Abstract: The invention relates to a method for measuring the level of a preselected analyte in a sample of blood of a human or animal patient by incubating the test sample with an antibody specific to the analyte to form an immunocomplex, which then interacts with the white blood cell fractions and result in the production of oxidants. Oxidants are detected using chemiluminescent reagents. The assay is performed on the sample and in addition includes a measurement of the oxidant production resulting from a maximal stimulatory dose of immunocomplexes, providing a ratio to indicate the level of analyte in the sample. The white blood cell oxidant response may be enhanced by the inclusion of certain agents such as zymosan.

Abstract: A method for the preparation of a fluorescence resonance energy transfer (FRET) substrate having donor and acceptor species on opposite sides of a proteolytic cleavage site and wherein the donor and/or acceptor species are attached via the side chain(s) of amino acid(s) therein. The method comprises contacting a reactive donor or acceptor species with a polypeptide substrate having the side chain(s) of amino acid(s) therein adapted for reaction with the reactive species and then contacting the substrate so obtained with a corresponding reactive donor or acceptor species. Novel FRET substrates so prepared and their use in assays to identify modulators of protease activity.

Abstract: A chemiluminescent assays for the determination of the presence or amount of a biopolymer in bound assays using 1,2-dioxetanes in connection with AttoPhos™ as chemiluminescent substrates for enzyme-labeled targets or probes is provided. Further disclosed is a kit for conducting a bioassay for the presence or concentration of a biopolymer comprising a) an enzyme complex; b) a 1,2-dioxetane; and c) AttoPhos™.

Abstract: This invention relates to an improved method for the multi-parameter analysis of cells from peripheral blood or bone marrow. The method uses a nucleic acid dye, at least two fluorescently labelled monoclonal antibodies and at least two light scatter parameters to differentiate and discriminate between and among different cells in the blood or bone marrow.

Abstract: The present invention provides high throughput screening systems for identifying antifungal compounds. The method can be performed in plurality simultaneously with fluorescence or absorbance readouts.

Abstract: The present invention provides methods for diagnosing and/or monitoring thrombophilic disease in a patient that can result from the antiphospholipid antibody syndrome (aPL syndrome). The methods of the invention are premised on the inhibition of binding of an anticoagulant protein, annexin, preferably annexin-V, to phospholipids by antiphospholipid (aPL) antibodies in a patient blood sample.

Abstract: An apparatus and method for real-time measurement of a cellular response of a test compound or series of test compounds (303) on a flowing suspension of cells (349), in which a homogeneous suspension of each member of a series of cell types (349) is combined with a concentration of a test compound (303), directed through a detection zone (355), and a cellular response of the living cells is measured in real time as the cells in the test mixture are flowing through the detection zone (355). The apparatus may be used in automated screening of libraries of compounds, and is capable of real-time variation of concentrations of test and standard compounds and generation of dose/response profiles within a short time span.

Abstract: Disclosed herein are methods and reagents kits forth quantitative in vitro determination of the functional determination of multi-drug resistance in cells, as well as for the clinical screening of potential modulators of multi-drug resistant transport activity in cells. The method of the invention is based on the measurement of the accumulation rate of free calcein within the cells of the specimen (advantageously by flourescence measurement), after exposing the cells in vitro to a cell permeable form of calcein that is a good substrate for MDR proteins present in the sample. The cell permeable form of calcein is converted within the cell by intracellular enzymes to free calcein. Comparison of free calcein accumulation in the presence and absence of a potential inhibitor of transport activity permits the rapid screening of such inhibitors.

Abstract: The present invention relates to processes for the direct detection of biochemically functional retroviruses in biological samples. The processes of the invention are characterized by the structure-specific extraction of retrovirus particles and a subsequent analysis and detection of retrovirus-specific enzymatic reactions. The processes of the invention have broad application in the diagnosis of retroviral infection and virological research.

Abstract: Inorganic phosphate may be detected and optionally quantified via the coupling of a phosphate-dependent enzymatic reaction with an enzyme system that generates hydrogen peroxide in the presence of a chromogenic or fluorogenic peroxidase substrate. Phosphate consuming or phosphate-producing enzymes or their substrates may also be detected and/or quantified, including pyrophosphatase enzymes or pyrophosphatase.

