Abstract: The present invention relates to a dry immunoassay analytical element for assaying a ligand, comprising a support bearing: (a) a label zone comprising an enzyme labeled ligand or an enzyme labeled receptor; (b) a spreading zone; (c) a receptor zone comprising a fixed concentration of an immobilized receptor for the ligand; and (d) a gravure zone comprising a diaryl telluride. A prefered embodiment of the present invention further comprises a vanadyl salt. The present invention further relates to a method for performing an assay using an element as described above.

Abstract: A method and a reagent for highly accurate measuring of trace amount of metals in a sample of living body. The method and the reagent for measuring metals in a sample of living body includes:(1) a method for measuring metals in sample of living body, in which metals other than the objective metals of measurement are added to the measurement system to release the objective metals bonding to co-existing substances in the sample from the co-existing substances; and(2) a reagent for the measurement, in which metals other than the objective metals of the measurement are contained in a reagent for measuring metals in a sample of living body in order to release the objective metals bonding to the co-existing substances in the sample from the co-existing substances.

Abstract: Simplified calibration and improved dose-response linearity of the standard curve of competitive assays are obtained by adding a predetermined amount of unlabeled competitive binding compound (usually just the analyte itself) to a first reagent containing anti-analyte antibodies before the reagent is reacted with a sample containing an unknown amount of analyte.

Abstract: This invention relates to an improved method for detecting and quantifying the presence of a target molecule, such as an antigen, an antibody or a polynucleotide, in a sample which method uses alkaline phosphatase as the reporter enzyme and the reduction of a tetrazolium salt to a formazan as part of the detection/signaling system.

Abstract: The invention relates to a process for the immunochemical determination of an analyte in a sample by means of a first specific binding partner, where the specific binding partner being immobilized on a support and the extent of the binding of the analyte to the specific binding partner being determined by means of a further specific binding partner which directly or indirectly bears a label, wherein there is additionally added to the process a binding factor, which is not labelled.

Abstract: A reagent is suitable for measuring the concentration of an analyte in a hemoglobin-containing biological fluid, such as whole blood. The reagent comprises dehydrogenase enzyme that has specificity for the analyte, NAD or an NAD derivative, a tetrazolium dye precursor, a diaphorase enzyme or an analog thereof, and a nitrite salt. The reagent causes dye formation that is a measure of the analyte concentration. The nitrite salt suppresses interfering dye formation caused non-enzymatically by the hemoglobin. Preferably, the reagent is used in a dry strip for measuring ketone bodies, such as beta-hydroxybutyrate.

Abstract: Disclosed is an improved method for the detection of an analyte in a fluid test sample using a strip of a negatively charged matrix material having a zone containing mobile, labeled binding partner for the analyte and a separate zone for capturing the labeled binding partner as it is carried through this zone by the fluid test sample. The improvement involves combining the fluid test sample with a polyalkoxylated amine surfactant to control non-specific binding of the labeled binding partner to the negatively charged matrix material.

Abstract: The present invention provides a heretofore unknown use for a aqueous reagent composition that serves as a universal rinse for performing and/or improving a variety of hematological analyses on automated analyzers. The universal rinse reagent comprises a phosphate buffer to maintain the rinse solution pH at from about 7.0 to about 7.6; a nonhemolytic nonionic surfactant, such as a Pluronic.RTM. an alkali metal salt, such as NaCi; antimicrobial and anti-oxidant compounds; and has an osmolality of about 285 to 305 mOsmol/kg. The universal rinse reagent composition is highly suitable for use in the rinse phases or cycles of all types of blood cell analysis methods and processes performed on semi- and fully-automated systems. The invention allows the replacement of mutiple and specific rinse solutions with the disclosed universal rinse to obtain accurate and acceptable results, independent of the types of blood cell analyses that are performed.

Abstract: The present invention provides a method for measuring an amount of LDL-cholesterol in samples specifically and accurately and also a reagent used for the method, and by using the present invention, such effect can be attained that LDL-cholesterol can be measured directly using so far widely used automatic analyzer, which has been impossible in known measurement methods.

Abstract: A method for avoiding influence of hemoglobin, in which the influence is caused by the changes with time in the absorption wavelength of hemoglobin when light absorption of a color reaction of a sample is measured by a rate assay, wherein the method comprises measuring the light absorption at a wavelength of from 517 to 529 nm or from 580 to 592 nm.

Abstract: A chromatographic strip positive readout binding assay device and method suitable for quick, sensitive and reliable field testing for small molecules such as environmental contaminants, drugs of abuse, therapeutic drugs and hormones. The detectable label is not attached to either the analyte or to the analyte receptor. Low affinity binding pairs can be used in the positive readout binding assay.

