Abstract: A "one step" device for the detection of analytein clinical assays is disclosed. A visible label comprising a visible moiety and a ligand that binds to or competes with analyte is contained in a fluid conducting membrane or prefilter; and the analyte and associated ligand are conducted by the flow of sample into a detection zone. The detection zone contains a capture reagent that binds to label or to analyte bound to label.

Abstract: A method and apparatus for carrying out affinity purification of a ligate. The method comprising, (a) providing a ligate containing liquid to a first side of at least one porous hollow fiber membrane with a ligand immobilized thereto that binds and separates the ligate from the liquid, (b) withdrawing a first portion of the liquid from the first side of the porous hollow fiber membrane, (c) recirculating the first portion of liquid to the first side of the porous hollow fiber membrane, (d) repeating steps (a) to (c) until a majority of the liquid has flowed through the porous hollow fiber membrane, and (e) providing an elution solution to one side of the porous hollow fiber membrane under a pressure sufficient to cause the elution solution to flow into and through the membrane to effect disassociation of any ligate-ligand bonds wherein any ligate bound to the ligand is eluted with the elution solution.

Abstract: A method for increasing the binding activity of specific binding members bound to a solid phase material, e.g., a particle, that has been sterically stabilized. This increase in binding activity is brought about by degrading a steric stabilizer on the surface of the solid phase material. The method involves both immobilizing a specific binding member on the surface of a solid phase material and degrading a steric stabilizer on the surface of that solid phase material. In the preferred embodiment, the method involves the immobilization of a specific binding member on the surface of the sterically stabilized solid phase material, with subsequent degradation of the steric stabilizer.

Abstract: A method of immunologically assaying intact human osteocalcin in a human assay sample is provided by using an antibody having an epitope in a region of an amino acid sequence 1 to 20 on the N-terminal side of human osteocalcinand an antibody having an epitope in a region of an amino acid sequence 36 to 49 on the C-terminal side of human osteocalcin. Furthermore, a method of immunologically assaying the total amount of human intact osteocalcin in a human assay sample is provided. A reagent and a kit therefor are also described, as well as a monclonal antibody and a polyclonal antibody used for the assay a process for producing these antibodies; and a process for use of these antibodies.

Abstract: Insoluble supports are prepared which possess high surface areas and efficiently dispersed isocyanate groups. These reactive supports are useful for covalently binding proteins which preferably are enzymes and provide catalysts for conducting organic reactions.

Abstract: There are provided a method and an apparatus for measuring an immunologically active material by physically or chemically immobilizing material immunologically active to a material to be measured of a specimen to the dehydrated solid fine particles, providing a desirable dispersion comprising said immunologically active material immobilized to said solid fine particles and said specimen in a liquid medium, reacting them to cause a reaction mixture in an agglutinated state and optically measuring said agglutinated state of the reaction mixture to thereby quantitatively determine the content of said material to be measured with an improved accuracy.

Abstract: Polyolefin particles are chemically modified by oxidation to provide a large surface area and high loading. The particles result in low back pressure in column systems, and are economical to manufacture. The particles are useful as supports in a wide range of applications including general organic as well as biopolymer synthesis, library methods, purification processes and enzyme mediated processes. In a preferred embodiment, polyethylene or polypropylene particles are oxidized in a solution containing trifluoroacetic acid or trifluoromethane sulfonic acid, chromium trioxide and sulfuric acid to provide the particles with a chemically reactive irregular surface and open channels that extend below the surface and up to essentially the length of the radius of the particles resulting in increased surface area and decreased density. The particles have pendant functional groups produced by the oxidation and/or by subsequent chemical reaction.

Abstract: The present invention provides a method of identifying immunocompromised individuals who are at high risk for development of cryptococcosis, comprising the step of measuring the levels of IgG antibody to glucuronoxylomannan in the sera of said individuals. Also provided are various methods of detecting antibody to glucuronoxylomannan in sera and a kit for assaying the presence of antibody to glucuronoxylomannan in the sera of immunocompromised individuals.

