Abstract: Embodiments described herein include methods and assays for detecting an analyte in a sample using a plurality of control zone capture agents. Some embodiments include detection of multiple analytes in a sample utilizing a plurality of analyte binders and a control zone containing multiple control zone capture agents. In some embodiments, the multiple control zone capture agents capture a plurality of binders within one control zone. Test results are determined by comparison of the control zone signal to a test zone signal.

Abstract: Embodiments described herein include methods and assays for detecting an analyte in a sample using a plurality of control zone capture agents. Some embodiments include detection of multiple analytes in a sample utilizing a plurality of analyte binders and a control zone containing multiple control zone capture agents. In some embodiments, the multiple control zone capture agents capture a plurality of binders within one control zone. Test results are determined by comparison of the control zone signal to a test zone signal.

Abstract: Diagnostic devices and methods are provided. The diagnostic devices preferably comprise a test strip with a test area. The sample to be analyzed contacts the test area, which comprises a specific binding partner for the analyte of interest. Analyte, if present in the sample, binds to the immobilized binding partner in the test area and is subsequently contacted with a conjugate. The conjugate specifically binds the analyte and provides a visual indication of the presence of the analyte. The devices may be used for the diagnosis of particular diseases or disorders in a patient, such as HIV or hepatitis. They may also be used to determine if an individual is pregnant.

Abstract: The present invention provides a process for dissociating Fc-containing molecules from complexes of Protein A/Fc-containing molecules or mixtures containing Fc-containing molecules and Protein A. The association, e.g., by hydrophobic interactions, between the Fc-containing molecules and Protein A can be reduced or inhibited by raising the pH of dissociation. The pH of dissociation can be raised by addition of agents capable of inhibiting hydrophobic interactions, including buffers containing arginine and/or ethylene glycol, to the mixture, either prior to adding the mixture to the column chromatography substrate, after adding the mixture to the column chromatography substrate, or both prior to and after adding the mixture to the column chromatography substrate.

Abstract: Carriers hold remote-acting bodies which can be manipulated by a remote force, and also hold a micro-substance which is a target substance of an assay. The remote-acting bodies are manipulated in order to control the positions of the micro-substances, so as to execute assays for various target substances efficiently, at low cost, easily, and reliably. Various aspects of interest include the carriers which hold the micro-substances, a system suspending the carriers, an apparatus for manipulating the carriers, and a method of controlling the position of the carriers.

Abstract: Separation apparatus and method for separating magnetic and/or magnetically-labeled particles from a test medium. Test medium within a reaction chamber is caused to flow past a collecting surface, and a high-gradient magnetic field is applied to the surface to capture magnetically responsive particles in the test medium. The particles are deflected toward the collection surface by baffles, a spinner, or a sprayer, or are funneled past the surface by a plunger operable to be displaced into close proximity to the surface to provide a narrow flow path for the particle-laden test medium. The particles normally suspended in the medium are separated out of suspension by adhesion to the collection surface. The particles may be resuspended by removal of the surface from the high-gradient field, or removal of the high-gradient field from the surface. The collection surface is a thin-walled non-magnetic material having a plurality of magnetic pole faces positioned therearound.

Abstract: The invention provides improved methods of regenerating and using affinity chromatography material, in particular Protein A affinity chromatography resins.

Abstract: The invention relates to a method for detecting specific target-cells in a simple and time saving way, using paramagnetic particles, antibodies recognizing the Fc portions of target-cell associating antibodies and target-cell associating antibodies directed to specific antigen determinants in the target-cell membranes. Incubation of the cell suspension with a mild detergent and/or second set of antibodies or antibody fragments, prelabeled or not with fluorescent agents, metallocolloids, radioisotopes, biotincomplexes or certain enzymes allowing visualization, with dramatically increase the specificity of the method. The method can further be used for isolation of the target-cells by magnetic field application and kit for performing the method according to the invention is described.

Abstract: A method for purifying proteins by Protein A chromatography is described which comprises removing contaminants by washing the solid phase with various intermediate wash buffers.

Abstract: A method for purifying proteins by Protein A chromatography is described which comprises the steps of: (a) adsorbing the protein to Protein A immobilized on a solid phase comprising silica or glass; (b) removing contaminants bound to the solid phase by washing the solid phase with a hydrophobic electrolyte solvent; and (c) recovering the protein from the solid phase.

