Abstract: This invention relates to a method and apparatus for detecting a biological molecule associated with enzyme activity in a sample. The invention is applicable to detecting a microorganism associated with an enzyme in a sample such as water, food, soil, or a biological sample. According to a preferred embodiment of the method of the invention, a sample containing an enzyme of interest or a microorganism associated with the enzyme is combined with a suitable substrate, and a fluorescent product of the enzyme-substrate reaction is selectively detected. The fluorescent product is detected with a partitioning element or optical probe/partitioning element of the invention. In one embodiment the partitioning element provides for partitioning of only the fluorescent product molecule into the probe. The invention also provides an automated system for monitoring for biological contamination of water or other samples.

Abstract: Provided herein are microfluidic devices and methods useful for sensitive detection of analytes. The methods and devices described herein are also useful for detecting direct or indirect binding of enzymes or catalysts to a surface, for example a surface having analytes bound thereon. Methods disclosed herein include embodiments utilizing a pre-concentration scheme to improve signal levels of corresponding reporter moieties.

Abstract: The invention describes a method for assaying HCV NS3 protease activity using an NS3•4A protease molecule. The invention also provides a method for screening and identifying modulators of NS3 protease.

Abstract: The disclosure provides compositions, methods and kits for assessing immunogenicity, potency, or both, of meningococcal capsular saccharide vaccines. The assessment is based upon measurement of binding of a bactericidal antibody to a capsular saccharide component in the vaccine.

Abstract: The invention relates to a method of measuring the immunological activity of a vaccine preparation in the form of a mixture of one or more allergenic enzyme(s) and an oxygen-containing metal salt adjuvant, wherein the mixture comprises a liquid phase and a solid phase, and wherein at least a part of the allergenic enzyme(s) is adsorbed to the solid phase, the method comprising the steps of measuring the enzymatic activity of the mixture in an enzyme activity assay, and using the measurement obtained as an indication of the immunological activity of the vaccine preparation, or using the measurement obtained for quantifying the amount of allergenic enzyme.

Abstract: The present invention provides methods of detecting ovarian cancer using biomarkers.

Abstract: A method to detect the presence or amount of at least one molecule for an enzyme-mediated reaction in a multiplex luminogenic/nonluminogenic assay is provided.

Abstract: Embodiments of the invention are generally directed to compositions and methods of delivering one or more transgene to a target cell, such as a tumor cell, in a site-specific manner to achieve enhanced expression and to constructs and compositions useful in such applications. In certain aspects, expression from a therapeutic nucleic acid may be assessed prior to administration of a treatment or diagnostic procedure to or on a subject.

Abstract: A diagnostic test kit for detecting the presence or quantity of an enzyme or enzyme inhibitor is provided. The diagnostic kit utilizes reactive complexes to facilitate the detection of the enzyme or enzyme inhibitor. The reactive complexes include a substrate joined (e.g., covalently bonded, physically adsorbed, etc.) to a reporter and specific binding member. In one embodiment, for example, a peptide, protein, or glycoprotein substrate is joined to a reporter (e.g., dyed latex particle) and specific binding member (e.g., biotinylated compound). In this embodiment, the substrate provides a cleavage target for a proteolytic enzyme. Specifically, upon contacting the reactive complexes, the proteolytic enzyme cleaves the substrate and releases the reporter and/or specific binding member. The signal exhibited by the released reporters may then be used to indicate the presence or quantity of an enzyme or enzyme inhibitor within the test sample.

Abstract: The present invention is directed to a method of detecting intact fibrinogen, comprising the steps of: a) providing a sample containing at least some fibrinogen optionally converted at least in part to fibrin, and optionally containing thrombin; b) solubilizing the sample in a solubilizing solution that inhibits thrombin activity; c) after optional SDS-PAGE transferring/applying a portion of said sample to a protein-binding membrane; d) reacting the fibrinogen with a primary monoclonal antibody capable of binding to fibrinopeptide A moiety; and e) detecting the quantity of intact fibrinogen in the sample by quantifying the amount of the bound primary monoclonal antibody.

Abstract: The application relates to a substrate for measuring the activity of a deubiquitinating enzyme (DUB), comprising a diubiquitin molecule, wherein an ubiquitin monomer is labeled with a fluorescent label, as well as an assay for DUB enzymes using such substrates.

