with subsequent sequence identification by mass spectroscopic finger printing of tryptic digests.

Abstract: Growth differentiation factor-14 (GDF-14) is disclosed along with its polynucleotide sequence and amino acid sequence. Also disclosed are diagnostic and therapeutic methods of using the GDF-14 polypeptide and polynucleotide sequences.

Abstract: This application relates to a method and apparatus for the electrophoresis, blotting and detection of nucleic acids, proteins and other biological molecules using a porous polymer gel carrier and assembly. The combination of a porous polymer gel carrier and attached laminate layer facilitates the electrophoresis and subsequent blotting analysis of nucleic acids, proteins and other biological molecules. The porous polymer gel carrier also facilitates the use, handling and processing of fragile separation materials such as polyacrylimide and agarose gels. The method and apparatus also permits the convenient packaging and distribution of standardized electrophoresis and blotting products.

Abstract: Immunoassay methods for measuring the concentration of an analyte in a test specimen are described. The methods use an immunoreagent, where one of the analyte and the immunoreagent is an antigen, and the other of the analyte and the immunoreagent is an antibody which specifically binds to the antigen. An important feature distinguishing these immunoassays over conventional immunoassays is that the standard sample containing a known concentration of analyte is measured in the same reaction vessel as the test specimen containing an unknown amount of analyte.

Abstract: Throughput rates for microfluidic serial analysis systems are optimized by maximizing the proximity and speed with which multiple different samples may be serially introduced into a microfluidic channel network. Devices are included that include optimized parameters based upon desired throughput rates for a given set of reagents, reaction times and the like.

Abstract: The invention is based on the discovery that certain variants of &bgr;-casein may induce Type-1 diabetes in susceptible individuals while other variants do not. The invention consists of the selection of non-diabetogenic milk producing cows and recovering and processing their milk and milk products. Another aspect of the invention is selectively breeding cows which produce the non-diabetogenic milk.

Abstract: A method of distinguishing between fluorescent light emitted by green fluorescent protein and fluorescent light emitted by auto-fluorescent molecules, the method comprising: using plane-polarised light to illuminate a sample containing green fluorescent protein and auto-fluorescent molecules; detecting the intensity of fluorescent light that is emitted with a first polarisation from the sample; detecting the intensity of fluorescent light that is emitted with a second polarisation from the sample; and subtracting a first of said detected intensities from a second of said detected intensities to obtain a difference signal.

Abstract: The invention concerns a device and a method for ultrasensitive detection comprising an electrically inert porous sheet sandwiched between two positively charged fine electrodes. The invention uses the capacity of the electrodes, resulting from their powerful electrostatic properties, for sensing marked antibody complexes related to antigens detected in the sample, and to leave marked antibodies non-complexed with said antigens free. The method consists in depositing a sample droplet on the porous material sheet upstream of the electrodes and in detecting downstream of the electrodes, in a signal-zone, the possible presence of an antigen-antibody complex which has diffused in the porous material. The invention is useful for detecting some millipicogrammes of antigen per milliliter of sample.

Abstract: Methods, compositions, kits, and a system are disclosed for detecting one or more analytes in a sample. A mixture comprising the (i) sample, (ii) a first binding reagent comprising a cleavage-inducing moiety and a first binding agent specific for an analyte, and (ii) one or more electrophoretic probes each having a second binding agent is subjected to conditions under which binding of respective binding agents occurs. The interaction between the binding agents and the analyte brings the cleavage-inducing moiety within a proximity effective for cleaving a cleavable linkage tethering an electrophoretic tag to the second binding agent, thereby releasing the tag for electrophoretic separation. Separation of different tags occurs by virtue of their distinct electrophoretic mobilities. After separation, a signal amplification moiety on each tag is activated to generate a signal to indicate the presence of a particular analyte in the sample.

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: The invention concerns an alkaline pH, free solution capillary electrophoresis method for analysing clinical samples comprising protein constituents, characterized in that it comprises at least one step in which the sample is introduced into a capillary tube containing a buffer system comprising, as the buffer, a biological buffer with a pKa at 25° C. in the range 8.8 to 10.7 and at least one additive that can increase the ionic strength of the buffer system.

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: Methods using gel electrophoresis and mass spectrometry for the rapid, quantitative analysis of proteins or protein function in mixtures of proteins derived from two or more samples in one unit operation are disclosed.

Abstract: A bio-assay test system includes a test fixture, a measurement system, and a computer. The test fixture includes a bio-assay device having a signal path and a retaining structure configured to place a sample containing molecular structures in electromagnetic communication with the signal path. The measurement system is configured to transmit test signals to and to receive test signals from the signal path at one or more predefined frequencies. The computer is configured to control the transmission and reception of the test signals to and from the measurement system.

Abstract: The invention concerns a method for the immunological determination of an analyte in a sample using magnetic particles coated with the analyte to be determined or analyte-specific bonding partners and directly detectable non-magnetic particles coated with analyte-specific bonding partners or the analyte to be determined or using a non-magnetic substance which is indirectly detectable, and incubation of the reaction mixture. The method is characterized in that the magnetic particles are subsequently separated from the reaction mixture using a magnetic test strip and the analyte concentration is determined directly.

