Abstract: This invention relates to improved methods and novel compositions for enzyme complementation assays for qualitative and quantitative determination of a suspected analyte in a sample. The use of enzyme-acceptor and enzyme-donor polypeptides prepared by recombinant DNA techniques or chemical polypeptide synthesis techniques which are capable of interacting to form an active enzyme complex having catalytic activity characteristic of .beta.-galactosidase is described. Both homogeneous and heterogeneous assays utilizing these polypeptides are described.

Abstract: Vesicles are formed, in part, from an amphiphilic chelating agent whereby a detectable metal marker can be complexed to the vesicle. A portion of the vesicle may also be formed from an amphiphilic compound derivatized with a ligand, whereby the vesicle may be used as a tracer in an assay. If the detectable metal marker is a fluorescent rare earth metal, the assay may be a time delay fluorescent assay. The vesicles may be dried, and reformed by addition of water for use in an assay.

Abstract: Early pregnancy can be determined by detection of a protein in milk, which protein is associated with the placental membrane. The protein is characterized by having an approximate molecular weight of 47,000 to 53,000 and an isoelectric point of from about 4.0-4.4.

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: 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: Bifunctional aromatic compounds are employed as rigid coupling compounds for coupling one organic compound to another. For example, paranitrophenylisocyanate may be employed as a coupling compound for coupling thyroxine to a fluorescent dye to form a tracer for use in an assay.

Abstract: Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological activity for bonding to a particular biologically active group of a material and method in which the luciferin and biologically active molecule conjugate is added to a material having a group for combination with the biological activity of the conjugated molecule, the mixture is incubated to bond the material to the reagent through the biological activity and the product of the bonding is reacted with luciferase to produce bioluminescence having intensity dependent on the concentration of the material being assayed.

Abstract: Compounds and methods featuring, in one aspect, compositions containing an organic boronic acid and one or more reporter groups.

Abstract: Immunogens for preparing antibodies against the drug lidocaine and related compounds, labeled conjugates, synthetic intermediates, and the use of such antibodies and labeled conjugates in immunoassays for determining lidocaine and such related compounds. The immunogens comprise lidocaine or an analog thereof coupled through one of the aromatic methyl groups to a conventional immunogenic carrier. The antibodies and labeled conjugates are particularly useful in homogeneous nonradioisotopic immunoassays for measuring lidocaine or its analogs in biological fluids such as serum.

Abstract: A method of chemical analysis is disclosed which employs a release tag compound of general formulaRx--Re--Sin which Rx is a reactivity group capable of forming a covalent bond with another molecule, Re is a release group capable of being cleaved, and S is a signal group. The release tag is covalently bonded to a substance of interest which is to be determined in the course of a chemical analysis for an analyte, release group Re is cleaved at an appropriate point in the analytical procedure, and signal group S is determined, thereby determining the substance of interest. Where the substance of interest is the analyte, determination of S determines the analyte. Where the substance of interest is not the analyte but is related to the analyte concentration, determination of S allows indirect determination of analyte.

Abstract: A method and system for detecting and preferably measuring the presence of an activated complement complex in a sample is discussed. The presence of such an activated complex is indicative of complement pathway activation and includes a first complement component and a second complement component. The method uses a first binding agent specific to the first complement component and a second binding agent specific to the second complement component which when bound with the complex forms an aggregate. The second specific binding agent includes a label whose presence is used to detect and measure the amount of aggregate and therefore activated complex in a sample. An assay system and aggregate for use in an assay system are also discussed.

Abstract: A test composition, device, and method for their use in a homogeneous specific binding assay which employs a substance having reactant activity, i.e., a reactant, as a labeling substance in the detection of a ligand in a liquid medium. The test composition and device comprise a conjugate formed of a specific binding substance coupled to the reactant. The reactant advantageously is an enzymatic reactant such as an enzyme substrate or coenzyme. The activity of the conjugated reactant as a constituent of a predetermined reaction is affected by reaction between the specific binding substance in the conjugate and a specific binding counterpart thereto. The presence of a ligand in a liquid medium may be determined using competitive or displacement binding or sequential saturation techniques wherein the specific binding substance in the conjugate is the ligand or a specific binding analog thereof, or using a direct binding technique wherein the specific binding substance is a specific binding partner of the ligand.

