Abstract: The present invention provides a process for the preparation of an immuno-reactive, porous carrier material by application of a solution of a first reaction component of an immuno-reaction and of a solution of a second component of an immuno-reaction coprecipitating therewith, incubation of the carrier material impregnated with the solutions for the immuno-precipitation, optional washing and subsequent drying of the impregnated carrier material, wherein a solution of both components of the immuno-reaction is prepared, which solution contains an inhibitor for the immuno-precipitation, the carrier material is impregnated with this solution and then the immuno-precipitation is initiated by removal of the inhibitor or by removal of the inhibiting action.

Abstract: An affinity matrix and a method for the detection of low molecular weight compositions such as aflatoxins are provided utilizing specific monoclonal IgM antibody having an affinity constant not less than about 1.times.10.sup.9 liters per mole. Methods for the preparation and use of such affinity matrices are also given. The detection is rapid, accurate, reproducible, and allows for quantitative recovery of the composition of interest.

Abstract: A method and composition for use in preparing red blood cells for use as indicator cells to detect the presence of antigens and antibodies in biological fluids is the subject of the present invention. Freshly collected human or animal erythrocytes are washed six times with isotonic saline, and suspended to a hematocrit of four percent (volume/volume) with isotonic saline. An incomplete antibody, specific for the antigen or antibody to be detected, in a quantity less than that which would cause irreversible agglutination, is added to the erythrocyte solution. The mixture is incubated, washed, and suspended to a hematocrit of 3-5 (v/v) with isotonic saline. A second antibody, immuno specific for the first antibody and in an amount sufficiently great to immunologically bind substantially all of the antibody carrying red blood cells but not so great as to cause irreversible agglutination, is attached to the erythrocytes. This mixture is incubated, washed, and suspended to a hematocrit of 0.

Abstract: Bone marrow or blood containing tumor cells can be freed from the tumor cells by bringing it into contact with an water-insoluble antitumor monoclonal antibody which is prepared according to the enzyme insolubilizing method and thereby combining the tumor cells with the insolublized antibody and destroying the tumor cells along with human serum complement on the insolublized antibody. The thus obtain bone marrow or blood free from tumor cells is used favorably in the autologous-bone-marrow transplantation method for treatment of tumor.

Abstract: A composition of matter and method for using the composition of matter to isolate various cell elements from a heterogeneous cell suspension in a simple one step procedure. The composition is a monoclonal antibody mixture including complement and a non-toxic density gradient media. The composition provides a negative selection technique in which undesired subpopulation cells are lysed and separated from the desired cell subpopulation. The invention has particular application to isolation of white cell elements.

Abstract: A diagnostic assay support matrix is prepared by coating a support matrix with a polymer resistant to nonspecific protein binding and immobilizing the desired biologically active agent on the polymer surface.

Abstract: A method for performing a diagnostic immunoassay by solid phase separation for digoxin. To a reaction mixture of a test sample and labeled anti-digoxin antibody, which forms a complex of any digoxin present in the test sample, is added a solid phase material having an immobilized ouabain triacetate derivative compound capable of binding any excess labeled antibody. The solid phase material is chosen to rapidly settle whereby a solid and liquid phase is formed. The liquid phase can then be extracted to measure the amount of digoxin-labeled antibody present therein. Ouabain triacetate derivative compounds possess sufficient affinity for anti-digoxin antibodies, and are therefore useful in a solid phase separation based digoxin immunoassay for settling out such antibodies without contributing to undesired background interference.

Abstract: A method for conducting a ligand assay in an inert porous medium wherein a binding material is immunologically immobilized within the medium, which includes the steps of immunologically immobilizing a binding material within a finite zone of the medium, applying an analyte to the zone containing the immobilized binding material, applying a labeled indicator to the zone which becomes immobilized within the zone in an amount which can be correlated to the amount of analyte in the zone, applying a solvent to substantially the center of the zone to chromatographically separate the unbound labeled indicator from the zone, and measuring the amount of labeled indicator remaining in the zone.

