Abstract: The invention relates to a method of preparing heteromultimeric polypeptides such as bispecific antibodies, bispecific immunoadhesins and antibody-immunoadhesin chimeras. The invention also relates to the heteromultimers prepared using the method. Generally, the method provides a multispecific antibody having a common light chain associated with each heteromeric polypeptide having an antibody binding domain. Additionally the method further involves introducing into the multispecific antibody a specific and complementary interaction at the interface of a first polypeptide and the interface of a second polypeptide, so as to promote heteromultimer formation and hinder homomultimer formation; and/or a free thiol-containing residue at the interface of a first polypeptide and a corresponding free thiol-containing residue in the interface of a second polypeptide, such that a non-naturally occurring disulfide bond is formed between the first and second polypeptide.

Abstract: The invention relates to a method of preparing heteromultimeric polypeptides such as bispecific antibodies, bispecific immunoadhesins and antibody-immunoadhesin chimeras. The invention also relates to the heteromultimers prepared using the method. Generally, the method provides a multispecific antibody having a common light chain associated with each heteromeric polypeptide having an antibody binding domain. Additionally the method further involves introducing into the multispecific antibody a specific and complementary interaction at the interface of a first polypeptide and the interface of a second polypeptide, so as to promote heteromultimer formation and hinder homomultimer formation; and/or a free thiol-containing residue at the interface of a first polypeptide and a corresponding free thiol-containing residue in the interface of a second polypeptide, such that a non-naturally occurring disulfide bond is formed between the first and second polypeptide.

Abstract: The present invention relates to at least one novel human EPO mimetic hinge core mimetibody or specified portion or variant, including isolated nucleic acids that encode at least one EPO mimetic hinge core mimetibody or specified portion or variant, EPO mimetic hinge core mimetibody or specified portion or variants, vectors, host cells, transgenic animals or plants, and methods of making and using thereof, including therapeutic compositions, methods and devices.

Abstract: Cells can be labeled with products which they secrete and release in an efficient manner by coupling the cells at their surface to a specific binding partner for the product and allowing the product to be captured by the specific binding partner as it is secreted and released. The product-labeled cells can then be further coupled to suitable labels, if desired, and separated according to the presence, absence, or amount of product.

Abstract: Methods for the high yield production of antibody Fv-containing polypeptides, especially Fab? and F(ab?)2 antibody fragments are provided. Expression of heavy and light chain Fv in a microbial secretory system is followed by recovery of Fv from the periplasm under conditions that maintain a cysteine residue as a free thiol. The free thiol is reacted with free thiol of an antibody fragment of the same or differing specificity, or with agents such as diagnostic labels or therapeutic moieties. The products offer advantages of homogeneity and purity not available through the use of known methods for preparing such derivatives.

Abstract: Effector functions are provided to a desired target with improved specificity by use of two or more complementary targeting components. The targeting components assemble a functional moiety at the target. At the target, the functional moiety itself provides an effector function or binds to additional components which provide an effector function. The effector function may be an enzymatic activity, a label or a signal. The binary or polynary targeting system may be used for analyte determination as well.

Abstract: A diagnostic assay, preferably immunoassay, is provided for the detection and determination of MGP in a human serum sample, which comprises the use of one or more antibodies, in particular monoclonal antibodies, specifically recognising epitopes on and/or conformations of human Matrix Gla-Protein. Also, a method is provided for using MGP-related antigens as biomarkers for certain diseases, for example, atherosclerosis and other vascular diseases, and angiogenesis/neogenesis in tumor development. Further, monoclonal antibodies of class IgG are provided for use in the assay, which are defined herein as mAb3-15 and mAb35-49.

Abstract: The invention provides assay methods and kits that in general measure the level of a first analyte in a sample reduced by the level of a second analyte present in the same sample. In one embodiment, where levels of a first analyte from a first source is desirably determined and first analyte in the sample released from a second source is accompanied by proportional co-release of a second analyte, the assay identifies the level of first analyte released only from the first source. For analytes within bodily fluids, the assay can differentiate between elevated levels of analyte specific to the particular physiological or pathological state and elevated levels not specific to the particular state, providing single tests with diagnostic utility.