Abstract: Antibodies, polypeptides, and polynucleotides are provided for the detection, prevention, amelioration and treatment of diseases caused by Actinobacillus actinomycetemcomitans.

Abstract: A method of assessing arachidonic acid (AA) metabolites-dependent hypertension by measuring glucuronidated dihydroxyeicosatrienoic acids (DHETs) and DHET metabolites in a biological sample which contains the epitopes unique to DHET (using any methods including GC/MS, LC/MS or ELISA). An example of the glucuronidated DHET metabolite is DHET-alcohols such as omega or omega-1 oxidated DHET and DHET esterified glycerol. The method further includes determining the amount of glucuronidated molecules containing a DHET-specific epitope which is immunoreactive with antibodies produced against DHETs.

Abstract: The present invention is related to accurate detection methods for the measurement only of myeloperoxidase (MPO) levels or neutrophils, preferably equine neutrophils, in complex biological samples. The present invention is further related to ELISA and SIEFED assays for such detection. SIEFED detection sensitivity of active peroxidase activity was found to be enhanced by the addition of nitrite. Such MPO measurement finds its use in many applications such as the prediction, diagnosis and/or monitoring of pathologies correlated with neutrophil activation and/or destruction; the evaluation of drugs and/or immunomodulators; the assessment of immune responses, either natural and/or after treatment with immunomodulators and/or drugs; and the study of cells and their ability to fight microorganisms and/or to destroy them.

Abstract: The invention is directed to droplet actuator devices and assay methods. The method may include immobilization of the enzymatic substrate including forming an inclusion complex with the substrate within an aqueous environment in contact with an oil.

Abstract: The present invention provides compounds, compositions, and methods for detecting, diagnosing and treating cancers such as glioblastoma multiforme.

Abstract: This invention provides: novel protein homologous of a Kunitz domain, which are capable of binding kallikrein; polynucleotides that encode such novel proteins; and vectors and transformed host cells containing these polynucleotides.

Abstract: The invention provides enzymatic methods for direct determination of percentage of glycated hemoglobin in blood samples without the need of a separated measurement of total hemoglobin content in blood samples. The methods utilizes one or two different types of oxidizing agents which selectively oxidize low-molecular weight reducing substances and high-molecular weight (mainly hemoglobin) reducing substances in blood samples, coupled with enzymatic reactions catalyzed by proteases, fructosyl amino acid oxidase. The amount of hydrogen peroxide generated in the reaction is measured for determination of percentage of glycated hemoglobin in blood samples. The invention provides kits for performing the methods of the invention.

Abstract: The invention provides enzymatic methods for direct determination of percentage of glycated hemoglobin in blood samples without the need of a separated measurement of total hemoglobin content in blood samples. The methods utilizes one or two different types of oxidizing agents which selectively oxidize low-molecular weight reducing substances and high-molecular weight (mainly hemoglobin) reducing substances in blood samples, coupled with enzymatic reactions catalyzed by proteases, fructosyl amino acid oxidase, and peroxidase. The invention provides kits for performing the methods of the invention.

Abstract: The invention relates to methods for determining the activity of a proteolytic coagulation factor of the blood coagulation cascade in a body fluid such as whole blood or plasma. A combination is provided in a reaction mixture. The combination comprises the sample and an activation agent for activating a proteolytic coagulation factor of the blood coagulation cascade or for activating the blood coagulation cascade. The effect of the activating on a reagent system comprising a cleavable moiety is evaluated. The cleavable moiety is or becomes bound to a chemiluminescent agent or a sensitizer agent or both. The chemiluminescent agent and the sensitizer agent are related in that, when in close proximity, energization of the sensitizer agent results in energization of the chemiluminescent agent. The effect of the activating is related to the activity of a proteolytic coagulation factor of the blood coagulation cascade wherein the effect is the extent of cleavage of the cleavable moiety.

Abstract: The invention provides methods for predicting, diagnosing or monitoring acute cardiac disorders, cardiac transplant rejection, or distinguishing acute cardiac disorders from pulmonary disorders, by measuring BNP signal peptide levels in a sample taken from a subject shortly after onset of, or presentation with the disorder or transplant rejection.