Abstract: An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

Abstract: Libraries of compounds such as nucleic acids or peptides are contacted with a target molecule and libraries that have at least one compound that bind with at least a minimum activity are determined by a reiterative process in which a change in the rate of recovery (elimination) of compounds that bind to the target indicates that the library contains such a compound. The procedure may also be used to determine indirectly the sequence of such compound by employing sublibraries, each of which have a known entity at a known position of the compound.

Abstract: The invention concerns interference-eliminating agents for avoiding unspecific interactions in immunoassays in which avidin or streptavidin or a derivative thereof are used as the interference-eliminating agents.

Abstract: A method of inhibiting proteolytic degradation of a thermally-stable intracellular protein is described. The method involves adding 1 or more denaturing agents to a sample which contains the protease and the protein of interest and heating the resulting solution at a temperature and for period of time sufficient to denature the protease. The method optionally includes a step for lysing the cell if the protein of interest is contained in a cell in order to release said protein. Additionally, a method of determining Mx protein induced by interferon in a blood sample is described. The method involves adding to a blood sample a lysing agent, a denaturing agent, and a detergent selected to solubilize Mx protein. The sample containing Mx protein is then heated at a temperature of from about 50.degree. C. to about 60.degree. C. for a period of time of from about 1 minute to about 30 minutes, and the Mx protein in the solution then is determined.

Abstract: The present invention is an improvement in a method using a reporter dehydrogenase for detecting the presence of an analyte in a sample in which endogenous dehydrogenases present in the sample interfere with the detection of the analyte. The improvement is the inclusion of an inhibitor of the endogenous dehydrogenase in the assay medium. The improvement finds particular utility in the measurement of ethyl alcohol using alcohol dehydrogenase as the reporter enzyme when lactate and lactate dehydrogenase present in the sample interfere with the detection of the analyte. Particulary effective inhibitors which prevent this interference are fatty acids, such as, agaric acid and lauric acid, and alkyl sulfate salts, such as, sodium dodecyl sulfate and sodium decyl sulfate.

Abstract: A device for performing an enzyme-labelled binding assay comprises an absorbent material and a developing solution containing a substrate for said enzyme, wherein the absorbent material is provided with a plurality of reagent zones including an indicator reagent zone and is capable of transporting the developing solution by capillary action sequentially through the reagent zones, and wherein the indicator reagent zone includes a reagent capable, directly or indirectly, of immobilizing an enzyme-labelled reagent in an amount dependent on the assay result, characterized in that the absorbent material includes a reagent that prevents a signal formation except where enzyme-labelled reagent is immobilized at the indicator reagent zone. The absorbent material is suitably in the form of an elongate strip provided with transverse reagent zones. The device is useful for performing immunoassays, including immunometric assays and dual analyte assays.

Abstract: Methods of maintaining or providing sterile or specific pathogen free environments are disclosed. The methods involve the use of semi-permeable membrane materials between the sterile or pathogen free environment and surrounding potentially hostile environments.

Abstract: Disclosed are a composition and method for determining the levels of specific immune responsiveness to a glycoprotein in an individual being treated therewith by (i) contacting a body fluid sample obtained from the individual prior to glycoprotein treatment with glycoprotein that has been modified to have an oxidized carbohydrate portion; (ii) contacting a body fluid sample obtained from the individual subsequent to glycoprotein treatment with the glycoprotein that has been modified to have an oxidized carbohydrate portion; and (iii) observing the degree of difference in the specific immune response to the oxidized glycoprotein in the pre- and post-treatment samples.

Abstract: A method is provided for enhancing the stability and increasing the activity of polymer particle reagents by incubating a polymer particle which has been covalently bound to a compound of biological interest, such as a drug, with a modifying agent that will place a negative charge on the surface of the polymer particle and reduce residual functional groups. The modifying agent is preferably an anionic nucleophile selected from the group consisting of .beta.- mercaptoacetic acid and the thiosulfates.

Abstract: The present invention discloses novel immunogens, antibodies prepared from such immunogens, and labeled reagents useful in immunoassays for the detection and quantification of testosterone in a test sample. Also disclosed are immunoassays using these reagents and methods for synthesizing these reagents. The immunoassays are preferably microparticle enzyme immunoassays (MEIAs) and fluorescence polarization immunoassays (FPIAs). Further disclosed are novel starting materials for making the above novel immunogens and labeled reagents. Methods for making the novel immunogens and labeled reagents from the novel starting materials are also disclosed.