Abstract: An assay system for detecting the binding of a mobile reactant to an immobilized reactant on an assay plate. The assay plate includes a substrate having an assay spot deposited thereon. The assay spot includes the immobilized reactant. In the present invention, a carrier dye is included in the assay spot, the amount of the carrier dye indicating the amount of the immobilized reactant that is present in the assay spot. In the preferred embodiment of the present invention, the assay spot is generated by depositing liquid on the assay plate at a location corresponding to the assay spot. The carrier dye is dissolved in the liquid and remains after the liquid evaporates. The amount of carrier dye in each spot may be measured spectroscopically and provides a means for identifying defective assay plates.

Abstract: The invention relates to activated support materials which comprise epoxy-substituted poly(meth)acrylamides, characterized in that:a) the base support contains aliphatic hydroxyl groups,b) the covalently bonded polymers are bonded to the support via a terminal monomer unit,c) the polymers contain monomer units of the formula I,d) the monomer units are linearly linked, ##STR1## in which R.sup.1, R.sup.2 and R.sup.3 independently of one another are H or CH.sub.3,R.sup.4 is H or alkyl having 1-5 carbon atomsandn is an integer between 1 and 5 and to the use thereof.

Abstract: There is disclosed an apparatus for the determination of a trace amount of an analyte substance or organism. A substrate solution is brought into contact with a small diameter tube which has captured, at least on its inner surface, a trace amount of an analyte substance or organism. The trace amount has the ability to change the pH of the substrate solution.

Abstract: The present invention provides a new analytical method that an antigen, an antibody or a nucleic acid can be assayed at a high sensitivity in the immunoassay.The present invention provides a method for assaying a substance which comprises the steps of (a) reacting an antigen, an antibody or a nucleic acid which is the substance to be measured with an antibody, an antigen or a nucleic acid to which a polyacridinium compound is bound and which can be bound to the substance to be measured, (b) carrying out a luminescence treatment, and (c) measuring the luminescent intensity of the reaction solution.

Abstract: A multi-purpose on-line or field-portable system and method for monitoring the presence and concentration of selected antigen-antibody reactions singly or in combination that result from the presence of specific microorganisms or free antigens present or suspended in aqueous solutions, during a given time period. The detection system comprises a detection column and two sensors mounted around the detection column. Each sensor consists of an electromagnetic radiation source and an appropriate detector for the electromagnetic radiation. The reacted analyte tends to accumulate at the sensor located at the bottom detection column. The lower sensor continually nulls against the upper sensor to subtract any optical effects due to non-reactants in the aqueous process or environmental stream. The response from the detector sensors drive an electric circuit, which provides an output signal. In the on-line automatic version, the signal can drive elements of a process system by switching automated valves.

Abstract: Directly dividing the contents of each sub-pool into the sub-pools for the next step in the synthetic scheme for producing a combinatorial library reduces the standard deviation, .sigma., relative to the standard deviation of the split synthesis method for producing such libraries.

Abstract: The instant invention provides a highly fluorescent conjugate which is useful in specific binding assays, and which comprises a specific binding member bound to a fluorescent polymer. The fluorescent polymer comprises a backbone polymer having multiple signal generating groups immobilized thereon and, optionally, cyclodextrin moieties in association with the polymer. Also provided is a novel process for creating 6-cyclodextrin monoaldehyde.

Abstract: Red blood cells are removed from whole blood or a fraction thereof by contacting whole blood with a combination of an agglutinating agent and nucleating particles to form clusters of red blood cells. High molecular weight polyethylene glycol may be added further to enhance agglutination. The clusters of red blood cells are much larger than the size of individual red blood cells, so that the clusters can easily be filtered through a porous medium. The plasma which is substantially free of red blood cells is further passed through a filter that optionally contains an additional agglutinating agent. Flow-delay means may be provided to return the fluid sample in contact with a regard for a predetermined time.