Abstract: The present invention relates to immunochemical visualization and quantification of single target entities, such as single molecules, single molecular structures, single particles, etc. in samples wherein said single entities are immobilized. In particular, the invention relates to methods for visualization and quantification of single units of biological or chemical targets, in particular to immunochemical visualization of single molecules of biological targets in histological samples. The methods of the invention comprise a step of forming discrete deposits of detectable molecules at single target sites of sample mediated by an enzyme with oxydoreductase activity, wherein a single target site comprises a single unit of a target. The invention also relates to assays comprising the present visualization and quantification methods and diagnostic applications of said methods.

Abstract: Dual colored fluorescent indicators of specific tyrosine kinase activity and methods of use thereof are disclosed.

Abstract: The present invention provides a new method for engineering or evolving enzymes to have desirable characteristics. Among the desirable characteristics is the ability to control catalytic activity through the use of a trigger molecule that rescues a catalytic site defect introduced during the engineering process. The method includes co-evolving enzyme and substrate to retain or improve substrate binding activity in the absence of catalytic activity.

Abstract: The present invention relates to peptides and polypeptides having the sequence SAVTFAVCAL or variants thereof, capable of binding to Calcineurin and/or to NS5A-TP2 and to their use in therapy, as well as to nucleic acid sequences and vectors encoding these peptides and polypeptides, and to cells comprising said polypeptides, nucleic acid sequences or vectors. The invention further relates to the use of the peptides, polypeptides or their derivatives to bring about phenotypic changes in mammalian cells, particularly to up-regulate calcineurin activity. The invention finally relates to a method for intracellular identification of substances which bind to calcineurin and which modulate the physiological effects of calcineurin.

Abstract: The present invention relates to antibodies with a specificity for BACE1. More specifically, the invention provides monoclonal antibodies that bind to BACE1 and are capable of inhibiting the activity of BACE1 and methods producing these antibodies. The antibodies can be used for research and medical applications. Specific applications include the use of BACE1-specific antibodies for the treatment of Alzheimer's disease.

Abstract: A system and method for identifying a botulinum neurotoxin inhibitor employing a botulinum neurotoxin substrate complex having a peptide substrate, preferably SNAP-25, a reporter domain on one side of said peptide substrate and an immobilization domain on the opposite side of said peptide substrate. The botulinum neurotoxin inhibitor is identified by its ability to decrease the relative amount of cleaved complex, detected through measuring a decrease in complex bound to a solid support. The method of the present invention also utilizes novel cells that express a botulinum neurotoxin substrate complex. Also provided are novel stable cell lines that express the botulinum toxin substrate complex and viral vectors capable of efficiently expressing an active light chain of the BoNT within mammalian cells.

Abstract: A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

Abstract: This invention provides methods and systems for identifying and typing toxicity of chemical compositions, as well as for screening new compositions for toxicity. The invention involves detecting alterations in gene or protein expression and hence establishing molecular profiles in isolated mammalian LSCs contacted with various chemical compositions of known and unknown toxicities, and correlating the molecular profiles with toxicities of the chemical compositions.

Abstract: An adhesive composition for use in devices and methods for measuring a presence or a concentration of a particular component, such as an antigen, in a sample, such as blood, are provided. In one exemplary embodiment of an adhesive composition, the composition includes an adhesive, water, a poloxamer, and an anticoagulant. The adhesive can include particular properties, such as being hydrophilic, pressure-sensitive, heat-activated, and/or water soluble. The adhesive is particularly useful because it can help improve the flow of sample a device. For example, when the device is an immunosensor, the adhesive can help prevent the blood from clotting in chambers of the immunosensor. This results in a more efficient and accurate determination of the concentration of the sample. Methods of making the composition and device in which the composition can be used are provided, as are methods of using the same.

Abstract: A homogeneous method of determining inhibitors of proteolytically active coagulation factors (anticoagulants) in a sample, in particular direct thrombin and factor Xa inhibitors, and also a test kit to be used in such a method. Use is made of ligands which bind to the proteolytically active coagulation factor but are not cleaved by the latter and compete with the anticoagulant to be determined.

Abstract: A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

Abstract: The invention features methods for the identification of leucine-rich kinase 2 (LRRK2) inhibitors using indol ligand 91. Generally, these methods include identifying compounds that compete with indol ligand 91 for binding to LRRK2.