Abstract: A process is provided for the preparation of magnetic particles to which a wide variety of molecules may be coupled. The magnetic particles can be dispersed in aqueous media without rapid settling and conveniently reclaimed from media with a magnetic field. Preferred particles do not become magnetic after application of a magnetic field and can be redispersed and reused. The magnetic particles are useful in biological systems involving separations.

Abstract: Immunoassays which utilize an enzyme linked ligand or receptor wherein the enzyme is bacterial luciferase; mercantile kit useful in performing said immunoassay; and compounds utilized in performing said assay.

Abstract: Immunoassay method and reagent means for determining theophylline in biological fluids such as serum. The antibody employed in the method is prepared from an immunogen comprising theophylline coupled at its 9-position to an immunogenic carrier material.

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 tracer composition is described whereby a nonradioactive photon emitter is coupled to a ligand, antigen or antibody for use in various immunoassay methods of analysis. The photon emitters employed are bioluminescent proteins such as firefly or bacterial luciferase as well as other luciferases from various species. To synthesize the new tracer, these photon emitters are coupled to antigens or antibodies using coupling agents such as glutaraldehyde, CNBr, carbodiimide and others. The coupling method may also include intermediate materials such as polysaccharides, polypeptides, polyacrylamides and others coupled between the photon emitter and the antigen or antibody.Methods are described for using this tracer in both competitive and noncompetitive immunoassays to detect proteins, hormones, drugs, viruses and the like. Detection is accomplished by activating the tracer in a bioluminescent reaction and measuring the emitted light by photometric instruments or by photographic film.

Abstract: Conjugates of penicilloic acid derivatives and certain poly(amino acids), which are either antigenic or enzymatic, are provided. Antibodies raised against the antigenic poly(amino acids) and the enzyme conjugates are used as reagents in immunoassays. In particular, the compounds of the present invention can be used for measuring the presence of a .beta.-lactamase in a patient serum sample by adding a known amount of penicillin to the sample and observing the production of penicilloic acid over a fixed period of time.

Abstract: This invention is directed to AUT-PK 500, a novel autophosphorylating protein kinase, to the purification and characterization of AUT-PK 500 from rat adrenocortical carcinoma, to the use of AUT-PK 500 as a marker for neoplasia cells, and to a radioimmunoassay for detecting AUT-PK 500 in neoplasia cells.

Abstract: The present invention concerns an autologous precipitating antibody and the gp70 pigment-associated antigen on melanoma cells which it recognizes. The antibody is useful in detecting pigmented melanoma cells in excised specimen, serum or urine.

Abstract: An antibody of a human leukemia virus-related peptide obtained by collecting an antibody produced in a mammal body by administering to the mammal an antigen prepared by reacting a human leukemia virus-related peptide represented by formula (1):R-Pro-Val-Met-His-Pro-His-Gly-Ala-Pro-OH (1)whereinR is a hydrogen atom or a group shown by formula, H-Tyr-;as a hapten, with a carrier in the presence of a hapten-carrier binding agent.

Abstract: Early pregnancy can be determined by detection of a protein in serum, which protein is associated with the placental membrane. The protein is characterized by having an approximate molecular weight of 47,000 to 53,000 and an isoelectric point of from about 4.0-4.4.

Abstract: Fluorescent conjugates are employed providing combinations of a fluorescent sensitizer and a fluorescent phycobiliprotein. The conjugates find use in applications where large Stokes shifts, high absorption coefficients and high fluorescence quantum yields are desired. Particularly, combinations of phycobiliproteins are employed where the wavelength of excitation may be 50 nm or more different from the emission wavelength.