Abstract: Reagents and methods for the detection of Staphylococcus aureus are provided. The reagents contain an antibody that binds to a capsular polysaccharide of type 5 of Staphylococcus aureus, and can be used in methods for detection of oxacillin resistant Staphylococcus aureus that escapes detection by agglutination in the presence of fibrinogen and antibodies directed against protein A of Staphylococcus.

Abstract: Methods for rapid detection and/or semi-quantitation of anti-adenovirus antibody are disclosed. Anti-adenovirus antibody is detected using a device comprising a membrane with adsorbed antigen which specifically binds anti-adenovirus antibody and an absorbent pad which is contacted with the membrane. By consolidating detection reactions in a confined location and eliminating the need to manually remove input reagents, detection and semi-quantitation is achieved rapidly and conveniently. The invention also provides kits and devices for detection and/or semi-quantitation of anti-adenovirus antibodies.

Abstract: The present invention is directed to a single test system and method for determining the integrated glycemic condition of a subject by measuring the concentration of glucose and the level of protein-bound glucose in a subject's body fluid, such as whole blood. The glucose concentration is indicative of the subject's immediate glycemic condition, whereas the protein-bound glucose concentration is indicative of either intermediate or long-term glycemic condition. Optionally, other analytes indicative of glycemic condition, such as ketone bodies or fatty acid derivatives, can also be measured. The present invention also provides a method of diagnosing diabetes. The invention additionally provides a method for analyzing the concentration of fructosamine in less than or equal to five minutes without the use of a reaction accelerator.

Abstract: A method of identifying interface residues in a biomolecular complex are identified by selecting one molecule with residues to be identified, and the nonexchangeable hydrogens (protons) and not less than 70% of exchangeable hydrogens (protons) in the one molecule are exchanged to deuteriums, followed by positional identification of the exchangeable hydrogens (protons) which are located on this molecule within 10 angstrom (Å) from hydrogen (s) (proton(s)) in a neighboring biomolecule in the complex and are cross-saturated by cross-saturation phenomena through the interface in the complex. Using the inventive method, the contact interfaces of biomolecular complex such as protein—protein complexes can be identified more accurately and easily than traditional methods.

Abstract: The present invention is directed to novel methods for enhancing the ability to separate a species of interest from other different species present in a free solution mixture thereof which takes advantage of interactions that occur between soluble, small molecular weight ligands and the species of interest. The small molecular weight ligands employed in the present invention function to interact with a species of interest in a mixture of different species through either affinity, hydrophobic and/or ionic interactions, thereby altering the molecular weight, hydrodynamic volume and/or isoelectric point of the species of interest and rendering it separable from other component(s) in the mixture.

Abstract: A method is disclosed for producing a soluble conjugate vaccine, and preferably protein/polysaccharide conjugates. In this process, the polysaccharide is reacted with a reagent so as to provide a functional group on the polysaccharide molecule. Once the functional group is in place, the polysaccharide is reacted with a homobifunctional or heterobifunctional vinylsulfone to produce a vinylsulfone derivatized polysaccharide. Thereafter, the vinylsulfone derivatized polysaccharide is reacted with a protein to produce the conjugate. If desired, the protein may be derivatized with a functional group prior to the conjugation reaction step. In an alternative embodiment, the protein may be functionalized with a reactive group and then derivatized with the vinylsulfone group to produce a vinylsulfone derivatized protein. This protein may then be reacted with a polysaccharide to produce the conjugate. Optionally, the polysaccharide may be functionalized with a reactive group prior to the conjugation reaction.

Abstract: The invention relates to methods of determining the presence or amount of an analyte in a sample suspected of containing the analyte, said method comprising the steps of: (a) bringing together in an aqueous medium to form a mixture: (i) the sample; (ii) at least one specific binder for the analyte; (iii) a first binding agent coupled to either (1) exogenous analyte or (2) the specific binder for the analyte; (iv) a support comprising a second binding agent; b) adding an activator to the mixture, wherein the activator binds the first binding agent and the second binding agent of the support to immobilize the first binding agent; c)determining the amount of the analyte in the sample by detecting the immobilized first binding agent, the presence or amount thereof being related to the presence or amount of the analyte in the sample.