Abstract: A novel technique is disclosed for the prevention of false positive reactions in immunological testing which are caused by interference of C.sub.1 and C.sub.1q. The method is based on heating a sample of a body fluid at a temperature of 59.degree.-64.degree. C. in the presence of a particular neutral salt. A method for screening for rheumatoid factor is also disclosed.

Abstract: An in vitro immunoassay to detect and quantitate soluble crosslinked and non-crosslinked DesAABB fibrin polymers in a sample from a subject. The assay can be used to support a diagnosis of, to evaluate, and to monitor, in a mammalian subject, a thrombotic event, including, but not limited to, myocardial infarction, pulmonary embolism, stroke and deep vein thrombosis.

Abstract: Compounds having detergent properties are disclosed. When a modifying reagent is brought into contact with these compounds, the detergent properties are decreased. These compounds are useful, for example, as solubilizing agents for microbial antigens and/or antibodies and for reversibly wetting hydrophobic surfaces. Accordingly, methods are disclosed for increasing the hydrophilic properties of a material, such as a microbial antigen and/or antibody, the methods generally comprising the steps of contacting the material with the compound having detergent properties and a modifiable group, and modifying the compound with a modifying reagent. Kits are also disclosed for use in accordance with this methodology.

Abstract: The present invention provides methods for determining the concentration of analytes in liquid samples in which the amount of binding agent having binding sites specific for a given analyte in the liquid sample is immobilized in a test zone on a solid support, the binding agent being divided into an array of spatially separated locations in the test zone. The concentration of the analyte is obtained by back-titrating the occupied binding agent with a developing agent having a marker and integrating the signal from each location in the array. The present invention also provides a method for determining a value representative of a fraction of binding sites of the binding agent which are occupied by the analyte, comprising immobilizing the specific binding agent on a solid support, wherein the specific binding agent used for the fractional occupancy is present in an amount less than 0.

Abstract: Ligands that interact with a target can be more easily identified if false positive interactions (either specific or non-specific) from the detecting system are differentiated from the target-specific interaction. An improved method of identifying peptides which bind with a target protein is presented. The steps are: binding a random library of peptides to a support material, allowing detection reagents to contact the peptides and the support material then identifying these interactions, then allowing the target protein to selectively bind to the peptides, allowing detection reagents to contact the bound target protein, and characterizing the peptide bound to the identified support material. Interaction of a ligand or the support material with the detection reagents will cause a distinct color change which distinguishes those ligands which selectively bind to target protein. The characterized peptide can then be used in affinity purification of the target protein.

Abstract: The present invention provides a method of improving the sensitivity and accuracy of a lead assay. The method enhances the recovery of lead during isolation of the lead from interfering compounds by maintaining the lead in a sample solution and making the recovered lead available for detection by the assay. An enhancing reagent complexes with the lead isolated in the sample solution. The enhancer includes a chelator having a lead equilibrium binding constant in the range of about 4 log K to about 13 log K. A kit for performing such a lead assay is also provided.

Abstract: N.sup..epsilon. -carboxymethyllysine, a carboxymethylated protein in which at least part of the side-chain amino groups of amino acid residues constituting the protein is carboxymethylated, or a carboxymethylated peptide in which at least part of the side-chain amino groups of amino acid residues constituting the peptide is carboxymethylated can be used as a marker for diabetes mellitus or diabetes mellitus complications.

Abstract: The present invention provides devices and methods for the detection of creatinine in biological fluids using lateral flow methodologies. The invention is particularly useful in providing one step creatinine assays for correcting urinary steroid hormone assays.

Abstract: The invention relates to a method for determining the extent of sepsis and/or an infection in a human or animal patient by detecting the amount of an antigen indicative of such infection. The amount of the antigen is detected in a patient blood derived test sample containing blood cell fractions.

Abstract: The invention concerns the use of a composition which is composed of several different antibodies or/and antibody fragments which serves as a reagent to eliminate rheumatoid factor interference in an immunochemical method.

Abstract: A method and device is described for direct removal of heparin from whole blood during extracorporeal therapy. In clinical situations where the blood is heparinized to prevent clotting in the extracorporeal circuit containing e.g., a hemodialysis cartridge or a heart-lung machine, it would be desirable to eliminate the systemic heparinization of the patients. The extracorporeal circuit contains a device having antithrombin III immobilized and inserted between the outlet port of the primary extracorporeal device, such as the hemodialysis cartridge or heart-lung machine and the patient. When the heparinized blood comes in contact with the immobilized antithrombin III, heparin is removed and the reperfused blood is substantially free of heparin. The advantage of the system lies in the fact that heparin is removed from the blood rather than neutralized. This helps overcome the adverse reactions and side effects associated with the use of heparin anticoagulation.