Abstract: Assay methods are disclosed involving specific binding reactions which are simplified compared to known methods. A compound capable of producing chemiluminescence is immobilized on a solid support as is a member of a specific binding pair for capturing an analyte from a sample. An activator compound that activates the chemiluminescent compound and is conjugated to a specific binding pair member is added in excess along with the sample to the solid support. Addition of a trigger solution causes a chemiluminescent reaction at the sites where the activator conjugate has been specifically bound. The assay methods are termed non-separation assays because they do not require removal or separation of excess detection label (activator conjugate) prior to the detection step. The methods are applicable to various types of assays including immunoassays, receptor-ligand assays and nucleic acid hybridization assays.

Abstract: The present invention concerns an array-based analytical system and method having an enhanced sensitivity which allows for simple and rapid analysis of relative unmodified samples which comprises an analytical system of the type having a plurality of different first members of a specific binding pair affixed in an array thereupon, a mixture including at least one second member of a specific binding pair capable of binding to one of the first members so as to form a specific binding pair which is affixed to the support member, and a reporter system that produces a detectable signal indicative of the presence of the specific binding pair on the support member and wherein the reporter system includes an amplified reporter system that is independent of layering.

Abstract: A method for the multiplexed diagnostic and genetic analysis of enzymes, DNA fragments, antibodies, and other biomolecules comprises the steps of constructing an appropriately labeled beadset, exposing the beadset to a clinical sample, and analyzing the combined sample/beadset by flow cytometry. Flow cytometric measurements are used to classify, in real-time, beads within an exposed beadset and textual explanations, based on the accumulated data obtained during real-time analysis, are generated for the user. A secondary reagent, such as a metal or magnetic particle, is added to the beadset to assist in the analysis. Detection techniques, such as such as light scatter, Rayleigh scatter, Raman scatter, surface plasmon resonance, magnetic induction, or magnetoresistance are used to detect the particle labels.

Abstract: The present invention relates to an assay for transferases. The assay comprises a first moiety comprising a transferase substrate and a second moiety capable of binding to the transferase substrate after it has been acted on by the transferase. One of the first and second moieties comprises a fluorophore and the other of the first and second moieties causes a change in fluorescence of the fluorophore. Thus, when the second moiety binds the transferase substrate after it has been acted on by the transferase, a change in fluorescence can be detected. The assay allows for agents that modulate the activity of the transferase to be screened.

Abstract: The invention provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs recognized by lipoic acid ligase and lipoic acid ligase mutants.

Abstract: Methods and reagents are disclosed for pretreating a sample suspected of containing a hydrophobic drug for conducting an assay method for detecting the hydrophobic drug. A combination is provided in a medium that includes the sample, a releasing agent for releasing the hydrophobic drug and the metabolites from endogenous binding moieties, and a selective solubility agent that provides for substantially equal solubility of the hydrophobic drug and the metabolites in the medium. The selective solubility agent includes a water miscible, non-volatile organic solvent and is present in the medium in a concentration sufficient to provide for substantially equal solubility of the hydrophobic drug and the metabolites in the medium. The medium, which may further include a hemolytic agent, is incubated under conditions for releasing the hydrophobic drug and the metabolites from endogenous binding moieties. The pretreated sample may be subjected to an assay for determining the hydrophobic drug.

Abstract: An analytical strip and a detecting method using the analytical strip are provided. The analytical strip includes a substrate having a channel thereon. The channel has a first region, a second region and a third region, which are arranged successively. A first antibody is localized in the first region. A saccharide and a peroxidase are localized in the first or second region. A second antibody for recognizing a different epitope of an identical antigen with the first antibody is immobilized in the second region. A substrate reagent including a saccharide oxidase is localized in the third region.

Abstract: The present invention relates to agents, and methods for identifying compounds, which agents and compounds result in the stabilization of mast cells, in particular that inhibit mast cell degranulation. In addition, the invention relates to compositions and methods for the use thereof in treating conditions that are characterized by mast cell degranulation and/or inflammation, including allergic rhinitis.

Abstract: A homogeneous method for determining the enzymatic activity of an analyte in a sample permits simultaneous determination of the amount of enzyme in the same reaction mixture and determination of enzyme activators or inhibitors.