Abstract: Method and test kit for assaying in a sample an analyte which is a bloodgroup antigen present on erythrocytes or an antibody binding to such a bloodgroup antigen. To that end, the sample is treated with a reagent containing a binding partner for the analyte, so that a complex of bloodgroup antigen present on erythrocytes and antibody bound thereto is formed if the sample contains analyte. The analyte is a bloodgroup antigen present on erythrocytes, the analyte binding partner is an antibody capable of binding to the bloodgroup antigen and if the analyte is an antibody binding to a bloodgroup antigen, the analyte binding partner is the bloodgroup antigen present on erythrocytes. Erythrocytes, complex or non-complexed, are then separated from non-bound antibodies using a separation medium with a density higher than that of the liquid containing the antibodies but lower than the density of crythrocytes.

Abstract: This invention provides fluoroassay which comprises labeling analyte molecules with a fluorescent material having a nucleic acid portion stained with a sufficient number of fluorochrome molecules so as to be measurable as fluorescent spots, and a reactive group binding to the analyte molecule specifically, immobilizing the labeled analyte on a solid phase, and counting the number of fluorescent spots. The nucleic acid portion of the fluorescent labeling material is a double-stranded or single-stranded nucleic acid, and the staining with the fluorochrome molecules is performed with intercalating type, minor groove binding type, or covalently binding to the nucleic acid type.

Abstract: The antitope of an antibody is masked with a masking agent, followed by immobilization on a support. The masking agent is then eluted to produce an improved immunosorbent, which is capable of binding more than double the amount of an antigen than existing immunosorbents having the same antibody bound at the same density. Preferably, the masking agent is an antigen or other compound having an epitope for which the antitope of the bound antibody has an avidity. In a preferred embodiment, greater than 30% of the bound antibodies maintain the same vicinity as when unbound for specific antigen or hapten molecules. Preferably, the support is formed of any conventional immunosorbent support material which allows the bound and unbound antibody to maintain an avidity for the same compounds or antigens.

Abstract: The invention relates to a method for detecting specific target-cells in a simple and time saving way, using paramagnetic particles, antibodies recognizing the Fc portions of target-cell associating antibodies and target-cell associating antibodies directed to specific antigen determinants in the target-cell membranes. Incubation of the cell suspension with a mild detergent and/or second set of antibodies or antibody fragments, prelabeled or not with fluorescent agents, metallocolloids, radioisotopes, biotincomplexes or certain enzymes allowing visualization, with dramatically increase the specificity of the method. The method can further be used for isolation of the target-cells by magnetic field application and kit for performing the method according to the invention is described.

Abstract: Reagents and methods for the detection of Staphylococcus aureus are provided. The reagents contain an antibody that binds to a capsular polysaccharide of type 5 of Staphylococcus aureus, and can be used in methods for detection of oxacillin resistant Staphylococcus aureus that escapes detection by agglutination in the presence of fibrinogen and antibodies directed against protein A of Staphylococcus.

Abstract: A novel signal amplification system for immunoassays that minimizes non-specific signals is disclosed. Immunoassay methods, reagents and test kits are described for obtaining immunoassays of enhanced sensitivity. The reagents include antibody-variant DNA conjugates, wherein the variant DNA is a substrate for an RNA-dependent RNA polymerase, such as, QB replicase. Immunoassay methods to detect, or to detect and quantitate, analyte in test samples comprise transcribing the variant DNA of said antibody-DNA conjugates that are bound to analyte, to RNA, and replicating the RNA transcripts, wherein the presence or quantity of variant RNA replication products can be correlated with the presence or quantity of analyte in the test samples. Further, methods are provided to detect, or to detect and quantitate, simultaneously two or more analytes in a test sample.

Abstract: The present invention is directed to novel methods for enhancing the ability to separate a species of interest from other different species present in a free solution mixture thereof which takes advantage of interactions that occur between soluble, small molecular weight ligands and the species of interest. The small molecular weight ligands employed in the present invention function to interact with a species of interest in a mixture of different species through either affinity, hydrophobic and/or ionic interactions, thereby altering the molecular weight, hydrodynamic volume and/or isoelectric point of the species of interest and rendering it separable from other component(s) in the mixture.