Abstract: The excitation of label molecules usable in chemical and biochemical analysis by electrical pulses at electrodes covered with a thin insulating film, and the use of such electrodes in chemical, clinical and biochemical analysis. The electrodes include a conducting base material that has been coated with an organic or inorganic insulating film or multiple layers of such films, so that either one or several label compounds can be excited to an excited state which is deexcited by emission of ultraviolet, visible or infrared light, in aqueous solution providing the basis for reproducible analytical applications in bioaffinity assays such as immunoassays and DNA-probing assays.

Abstract: Methods are disclosed for determining an analyte in a medium suspected of containing the analyte. One method comprises treating a medium suspected of containing an analyte under conditions such that the analyte, if present, causes a photosensitizer and a chemiluminescent compound to come into close proximity. The photosensitizer generates singlet oxygen and activates the chemiluminescent compound when it is in close proximity. The activated chemiluminescent compound subsequently produces light. The amount of light produced is related to the amount of analyte in the medium. Preferably, at least one of the photosensitizer and chemiluminescent compound is associated with a surface which is usually a suspendible particle, and a specific binding pair member is bound thereto. Compositions and kits are also disclosed.

Abstract: Provided herein are assays for measuring in vivo levels of activated mannan-binding protein-associated serine protease (MASP-1 and MASP-2) activity. Also provided are compounds that are useful for assessing the in vivo levels and for monitoring in vitro and in vivo complement-activation (C-activation).

Abstract: The present invention relates to a method of using a storage-stable, pre-stained 3,3′,5,5′-tetramethylbenzidine (TMB) substrate in an enzyme substrate. The TMB substrates comprise: (a) a water-based TMB substrate composition which includes a peroxide, e.g., hydrogen peroxide or urea hydrogen peroxide, and (b) a dye which is soluble therein. Suitable dyes to be included in the substrate preferably have an absorbance at 450 nm of at the most milli absorbance units. Examples of dues are Pholixine B, Eosin B and Quinaldine Red. The substrates are advantageous in that they include a dye which is visible to the human eye. This is especially valuable when the substrates are handled and measured. The substrates are especially suitable for enzyme assays such as enzyme-linked-immunosorbent-assays (ELISA), e.g. horse radish peroxide (HRP) is used. The substrates may advantageously include less than 5% organic solvents and a solubility increasing agent such as polyvinylalcohol.

Abstract: Methods and compositions for detecting and localizing light originating from a mammal are disclosed. Also disclosed are methods for tracking light emission to selected regions, as well as for tracking entities within the mammal. In addition, animal models for disease states are disclosed, as are methods for localizing and tracking the progression of disease or a pathogen within the animal, and for screening putative therapeutic compounds effective to inhibit the disease or pathogen.

Abstract: In accordance with the present invention, a method of conducting an assay of a sample containing an analyte of interest includes the step of forming a mixture so as to bring a metal-ligand complex into interactive proximity with the sample containing the analyte of interest. The mixture is irradiated with electromagnetic light energy so as to cause emission of light indicative of the analyte of interest. The emitted light is measured, and the measurement of the emitted light is utilized to measure the analyte of interest. The metal-ligand complex can be [Re(bcp)(CO)3(4-COOHPy)]+, [Os(phen)2(aphen)]2+, [Os(tpy)(triphos)]2+, [Os(tppz)2]2+, and [Os(ttpy)2]2+, or the like. Also, the present invention is directed to a metal-ligand complex of the formula [Re(bcp)(CO)3(4-COOHPy)]+.

Abstract: The present invention provides a process for determining the efficacy of anti-viral therapy in an HIV-infected subject receiving such therapy. The process includes the steps of a) detecting the level of transcriptionally active HIV in monocytes of the subject at a plurality of different times, b) comparing the detected HIV levels, and c) correlating changes in the detected HIV levels over time with the therapy. The process can be used to monitor the efficacy of treatment with any anti-HIV agent such as AZT, 3TC, DDC, Indivar, or Saquinavir. Decreases in HIV levels over time indicate an efficacious treatment. Increases in detected HIV levels over time indicate resistance to treatment.

Abstract: Novel fluorescent substrates of human cytochrome P450 enzymes are provided. Also provided are methods for their manufacture and use. These substrates are useful in assessing cytochrome P450 enzyme activity and in selecting compounds which inhibit cytochrome P450 enzyme activity and, in particular, for identifying potential adverse drug interactions which are mediated by inhibition of cytochrome P450 enzyme activity. The compounds are particularly useful for assessing cytochrome P450 CYP3A enzyme activity.