Abstract: This invention relates to a cell enumeration immunoassay which uses a calibrated standard, i.e., a substance which behaves like the sample under study and the concentration of which can be correlated to the concentration of the cells. This immunoassay is an efficient alternative to flow cytometry.

Abstract: A biosensor detector method for detecting biological targets, using specific binding, or hybridization techniques coupled with enzymatic amplification and the mass sensing capability of a piezoelectric oscillator. An optical biosensor is also contemplated.

Abstract: An analytical system for performing an immunoassay is provided wherein the system contains an element that uses capillary action to draw a sample into a reaction chamber charged with reagent containing an immobilized antibody or immobilized antigen, where reaction between the liquid sample and the reagent is monitored, and optionally contains a method for controlling the moment that transport of the sample from the sample well to the reaction chamber is initiated.

Abstract: The invention pertains to dipstick immunoassay devices. The device comprises a base member and a single, combined sample contact zone and test zone, wherein the test zone incorporates the use of symbols to detect analytes in a sample of biological fluid. A first immunological component, an anti-immunoglobulin capable of binding to an enzyme-labeled antibody, is immobilized in a control indicia portion. A second immunological component, capable of specifically binding to a target analyte which is bound to the enzyme-labeled antibody to form a sandwich complex, is immobilized in a test indicia portion. The enzyme-labeled antibody produces a visual color differential between a control indicia portion and a non-indicia portion in the test zone upon contact with a substrate.

Abstract: A dry detection reagent for detecting an immunologically active substance is prepared containing solid fine polymer particles immobilizing a substance immunologically active for the substance detected. The polymer particles are prepared by one of the following methods: a) ternary copolymerization of a monomer having carboxyl groups, a monomer having amino groups and styrene or its derivatives; b) binary copolymerization of a monomer having carboxyl groups and styrene or its derivatives, and reacting part of the carboxyl groups with a bifunctional amine such as ethylene diamine to provide amino groups; or c) copolymerizing styrene or its derivatives with a monomer having amide groups, and converting part of the amide groups to amino groups and carboxyl groups with the Hofmann reaction, or hydrolyzing part of the amide groups to form carboxyl groups.

Abstract: Red blood cells are removed from whole blood or a fraction thereof by contacting whole blood with a combination of an agglutinating agent and nucleating particles to form clusters of red blood cells. High molecular weight polyethylene glycol may be added further to enhance agglutination. The clusters of red blood cells are much larger than the size of individual red blood cells, so that the clusters can easily be filtered through a porous medium. The plasma which is substantially free of red blood cells is further passed through a filter that optionally contains an additional agglutinating agent.

Abstract: An assay device for detecting the presence of analytes in an unknown sample includes a reaction system wherein resilient storage reservoirs containing reagents are fluidly connected to a track containing the sample. An actuation mechanism forces the reagent from each of the reservoirs into the track where the reagents mix together and with the sample. The mechanism produces a first flow rate and the mechanism is operable to reverse the pressure applied to the reservoirs to reverse the direction of flow of the fluids in the track for a predetermined period of time after which the flow is again reversed. The mechanism then reduces the force applied to allow a second flow rate less than the first flow rate so that reaction can occur whereby a determination may be made as to whether the target analyte is present in the sample.

Abstract: Monoclonal antibodies have been prepared which are of the IgG isotype and are highly specific for horseradish peroxidase. One group of antibodies inhibits at least about 95% of the normal activity of horseradish peroxidase when bound to the enzyme. A second group of antibodies inhibits less than about 20% of the enzymatic activity when bound to the enzyme, but prevents the binding of the antibodies from the first group. The antibodies in either group can be conjugated to specific binding ligands such as drugs or hormones.

Abstract: A method for detecting and/or assaying viral compounds, wherein the viral compounds are bound with .beta.2'-glycoprotein I which is a glycoprotein having a molecular weight of 50,000.+-.3,000 daltons from the method for producing albumin obtained as a blood plasma by-product according to FR-A 2 690 444. Viral compound separation may be simplified by binding the .beta.2'-glycoprotein I to a preferably solid support or by binding the viral compounds to a support and exposing same with the .beta.2'-glycoprotein I conjugated with a marker. A solid support to which at least the .beta.2'-glycoprotein I is bound, is also disclosed.