Abstract: Disclosed is a method of inhibiting the binding of a cell bearing a cell adhesion protein to a molecule or cell bearing a carbohydrate determinant specific for the cell adhesion molecule. The method involves contacting the cell adhesion protein-bearing cell with an AGP-antibody bearing the carbohydrate determinant. Also disclosed are AGP-antibody fusion proteins to which are covalently bonded carbohydrate moieties which interfere with the antibody portion's ability to fix complement and bind an Fc receptor. The methods of the invention may be used, for example, in any antibody-based therapy, for example, to reduce inflammation.

Abstract: A method for the direct analysis of analyte in keratinized structures, e.g., hair, fingernails and toenails, which comprises preparing a mixture containing a low redox potential compound such as dithiothreitol or dithioerythritol, an enzyme suitable for the degradation of the keratin structure and a sample of the keratin structure; permitting the enzyme to at least substantially degrade the sample of keratin structure, filtering the digest solution to remove substances which may interfere with ligand based analytical methods and subjecting the filtered digest solution to analysis to determine the identity and amount of analyte in the keratin substance sample. To accelerate the method, cupric sulfate may be added to the mixture after degradation of the keratin sample. The enzyme may be a peptidase, endopeptidase or proteinase, with papain, chymopapain, and proteinase K being preferred for use in the invention.

Abstract: Compositions and methods of use thereof for capture and detection of selected molecules are described. In one embodiment, a first composition includes a ligand component, such as an antibody coupled to a nucleic acid component. An a preferred embodiment, the nucleic acid is labeled with a fluorescent marker to facilitate detection. Another aspect of the invention is the ligand component bound to a solid support via a complementary nucleic acid component and a linker moiety. The method involves binding the target with the first composition in free solution, then binding the target to the solid support by means of both DNA hybridization and antibody-antigen affinity binding. Unbound molecules are washed away, and then the bound targets are detected by fluorescence detection. Vital stains can also be used to detect viable cells.

Abstract: Bis-maleimido cross-linking reagents join labelled antibodies having at least one sulphide interchain bridge. The conjugates have an enhanced binding capacity and have good blood clearance in vivo. The conjugates are of use in the diagnosis and therapy of tumors and may be prepared by reaction of the cross-linking reagent with the antibody.

Abstract: The present invention relates to methods for the production of a stable troponin preparation and its use as a calibrator and/or control in immunoassays. The formulation is prepared from mammalian, preferably bovine, heart tissue which provides a calibrator/control composition which remains stable over a long period of time.

Abstract: Provided are a monoclonal antibody making it possible to detect a native fibrin monomer, which is produced at the initial state of blood coagulation, and soluble fibrin; a hybridoma; and an immunoassay for detecting the initial stage of blood coagulation with high sensitivity, quickly, using the monoclonal antibody. Using a fibrinogen analog in blood as an immune source, cell fusion is carried out to prepare a monoclonal antibody which is not reactive with fibrinogen and is specifically and simultaneously reactive with a native fibrin monomer (that is, a fibrin monomer which is present in a body fluid, in particular in blood, and is not solubilized) and soluble fibrin. The fibrin monomer analog is preferably fibrinogen treated with bathroxobin, which is a snake venom.

Abstract: A method for detecting or assaying one constituting member in a specific binding pair, for example, the antigen in an antigen/antibody pair, by utilizing specific binding such as binding between an antigen and an antibody, together with redox reaction for detecting a label, wherein an oxygen micro-electrode with a sensing surface area of 1 mm2 or less is used; and an apparatus to which the method is applicable. According to the method and by using the apparatus, redox reaction for assaying the label can be completed in such a short time as several minutes. Therefore, an inexpensive disposable apparatus for household use can be realized.

Abstract: Conjugate probes are prepared in a one step process by incubating a macromolecule and a labeling group with an unsaturated polyaldehyde as the conjugating agent. The conjugating agent is capable of bonding virtually any labeling group to a macromolecule. Conjugate probes have been shown to have a high degree of specificity and exhibit a strong signal with minimal background.

Abstract: An improved homogeneous enzyme immunoassay process for quantitatively analyzing an antigen by determining the change in the enzymatic activity caused by a reaction between the antigen and an enzyme-labeled antibody. The antigen is reacted with an enzyme-labeled antibody, followed by the reaction with a second antibody capable of recognizing and binding to a different epitope and then with a third antibody capable of recognizing and binding to the second antibody. The enzymatic activity of the labeling enzyme is determined by a water-insoluble substrate. Using the water-insoluble substrate, steric hindrance is enhanced. A highly-sensitive analysis can be carried out by a simple operation even when the antigen has a molecular weight falling within an intermediate range, for example, a range of M.W. 10,000 to 70,000.