Abstract: Preparations useful in the diagnosis of infectious mononucleosis are disclosed. Horse and sheep erythrocytes are purified and essentially homogenous glycoproteins are extracted. The purified glycoproteins are useful as antigens in testing for the presence of the heterophile antibodies of infectious mononucleosis and in the preparation of reagents for the enumeration of rosetting lymphocytes without having to have available fresh sheep blood.

Abstract: The invention comprises a method for the purification of a human lung tumor-associated antigen (hLTAA) specific to human lung tumors of diverse histological characteristics; serum levels of hLTAA correlate with lung tumor incidence, and appear to usefully discriminate between various stages of the malignancies. The invention further comprises an immunoassay predicated on purified hLTAA for the detection and quantitative determination of hLTAA in biological fluids, particularly blood serum, and diagnostic systems for clinical immunoassay procedures.

Abstract: In a method for immunochemical assay of human chorionic gonadotropin involving use of an antibody immobilized on a carrier, an antigen and an antibody to which a labeling agent has been attached, when the antibody supported on the carrier and the antibody coupled to a labeling agent are different antibodies which are not overlapping in antigen-determining site and one of said different antibodies is specifically reactive to human chorionic gonadotropin, a high reproducibility of the result of the immunochemical assay is obtained.

Abstract: A process for detecting the presence of an antigen in a specimen is described, which process comprises:(A) contacting said specimen with a substrate coated with antibodies of said antigen, incubating the contacted substrate and washing the substrate;(B) contacting the washed material of step (A) with a hapten conjugated antibody against said antigen, incubating the so-contacted material and washing the so-incubated material;(C) contacting the washed material of step (B) with a radioactive material labeled or enzyme containing anti-hapten antibody, incubating the so-contacted material and washing the same; and(D) effecting radioimmunoassay if said antibody is radioactive or enzyme labeled immunoassay if said antibody contains an enzyme moiety.Quantitative determination of the antigen in the specimen is effected by comparing the counts of the radioimmunoassay or the concentration of enzyme against a standard as by photocolormetric methods.

Abstract: Sandwich immunoassays are provided wherein an anti-analyte antibody is coupled with a detector labeled antibody, the anti-analyte antibody being substituted with a N,N,N-trimethylammoniumphenyl group and the detector labeled antibody is an antibody specific to the N,N,N-trimethylammoniumphenyl group. The invention includes novel assays, antigens and antibodies.

Abstract: Carcinoembryonic antigen contains immune determinates in common with .alpha.-acid glycoprotein. Carcinoembryonic antigen-containing compositions are purified by adsorption onto antibody to .alpha.-acid glycoprotein. The purified compositions may be employed as standards in carcinoembryonic antigen assays or in the labelled form as tracers. Intact carcinoembryonic antigen is assayed by a modified sandwich-type immunoassay. Other cancer-associated substances may be identified by searching for high molecular weight analogues of normal proteins.

Abstract: A process for the preparation of immunologically active tagged conjugates by covalent linking of an enzyme with an antigen or haptene, which process comprises placing a solution of a mixture of immunologically active and inactive conjugates in contact with an insoluble complex former able to form a non-immunological complex reversibly with the untagged antigen or haptene component of the conjugate, separating the insoluble complex former, and eluting with a desorbent for untagged antigen or haptene.

Abstract: Immunogen conjugates comprising N-substituted-amino-3-desoxydigoxigenin derivatives coupled to conventional immunogenic carrier materials, and antibodies raised against such conjugates. Also provided are labeled digoxigenin conjugates for use with the digoxigenin antibodies in preferred immunoassay techniques for determining digoxin in biological fluids.

Abstract: Vitamin B.sub.12 in liquid samples, such as human serum or human plasma, is assayed by a competitive binding technique using intrinsic factor and certain labelled vitamin B.sub.12 derivatives. The labelled derivatives are formed from the (d)-monocarboxylic acid isomer of vitamin B.sub.12, which isomer is free from other monocarboxylic acid isomers (and derivatives thereof) of vitamin B.sub.12, by binding to the (d)-isomer, via the carboxylic group, a compound which is itself a label (e.g. an enzyme) or which comprises a label (e.g. a fluorophore), or to which a label is attached (e.g. a histidine ester to which .sup.125 I is attached). The labelled derivatives of the (d)-monocarboxylic acid of vitamin B.sub.12, free from other isomeric monocarboxylic acids and derivatives, are novel and constitute one aspect of the invention.