Abstract: Materials and methods are provided for producing patterned multi-array, multi-specific surfaces for use in diagnostics. The invention provides for electrochemiluminescence methods for detecting or measuring an analyte of interest. It also provides for novel electrodes for ECL assays. Materials and methods are provided for the chemical and/or physical control of conducting domains and reagent deposition for use multiply specific testing procedures.

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: The invention relates to a method for quantitating the level of a preselected analyte in a sample of blood of a human or animal patient by incubating the test sample with an antibody specific to the analyte to form an immunocomplex, which then interacts with the white blood cell fractions and result in the production of oxidants. Oxidants are detected using chemiluminescent reagents. In addition, the white blood cell oxidant response may be enhanced by the inclusion of certain agents such as opsonized zymosan. As part of the assay, separate blood samples are also maximally stimulated with a maximal stimulatory amount of exogenously-added antigen, and corresponding antibody, to form immunocomplexes, to provide a response factor used in the quantitation of analyte.

Abstract: The subject invention relates generally to immunoassays for detection of antibodies by use chemiluminescent compounds. More particularly, the subject invention relates to chemiluminescent immunoassays to detect antibodies wherein a precomplex mixture is created and a two-step assay is performed resulting in a greater signal.

Abstract: Disclosed are methods and sensors for detecting the presence or concentration of an analyte in a sample, preferably a sugar such as glucose, which preferably utilizes a labeled engineered protein, or fragment thereof, that is capable of binding the analyte to be detected.

Abstract: A method and apparatus for ascertaining and measuring the quality of milk by analyzing reflected or absorbed colors or both with which the milk is irradiated. The milk is thus irradiated with three colors of light in wavelengths of red, green and blue and the intensity and color of the light reflected by the milk or absorbed thereby, or both, is measured. The light sources are periodically switched off whereby the receiving sensors measure the same red, green and blue wavelengths due to background sources to the extent that they are reflected or absorbed or both by the milk. This data is then compared with data with the light sources switched on to produce the net data which is relevant to the milk as it is being received from the animal being milked.

Abstract: Provided is a method of fluorescence detection of lipid membranes using lipophilic, functionalized nanocrystals; and lipid membranes labeled by the lipophilic, functionalized nanocrystals. The method comprises contacting an effective amount of lipophilic, functionalized nanocrystals with the lipid membranes; exposing the labeled lipid membranes to an excitation light suitable for exciting the functionalized nanocrystals present in the labeled lipid membranes to emit a fluorescence emission. Also provided is a composition comprising a functionalized nanocrystal which is bound to a substrate in a living tissue.

Abstract: The method of cancer screening allows for a safe, reliable, inexpensive and minimally invasive diagnosis. Cells are first collected through non-invasive or minimally invasive means. The collected cells are stored in a cell culture media to keep them viable. A chemical compound such as 5-ALA is introduced to the cultured cells. The cells are incubated a period of time in order for the interaction to occur between the introduced chemical compound and the collected cells. The cells are then studied under a fluorescence microscope which emits a specific frequency of light which matches the excitation frequency of light which would be required to induce flourescence of abnormal cells. Pre-malignant and malignant cells will fluoresce in response to the interaction with the introduced compound and the specific frequency of light, while normal healthy cells generally will not fluoresce. If necessary, the collected cells may be passed through a flow cytometer to more easily identify the fluorescing cells.

Abstract: A method and compositions for the detection and/or quantification of an analyte through the use of a plurality of labeled probes, with two or more said probes targeted to different regions of said analyte. In specific embodiments, the labels are separately distinguishable, and/or are present at different specific activities on the labels.

Abstract: The method is based on the carbon-13 labelled urea breath test, which comprises a) administering to the patient an aqueous solution of citric acid at pH comprised between 2 and 2.5; b) collecting a sample of the patient's breath with the object of determining the baseline content of carbon-13; c) administering to the patient a suitable amount of carbon-13 labelled urea; d) collecting a sample of the patient's breath with the object of determining the content of carbon-13 after the administration of the labelled urea; and e) analyzing the breath samples collected to determine the carbon-13 content before and after the administration of urea labelled with carbon-13. The kit has a container for the citric acid, another for the carbon-13 labelled urea, exetainers destined to containing the patient's breath, and means to blow the breath into the exetainers.