Abstract: A method is disclosed for determining an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp) consisting of ligand and complementary receptor. The method comprises combining in an aqueous medium (1) said sample, (2) a first reagent comprising a non-dispersed surface to which is bound an sbp member that becomes bound to the surface in relation to the amount of analyte in the sample. The volume of the aqueous medium containing the above reagents is sufficiently large to allow complete immersion of the first reagent therein and determination of the analyte but not so large to result in substantial interference in the determination upon addition of a second reagent reactive with the conjugate of enzyme and sbp member and capable of generating a signal. The second reagent is present in a medium of sufficient volume to substantially increase the volume of the liquid medium and reduce interference in the determination.

Abstract: An immunoassay in which a thermally induced phase separation is used to effect the separation of specifically bound reactants from free reactants is disclosed. A first reactant is conjugated to a temperature-sensitive polymer to form a polymer/reactant conjugate, and a second reactant is conjugated to a reporter to form a reporter/reactant conjugate. The polymer/reactant, reporter/reactant, and biological fluid samples suspected of containing the analyte are admixed in solution at a temperature other than that at which the polymer will precipitate. Specific binding is allowed to occur, thereby forming a ternary complex. The salt concentration of the adjusted solution is then adjusted to a concentration sufficient to cause the complex to precipitate from the solution, the amount of reporter activity in the precipitated complex or in the solution measured and the presence and/or concentration of the analyte therefrom determined.

Abstract: An improvement in an assay method which relies on the detection of a labeled, solubilized specific binding complex is achieved by linking the label to a component of the complex through a bond cleavable under conditions compatible with the assay and with the complex in solution.

Abstract: A solid phase immunoassay comprises the steps of(a) immobilizing an immunoreagent on the surface of a carrier comprised of an inert synthetic resin selected from the group consisting of polyimides and polyfluorinated synthetic resins,(b) contacting the immunoreagent with a complementary immunoreagent whereby an immunocomplex immobilized on said carrier is formed,(c) quantitating the immobilized immunocomplex.An element useful in conducting this solid phase immunoassay is prepared by a process of treating the surface of an article comprised of a synthetic polymer selected from the group consisting of polyimides and polyfluorinated synthetic resins to make it adsorptive of an immunoreagent which comprises the steps of(a) thoroughly rinsing the surface with a water-miscible organic solvent,(b) thoroughly rinsing the surface with water.

Abstract: Immunological fluorometric assay method, wherein an antigen (1) attached to magnetic particles (3) reacts in a liquid with an antibody (2) present in the sample and with an antibody (4) marked with a fluorescent substance. After the reaction, the particles are pulled by means of a magnetic field through a second (6) liquid layer onto the bottom of the measurement vessel, whereupon the particles are excited, and emitted radiation is collected through the wall of the vessel.

Abstract: A method for detecting and/or determining an agglomerate formed by the reaction between a specific binding agent and the corresponding acceptor substance according to the general methodology of blot overlay assays by using colloidal metal particles labelled components which may be visualized as a coloured signal at the surface of the blotting medium characteristric for the colloidal metal particles used or quantitatively determined following art-known spectrophotometric procedures such as densitometry.

Abstract: A heterogeneous specific binding assay method for determining the amount of a suspected analyte in an aqueous test medium wherein a reaction mixture is formed by combining the test medium with assay reagents including a labeled reagent, an immobilizable component, and a binding substance which causes the immobilizable component to precipitate. Free and bound species of the labeled reagent are formed as a function of the amount of the analyte in the test medium. One of the free and bound species of the labeled reagent is immobilized by binding of the immobilizable component with the binding substance. The immobilized labeled reagent is seperated from labeled reagent which has not been immobilized, and the amount of label in the labeled reagent in one of the separated fractions is determined and related to the amount of analyte in the test medium.

Abstract: Fragments of human Factor VIII:C which bind to antibody inhibitors of Factor VIII in patients afflicted with such inhibitors are disclosed. The method of treating Factor VIII inhibitors, by administering one or more of these fragments, is also disclosed.

Abstract: Method for carrying out immunoassays, in which method an antibody (or antigen) labeled with a tracer so as to be fluorescent is attached onto an antigen (or antibody, respectively) present at the inside wall of the measurement vessel, a liquid denser than the sample is added to the measurement vessel, which said denser liquid displaces the liquid from underneath, and both the excitation radiation is passed into the sample and the fluorescent radiation is collected to the detector through the wall or the bottom of the measurement vessel.