Abstract: Specific peptide fragment from the feline immunodeficiency virus (FIV), and its use as a diagnostic reagent.The said peptide fragment, derived from the Env protein of the Wo strain of the feline immunodeficiency virus (FIV) (peptide P253), corresponds to positions 693-709 of the said Env protein and exhibits the following sequence:Leu-Gly-X-Asn-Gln-Asn-Gln-Phe-Phe-X-Lys-Val-Pro-Ser-Ala-, in which X represents a cysteine or a serine, as follows:Leu-Gly-Cys-Asn-Gln-Asn-Gln-Phe-Phe-Cys-Lys-Val-Pro-Ser-Ala (SEQ ID NO: 1),Leu-Gly-Ser-Asn-Gln-Asn-Gln-Phe-Phe-Ser-Lys-Val-Pro-Ser-Ala (SEQ ID NO: 2),Leu-Gly-Cys-Asn-Gln-Asn-Gln-Phe-Phe-Ser-Lys-Val-Pro-Ser-Ala (SEQ ID NO: 3),Leu-Gly-Ser-Asn-Gln-Asn-Gln-Phe-Phe-Cys-Lys-Val-Pro-Ser-Ala (SEQ ID NO: 4).

Abstract: Sets and libraries of sets of polypeptides that are related in sequence to melittin are disclosed that have antimicrobial, hemolytic and hydrolyrically catalytic activities, as are processes for making and using the same. A contemplated set is a mixture of equimolar amounts of a polypeptide of SEQ ID NO:2, and more preferably SEQ ID NO:3.

Abstract: The present invention is a sample preparation system and method that can be used with all types of analyte materials, that produces homogeneously deposited crystals across a sample surface, and that lends itself to automation. In this system and method, analyte crystallization is caused by lyophilization. A homogeneous analyte/solvent mixture is placed on a sample surface. The mixture is frozen, then the solvent is sublimated through the application of a vacuum. A homogenous distribution of analyte crystals across the sample surface results.

Abstract: The present invention provides a method for determining the amount of HIV-1 p24 antigen or anti-HIV-1 p24 antibody present in a suitable bodily fluid sample from an HIV-1-infected subject. This invention also provides a kit for determining the amount of HIV-1 p24 antigen or anti-HIV-1 p24 antibody present in an HIV-1-infected subject.

Abstract: Gelatin and aminodextran coated polymer core particles useful in immunoassays and methods of making the same are disclosed. The preparation of aminodextrans having varying amounts of amine groups is also described, as is a method of crosslinking gelatin and aminodextran without the use of a stabilizer.

Abstract: A novel method of detecting antibodies to thrombomodulin in plasma or serum as an indication of an individual's propensity to thrombosis or inflammation is disclosed. A method for identifiying patients at risks of thrombosis or glomerular nephritis by monitoring autoantibodies to truncated soluble thrombomodulin is revealed. A preferred method is an ELISA assay to detect antibodies to thrombomodulin.

Abstract: Disclosed are devices for detecting the presence of a preselected analyte in a fluid sample. The devices comprise a substrate microfabricated to define a sample inlet port, and a mesoscale flow system that includes a sample flow channel extending from the inlet port. The mesoscale flow system further includes an analyte detection region in fluid communication with the flow channel comprised of a binding moiety for specifically binding the analyte. The detection region is constructed with a mesoscale dimension sufficiently small to enhance binding of the binding moiety and the analyte. The binding moiety may be immobilized in the detection region. The mesoscale detection systems of the invention may be used in a wide range of applications, including the detection of cells or macromolecules, or for monitoring reactions or cell culture growth.