Abstract: The present invention relates to isolated and pure Toxoplasma gondii antigenic fragments, recombinant polypeptides, nucleic acids encoding them, primers and probes derived from the same, as well as the use of these polypeptides, nucleic acids, primers and probes in methods and kits for the diagnosis and prevention of T. gondii infection in mammals (humans and animals).

Abstract: One aspect of the present invention is a multi-well platform for fluorescence measurements, comprising a plurality of wells within a frame, wherein the multi-well platform has low fluorescence background. Another aspect of the present invention is a system for spectroscopic measurements, comprising reagents for an assay and a multi-well platform for fluorescence measurements. A further aspect of the present invention is a method for detecting the presence of an analyte in a sample contained in a multi-well platform by detecting light emitted from the sample. Another aspect of the present invention is a method from identifying a modulator of a biological process or target in a sample contained in a multi-well platform by detecting light emitted from the sample. Another aspect of the present invention is a composition identified by this method. A further aspect of the present invention is a method to identify a therapeutic.

Abstract: A method for detecting and measuring exoglycosidase activity is presented. The method employs derivatives containing the fluorescent group 4-methylumbelliferyl (“4-Mu”) at a pH lower than that conventionally employed. While the fluorescence intensity due to the 4-Mu group is considerably diminished at the lower pHs employed, the fluorescence intensity is still sufficient to continuously measure exoglycosidase activity in the activity range commonly assayed. The method is easily adaptable to high throughput enzyme assay systems and automated data analysis method. The method also provides a means to detect alterations in exoglycosidase activity that are independent of expression levels.

Abstract: Electrochemiluminescent assay methods for determining an analyte of interest, which methods include forming one or more compositions which include, in the aggregate, (a) a sample to be tested for the analyte of interest, (b) a component capable of specifically binding with the analyte, (c) a label reagent which, when oxidized, is capable of electrochemiluminescence, (d) an amine which, when oxidized, forms a reducing agent, and (e) an electrolyte solution; subjecting such a composition containing the sample to conditions sufficient for specific binding to occur between the analyte of interest, if present, and one or more of the other components in the composition; thereafter, a composition containing the label reagent and amine, and reflecting the outcome of applying the aforementioned binding conditions, is exposed to conditions, such that both the label reagent and the amine are oxidized, the amine forms a reducing agent which interacts with the label reagent, electrochemiluminescence occurs, and the lumine

Abstract: A rapid method for detecting the presence or absence of coliform bacteria in a liquid or liquified dairy sample, for example skimmed milk. A growth medium containing a fluorogenic substrate is combined with the sample and is incubated for a brief period of about 7-9 hours after which a single fluorescence value is measured. Total or thermotolerant coliform bacteria are determined to be present in the sample if the single fluorescent measurement exceeds a predetermined threshold value.

Abstract: A method of in situ analysis of a biological sample comprising the steps of (a) staining the biological sample with N stains of which a first stain is selected from the group consisting of a first immunohistochemical stain, a first histological stain and a first DNA ploidy stain, and a second stain is selected from the group consisting of a second immunohistochemical stain, a second histological stain and a second DNA ploidy stain, with provisions that N is an integer greater than three and further that (i) if the first stain is the first immunohistochemical stain then the second stain is either the second histological stain or the second DNA ploidy stain; (ii) if the first stain is the first histological stain then the second stain is either the second immunohistochemical stain or the second DNA ploidy stain; whereas (iii) if the first stain is the first DNA ploidy stain then the second stain is either the second immunohistochemical stain or the second histological stain; and (b) using a spectral data collec

Abstract: Xanthan esters and acridans are substrates for horseradish peroxidase. These stable, enzymatically cleavable chemiluminescent esters are substrates for horseradish peroxidase which, together with peroxide is among the extensively used enzyme in enzyme-linked detection methods, including immunoassays, oligonucleotide detection and nucleic acid hybridization. The novel compounds are used, together with peroxide, alkali and the peroxidase, to indicate the presence and/or concentration of target compounds. The assays may be enhanced by the use of polymeric quaternary onium enhancement compounds or similar compounds selected to enhance the chemiluminescence emitted.