Abstract: The use of immunologically non-specific peptide linked amino acids containing compounds that are capable of immobilizing circulating immune complexes for the purpose of detection or removal from serum or blood, such compounds including oligopeptides, modified oligopeptides, polypeptides, modified polypeptides, proteins, modified proteins, and in particular glycosylated polypeptides and proteins.

Abstract: A method for the isolation and quantitative detection of a selected single-stranded target polynucleotide from solution. The target polynucleotide is hybridized in solution to a single-stranded mediator polynucleotide, a probe polynucleotide, and an immobilized polynucleotide sequence. The sequence of the mediator polynucleotide comprises a first sequence complementary to a first portion of the target polynucleotide sequence and a second nucleotide sequence complementary to a portion of a single-stranded immobilized polynucleotide sequence. The probe polynucleotide, which carries a detectable label, is complementary to a second portion of the necleotide sequence of the target. The immobilized polynucleotide is immobilized by attachment to a solid support, and, through hybridization to the mediator polynucleotide, functions to immobilize the entire immobilized polynucleotide/target polynucleotide/probe polynucleotide "sandwich".

Abstract: A novel method is disclosed for the purification of monoclonal antibodies from antibodies of the same isotype but different light chain composition which are frequently present in the fluids in which such antibodies are generated. According to the method, the mixture is placed in a hydroxylapatite chromatography column and eluted with a buffer solution in a concentration gradient elution. With the appropriate optimization of system parameters, a separation of antibodies effective for both analytical and preparative purposes is obtained.

Abstract: A cap for a container in which immunoassays are performed includes a liquid absorbing material accessible to liquid in the container.

Abstract: Method for the determination of a component of the reaction between a specific binding protein and a substance bindable therewith which process comprises incubating a sample solution with a determined amount of binding component in insoluble form in the presence of a reagent containing a known amount of one of the binding components in enzyme-labeled form, separating the liquid phase from the solid phase by centrifuging and determining the activity of the enzyme remaining in the liquid phase in a centrifuge rotor during or after the action of increased gravitional force whereby the solid phase and the liquid phase are still in contact with one another.

Abstract: Fluorometric immunological assay method in which an antigen is attached onto magnetic particles, which are, for the time of measurement, pulled against the wall of the measurement vessel by means of the magnetic field. The magnetic particles adhering to the vessel walls are irradiated and the resultant emitted fluorescence is measured.

Abstract: Disclosed herein is an apparatus and process for conducting ligand receptor assays. The apparatus comprises a first member which is a membrane or a filter to which is bound a receptor for the ligand or which is capable of extracting cells carrying the ligand from a fluid sample. The apparatus further comprises a second member which is composed of absorbent material which acts when in contact with the first member to induce flow through the first member when a fluid sample is added to it. The apparatus is used to conduct assays by applying a sample to the upper surface of the first member to bind ligand in the sample by means of receptor fixed to the first member or, in certain cases, by extracting cellular material which has ligand associated with it. Addition of the sample is typically followed by addition of labeled antibody against the antigen being assayed followed by a washing step to remove unbound labeled receptor.

Abstract: Method to detect and/or determine the presence of cancer cells in mammals, including humans, and to monitor the progress of treatment for cancer. A human protein marker, the assay-marker, has been ascertained of the molecular weight 70,000-74,000, which is secreted by cancer cells at levels 10-fold or greater than that observed with normal cells. An assay is described where the assay-marker or an antigenically analogous protein, the analog-marker, is used to prepare antibodies to the assay-marker. The antibodies are then reacted with blood or serum samples to determine the level of the assay-marker in the sample. Thus, the assay essentially comprises (1) providing an antibody to the protein described above (2) reacting the antibody of step 1 with the sample to be tested and (3) measuring the level of the reacted antibody to detect and/or determine the presence and/or quantity of cancer cells.