Abstract: Assay methods are disclosed involving specific binding reactions which are simplified compared to known methods. A compound capable of producing chemiluminescence is immobilized on a solid support as is a member of a specific binding pair for capturing an analyte from a sample. An activator compound that activates the chemiluminescent compound and is conjugated to a specific binding pair member is added in excess along with the sample to the solid support. Addition of a trigger solution causes a chemiluminescent reaction at the sites where the activator conjugate has been specifically bound. The assay methods are termed non-separation assays because they do not require removal or separation of excess detection label (activator conjugate) prior to the detection step. The methods are applicable to various types of assays including immunoassays, receptor-ligand assays and nucleic acid hybridization assays.

Abstract: Methods of activating an apo-peroxidase are provided. The methods include the steps of providing a solution comprising an apo-peroxidase and a heme choline ester, hydrolyzing the heme choline ester with a choline esterase, and converting the apo-peroxidase to active peroxidase. The methods disclosed herein also provide for detecting the presence of an analyte in a sample. The methods include providing a binder capable of specifically binding the analyte wherein the binder is attached to a solid surface, applying the sample to the solid surface under conditions that permit binder-analyte binding, adding a choline esterase-conjugated binder that binds to the analyte, adding an apo-peroxidase, a heme choline ester, a peroxide, and a peroxide-activated signal generator, and detecting the signal produced by the peroxide-activated signal generator. An associated kit and components are also provided.

Abstract: A method of using a biological indicator to detect and identify sources of fecal contamination such as leaking sewer lines, septic tanks, and holding lagoons, using a microorganism having a biological indicator including an exogenous DNA indicator sequence incorporated into the genome of the microorganism. The exogenous DNA indicator sequence functions as a detectable signal that may be detected by visual detection methods or direct detection methods of the indicator sequence itself.

Abstract: The present invention provides a method for predicting the likely timing of the onset of menopause for a perimenopausal female subject by determining the amount of hLH?cf in a sample from the subject comprising the steps of: (a) contacting a sample from the subject with an antibody which specifically binds to hLH?cf without substantially cross-reacting with hLH, hLH? or hLH?cf, under conditions permitting formation of a complex between the antibody and hLH?cf; (b) measuring the amount of complex formed, so as to thereby determine the amount of hLH?cf in the sample; and (c) comprising the amount of hLH?cf in the subject's sample determined in step (b) with either (i) the amount determined for known postmenopausal female subject or (ii) the amount determined for a sample from a known premenopausal female subject, wherein an amount of hLH?cf in the sample similar to the amount of hLH?cf in the known postmenopausal sample indicates temporal proximity to the onset of menopause, and an amount of hLH?cf in the sampl

Abstract: The invention concerns the field of hemostasis, in particular blood coagulation disorders linked to an abnormal expression of tissue factor, and to physiopathological phenomena correlated with over-expression of the factor. The present invention provides a method for assaying the activity of circulating tissue factor in a biological sample. The method of the invention is carried out in vitro, in particular on a blood sample collected from a patient.

Abstract: This invention is in the field of medical devices. Specifically, the present invention provides portable medical devices that allow real-time detection of analytes from a biological fluid. The methods and devices are particularly useful for providing point-of-care testing for a variety of medical applications. In particular, the medical device reduces interference with an optical signal which is indicative of the presence of an analyte in a bodily sample.

Abstract: Compositions, devices, systems and methods for reducing and/or preventing photodamage of one or more reactants in illuminated analytical reactions by one or more of incorporating photodamage mitigating agents within the reaction mixture and/or interrogating different observation regions of the reaction mixture for a period that is less than a photodamage threshold period.

Abstract: Assay methods are disclosed involving specific binding reactions which are simplified compared to known methods. A compound capable of producing chemiluminescence is immobilized on a solid support as is a member of a specific binding pair for capturing an analyte from a sample. An activator compound that activates the chemiluminescent compound and is conjugated to a specific binding pair member is added in excess along with the sample to the solid support. Addition of a trigger solution causes a chemiluminescent reaction at the sites where the activator conjugate has been specifically bound. The assay methods are termed non-separation assays because they do not require removal or separation of excess detection label (activator conjugate) prior to the detection step. The methods are applicable to various types of assays including immunoassays, receptor-ligand assays and nucleic acid hybridization assays.