Abstract: For the serodiagnosis of a fluid sample, it is particularly advantageous to use a device comprising (A) a supported, porous membrane wherein a first immunoreagent is bound so as to be capable of binding a foreign analyte and forming a complex when the foreign analyte is brought into contact therewith; and (B) a matrix which presents a first surface and an opposing second surface and which contains a second, labeled immunoreagent that is capable of binding the foreign analyte to form a labeled complex when the foreign analyte is sandwiched between the first immunoreagent and the second immunoreagent. In such a device, the first surface of the matrix is adjacent to a surface of the membrane, (ii) the matrix is wettable by or soluble in an aqueous fluid, and (iii) the second immunoreagent is mobilized when the matrix is wetted.

Abstract: Assays for an anti-pre-S antibody in a human serum by an enzyme immunoassay in which a biotin-avidin system is used are disclosed. The assays include inhibition method, sandwich method, competitive method and Protein A or anti-human IgG antibody method in a solid phase.The assays can determine the anti-pre-S antibody titer in human sera highly sensitively and simply without use of radioactive materials.

Abstract: An assay to detect the presence of live non-A, non-B hepatitis virus in vitro. Bone marrow cells are exposed to a body fluid or biological preparation to be tested and the cells are placed in suspension. When using bone marrow cells, growth factors are added to the bone marrow stem cells. Therefore, if the number of colonies growing in the mixture are less than that number present in the culture of cells exposed to a sample that has been determined to contain no live virus, live non-A, non-B hepatitis virus is present in the sample tested. The assay is particularly useful to determine the presence of live non-A, non-B, hepatitis virus in a vaccine.

Abstract: Samples e.g. transfusion blood, are assayed for antibodies to retroviruses, e.g. AIDS virus, using an insolubilized antigen comprising retrovirus antigens bound to globulin, the globulin itself being bound to an inert solid support; and an immunoglobulin which contains specific antibody to the retrovirus antigens and which is labelled with a revealing label, and the soluble phase is then separated from the insoluble phase and the quantity of revealing label associated with either the soluble or the insoluble phase determined.The sue of labelled antibody in competition with test sera for binding on the insolublized antigen permits better identification of antibody containing specimens. The retroviruses may be a human T-lymphotropic retrovirus HTLV-I, II or III or a new retrovirus isolate CBL-1 etiologically related to AIDS.

Abstract: Hybridomally produced monoclonal IgM antibodies having high affinity are useful for the immunoassay and purification of viral antigens.

Abstract: Anti-NANBH (anti-non-A, non-B hepatitis) antibody comprises IgM isolated from sera of monkeys artificially infected with extracts of feces from patients known to be suffering from epidemic NANB hepatitis. A process for isolating an antibody of this type is provided. A reagent for detecting epidemic NANB hepatitis comprises the anti-NANBH IgM fixed to a solid support. The reagent is useful for diagnosing epidemic NANB viral hepatitis by forming an immunological complex between the anti-NANBH IgM and antigen in a fecal extract from a patient being diagnosed, and then incubating the immunological complex with enzyme-labeled IgG from serum of a human convalescing from epidemic NANBH. The complex is detected by developing the enzyme.

Abstract: Novel and improved methods for diagnosis, prognosis, prophylaxis and therapy of viral infections are described. The novel methods employ a virus, viral antigen or fragment thereof in which "perturbation" of an oligosaccharide moiety renders the virus, viral antigen or fragment thereof more specifically recognizable or reactive with neutralizing antibody. As described, "perturbation" of an oligosaccharide moiety encompasses any modification that (1) alters the chemical or physical structure of a carbohydrate residue that is naturally present; (2) that removes, wholly or in part, a carbohydrate residue; and/or (3) that prevents or alters addition of a carbohydrate residue. A variety of methods for oligosaccharide "perpetuation" are also described.

Abstract: Diluent compositions for preparing specimens for immunoassay contain effective amounts of salt and non-ionic surfactants to inactivate viruses in the specimens and improve the sensitivity and specificity of the immunoassays; said diluents having some strengths of from about 21 to about 35 mS/cm and comprising 0.05 to 1% non-ionic surfactants, along with other conventional ingredients. The invention relates as well to immunoassay procedures using the novel diluents.