Abstract: The invention relates to a process for the immunoassay of a substance, such as an antigen, a hapten or an antibody.This process comprises the following stages:(1) contacting a sample containing the substance to be assayed with a labelled immunoactive reagent specific to said substance, the immunoactive reagent quantity being such that the substance to be assayed is in excess compared with that labelled immunoactive reagent;(2) contacting at least part of the thus obtained reaction medium with a solid phase, to which is fixed an immunoactive reagent specific to the substance to be assayed;(3) separating the reaction medium from the solid phase; and(4) determining the labelled immunoactive reagent content of the solid phase.This process is more particularly applicable to the assaying of lipoprotein and in this case use is made of an antibody labelled by an enzyme, such as the Raifort peroxidase. It is also possible to assay .alpha.-foetoprotein by using monoclonal antibodies.

Abstract: A chemically active membrane having a large surface area is provided in which a hydrophilic, microporous, skinless, polyamide membrane is chemically bound to a residue of an activating agent which is capable of reacting with a biologically active material.The chemically active membrane, formed by reacting a hydrophilic, microporous, skinless, polyamide membrane with an activating agent may be used to prepare a biologically active membrane having a large surface area which comprises an acceptor molecule such as a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulin, a carrier protein, a receptor, heparin, a coagulation factor, or a histone covalently bound to the hydrophilic, microporous, skinless, polyamide membrane by reacting the chemically activate membrane with the acceptor molecule.

Abstract: Apparatus and methods for selective removal of specific biological cells or specific antigens or antibodies, from fluid containing their mixture with other biological cells and particulates are described. Filtration by this biochemical filter system is effected in a continuous closed-loop fluid flow path. The apparatus described herein comprises a source of fluid containing specific biological cells, antigens or antibodies to be removed; a source of complementary cells or complementary antibodies for the antigens and complementary antigens for the antibodies which can form large agglutinates following a biochemical reaction a reaction chamber providing conditions favorable for fast clump formation following the reaction; a filter for trapping large agglutinates; one or more pumps to regulate various flow rates; and necessary connecting links to form a closed-loop fluid flow path that includes the sources of biological cells, antigens or antibodies and complementary cells, reaction chamber, and filter chamber.

Abstract: The invention relates to monoclonal anti-idiotypic antibodies to human anti-DNA antibodies. Monoclonal, anti-idiotypic antibodies are produced using hybridoma technology. The antibodies are used as diagnostic reagents in methods to determine the presence of anti-native DNA antibodies in serum from patients suspected of having systemic lupus erythematosus.

Abstract: The invention relates to monoclonal anti-idiotypic antibodies to human anti-DNA antibodies. Monoclonal, anti-idiotypic antibodies are produced using hybridoma technology. The antibodies are used as diagnostic reagents in methods to determine the presence of anti-native DNA antibodies in serum from patients suspected of having systemic lupus erythematosus, and as therapeutic reagents in methods to remove the anti-native DNA antibodies from the serum of patients with systemic lupus erythematosus.

Abstract: This invention relates to the use of differential separative properties and modified assay containers in improving the convenience of separative scintillation proximity assay (SSPA) and extending its range of practical application.

Abstract: A procedure for detecting malignancy includes culturing human colon tumor cells in a capillary system. A rabbit is immunized with byproducts of the culture. An antibody produced in the rabbit is labeled with .sup.125 I using lactoperoxidase according to a known method. Blood samples are drawn from a being to be tested. The drawn blood is processed to produce serum. The immune complexes are removed from the serum with purified protein A from the Staphlococcus Aureus Cowan strain. The removed immunocomplexes are dissociated with 0.2M glycene/HCl pH 2.8. The labeled antibody is combined with the antigen component of the immunocomplex to produce a new labeled immunocomplex. The newly formed immunocomplex is precipitated with PEG 6000. The newly formed labeled immunocomplexes are counted in a gamma auto counter.

Abstract: A method for detecting or removing a substance in a medium is presented. Magnetic material, particularly magnetic bacteria or magnetic particles contained therein, are treated to render them receptive to binding or attachment to the substance sought to be detected or removed. Following binding or attachment to the treated magnetic material, the medium is subjected to a magnetic field, which results in removal of the magnetic material and the substance bound or attached thereto.