Abstract: The invention relates to a method for staging sepsis in a patient by concurrently measuring in a sample of the patient's blood four parameters which are indicative of the stage of sepsis: 1) the level of microbial product level in the blood; 2) the level of tumor necrosis factor (TNF) response reserve; 3) the maximum oxidant production by neutrophils, and 4) the responsiveness of the patient's neutrophils to immunocomplexes. Based on determining the stage of sepsis, appropriate treatment for the patient may be identified.

Abstract: A method is provided for determining in a medium the presence or amount of an analyte which is capable of binding to a ligand partner to form a ligand complex, which method comprises: (a) contacting the medium with either: (i) a ligand partner conjugated with an enzyme being capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, or (ii) a ligand partner and either a competing analyte or an analog of said analyte, a competing analyte or analyte analog being capable of forming a ligand complex with the ligand partner and the competing analyte or analyte analog being conjugated with an enzyme capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, (b) optionally separating complexed and uncomplexed enzyme conjugates, (c) causing or allowing the reaction or cascade of reactions to occur to produce hydrogen peroxide by contacting the complexed or uncomplexed enzyme conjugate with a corresponding enzyme su

Abstract: The present invention relates to a method for the specific labeling of a protein containing selenocyst(e)ine and/or cyst(e)ine groups, comprising the following steps:at least one incubation of a protein-containing sample with at least one modifying agent specific for selenocyst(e)ine and/or cyst(e)ine groups, followed byat least one further incubation of said protein-containing sample with at least one labeling agent specific for selenocyst(e)ine and/or cyst(e)ine groups;wherein at least one substance interacting with said protein is added prior to and/or during and/or after at least one of said incubations.

Abstract: A sentinel chamber for containing microorganisms in a matrix, and methods for the use of the chamber in studying organisms in the field. The matrix is typically the same material as that within which the chamber will be placed. The chamber is made of a cylindrical body having a porous filter at each end. The filter at one end is contained under a snap-on cap, making the filling of the chamber quick and easy. The method of the invention uses the sentinel chambers to study the characteristics and survival of a sentinel organism in a particular environment. The medium present in the environment (i.e. soil, manure, sewage sludge, etc.) is placed in the sentinel chamber and a quantity of the sentinel organism (i.e. C. parvum oocysts) is injected into the medium. A filter is placed across the open end of the chamber, and held in place by the cap. The sentinel chamber is then buried or immersed in the environment (i.e. a field, manure pile, septic tank).

Abstract: A method for the detection or quantitation of a water-borne parasite, such as Cryptosporidia. The detection or quantitation is accomplished by an electrochemiluminescence assay comprising the steps of filtering water to obtain a sludge thought to contain the parasite or a fragment thereof; extracting a sample of said sludge in a extraction medium to form antigenic derivatives of said parasite; forming an assay mixture comprising a sample of said extracted sludge and an antibody specific to said antigenic derivative; incubating said assay mixture to permit binding of said antibody and said antigenic derivative; and conducting an electrochemiluminescence assay for the bound complex of antibody and antigenic derivative, thereby detecting or quantitating the Cryptosporidia in said water.

Abstract: Novel screening methods for identifying antimicrobial agents involving use of membrane potential indicator dyes are provided.

Abstract: The invention provides a compound, useful as an optical probe or sensor of the activity of at least one cytochrome P450 enzyme, and methods of using the compound to screen candidate drugs, and candidate drugs identified by these methods. The optical probe of the invention is a compound having the generic structure Y-L-Q, wherein Y is selected from the group consisting of Q as herein defined, saturated C.sub.1 -C.sub.20 alkyl, unsaturated C.sub.1 -C.sub.20 alkenyl, unsaturated C.sub.1 -C.sub.20 alkynyl, substituted saturated C.sub.1 -C.sub.20 alkyl, substituted unsaturated C.sub.1 -C.sub.20 alkenyl, substituted unsaturated C.sub.1 -C.sub.20 alkynyl, C.sub.1 -C.sub.20 cycloalkyl, C.sub.1 -C.sub.20 cycloalkenyl, substituted saturated C.sub.1 -C.sub.20 cycloalkyl, substituted unsaturated C.sub.1 -C.sub.20 cycloalkenyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl; L is selected from the group of (--O(substituted ortho-phenyl)CR.sup.2 H).sub.p --, (--O(substituted meta-phenyl)CR.sup.2 H).sub.