Abstract: New polymers are disclosed which absorb or emit photons in the visible or ultraviolet spectrum. The polymers when bound to ligands or receptors are useful as reagents for the detection of substances in physiological fluids. Also disclosed are methods of preparing said polymers and reagents and methods of employing said reagents in immunoassay systems.

Abstract: The reticulocyte content present in a specimen of red blood cells is quantitatively measured based upon the selective immunoreactivity of the reticulocyte portion of the specimen with a reticulocyte-specific antibody which is immunoreactive with proteinaceous material associated with reticulocytes but not associated with mature red blood cells. Such immunoreactive proteinaceous material may be transferrin, transferrin receptor, transcobalamin II, or transcobalamin II receptor. Various procedures are described for quantitating such selective immunoreactivity, including fluorescent and radioactive detection techniques employing direct or indirect fluorescent or radioactive labeling of the reticulocyte-specific antibody.

Abstract: Pancreas specific protein systems, including Pan Ag purified antigen having molecular mass of about 2.25.times.10.sup.5 daltons and pancreas specific antibodies to such antigen, and methods for providing and utilizing such antigen and antibodies.

Abstract: This disclosure relates to assay of a glycoprotein component of human breast gross cystic disease fluid which has been designated GCDFP-15. This material is a useful marker in monitoring the efficacy of therapy in women with metastatic breast carcinoma and also in determining the maturity of the fetus in pregnant women. The assay for GCDEP-15 can also be used in conjunction with other assays for breast carcinoma such as an assay for carcinoembryonic antigen (CEA) whereby the utilization of both tests is more effective in monitoring for recurrence of disease than using either assay alone.

Abstract: An improved immunoassay for detecting cannabinoids is described. This immunoassay is characterized by utilizing, as reagents, antibody, .sup.125 I-radiolabeled antigen, an unlabeled antigen and an anion exchange resin for separation of labeled and unlabeled antigen. The iodinated tetrahydrocannabinol moiety is stabilized by the addition of small quantities of antioxidants such as butylated hydroxy toluene.

Abstract: Novel oligopeptide used as reagents or in the preparation of reagents in relation to gamma-carboxyglutamic acid-containing protein of bone. Particularly, labeled oligopeptides and antibodies prepared from immunogen conjugates are disclosed for use in immunoassays.

Abstract: The method of producing clinical assays for use of monoclonal antibodies in the diagnosis of Herpes simplex virus (HSV) infections and the differentiation of Herpes Simplex virus types 1 and 2 as a diagnostic kit for differentiating HSV-1 and HSV-2 utilizing clone 1D4 against HSV-1 and clone 3E1 against HSV-2.

Abstract: A method for the simultaneous immunochemical assay of trace components in a sample involving competitively reacting the antigens or antibodies in a sample and labeled antigens or labeled antibodies for limited binding sites. The labels are spectral sensitizing dyes. The reaction products are contacted with silver halide and exposed to light having wavelengths corresponding to the absorption spectra of the spectral sensitizing dyes.

Abstract: An improved immunoassay for the polypeptide hormone thymosin .alpha..sub.1 is described. The assay employs an improved antibody which is elicited by an immunogen which has been prepared by coupling [Tyr.sup.1 ]-thymosin .alpha..sub.1 to an immunogenic carrier protein using a bifunctional diazonium coupling agent.