Abstract: This invention describes a process for immuno-chemical quantitative determination of immunologically active substances. The method involves a first incubation of a sample containing the active substance with a labelled binder, which contains an antibody or antibody fragment. After complexing has taken place, solid phase bound active substance identical to the substance being quantitatively determined is added. The solid phase bound substance binds with free binder, and the solid and liquid phases are separated. A second antibody which is specific either to antibody or the solid phase antibody-substance complex is then added to the liquid phase. This second antibody is non cross-reactive with individual complex components. The amount of labelled first antibody bound to second antibody is then determined.

Abstract: Assay for a hapten or antigen wherein supported hapten or antigen is used to separate "bound" and "free" fractions. The sensitivity of the assay is increased.

Abstract: Viruses are detected by means of an immunoassay method in which an extended solid phase coated with antiviral antibody is employed to bind and remove virions from a specimen by forming an immuno-complex with antigens of said virions, a mobile solid phase comprising a dispersion of microspheres coated with the antiviral antibody is used to bind said microspheres to antigens associated with said immuno-complex, and the presence of bound microspheres is detected. The detection sensitivity is amplified by the ability to more readily detect the microspheres, which may be dyed or labelled. The extended solid phase advantageously may be in the form of a dipstick which can be easily contacted with the specimen. A virus detection kit provides the extended solid phase and mobile solid phases, each coated with antiviral antibodies.

Abstract: A method of specimen treatment preparatory to conducting an immunoassay is disclosed whereby a microbial protein is solubilized by a detergent at elevated temperatures and in the presence of an alkali or alkaline earth metal ion. At elevated temperatures, the detergent is soluble. However, at lower temperatures, the presence of the metal ion renders the detergent insoluble so that it is prevented from interacting in the immunoassay procedure. A specific application is in the solubilization of the principal outer membrane protein of Chlamydia trachomatis.

Abstract: The invention relates to techniques for isolating from a mixed population of cells disired living cells either producing and releasing a particular product or having a characteristic molecule on their surface. The isolation techniques depend upon the localized interaction between the product (or molecule) and other agents added to the system such that distinguishable conditions can be caused to occur (or not occur) only in the immediate vicinity of desired cells which produced and released the product or which contain the molecule on their surface.

Abstract: A heterogeneous binding assay in which a liquid component and a granular particulate solid phase are incubated together for a predetermined period of time. To prevent unwanted unit gravity sedimentation of the granular particulate solid phase during incubation the density of the liquid component is controlled such that it is substantially equal to the density of the granular particulate solid phase, thus preventing sedimentation. The density may be controlled by adding a density modifying medium such as a colloidal suspension of silica particles coated with polyvinylpyrrolidine. After incubation, the density of the liquid component may be reduced allowing unit gravity sedimentation and hence separation of the solid phase from the liquid component. The assay is particularly applicable to immunoassay (for example radioimmunoassay and immunoradiometric assay). Immunoradiometric assays for human growth hormone, thyroid stimulating hormone and alphafetoprotein are described.

Abstract: There are provided bifunctional chelating agents, which are analogues of EDTA, conjugates of same with a variety of haptens and conjugates of such conjugates with haptens with certain metals, forming metal complexes. The metal complexes have a variety of uses, among these various assays for the determination of haptens and macromolecules. There is also provided a process for the production of the above. There are provided radioimmunoassays based on the above conjugates with metal cations.

Abstract: Microcapsules for effecting an immune response, especially through agglutination reaction, comprise an antigen or antibody bound to functional groups, such as amino groups, carboxy groups, etc., on a wall surface of the microcapsules via cross linking agents, such as polyfunctional isocyanates, isothiocyanates, etc. The antigen or antibody is strongly bound to the wall surface of the microcapsules to thereby achieve excellent detection sensitivity.

Abstract: The present invention relates to a simple and accurate method to be applied in immunoassay technique for separating the free ligand from the antibody-bound ligand.The method involves the use of a solvent extraction operation which at equilibrium provides the formation of two distinct phases the free- and bound-fraction. The solvent to be utilized should be slightly water miscible or completely water immiscible, having the property of marked selective extraction power toward the gamma-labelled constituent to be determined. The method is applicable in a liquid-liquid system, a solid-liquid system and solid-solid system.The method is useful for radioimmunoassay, free radical assay or metallo-immunoassay, being most versatile having also the advantage that as a result of the free ligand being extracted into the solvent, the determination of the quantity of gamma-labelled substance in the free and/or bound fraction compares very favorably with the known prior art methods in immunoassays.