Abstract: The invention is generally directed toward an analytical method to determine the concentration of an analyte or free analyte fraction in a sample. More particularly, the method encompasses applying a sample containing an analyte to an immunoaffinity column capable of selectively extracting the analyte in the millisecond time domain. The concentration of the analyte is then determined by chromatographic immunoassay.

Abstract: According to the biosensor and the blood component analytical method of the present invention, in a biosensor that is made of a single layer or plural layers of a porous material as shown in FIG. 1, having a reagent holding part and utilizing chromatography, a cell shrinkage reagent is carried on at least part of the reagent holding part, or at least part of a chromatographically developed part that is upstream of the reagent holding part.

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

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: In a process for the quantitative optical analysis of fluorescently labelled biological cells 5, a cell layer on a transparent support at the bottom 2 of a reaction vessel 1 is in contact with a solution 3 containing the fluorescent dye 4. The sensitivity of analytical detection can be considerably improved if to the fluorescent dye 4 already present in addition a masking dye 9, which absorbs the excitation light 6 for the fluorescent dye 4 and/or its emission light 7, is added to the solution 3 and/or if a separating layer 10 permeable to the solution and absorbing and/or reflecting the excitation light 6 or the emission light 7 is applied to the cell layer at the bottom 2. The separating layer 10 must be composed such that it has a high power of reflection for the luminescent light 11.

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: 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: Systems and methods are presented for detecting molecular binding events and other environmental effects using the unique dielectric properties of the bound molecular structure or structures. A molecular binding region is coupled along the surface of a signal path. A test signal is propagated along the signal path, whereby the test signal couples to the molecular binding region, and in response, exhibits a signal response.

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: Systems and methods for detecting molecular binding events and other environmental effects using the unique dielectric properties of the bound molecular structure or structures are presented. A molecular binding layer is coupled along the surface of a signal path. A test signal is propagated along the signal path, whereby the test signal couples to the molecular binding layer, and in response, exhibits a signal response.

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: In a process for the combinatorial production of material samples in the form of a two-dimensional matrix in the surface region of a sheet-like substrate, at least two different dispensable material components are dispensed as a suspension from one or more dispensing devices which allow the release of individual suspension drops onto the same point of the substrate, so that materials of different composition are obtained in different surface regions of the substrate.

Abstract: Novel bipyridyl-osmium complex conjugates, their preparation, and their use in electrochemical assays are described. The redox reversible-osmium complexes can be prepared to exhibit unique reversible redox potentials and can thus be used in combination with other electroactive redox reversible species having redox potentials differing by at least 50 millivolts in electrochemical assays designed for use of multiple electroactive species in the same cell and in the same sample without interference between the two or more redox coupled conjugate systems.

Abstract: The present invention provides for amplification methods for cloning and copying genetic material on surfaces as well as copying biological material insofar as, in a broader sense, it can be classified as a ligand-receptor system. The invention therefore relates in particular to a method for propagating ligands and receptors on at least two surfaces, comprising (a) immobilizing a first ligand on a first surface of a substantially solid phase; (b) adding a solution of receptors and binding complementary receptors to the first ligand; (c) transferring the receptor to a second surface and immobilizing the receptor at that location; (d) attaching an additional ligand to the immobilized receptor; and (e) transferring the additional ligand to the first surface and immobilizing it at that location, wherein the steps set forth above may be repeated multiple times.

Abstract: Immunoassay reagents, methods and test kits for the specific quantification of vancomycin in a test sample are disclosed. The reagent comprises antibodies prepared with immunogens of FIG. 6 wherein P is an immunogenic carrier material and X is a linking moiety.

Abstract: Systems and methods are presented for detecting molecular binding events and other environmental effects using the unique dielectric properties of the bound molecular structure or structures. A molecular binding region is coupled along the surface of a signal path. A test signal is propagated along the signal path, whereby the test signal couples to the molecular binding region, and in response, exhibits a signal response.

Abstract: The present invention comprises method of detecting biomolecules in situ in gel separation medium following electrophoresis that provides sensitivity similar to blotting. This method is applicable to in situ detection of a wide range of biomolecules including proteins and nucleic acids. After electrophoretic separation of a sample in gel separation media comprising a gellable polymeric material other than cross-linked polyacrylamide, the gel is contacted with a solution comprising at least one detectably labeled reagent directed to a biomolecule under conditions suitable for binding of the reagent to the biomolecule. Alternatively, the gel is contacted with a solution comprising at least one non-detectably labeled reagent directed to a biomolecule and a solution comprising at least on detectably labeled reagent directed to a biomolecule. In either case, binding of the detectably labeled reagent is assessed and indicates detection of biomolecules in the gel.