Abstract: A method for the purification of HBc antigen, particularly HBc antigen produced by recombinant organisms by means of DNA recombination technique, which comprises subjecting a solution containing HBc antigen to acid-treatment, wherein the solution containing HBc antigen is acidified to a pH range of not higher than 6 by adding an acid and then resulting precipitates of lipid and contaminant proteins are removed, and then subjecting the acid-treated solution containing HBc antigen to an ion exchange chromatography with an anion exchanger, and a method for measuring HBc antibody by using said purified HBc antigen in a passive hemagglutination method, EIA method or RIA method.

Abstract: A simultaneous sandwich immunoassay employing high-affinity monoclonal antibodies is disclosed. This simultaneous sandwich assembly has surprising sensitivity compared to forward and reverse sandwich assays for the detection of multi-determinant antigens such as hepatitis B surface antigen.

Abstract: A single-antibody inhibition assay for detecting antibody to HBcAg using a labeled monoclonal antibody. A competitive ELISA method of screening hybridoma supernatants for monoclonal antibodies to HBcAg. A method of producing high affinity monoclonal antibody to rHBcAg by immunizing a mouse with rHRcAg using a short immunization schedule. Monoclonal antibodies to immunodominant epitope of HBcAg.

Abstract: A method for obtaining an actual linear standard curve in a sandwich type of immunoassay where a first antibody bound to an insoluble support and a second unbound labelled antibody complex with the antigen contained in a test sample to form an insoluble antibody:antigen:labelled antibody complex which is then detected. Unbound unlabelled first antibody and/or unbound unlabelled second antibody are added to the reaction mixture to divert excess antigen away from the desired end-product complex, thus rendering the antigen of interest the rate-limiting factor in the overall immunoreaction. This results in a pseudo first-order reaction which produces an actual linear standard curve.

Abstract: Disclosed is a method for detection for serum antibody and/or microbial surface antigen by radial partition immunoassay. The method of this invention is applicable to (a) the evaluation of a clinical specimen for identification of a microorganism; (b) antimicrobial sensitivity assays for determination of an efficacious antibiotic for use against a specific microorganism; (c) a semi-quantitative determination of viral surface antigen; and, (d) a quantitative method for the determination of the presence of serum antibodies to microbial antigens. In each of the foregoing applications, the analyte of interest can be immobilized within a porous matrix (solid phase) by simple pipetting of the sample onto the prepared matrix. Appropriate reagents are subsequently applied to the matrix to effect immunochemical interaction of a labeled binding material to the surface antigen (or antibody) of the analyte of interest. The portion of the matrix within which such interaction takes place is termed the "reaction zone".

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: The surface antigen protein of human Hepatitis B virus is synthesized in Saccharomyces cerevisiae as a 23,000-26,000 dalton polypeptide, essentially free of intermolecular disulfide bonds. This antigen is a poor immunogen in animals and man. No prior precedent or method exists for efficiently converting the non-disulfide bonded antigen to a fully intermolecular disulfide bonded particle. We describe the first example of such a conversion in vitro and show that the act of this conversion enhances the immunogenicity of the antigen about 10-fold. The in vitro conversion makes practical the production of hepatitis B surface antigen from microorganisms using recombinant DNA methods.

Abstract: The present invention discloses a screening test for detecting the presence of contaminating or infectious agents causing non-A, non-B hepatitis or AIDS in a blood donor setting. A kit for the detection of contaminating agents belonging to the group of retroviruses is also disclosed. Screening blood or blood related products so as to prevent spreading of infection or contamination due to retroviruses is now made possible by the present invention.

Abstract: Non-A, non-B hepatitis antigen, antigen compositions, vaccine and assay test for the detection of non-A, non-B hepatitis antigen. The antigen is characterized as having a particle size of about 2.0 nm to about 5.0 nm, a density of from about 1.24 to about 1.30 g/cm.sup.3, contains ribonucleic acid and occurs either free or bound to IgG molecules.The antigen can be isolated by isopycnic banding and chromatographic fractionation, and preferably by enzymatic digestion, isopycnic banding and chromatographic fractionation.

Abstract: A novel method for the determination of protecting anti-HBV immunoglobulins is based on the use of two different HBsAg reagents: one insolubilized and having the antigenic type AX and one labelled and having the antigenic type AY, wherein A represents the epitope or epitopes which are common to all HBsAg serotyes, wherein X and Y both represent combinations of the epitopes which are not common to all HBsAg serotypes, and where the combinations X and Y have no epitopes in common.