Abstract: Disclosed herein is an apparatus and process for conducting immunoassays. The apparatus comprises a first member which is a membrane or a filter to which is bound an antibody, typically a monoclonal antibody, or which is capable of extracting cells from a fluid sample. The apparatus further comprises a second member which is composed of absorbent material which acts when in contact with the first member to induce flow through the first member when a fluid sample is added to it. The apparatus is used to conduct immunoassays by applying a sample to the upper surface of the first member to bind antigen in the sample by means of antibody fixed to the first member or, in certain cases, by extracting cellular material which has antigen associated with it. Addition of the sample is followed by addition of labeled antibody against the antigen being assayed followed by a washing step to remove unbound labeled antibody.

Abstract: A process for determining the presence of polyvalent antigens by incubation with three receptors is presented. In addition, a kit for carrying out this process is provided as well. One of said receptors is bound to a solid support and the other two, in solution, derive from the same animal species.

Abstract: A reagent and merchandizing kit for use in the diagnosis of syphilis by hemagglutination of an anitgen obtained from a culture of pathogenic Treponema pallidum Nichols and which is sensitized on carrier particles in the presence of antibody wherein said antigen is substantially devoid of proteinic fractions of said culture having a specific gravity of less than 1.01, and methods of preparing the same.

Abstract: A ligand is dissociated from a binder specific for the ligand by directing a beam of ultrasound at the complex of ligand and binder. The procedure may be employed in a solid phase assay to separate a labeled form of ligand, employed as a tracer, from a binder supported on a solid support.

Abstract: The present invention relates to a method which eliminates centrifugation and decantation steps, to be performed in an automatic manner for carrying out specific binding assay tests, wherein liquid and solid phases are present.According to the invention, use is made of a specially designed device, consisting of a mixing reservoir into which is fitted snugly a mixer separator having a channel in the vertical axis of the mixer-separator. A rack holding a number of said mixing reservoirs containing the incubated reagents and analytes, capped with the mixer separators, is placed into a press-device designed to perform at a controlled rate a downward movement. The mixer separators are pushed downwards into the mixing reservoirs at a chosen rate for a preselected distance to complete the mass transport and separation operations. The separation devices are removed and either one of the separated phases can be measured in the desired analytical instrument for a quantitative or qualitative determination.

Abstract: Rates of binding between members of a specific binding pair, e.g., ligand-receptor, are greatly enhanced by short-term ultrasonication of an aqueous medium containing the specific binding pair. The enhanced rates find particular use in specific binding protein assays.

Abstract: A test kit and assay for detecting the presence of minute amounts of an organic reactant in a test sample includes a plurality of beads or other types of support structures that are impregnated with a fluorescer and coated with a ligand that specifically binds to the organic reactant being investigated. The beads are placed in an aqueous solution, together with the reactant which has been radiolabeled. The portion of the radiolabeled reactant that binds to the ligand is thereby brought in close enough proximity to the beads to activate the fluorescer to produce light energy. The radiolabeled reactant that does not bind to the ligand is, for the most part, disposed too far away from the beads to enable the radioactive energy emitted thereby to reach the fluorescer integrated into the beads. Thus, the level of light energy produced by the fluorescer is indicative of the amount of reactant present in the test sample.

Abstract: Allergenic factors are isolated from antigen extracts by preparing a column with pooled serum from patients with allergy to a given allergen system and passing over this column a pool of allergen extracts from different sources. Allergen specific immunoglobulin antibodies in purified form are obtained by forming an antigen-specific antibody complex from serum containing the antibodies and a material containing the antigen, recovering the complex and disassociating it, forming a second complex of the antigen with an excess of a second antibody so as to realize a combination of the second complex, free first specific antibody and free second specific antibody and then separating the second complex from the free first and second antibodies.