Abstract: The present invention relates to stable compositions useful as primary standards and calibrators and controls comprising a cardiac troponin I (cTnI) such as native, recombinant, addition and deletion forms thereof, whether or not complexed with other troponin subunits such as TnC and/or TnT, in an inactivated human serum. The compositions are obtained by incubating troponin complexes with human serum. The compositions are characterized by an immunodetectability ratio of epitopes on the N-terminal segment to epitopes on the C-terminal segment substantially equivalent to that of pooled, fresh serum from acute myocardial infarction patients.

Abstract: For the production of hetero-bispecific monoclonal antibodies at least the genes for the light chain and for the variable part of the heavy chain are isolated from a hybridoma cell line which secretes an antibody with a desired specificity and they are inserted into a eukaryotic plasmid vector which contains a marker capable of selection together with a strong promoter, this expression vector is transfected into a hybridoma cell line which secretes antibodies with a second desired specificity, the cell line is cultured, the antibodies are obtained and the bispecific antibody is isolated.

Abstract: Compositions and methods for avidin or protein having affinity for biotin immobilized on an inert support material, e.g., agarose, are disclosed. The compositions have high activity levels for binding biotin and may include a bulking agent, e.g., maltose, and a protectant to maintain the stability and integrity of the composition during lyophilization and terminal sterilization processes. The compositions have applicability in any instance where avidin agarose and/or the avidin/biotin technology are useful. In particular, the present compositions are useful in an enzyme capture system to prepare fibrin monomers useful for fibrin sealants.

Abstract: A conjugate consisting of a sequence of the analyte and an antibody against one of the antibodies used in the test can be employed, in aqueous solution and in precisely known quantity, as a stable calibrator in a sandwich immunoassay for detecting the analyte.

Abstract: The invention relates to an artificial positive control reagents based on antibody conjugates that are used in immunochemical detection methods and to processes for the preparation of these reagents.

Abstract: The invention provides monospecific antibodies that are specifically reactive with fibrinopeptide B (FPB) and with fibrinogen and fragments thereof containing the amino acid sequence defined by SEQ ID NO:1. The invention also provides anti-FPB probes, including monospecific anti-FPB antibodies that have been detectably labeled. In addition, the invention provides methods of using the monospecific antibodies for detection of fibrinopeptide B, as well as reagents and kits for performing the methods. For example, the invention provides a method for detecting fibrinopeptide B with specificity in biological samples such as blood samples, by using the antibody to immunometrically bind to the fibrinopeptide B. Diagnostic methods for determining information associated with atherogenesis and/or thrombogenesis. The invention further provides continuous cell lines (hybridomas) that produce monospecific anti-FPB antibodies.

Abstract: An in vitro immunoassay to detect and quantitate soluble crosslinked and non-crosslinked DesAABB fibrin polymers in a sample from a subject. The assay can be used to support a diagnosis of, to evaluate, and to monitor, in a mammalian subject, a thrombotic event, including, but not limited to, myocardial infarction, pulmonary embolism, stroke and deep vein thrombosis.

Abstract: The invention relates to the measurement of total leukocyte antigens, or fragments thereof, and the use of such measurements to enumerate cells, especially in whole blood. The term "total" leukocyte antigen used herein refers to the total amount of a leukocyte antigen in a sample, including that present in membrane and intracellular compartments and extracellular soluble compartments. Measurements of a total leukocyte antigen can be used to type cells, detect or diagnose disease or to monitor disease therapy. In a further embodiment, the invention relates to the measurement of both the amount of total leukocyte antigen and the amount of the soluble form of the leukocyte antigen and a comparison of the measured levels.

Abstract: The invention relates to a method of detection, a sensor and a test-kit which find application in immunological detection (e.g., immunoassay). The invention provides, inter alia, a method of detection, suitable for use in immunological detection of an entity, which method includes the use of a secondary species (as defined in the specification), the use of a first detectable species, and the use of a second detectable species. The method may include, for example, the use of a primary species, a secondary species, a first detectable species and a second detectable species. The primary species may be, for example, an antibody or a ligand. The secondary species may be, for example, an auxiliary species such as an auxiliary binder or an auxiliary ligand, or a species which has a part which is an auxiliary function. The entity to be detected may be an analyte species as such or may be an entity which carries or includes analytes species.