Abstract: The present invention concerns a method for detection of the core antigen of hepatitis B virus in a sample comprising a first incubation with carrier-bound antibodies directed against hepatitis B surface antigens and a second incubation with antibodies against the core antigen in the presence of a detergent.Also claimed are test kits for carrying out this method and an immunochemical reagent comprising antibodies directed against the surface antigen and against the core antigen conjointly bound to a carrier.

Abstract: The present invention relates to polyfunctional hydrophilic spherical microparticles of uniform size which contain from 5 to 95% of units originating from the copolymerization of an aldehyde of the formula ##STR1## R.sub.1 being an alkyl radical, from 5 to 90% of units originating from the copolymerization of an acrylic acid ester of the formula ##STR2## in which R.sub.2 is H or alkyl and R.sub.3 is a hydroxyalkyl group, less than 15% of units originating from an acrylic acid derivative chosen from amongst ##STR3## R.sub.4, R.sub.5 and R.sub.6 being alkyl radicals, and from 0.1 to 10% of units originating from a crosslinking product.The invention also relates to a process for the preparation of the said microparticles and to the use of these microparticles for the nephelometric determination of HB.sub.s antigen.

Abstract: A single polypeptide antigen that includes the amino acid residue sequence and epitope of a conformation-independent antigenic determinant and the amino acid residue sequence but lacks the epitope of a conformation-dependent antigenic determinant is disclosed as are methods of its manufacture and use and articles of manufacture using the same. The uses of the pre-S(2) region polypeptide encoded by the hepatitis B virus genome as a T cell proliferating agent and as a potentiator for enhancing the humoral immune response of animals that exhibit a low humoral response to an S region-containing immunogen are also disclosed.

Abstract: The present invention discloses an isolated and purified antigen specific to non-A, non-B hepatitis (NANBH) causing agent. The utility of the antigen as a diagnostic serologic marker and as a screening device for detecting the carrier or source of non-A, non-B hepatitis or infective factor thereof, particularly in a blood bank or plasmapheresis setting and preventing transmission of NANBH by isolating the source is described. Use of the antigen as vaccine to induce protective antibodies capable of neutralizing NANBH infectivity is also disclosed. A kit for detecting the presence or identifying the carriers or sources of non-A, non-B hepatitis or causative agent thereof is also disclosed.

Abstract: Compounds useful in a simultaneous multiple assay for analytes such as steroids, proteins, peptides, carbohydrates or drugs. The compound or compounds are prepared by labelling an individual analyte with a radioisotope through a chelating agent to form a coordinated compound. The assay uses one or more chelated labelled analytes with one or more labelled analytes wherein each radioisotope is different.

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 sensitive direct immunoassay system is provided for the detection of an antigen in body fluids. A single antibody which reacts with an antigen or antigens and which is bonded to an insoluble member, is incubated with a test sample. During this first period of incubation a portion of an antigen present in the test sample will combine with the antibody immobilized on the insoluble member. The antibody bonded member, to which antigen is attached, is then washed and incubated with an enzyme tagged antibody reagent. During the second incubation, the tagged antibody reacts with antigen fixed to the antibody member in the first incubation. Thus, an immobilized "sandwich" is formed of an insoluble member- antibody-antigen-enzyme tagged antibody. After the second incubation, the member is washed again to remove unreacted enzyme antibody reagent. The member is then exposed to a substrate which is converted by the enzyme to produce an end product.

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 `unmasking` delta antigen in the blood of an animal, known or caused to be infected with the antigen. The process involves treating serum from the animal with a surfactant and optionally with an antibody--antigen dissociating agent. The blood derived delta antigen is used as a diagnostic agent in the detection and determination of different classes of antibodies to hepatitis D. virus (delta agent) by enzyme-immunoassay and radio-immunoassay.

Abstract: HBcAb in a biological fluid is adsorbed on a surface which is then coated with BSA. The coated surface is then incubated first in the sample and then in the presence of radiolabelled HBcAb.

Abstract: A composition having a protein binding solid support onto which is bound a mixture of antigens and antibodies which are both bound to the solid support individually and are not present in the form of an immune complex.

Abstract: A radiolabeled or enzyme labeled peptide having no more than 60 amino acids in the chain of the peptide; the peptide having covalently linked amino acids disposed in a steric configuration which is recognized by and bound by an antibody. The labeled peptides can be utilized in various processes to detect the presence of a given antibody or antigen in a sample. Hepatitis B surface antigen and antibody to same may be so detected.