Abstract: Materials and methods are provided for producing patterned multi-array, multi-specific surfaces which are electronically excited for use in electrochemiluminescence based tests. Materials and methods are provided for the chemical and/or physical control of conducting domains and reagent deposition for use in flat panel displays and multiply specific testing procedures.

Abstract: Fluorescent dyes possess reactive linkers for conjugating to nucleic acids, carbohydrates and peptides. The conjugates fluoresce in the visible and UV spectrum and have an excellent solvochromatic response as compared to other fluorescence or chromatic labels. The conjugates are stable but also have medium sensitive. The fluorescent dyes have little triplet state formation and are not photoreactive, making them an excellent substance for biological investigations. Uses for the dyes include protein labeling, DNA labeling, single molecule spectroscopy and fluorescence. A synthesis of the dyes is disclosed. Methods of use include the detection of carbohydrate-protein interactions.

Abstract: In a method of determining heparin content, a known amount of thrombin (FIIa) or activated coagulation factor X (FXa) is added to a mixture which comprises a known concentration of chromogenic substrate (S), a known concentration of antithrombin (AT) and a sample having an unknown heparin concentration. The amount of FIIa or FXa is chosen such that at most 20%, of the S present is reacted during the period which the AT needs to deactivate all the FIIa or FXa present. The final concentration of the reacted chromogenic substrate p-nitroanilide ([pNA].sub.final) is determined after completion of the reactions, and the [pNA].sub.final is used to determine the rate constant (k.sub.dec) of the reaction of FIIa or FXa with AT using the relationship: ##EQU1## The heparin concentration in the sample can then be determined using the value of k.sub.dec from a predetermined calibration curve of k.sub.dec against heparin concentration.

Abstract: The invention discloses a fluorescent bead that can be used for determining the temperature and/or metabolic state of a single cell contained in a cell/tissue sample. The fluorescent bead contains at least one temperature-sensitive fluorophore. The invention also includes methods of monitoring cellular metabolism and methods of diagnosing abnormal metabolism of a cell.

Abstract: Culture media for microorganisms containing blood or hemin, particularly Trypticase Soy Agar with blood, and chocolate agar, are combined with known chromogenic substrates to produce chromogenic media. Methods for preparing these chromogenic media include adding chromogenic substrates to the surface of previously prepared media, or incorporating the chromogenic substrate into the media as it is prepared. Methods for distinguishing microorganisms in a sample using these culture media are also described.

Abstract: A method for determining concentration of an analyte in a biological sample comprising the steps of:(a) combining the biological sample, at least one oxidizing enzyme for the analyte of interest, nicotinamide adenine dinucleotide (hereinafter NAD.sup.+), and a chemiluminescent label to form a reaction mixture;(b) allowing the analyte to undergo an oxidation-reduction reaction and NAD.sup.+ to be converted to the reduced form of nicotinamide adenine dinucleotide (hereinafter NADH) and further allowing the chemiluminescent label to react with NADH; and(c) determining the concentration of the analyte of interest in the biological sample by correlating the quantity of light emitted with the concentration of NADH.

Abstract: A method is disclosed for assaying a proteolytic enzyme comprising: (a) incubating an enzyme-containing sample with an immobilized fluorescence-quenched peptide having the formula Que-Sub-Flu-Spa-Car or Flu-Sub-Que-Spa-Car wherein Sub is a peptide chain containing a specific cleavage site for said proteolytic enzyme; Flu is a fluorophore; Que is a quencher capable of absorbing fluorescent radiation emitted by the fluorophore; Spa is a direct bond or a spacing chain; and Car is a water-insoluble and/or macromolecular carrier; (b) optionally separating the liquid from the carrier material; (c) irradiating said carrier material and measuring fluorescence. Also disclosed are immobilized substrates containing specific amino acid sequences for use in such an assay, especially in an assay for aggrecanase, and metalloproteinase-1, -3 and -13 activity.

Abstract: This invention provides methods and apparatus for applying a reagent to an analytical test device. The methods and apparatus employ a nozzle and a flexible restrictor to accurate control the amount of reagent applied to the test device. The analytical test devices prepared by the methods of this invention are used to determine the presence or the quantity of an analyte in a liquid sample, such as whole blood.