Abstract: The invention provides a method for enriching a sample in tumor cells and determining the DNA ploidy and proliferative index of the tumor cells present in the sample. The method uses a pan-leukocyte monoclonal antibody conjugated to a separable substrate such as magnetic microspheres to conjugate and remove leukocytes from the sample. A fluorescently labelled monoclonal antibody specific to a tumor associated antigen or an antigen arising from the presence of the tumor and a DNA staining reagent are added to the leukocyte depleted sample. The sample is then analyzed by dual color flow cytometry to determine cells having an abnormal DNA content by gating on fluorescently labelled cells.

Abstract: The invention provides a method of assessment of carbohydrate-deficient transferrin in a transferrin containing body fluid, said method comprising the steps of:i) obtaining a transferrin containing liquid sample of or derived from a said fluid;ii) contacting said sample with a source of iron ions;iii) subsequently contacting said sample with an anionic ion exchange resin at a pH such as to cause carbohydrate-deficient transferrin to be retained by said resin;iv) subsequently contacting said resin with an eluant serving to release carbohydrate-deficient transferrin into the eluate from said resin;v) collecting a volume of said eluate substantially free from tetra- and penta-sialo transferrin; andvi) assessing the transferrin variant content in said volume of eluate.By including at least a proportion of the trisialotransferrin in the eluate, it is possible to use relatively simple assessment techniques such as turbidimetry in the assay.

Abstract: Assays and antibodies are disclosed for the detection and quantitation of cardiac specific 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 and T and the binary and ternary complexes may be related to the metabolic state of the heart. More specifically, immunoassays for determining the presence or amount of free, binary and ternary complexes of troponin I and T are claimed.

Abstract: The invention provides a fragment of C1q which is characterized in that a plurality of such fragments selectively binds immune complexes or aggregated immunoglobulins in the presence of monomeric immunoglobulin. The invention also provides a synthetic peptide comprising the sequence: ##STR1## or variants thereof capable of binding immunoglobulin. Like the C1q fragment, a plurality of the peptides can selectively bind immune complexes or aggregated immunoglobulins in the presence of monomeric immunoglobulin. As a result of this property, the fragments and peptides are well-adapted for removing immune complexes and aggregated immunoglobulins from fluids containing monomeric immunoglobulin, and for detecting or quantitating immune complexes in such fluids. The invention also provides a binding material for removing immune complexes or aggregated immunoglobulins from a fluid.

Abstract: The assay reagent and kit of the present invention suppress non-specific binding of a labeled substance onto a solid phase, and can assay one or more species of antibodies or one or more species of antigens by means of a single reagent in a simple manner. The assay method involves reacting immunological ligands in a test sample with the assay reagent which contains a combination of components (A) and (B), thereby forming complexes, which complexes are captured onto the independently and separately present Solid phases (C), to assay the label contained in the complexes.

Abstract: Solid supports for solid phase immunoassays comprise polyethyleneimine-coated negatively charged polymeric materials. The combination of negatively-charged polymer support and polyethyleneimine coating serves to eliminate nonspecific adsorption of biological molecules such as high molecular weight kininogen, which have affinity for solid surfaces and which interfere with immunoassays. The negatively charged polymeric support to which the polyethyleneimine coating is applied may advantageously comprise, for example, a carboxylate-modified styrene microparticle. Antigens or antibodies are coupled by covalent coupling agents to the polyethyleneimine coating to form solid phase immunoreagents. Immunoassays for biological molecules such as high and low molecular weight kininogen are provided.

Abstract: Progressive degradation of the osteocalcin present in the blood, serum or plasma samples obtained from a patient is observed in such samples and leads to falsification of the osteocalcin values obtained in a subsequent determination. It has been found that this osteocalcin degradation can be prevented by adding an amount of divalent metal ions, in particularly calcium ions, sufficient to prevent osteocalcin degradation to the sample immediately after it has been obtained or while it is being obtained.

Abstract: A method is provided for preparing lipoprotein (a) from a volume of biological fluid that is substantially free of lipoproteins of another class. The method involves an ultracentrifugation step in which at least one fraction is recovered that contains Lp(a). This material is then reacted with immobilized ligand to remove non-Lp(a) interfering substances from the fraction, the Lp(a) remaining unbound. The non-reacted Lp(a) is subsequently obtained in a form that is suitable for use in the analysis of any of protein concentration, protein isoform determination or cholesterol assays. A method of identifying and measuring an amount of one or more isoforms of Lp(a) is further provided.