Abstract: New and useful methods of producing stabilized enzyme antibody conjugates are disclosed which are particularly useful in forming multi-layer immunoassay test devices. In particular, the invention concerns the formation of a manganese ion and enzyme-antibody conjugate in aqueous solution and drying the solution to produce a dry stabilized enzyme-antibody conjugate. Further, this stabilized enzyme-antibody conjugate can be formed on a continuous web and dried in a heat tunnel. This continuous manufacturing process allows for the more efficient production of multi-layer test strips.

Abstract: A patient's health may be diagnosed by centrifuging blood samples in a transparent tube, which tube contains one or more bodies or groups of bodies such as floats, inserts, liposomes, or plastic beads of different densities. Each density-defined body carries analyte-capture binding materials such as antigens or antibodies, which are specific to an epitope, or other specific high affinity binding site on a target analyte which target analyte may be in the blood or other sample being tested; and the level of which analyte is indicative of the patient's health. At least one labeled binding material which is also specific to an epitope, or other specific high affinity binding site on the target analyte is added to the sample so as to form labeled binding material/analyte/body complexes in the sample.

Abstract: Stabilized microspherical particles having hydrophobic liquid cores prepared as oil-in-water microemulsions. The particles are stabilized by a surface layer comprising an amphiphilic compound and may be functionalized to allow covalent coupling of a ligand to the surface of the particle. When used as tracers in assays, a water insoluble dye may be incorporated in the core liquid of the microparticles.

Abstract: An aqueous colloidal dispersion for diagnostic or immunodiagnostic tests, comprising non-polymer nuclei surrounded by a hydrophilic copolymer that contains functional groups, a method for the detection of a specifically binding substance or immunochemically active component in a test fluid, and test kit containing the aqueous colloidal dispersion.

Abstract: A method for the measurement of serum bile acids by ELISA comprising: preparing a bile acid active ester, reacting the ester with a bovine serum albumin solution, dialyzing, and immunizing a mammal other than human being with the dialyzate thus obtained as an antigen to thereby give an anti-bile acid antibody; reacting said active ester with an enzyme to thereby prepare an enzyme-labeled bile acid as an enzyme-labeled antigen; to a secondary antibody-coated plate, adding a dilution of the serum to be assayed, an anti-bile acid antibody solution and an enzyme-labeled antigen solution and reacting these substances followed by the addition of a substrate and the reaction therewith; measuring the absorbance of the reaction mixture and determining the concentration of each bile acid on the basis of the standard curve measured simultaneously; and referring the sum of the concentrations of these bile acids to the total bile acid concentration; and a method for the diagnosis of a liver disease comprising: calculating

Abstract: A new recombinant form of the plasmid-encoded protein pgp3 from C. trachomatis, serotype D, was purified by ion exchange column chromatography and shown to be suitable for quantitative immunoassy on clinical samples in an ELISA format.

Abstract: A method for forming an optical device for detecting the presence or amount of an analyte of interest comprising a substrate which supports an optically active layer, an attachment layer provided on the optically active layer, and a receptive layer specific the analyte provided on the attachment layer. The method comprises forming the optically active layer with a chosen refractive index on the substrate by curing the optically active layer on the substrate at a controlled temperature or for a controlled length of time and subsequently providing the attachment and receptive layers on the optically active layer.

Abstract: Permanently substituted oxyfluorinated surfaces can be formed on non-fluorinated substrates having a fluorinated surface or fluorocarbon coating applied by gas phase surface fluorination or plasma deposition. The oxyfluorinated surfaces can be refunctionalized by bonding organosilanes, isothiocyanate-containing fluorescent compounds and proteins, such as enzymes, antibodies and peptides directly to such surfaces. Surfaces refunctionalized with such protein based groups are useful in the fabrication of biological sensors, devices for separation of cell lines, filtration applications for selective binding of antigens. Masking techniques can be employed in forming a predetermined pattern of covered and exposed surfaces, for example, prior to oxyfluorination.

Abstract: HIV-1 peptides having at least one point mutation between position 593 and 611 of the HIV-1 gp160 amino acid sequence. The point mutation either is at position 604 or 610, or both positions. Immunoassays which utilize these peptides are provided, as well as, diagnostic test kits which contain these peptides.