Abstract: The present invention relates to a method for instrument determination of measured variable L(t) which changes with time, the intention being to determine the maximum rate value of L(t) in a region with a linear reaction profile, and the time as well as the duration of the reaction time window being variable with the linear region and depending on the nature of the reaction and the reaction conditions. Specifically, the present invention relates to the determination of protein concentrations with the aid of light scatter, which is produced by specific antibodies, in homogeneous solutions. In particular, the invention relates to reactions which take place slowly and have a largely linear profile over a relatively long time, the rate of formation of antigen-antibody complexes in the linear section of the reaction (VMaxLin) being determined as the measured variable.

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

Abstract: Calibrating an immunoassay by generating two reaction rate measuring curves, from samples having higher and lower relative levels of antigen, extrapolating a combination of the curves to cover sample concentrations known to contain an excess of antigen relative to an amount of capture reagent and combining the low end linear potion of the higher reaction rate measuring curve with the higher end portion of the extrapolated reaction rate measuring curve, thereby eliminating measuring inaccuracies otherwise arising from the hook effect. For antigen concentrations higher than the assay range, a high antigen signal utilizing the two rates avoids reporting false results.

Abstract: A method of determining the presence of chronic volume dependent hypertension is provided wherein a determination is made as to whether there has been a substantial reduction in phosphorylation of the blood-derived protein or renal proximal brush border membrane protein and if such reduction exists concluding that chronic volume dependent hypertension exists in a patient. The method may advantageously be practiced by employing blood serum or blood plasma as the body specimen containing the protein in determining whether a patient has chronic volume dependent hypertension, a cellular component of the blood, such as a blood-derived protein coming from the plasma membrane of lymphocytes. The method may include subsequent therapeutic patient treatment. Related diagnostic apparatus is also provided.

Abstract: Methods of detecting binding of a putative ligand to a 13C-enriched target molecule, methods of screening for compounds which bind to a 13C-enriched target molecule, methods for calculating the dissociation constant of a ligand compound which binds to a 13C-enriched target molecule, and methods employed in the determination of the specific amino acids in a 13C-enriched target molecule affected by the binding of a ligand, as well as compounds identified by these screening methods, are provided herewith.

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 detecting at least one substance (a ligand) present in a compound library using at least one additional substance (a receptor) that binds to the ligand comprises adding to the compound library such a receptor that has substantially higher molecular weight than the ligand to be identified, and performing of such a spectroscopic measurement technique with the mixture, without isolating the receptor-ligand complex, that can detect those dipolar resonance phenomena which occur upon a binding of a receptor to a ligand.

Abstract: The present invention relates to stable compositions useful as primary standards and calibrators and controls comprising a cardiac troponin I (cTnI) such as native, recombinant, addition and deletion forms thereof, whether or not complexed with other troponin subunits such as TnC and/or TnT, in an inactivated human serum. The compositions are obtained by incubating troponin complexes with human serum. The compositions are characterized by an immunodetectability ratio of epitopes on the N-terminal segment to epitopes on the C-terminal segment substantially equivalent to that of pooled, fresh serum from acute myocardial infarction patients.

Abstract: This invention relates to biologically-active polymers that are useful for analyte detection and isolation and delivery of substances. The biologically-active polymers are capable of specifically and reversibly binding to analytes, including molecules and cells. The biologically-active polymers are also capable of releasing substances upon electrical stimulation. The present invention provides compositions comprising biologically-active polymer membranes and methods for making these biologically-active polymer membranes that may be specifically designed to selectively bind cells and specific cell types, to affect cell growth characteristics, and to modulate cellular differentiation. These biologically-active polymer membranes may be controlled electrically to induce controlled cellular differentiation and modulate the cell growth cycle. These biologically-active polymers have many applications in biological and chemical fields.

Abstract: The present invention provides methods for determining the relative orientation of the individual components of a macromolecule in solution with respect to the global molecular coordinate frame of the macromolecule. The present invention further provides methods for applying this structural information, including for rational drug design.

Abstract: A method of determining affinity and kinetic properties for the solution interaction between an analyte and a binding partner therefor, which method comprises: (a) mixing a solution of said analyte with a solution of said binding partner, contacting the resulting reaction solution with (i) immobilised binding partner, or analogue, for said analyte, and/or (ii) immobilised analyte, or analogue, and monitoring the binding of said analyte and/or binding partner in said reaction solution to the respective immobilised species to determine the variation with time of the concentration of free analyte and/or binding partner in said solution; and/or (b) contacting a solution of the reaction complex of said analyte and a binding partner therefor with (i) immobilised binding partner, or analogue, and/or (ii) immobilised analyte, or analogue, and monitoring the binding of said analyte and/or binding partner in said reaction complex solution to the respective immobilised species to determine the variation with time of the

Abstract: Disclosed is a high throughput compatible assay that is useful for the identification of specific antagonists of TRAF-receptor interactions. The modular flexibility of the assay makes it possible to introduce simple modifications in order to measure the interaction of any TNF receptor cytoplasmic domain (or TRAF-binding protein) with any of the six TRAF proteins, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5 and TRAF6.