Abstract: Modified apo-peroxidases have been found useful in analytical methods to remove the bias caused by the presence of hemoglobin in the test specimens. While apo-peroxidases are known to remove serum interferents, they fail to overcome the hemoglobin bias, particularly when dry coated analytical elements are used in the assays. Blocking the nitrogen atoms of the imidzolyl groups of the histidine amino acids of the apo-peroxidase prevents catalytic activity of the protein in the presence of hemoglobin.

Abstract: A method and apparatus for immunoassays utilizes an improved collection technique of fluorescence induced emissions at the solid phase/liquid phase interface from surface plasmon resonance sensing devices. In a preferred embodiment, a solid phase substrate is coated with a thin film of a conducting material on which a first specific binding partner is directly or indirectly immobilized. The coated solid phase substrate is incubated with a liquid component comprised of a biological sample containing a specific ligand or analyte and a fluorescent labeled second specific binding partner in the case of immunometric assays, or a fluorescent labeled ligand or analog thereof in the case of competitive assays.

Abstract: A method for quantitatively determining cholesterol in high density lipoproteins, in which, prior to the determination of cholesterol by an enzymatic method, a surfactant and a substance which forms a complex with lipoproteins other than high density lipoproteins are added to a sample containing lipoproteins.The method does not require any pretreatments such as centrifugal separation. With a simple operation, cholesterol in HDLs can be measured effectively. Also, this method can be adopted in a variety of automated analyzers, and thus is very useful in the field of clinical assays.

Abstract: The present invention relates to a method of quantitative determination of sodium ions in a sample using .beta.-galactosidase, wherein the .beta.-galactosidase reaction is conducted in the presence of at least one chelating agent selected from of 1,2-cyclohexanediamine-N,N,N',N'-tetraacetic acid, ethylenediamine-tetraacetic acid, triethylenetetranine-hexaacetic acid, diethylenetriamine-N,N,N',N",N"-pentaacetic acid, 1,3 -diminopropan-2-ol-N,N',N'-tetraacetic acid, ethylenediamine-N,N'-dipropionic acid dihydrochloride, ethylenediamine-tetrakis(methylenesulfonic acid), iminodiacetic acid, hydroxyimino-diacetic acid and nitrilotriacetic acid.The method of quantitative determination of sodium ions of the present invention is useful in clinical examinations and the accuracy in the measurement by the method is high.

Abstract: In accordance with the present invention, there are provided novel methods and reagents for reducing background binding in antibody preparations having an unwanted affinity for intracellular protein(s). The invention method comprises treating an antibody preparation with permeabilized cells, then separating the antibody preparation from the permeabilized cells. The reagents are useful for reducing the level of background binding in antibody preparations. Also provided are antibody preparations that are substantially free of background binding.

Abstract: The invention is directed to methods and kits for detecting and measuring the presence or absence of perinuclear anti-neutrophil cytoplasmic autoantibody of ulcerative colitis or primary sclerosing cholangitis. The methods and kits of the present invention provide safe and reliable means for diagnosing ulcerative colitis and primary sclerosing cholangitis. The antigens reactive with perinuclear anti-neutrophil cytoplasmic autoantibody of ulcerative colitis and primary sclerosing cholangitis are also provided.

Abstract: A total reflection cell has a total reflection prism on at least one surface thereof. A mixed solution containing a gold colloid labelled antibody which is adsorbed on a gold colloid is stored in the total reflection cell, and a sample solution containing an antigen causing antigen-antibody reaction with the antibody is added thereto for forming a gold colloid labelled immune complex. A measuring beam is introduced into the total reflection prism from an incident optical system at an angle .theta. of incidence causing total reflection and an outgoing beam from the total reflection prism is received by a measuring optical system, thereby measuring absorption by the gold colloid labelled immune complex and carrying out qualification and determination of the antigen.

Abstract: An improved method for determining binding compounds from a mixture of similar compounds, particularly a phage peptide library, is provided. The method comprises contacting a mixture of candidate compounds with a target molecule presented on two or more different substrates.

Abstract: The present invention relates to a binary assay method capable of providing wide dynamic range, a high degree of precision and rapid processing times in which analyte is reacted successively with two independently determinable forms of solid-supported binding partner and also with a labelled ligand having affinity for the analyte or binding partner. The analyte concentration is determined from signals deriving from the resulting two forms of analyte-binding partner-labelled ligand systems by reference to a double standard calibration curve. A kit for use in the method is also disclosed.

Abstract: A method of assaying bone collagen breakdown levels in a human subject useful to screen for the presence of bone resorption disorders. Also disclosed is a method for monitoring the progression and/or treatment response of a cancer condition which involves or has the potential to progress to a metastatic condition which involves abnormalities in bone resorption rates.