Abstract: A method and test kit are disclosed for detecting the presence of hepatitis B virus in a test specimen containing at least a portion of the DNA of the virus. A test reagent comprises cloned hepatitis B virus-DNA that has been repurified by treatment with a restriction enzyme and labelled to high specific activity with a radioactive label. The sample to be tested is fixed to a solid matrix, incubated in the presence of the test reagent under hybridization conditions and detected by hybridization to the labelled DNA probe. The uncombined HBV-DNA (labelled) is removed from the substrate, and the hybridized HBV-DNA determined by scintillation counting or by autoradiography of the substrate.

Abstract: Ligand having at least two determinant or binding sites is contacted with a labeled first binder, an unlabeled second binder different than the first binder and a precipitating binder specific for the second binder, with such contacting precipitating a complex of the ligand and the first and second binders to thereby separate bound labeled binder from unbound labeled binder. The bound and/or unbound labeled binder is determined as a measure of the ligand.

Abstract: A process for the detection of an antigen or antibody in a specimen which process comprises:(a) contacting said specimen with a substrate having bound thereon a mixture of antigens and antibodies to said antigen or antibody in said specimen, said antibodies and said antigens bound to said substrate being separately bound to said substrate and not in the form of an immune complex, incubating the so-contacted substrate and washing the substrate;(b) contacting the washed material of step `a` with a radioactive material labeled or enzyme labeled antibody or antigen, incubating the so-contacted material and washing the same; and(c) effecting radioimmunoassay if said antibody or antigen is radioactive or enzyme labeled immunoassay is said antibody or antigen is enzyme labeled.

Abstract: In a process for qualitative and quantitative determination of antigens, antibodies, and antigen/antibody complexes, using a chemiluminescent marker in a liquid-phase or solid-phase assay the combination of chemiluminescent marker and activator for the marker is selected from the combinations(a) H.sub.2 O.sub.2 /chloramine (or functionally equivalent derivative) - fluorescein, methylene blue, thionine or functionally equivalent derivatives of these chemiluminescent markers; and(b) calcium hypochlorite-fluorescein or functionally equivalent derivative.The method is simpler and more sensitive than known methods and presents less danger to those performing the assay.

Abstract: Method and apparatus are provided for carrying out multiple simultaneous transfers of fluid. The method and apparatus are particularly directed toward immunoassays wherein immunologically active compounds, such as antigens and haptens, are detected through their associated antibodies. The device relies on the ability to transfer fluids, such as biological samples and reagents, between a reservoir and an associated receptacle. By providing a receptacle having a port at its lower end and which is otherwise hermetically sealed, such fluid transfer can be effected by immersing the port beneath the surface of the fluid in the reservoir and manipulating the pressure on the remaining surface area outside the port. The transfer of biological fluids at positive pressure provides enhanced fluids flow characteristics, particularly reduction or elimination of the tendency of these fluids to froth or bubble.

Abstract: A sensitive direct immunoassay system is provided for the detection of an antigen associated with hepatitis in body fluids. A single antibody which reacts with a hepatitis antigen or antigens and which is bonded to an insoluble member, is incubated with a test sample. During this first period of incubation a portion of an antigen present in the test sample will combine with the antibody immobilized on the insoluble member. The antibody bonded member, to which antigen is attached, is then washed and incubated with an enzyme tagged antibody reagent. During the second incubation, the tagged antibody reacts with antigen fixed to the antibody member in the first incubation. Thus, an immobilized "sandwich" is formed of an insoluble member- antibody-antigen-enzyme tagged antibody. After the second incubation, the member is washed again to remove unreacted enzyme antibody reagent. The member is then exposed to a substrated which is converted by the enzyme to produce an end product.

Abstract: A newly discovered particle in the urine and serum of non-A, non-B hepatitis patients has been associated with non-A, non-B hepatitis. The particle resembles a togavirus and is 50-60 nm in diameter with a discrete core of about 40 nm in diameter. The virus loses its infectivity for tissue culture upon heating at 25.degree. C. in aqueous suspension or by exposure to ether. The particle may be cultured in vitro or recovered from body fluids or tissues to make immunoassays and vaccines. The immunoassays may be employed to detect the particle antigens or antibodies thereto in test samples.