Abstract: Z domain variants of staphylococcal protein A have significantly reduced size but possess IgG-binding affinity equivalent to the wild type Z domain. These Z domain variants are suitable for use in affinity chromatography purification of proteins and in the treatment of staphylococcic diseases.

Abstract: A biologically recognizing layer on a solid phase consisting of biologically recognizing molecules comprising regions which recognize a substance to be analyzed and regions which do not recognize a substance to be analyzed.The biologically recognizing molecules are aligned on a suitably modified surface by means of molecular regions which do not recognize the substance to be analyzed. The biologically recognizing molecules are cross-linked with the aligning surface and are consequently covalently altered. The molecular regions recognizing the substance to be analyzed are not altered by the covalent bonding and retain their bonding activity.The layer is produced in a novel two-stage method. In the first step, the biologically recognizing molecules, the aligning molecules and carrier molecules are adsorbed. In the second step, the molecules are covalently anchored by cross-linking. Cross-linking is brought about by photolytic activation of a water soluble reagent bonded to the carrier molecules.

Abstract: The present invention relates to a method and kit for carrying out a qualitative or semi-quantitative assay. The invention uses a multi-layer device including an an uppermost cover layer of a water-impermeable material having a hole therein with a diameter of approximately 1-5 mm, an intermediate insoluble porous support layer having a first substance bound thereon in a reaction zone, the hole exposing at least a part of the reaction zone, and a layer of a hydrophilic material in contact with and positioned on the side of the insoluble porous support layer opposite the side with the cover layer. The device permits transverse, but not substantial radial, diffusion of liquid through the reaction zone. Approximately 1-50 .mu.l of a test sample and a second substance are applied to the reaction zone through the hole. A colloidal gold label is attached to the second substance.

Abstract: The present invention is a method useful for the detection of bloodgroup antigens and antibodies. The method of the present invention is envisioned for two types of assays: a direct assay and an indirect assay. The direct assay comprises adding a sample of erythrocytes to a reaction tube charged with two layers of immunoreactive particles, a first layer, preferably Sepharose, having Protein G coupled to the surface of those particles and a second layer, preferably Sephacryl S-200, having Protein A coupled to those particles in a buffer solution. Antibodies specific for the bloodgroup antigens tested for are coupled to the Protein G. The reaction tube is then centrifuged for a time sufficient to force to the bottom of the reaction tube erythrocytes that do not attach to the antibodies.

Abstract: A novel HIV type O immunodeficiency virus is disclosed which has the designation MVP-2901/94 and which has been deposited with the European Collection of Animal Cell Cultures (ECACC) under No. V 95012601. The characteristic antigens which can be obtained from the virus and which can be employed for detecting antibodies against retroviruses which are associated with immunodeficiency diseases are also disclosed, as are the partial DNA and amino acid sequences of the virus.

Abstract: A solid phase method of detection or assay of the presence or amount in a serum or plasma sample of a target antibody specific to a cell surface antigen. The sample is contacted with an immobilised preparation of cells bearing the antigen and antibody bound thereto is detected or assayed by means of an indicator comprising a binding partner for the antibody bound to labelled latex particles.

Abstract: Gliotoxic factor in the isolated or purified state, characterized in that it possesses toxic activity with respect to human or animal astrocytic cells, having the effect of a cytomorphological disorganization of their network of intermediate filaments and/or a degradation of the proteins of said intermediate filaments and/or cell death, in particular by apoptosis.

Abstract: A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.

Abstract: The present invention is a method and apparatus useful for the detection of bloodgroup antigens and antibodies. There are two preferred embodiments of the method: a direct assay and an indirect assay. The direct assay comprises adding a sample of erythrocytes to a reaction tube charged with a column of immunoreactive particles having an immunoglobulin binding ligand selected from the group consisting of Protein A, Protein G, Protein A/G or a universal kappa light chain binding protein coupled to the surface of the particles. Antibodies specific for bloodgroup antigens tested for are coupled to the ligand on the particles. The reaction tube is then centrifuged for a time sufficient to force to the bottom of the reaction tube erythrocytes that do not attach to the antibodies on the particles.