Abstract: An immunoassay process is provided for the detection of a target antigen in a fluid sample where an admixture of the target antigen and a labeled capture reagent against the target antigen is contacted by a solid polymeric carrier having bound to a portion of its surface a capture reagent against the target antigen and visual determination of a change in color of the bound labeled reaction product on the surface of the solid carrier member serves to readily detect the presence of the target antigen.

Abstract: A method of assaying bone collagen degradation activity in a human subject. In the method, a human urine sample is reacted with an antibody which (i) is capable of reacting immunospecifically with pyridinium crosslinks selected from the group consisting of native free pyridinoline and native free deoxypyridinoline, and (ii) has a ratio of reactivity toward the selected pyridinium crosslinks and urinary pyridinium peptides larger than 1,000 daltons in molecular weight, of greater than about 5:1. An immunocomplex forms between the antibody reagent and the selected pyridinium crosslinks which are present in the sample, and the amount of immunocomplex is measured. Also disclosed are antibody reagents and kits which can be used in the method.

Abstract: A rapid, simple method for preparing beads for calibrating flow cytometers which contain a known number of fluorophores per bead is presented. Briefly, the invention utilizes beads coated with a stable complex of a fluorophore and an enzyme. the enzymatic activity of a known number of beads gives an accurate measure of fluorophore density on those beads.

Abstract: Stabilized microspherical particles having hydrophobic liquid cores prepared as oil-in-water microemulsions. The particles are stabilized by a surface layer comprising an amphiphilic compound and may be functionalized to allow covalent coupling of a ligand to the surface of the particle. When used as tracers in assays, a water insoluble dye may be incorporated in the core liquid of the microparticles.

Abstract: An aqueous composition suitable for use when performing immunological procedures. The composition includes at least one biological buffer, dithiothreitol (alternatively referred to herein as "DTT"), and ethylene glycol. The composition can also include at least one biological detergent, at least one source of positive and negative counterions, e.g., salt, and at least one viscosity modifier, e.g., sugar. The buffer also can include at least one preservative, such as sodium azide. The pH of the composition preferably ranges from about 6.4 to 7.2. A kit containing the composition is also disclosed.

Abstract: Novel tethered hapten intermediates and related conjugates based on 3-phenyl-1-adamantaneacetic acid, as well as methods for making and using such conjugates. Haptens based on the above core structure may be substituted at any position on the phenyl ring, especially at the para position. Using tethered intermediates, immunogens, tracers, solid supports and labeled oligonucleotides are all described; as are methods for using the intermediates to prepare the conjugates, methods of using the conjugates to make and purify anitbodies, as assay tracers, and in nucleic acid hybridization assays. Kits containing haptenated oligonucleotides and anti-hapten conjugates are also described.

Abstract: Fluorescent latices, well suited for a wide variety of analytical techniques, comprise polymer particles having at least one hydrophobic fluorochrome encapsulated therein, notably at least one condensed polyaromatic compound or derivative thereof, or a tetraphenylporphine and/or organometallic complex thereof, and have a detection threshold for fluorescence which is less than or equal to 10.sup.-12 mol of particles per liter of latex.

Abstract: The present invention provides a reagent for assaying an endotoxin which comprises limulus amoebocyte lysate and an antibody to (1 .fwdarw. 3)-.beta.-D-glucan sensitive factor or a reagent which comprises a lysate substantially free from (1 .fwdarw. 3)-.beta.-D-glucan sensitive factor. The reagent of the present invention makes it possible to assay an endotoxin originating from gram-negative bacteria contained in a biological sample such as blood, urine and cerebrospinal fluid at an extremely high sensitivity without being affected by (1 .fwdarw. 3)-.beta.-D-glucan.

Abstract: A self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridization, antibody/antigen reaction, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micromachining techniques. The device can electronically control the transport and attachment of specific binding entities to specific micro-locations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific microlocations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.