Abstract: A device for determining the presence of antigens which comprises a first zone containing antigens and enzyme-linked antibodies which are capable of immunologically reacting with said antigens, said antibodies being positioned in said first zone such that they will be removed from said first zone when reacted with antigens passing through said first zone but not removed from said first zone in the absence of such antigens, and a second zone containing material capable of reacting with said enzyme-linked antibodies to produce a color forming reaction which indicates the presence of said antibodies.

Abstract: Process for the quantitative and qualitative determination of antigens, antibodies and their complexes by means of a chemiluminescing labelling substance activated or excited to chemiluminescence by an analytical reagent. By means of a serological reaction, initially an antigen/antibody complex is formed which is treated with a chemiluminescing conjugate containing chemiluminescing triphenylmethane dyes and the chemiluminescence of the chemiluminescing complex formed is measured. Qualitative or quantitative details on the substances to be measured are obtained on the basis of the resulting measured values. The dye labels are activated by either hypochlorites or mixtures of H.sub.2 O.sub.2 and a chloramine.

Abstract: Reagents and methods are described for an immunoassay test of increased sensitivity and decreased complexity employing an immunoglobulin specific for an antigen naturally or artificially placed upon the surface of an indicator particle coupled through the use of a hetero-bifunctional coupling reagent to a second antibody of differing specificity and specific for the antigen to be detected. In a preferred embodiment, a hetero-bifunctional coupling agent couples via a sulfhydryl group, a univalent immunoglobulin specific for the surface antigens on erythrocytes to a second multivalent immunoglobulin through an amide linkage with the latter immunoglobulin wherein said second immunoglobulin is specific for hepatitis-B surface antigen.

Abstract: Novel oligopeptides are provided having serologic activity for the a determinant of hepatitis B virus surface antigen. Included in the oligopeptide chain are a sequence of at least two cysteines and a lysine in proximity to the cysteines. The oligopeptides can find use in immunoassays, the formulation of vaccines, and the production of antisera.

Abstract: A stabilized radioimmunoassay product consisting of an antibody protein-bound to the cell wall of a selected bacterium whereby the antibody is irreversibly bound to the protein and remains specific for the antigen against which it was developed. The radioimmunoassay product is also characterized by the fact that the resultant complete product includes a phase of serum protein treatment to isolate the protein sites or other sites of nonspecific reactivity with the labelled antigen such that the non-specific binding of labelled antigen is significantly reduced. Also within the contemplation of the invention is the method of manufacturing the radioimmunoassay product which includes the initial phase of protein binding of the antibody to the selected bacterium and the subsequent serum treatment to isolate previously unutilized protein sites such that when said sites are subsequently exposed to labelled antigen it will be rejected.

Abstract: Human hepatoma cell lines, useful for metabolic studies such as screening potential carcinogens and mutagens, for cultivation of viruses, and for preparation of vaccines is obtained by culturing human hepatocarcinoma or hepatoblastoma on lethally irradiated cell feeder layers in the presence of a culture medium.

Abstract: Tree shrews (Tupaia belangeri) are used as an animal model to test the inactivation of vaccines, the harmlessness of blood products and the effectiveness of chemotherapeutic agents and disinfectants against viral hepatitis. The antibody determinations are performed at specific intervals for a period of 150 days in the case of viral hepatitis type A and 60 days in the case of viral hepatitis type B.

Abstract: A diagnostic reagent for hepatitis A antibody is prepared by adhering hepatitis A antibody to a surface by non-specific adsorption followed by specific coupling of hepatitis A antigen to the antibody. This reagent is useful in an in vitro assay for hepatitis A antibody.

Abstract: The present invention relates.Iadd., inter alia, .Iaddend.to improvements in the sandwich technique for the determination of a component of an antigen-antibody reaction in a liquid sample to be tested, utilizing as reagents (a) one component of said reaction bound to the surface of a water-insoluble, water-insuspensible, solid carrier, and (b) a component having the same immunological properties covalently linked to an enzyme. .Iadd.If and only if the component of the reagent that is water-insoluble and water-insuspensible has the same immunochemical properties as the component to be determined, is a predetermined amount of a binding partner for said first reagent (e.g., the reagent that is water-insoluble and water-insuspensible) added to the liquid sample. .Iaddend.The liquid sample is contacted and incubated with the reagent(s) to form a reaction mixture, the enzyme activity of either the liquid or solid phase of which is a measure of the presence and quantity of the component to be determined.