Abstract: Procedures are presented for isolating the major outer membrane protein of Chlamydia trachomatis. The isolated protein is a species specific antigen which comprises about 60% of the C. trachomatis cell outer membrane structure. The protein has a molecular weight ranging from about 38,000 to 44,000 daltons, with a mean molecular weight of about 39,500 daltons. The protein antigen is purified from C. trachomatis cells by first extracting the cell contents with a mild anionic detergent, preferably sarcosyl, to leave a residue of intact outer cell membranes. These outer cell membranes are then extracted with a strong anionic detergent, preferably sodium dodecyl sulfate, which solubilizes the 39,500 dalton antigen. The antigen is then purified by hydroxlapatite chromatography. The antigen is species specific for Chlamydia trachomatis and may be utilized in assaying Chlamydial infection in mammals.

Abstract: Novel amides of an iodothyronine compound such as tri-iodothyronine (T3) or thyroxine (T4) with an aminoacetic acid compound such as nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA) or diethylenetriamine pentaacetic acid (DTPA) may be labelled, e.g. radioactively labelled with I-125, to give labelled derivatives useful in assays of a free thyroid hormone in a biological fluid which also contains the thyroid hormone bound to one or more natural binders. The novel compound T4 EDTA is shown to bind much more strongly to antibodies to T4 than to the natural protein binders in the biological fluid.

Abstract: In a method of assay for a trace component such as antigen, antibody or enzyme utilizing immunochemical reaction or enzyme reaction in combination with photographic detection system comprising measuring optical density of a silver image formed in proportion to the antigen, antibody or enzyme to be measured, a novel analysis sheet comprising a support having provided thereon, in succession, a water absorbing layer and a silver halide emulsion layer is employed. The analysis sheet essentially increases the amount of water absorbed so that the analysis sheet is extremely effective for improving detection sensitivity.

Abstract: In an immunochemical measurement method of an antigen or antibody which comprises competitively reacting an antigen or antibody labelled with spectral sensitizer and an antigen or antibody to be measured, bringing either the reaction product of immune reaction or the unreacted component into contact with silver halide, exposing the same to light having a wavelength which the spectral sensitizer absorbs, developing the exposed silver, and, measuring optical density of the thus formed silver image and/or colored dye, the contact with silver halide is performed in the presence of a specific hydrazine compound. Thus, detection sensitivity is markedly improved.

Abstract: A method for performing multiple, simultaneous in vitro diagnostic tests is provided. The method utilizes a solid phase device comprising a receptacle and an insert. The receptacle has one or more fixed components immobilized on its inner surface. The insert has one or more fixed components--different from those immobilized on the receptacle--immobilized on its surface which is in contact with a fluid sample when inserted therein. The test is performed by placing a fluid sample, having two or more mobile components reactive with the fixed components, into the receptacle and in contact with the insert for a period of time and measuring the changes which are a function of the concentration of the mobile components.

Abstract: A highly sensitive, immunoassay method for determining the amount of an analyte in a sample containing a known analyte in an unknown concentration is provided. Sample; a polypeptide-labeled analog of the analyte, an antibody specific for said analyte, a polypeptide partner capable of non-covalently binding with the polypeptide-labeled analyte to form a complex having catalytic activity, and a substrate capable of being converted to a reporter molecule by the catalytic activity of said complex are brought together in a medium. The polypeptide-labeled analyte analog is capable of competitively binding to the antibody and the polypeptide partner, the antibody inhibiting the formation of a catalytically active complex in the absence of analyte, and the concentrations of the antibody, polypeptide partner and polypeptide-labeled analyte are such as to cause varying amounts of analyte to be directly related to the conversion of the substrate to the reporter molecule.

Abstract: A lectin is covalently bonded to an immunological conjugate such as an antibody-antigen or its equivalent. Then, the lectin-conjugate is isolated from the reaction product mixture by one of a number of alternative techniques involving one or more of the following types of reaction; (1) reversible reaction of the lectin with an insolubilized sugar to isolate lectin from the remainder of the mixture, (2) reaction of one immunological component (e.g., antibody) bonded to the lectin with an insolubilized corresponding component (e.g., antigen) to separate the antibody components from the remainder of the reaction mixture, and (3) filtration of the reaction components to separate on the basis of product molecular weight, size and/or shape of the components.