Abstract: A Dot-Blot assay ("spot test") with Bis-N,N,dioctadecylamide (BDA.TDA) as antigen was developed to detect anti-BDA.TDA antibodies in tuberculosis patients. To develop the antigen-antibody reaction, as a first step and in order to enhance the reaction, an anti-human rabbit serum was used followed by incubation with a protein A-colloidal gold conjugate. This assay showed almost equal sensitivity and specificity as the .beta.-galactosidase ELISA test which was conducted in parallel. This simple and fast assay could be used in places where ELISA equipment is not easily available.

Abstract: A sensitive chemiluminescence immunoassay method for field detection of the presence or the amount of low chlorinated biphenyl compounds in a solution is disclosed. The assay has a five minute analysis time and a working range of detection as low as about 1 part per billion chlorinated biphenyl. Kits for the detection of chlorinated biphenyls are disclosed.

Abstract: The binding site on an immunoglobulin for a protein is blocked by hydrophobically coupling the immunoglobulin to a blocking agent such as a label. This results in a detection reagent useful in a variety of test assays in which a protein and an immunoglobulin are advantageously used together, but separately. The reagent is useful in the simultaneous determination of IgG and one of IgA or IgM. Anti-IgA-IgG or anti-IgM-IgG is coupled to a hydrophobic label, particularly a pigment or dye, which label blocks the binding site on IgG for Protein A, and labeled Protein A is added.

Abstract: Water soluble reagents are claimed, comprising a water-soluble polymeric carrier molecule having attached thereto more than one connecting moiety wherein the connecting moiety is derived from divinyl sulfone, and wherein each connecting moiety is attached to a reactive functional group on the polymeric molecule, and wherein the reagents are capable of reaction with a molecular species having a functional group which is reactive towards the terminal vinyl group of the more than one connecting moiety and the molecular species is selected from the group consisting of labelling species, marking species, and targeting species.

Abstract: Processes are provided for pretreating body fluid compositions and subsequently analyzing the pretreated body fluid compositions for analytes of interest. Processes for pretreating the compositions include providing size exclusion gel having a molecular weight fractionation range or a molecular weight exclusion such that the size exclusion gel is capable of excluding or fractionating the analytes of interest, and then causing the composition to contact the size exclusion gel in order to separate the analytes from low molecular weight composition components which interfere with the separation and analysis of the analytes of interest. Processes for analyzing pretreated compositions include electrophoretic methods such as capillary zone electrophoresis which involve the separation and detection of analytes of interest.

Abstract: A magnetic separator for isolating magnetically-labeled substances of interest, such as immunological agents, from a non-magnetic test medium using a method of high gradient magnetic separation. The target substance is contacted with microscopic magnetic particles having a receptor for binding with the target substance. The test medium containing the magnetic particles is held in a non-magnetic container and placed into a gap within an arrangement of magnets for causing the magnetic particles to adhere to selected locations upon the interior wall of the container. The quantity of magnetic particles may be controlled to cause the magnetic particles collected upon the interior wall to form a monolayer. The magnets are arranged upon a yoke which may provide linear, surrounding multipolar, or partially surrounding multipolar configurations of magnetic pole faces about the gap.

Abstract: The present invention relates generally to test articles and assays for the detection of analytes in biological fluid samples. More particularly, the present invention relates to test articles an assays which employ dyed microorganisms as visual labels to detect suspected analytes.

Abstract: Disclosed is an assay system including a compound comprising an analyte-specific moiety having substituted thereon a polymer comprising plurality of self-quenching emitter moieties and a plurality of isocharged functionality separating the emitter moieties. The present invention provides compounds that overcome the undesirable effects of self-quenching when multiple emitter moieties are used for labelling of assay reagents. Avoidance of this self-quenching phenomenon by the compounds of the invention makes it possible to introduce a more concentrated degree of labelling on to analyte-specific molecules such as oligo nucleotide probes, antibodies and other specific binding proteins and analyte-specific polysaccharides. Therefor, it is possible to effect greater assay sensitivity because the number of labels per recognition molecule(analyte-specific moiety) can be increased beyond the point previously possible without the reduction in signal caused by self-quenching.