Abstract: Cross-linked antibody conjugates are described which have at least one interchain bridge containing a reporter or effector molecule. The bridge may be the residue of a homo-or heterofunctional cross-linking reagent, and is located away from the antigen binding domains of the antibody. The antibody conjugates have an enhanced binding capacity and in vivo have good blood clearance and, in the presence of a tumour high tumour; blood and tumour; bone ratios. The conjugates are of use in the diagnosis and therapy of e.g. tumours and may be prepared by reaction of a cross-linking reagent with an antibody.

Abstract: Methods for the high yield production of antibody Fv-containing polypeptides, especially Fab' and F(ab').sub.2 antibody fragments are provided. Expression of heavy and light chain Fv in a microbial secretory system is followed by recovery of Fv from the periplasm under conditions that maintain a cysteine residue as a free thiol. The free thiol is reacted with free thiol of an antibody fragment of the same or differing specificity, or with agents such as diagnostic labels or therapeutic moieties. The products offer advantages of homogeneity and purity not available through the use of known methods for preparing such derivatives.

Abstract: The assay off the present Invention is of particular use for detecting drugs, hormones, steroids, antibodies and other molecules circulating in the blood of a mammal or other animal.

Abstract: This invention relates to antibodies and is particularly, though not exclusively, concerned with diagnostic and therapeutic methods using monoclonal bi- or tri-specific antibodies. The invention also provides a method in which binding of a first antigen to a first antibody binding site causes release of a second antigen from an adjacent second antibody antigen binding site.

Abstract: The invention relates to artificial positive control reagents based on antibody conjugates that are used in immunochemical detection methods and to processes for the preparation of these reagents.

Abstract: There is described a positive calibrator/control composition for use in assays for the detection of antibodies to infectious disease agents. The composition includes a composite antibody of a nonspecific IgM immunoglobulin moiety covalently linked to a specific, non-IgM antibody moiety. Also described is an assay method which utilizes the positive calibrator or control composition.

Abstract: An enzyme-labelled antibody adapted for use in a homogeneous immunoassay is provided. The enzyme-labelled antibody is a conjugate of an enzyme with two or more different monoclonal antibodies, each of the monoclonal antibodies being capable of specifically recognizing and binding to a different epitope of the same antigen. By using the enzyme-labelled antibody in the homogeneous enzyme immunoassay process, an analyte can be quantitatively analyzed at a higher sensitivity through a simple operation. Also provided is a dry immunoassay element comprising an immunological reaction layer containing the enzyme-labelled antibody. By the provision of such an immunoassay element, a further simplified quick analysis of an analyte is realized to give an accurate result.

Abstract: The invention concerns a method of homogeneous immunoassay of a ligand in a liquid test sample which comprises: a)incubating a mixture of (1) the liquid test sample; (1i) where the ligand under assay has only one epitope, a covalent conjugate of the ligand with any mono- or poly-epitope molecule having at least one epitope distinct from the epitope of the ligand; (2) a labile enzyme; (3) a first bispecific monoclonal antibody protecting the activity of the encyme through binding to a first epitope on the encyme and capable of binding the ligand under assay; and (4) a second bispecific monoclonal antibody capable of bonding said enzyme and a second distinct epitope on the ligand under assay or an epitope of the said covalent conjugate; whereby a quaternary immunocomplex is formed; b) after incubation, subjecting the mixture to conditions whereby any enzyme not present in such a quaternary immunocomplex is inactivated; and c) determining the amount of detectable encyme, the determination being related to the am

Abstract: The present invention has multiple aspects. In its first aspect the present invention is directed to a trifunctional antibody-like compound that has tissue, organ, cell or molecule specificity and which is capable of being bifunctional when immobilized, via binding, at the tissue, organ, cell or molecule for which it has specificity. In particular, the present invention is directed to a trifunctional antibody-like compound of Formula I:F.sub.1 ab'--L--F.sub.2 ab'--L--F.sub.3 ab' (1)wherein L is the same or two different moieties for cross-linking F.sub.1 ab', F.sub.2 ab' and F.sub.3 ab';wherein F.sub.1 ab' is an Fab'-like fragment of a polyclonal or monoclonal antibody having specificity for an antigen expressed by the organ, tissue, cell or molecule of interest;wherein F.sub.2 ab' is an Fab'-like fragment of a polyclonal or monoclonal antibody having the same specificity as F.sub.