Abstract: A method is provided for removing a second ligand from a particle surface without substantially affecting the particle surface, comprising the step of exposing the particle to a first ligand immobilized onto a support, wherein the particle is exposed under conditions and for a residence time sufficient to allow the second ligand to desorb from the particle surface, and wherein the first ligand has an affinity for the second ligand that is at least two orders of magnitude greater than the affinity of the second ligand for the particle surface, such that the second ligand is removed from the particle surface without substantially affecting the particle surface.

Abstract: A method of testing for tuberculosis and leprosy in animals or humans and antigens used in the method. The method involves carrying out an assay on sera from humans or animals using an antigen to bind antibodies in sera. The antigens used in the method are synthetic pseudo cord factor-like glycolipids having the structures (I) or (II) below: ##STR1## wherein R represents a straight or branched chain alkyl group having 15 to 18 carbon atoms. The antigens can be produced by chemical synthesis, and can therefore be made available in suitable quantities, and are relatively stable at ambient temperatures while showing good sensitivity and specificity for tuberculosis and leprosy. The antigens can be used, for example, for enzyme-linked immunosorbent assays, and related "spot tests" for diagnosing tuberculosis and leprosy.

Abstract: A method for determining the amount of free ligand in a test ample where the ligand is also present bound to one or more endogenous receptors, comprising combining the test sample, ligand receptor and unlabelled, differential binding ligand analogue, incubating to permit the free ligand and unlabelled, differential binding ligand analogue to compete for the ligand receptor separating the ligand analogue, determining the amount of ligand receptor bound to the ligand or to the ligand analogue, and correlating the amount of bound ligand receptor to the amount of free ligand present in the test sample.

Abstract: The invention includes an agglutinant complex which is useful for the investigation of the antigens present on erythrocytes, and which results from affinity couplings between nonagglutinant IgG type antibodies specific for the antigen to be identified, a protein capable of binding to at least two sites on the Fc part of antibodies and an anti-immunoglobulin antibody or its fragments.

Abstract: An Fc receptor protein conjugated with Ferritin binds to an exposed Fc region of an antibody to form a cell/Ferritin complex. Magnetic particles are added to the medium and bind to the complex. The magnetic particles when bound to the complex significantly enhance the magnetic field gradient of the complex such that it may be separated magnetically from the medium.

Abstract: The invention relates to the use for the diagnosis of demyelinating neuropathies, in particular multiple sclerosis or MS, of endogenous lectins having an affinity for glycans rich in mannose or their protein subunits, these lectins or their subunits being immunologically related to the cerebellar soluble lectins (CSL or their protein subunits, respectively).

Abstract: The present invention involves a method for detecting bladder cancer in a subject. The method preferably comprises first collecting a urine sample from the subject. The presence of a proteinaceous substance having a molecular weight of about 180 kDa according to its relative electrophoretic migration rate through detergent-containing polyacrylamide gel is then measured. This substance reversibly binds concanavalin A and is complexed with gamma globulin while in the urine. The gamma globulin complex binds to Staphlococcal protein A. Said proteinaceous substance, when present in detectable amount, is an indicator of bladder cancer.The method of the present invention for diagnosing bladder cancer in a subject may also be described as comprising detection in a urine sample from said subject of a proteinaceous substance having a molecular weight of about 180 kDa and being unreactive with antibodies specifically binding to carcinoembryonic antigen or epidermal growth factor receptor.

Abstract: A procedure for finding and identifying red cell antibodies by means of the solid-phase method, comprising contacting erythrocytes having selected antigen patterns with sera or plasmas that are to be tested for antibodies and transferred along with a polyspecific anti-human globulin reagent onto a substrate coated with protein A or protein G, thereby to transfer the detection or identification cells or, in the case of autocontrols, inherent cells.

Abstract: The invention relates to a method for detecting specific target-cells in a simple and time saving way, using paramagnetic particles, antibodies recognizing the Fc portions of target-cell associating antibodies and target-cell associating antibodies directed to specific antigen determinants in the target-cell membranes. Incubation of the cell suspension with a mild detergent and/or second set of antibodies or antibody fragments, prelabeled or not with fluorescent agents, metallocolloids, radioisotopes, biotincomplexes or certain enzymes allowing visualization, with dramatically increase the specificity of the method. The method can further be used for isolation of the target-cells by magnetic field application and kit for performing the method according to the invention is described.