Abstract: A simultaneous dual analyte assay for determining the fertile period of the human menstrual cycle. The assay utilizes a capture reaction component consisting of P-3-G immobilized on a microporous membrane, a blocking reaction component consisting of anti E.sub.1 -3-G antibody, a labelled reaction component consisting of gold particle labelled anti E.sub.1 -3-G antibody, and an ambifunctional reaction component consisting of a hybrid immunoreactive substance having an anti P-3-G antibody binding site and a plurality of E.sub.1 -3-G determinant binding sites. An aqueous sample containing P-3-G and E.sub.1 -3-G is contacted with the components and the assay is calibrated to provide a positive assay result only when the concentration of P-3-G in the sample is less than a predetermined concentration and the concentration of E.sub.

Abstract: A rapid, competitive enzyme linked immunosorbent assay (cELISA) for the determination of Bluetongue virus antibodies in serum is described. This method utilizes either a biotinylated monoclonal antibody to Bluetongue virus and streptavadin-enzyme in conjunction with synthetic substrate, or an enzyme-conjugated monoclonal to detect antibodies specific for Bluetongue virus.

Abstract: The present invention provides cross-linked protein compositions consisting of two or more units of a target-specific protein joined by binding sulfhydryl groups on the target-specific protein units to a sulfhydryl-selective cross-linking agent and a method of making the compositions. These cross-linked protein compositions combine an increase in binding affinity due to the presence of multiple identical binding sites and stability to reduction conditions.

Abstract: A plasmid which contains a genetic code for an immunogenic portion which is conserved in many strains of nontypable Haemophilus influenzae and the bacterium containing this plasmid is disclosed. The immunogenic portion is preferably an epitope on an outer membrane protein of H. influenzae. A monoclonal antibody to the immunogenic portion and the hybridoma which will produce the monoclonal antibody is also included. The invention further includes a DNA probe constructed to correspond to the nucleic acids which code for the immunogenic portion. This probe may be labelled with a radioactive marker and may be used as a diagnostic tool to assay various clinical samples for the presence of H. influenzae.

Abstract: An antibody-enzyme conjugate is prepared having an enzyme to antibody ratio of approximately 3. The conjugate is produced by adding sulfhydryl groups to an antibody and maleimidyl groups to an enzyme to produce a modified antibody and enzyme, and reacting the modified antibody and enzyme to produce the conjugate. In producing the modified antibody and enzyme, about a 15 molar excess of reagents for introducing sulfhydryl and maleimidyl groups is used. When reacting the modified antibody and enzyme, a four molar excess of the modified enzyme is used, and reacting is stopped after a specified period of time by addition of selective reagents. The selective reagents may be cysteine and iodoacetamide.

Abstract: The present invention discloses, a bispecific monoclonal antibody to an ansamitocin derivative and a target antigen, particularly tumor-associated antigen, which can carry an anasamitocin derivative in a stable and inactive form at other sites than the target and release the active-form ansamitocin derivative at the target site, so that an anticancer agent having excellent durability and selectivity with little adverse action can be prepared using the bispecific monoclonal antibody and ansamitocin derivatives.

Abstract: A quantitative determination of a substance is performed in a homogeneous system based on a change in enzyme activity; differences between the activity of a free enzyme and that of an enzyme bound by an aggregation or chemical bonding are observed. A peroxidase-labeled antibody and antigen system is one of of the typical example. Since the reaction is effected in a homogeneous system, the amount of antigen can be easily measured by the difference of enzyme activity.

Abstract: New and useful chromophores have been isolated from the reaction mixture of proteins exposed to reducing sugars in the presence of sulfite over time. The chromophores are believed to be intermediates in nonenzymatic polypeptide glycosylation. The measurement of this chromophore makes possible both qualitative and quantitative assessment of the presence of nonenzymatic browning. Diagnostic and test kits are also disclosed.

Abstract: The present invention provides an enhancer DNA segment, a gene fragment having (a) the enhancer DNA segment and (b) a structural gene such as human D, V and J gene, and a promoter. A hybrid DNA (1) having a phage DNA fragment from Charon 4A phage vector and human DNA fragments, a Charon 4A phage containing the hybrid DNA (1), a hybrid DNA (2) having a PBR322 fragment and human DNA fragment, a transformant of E. coli C600 strain transformed with the hybrid DNA (2), plasmid pSV2-H1G1 obtained by inserting a human DNA fragment into a vector pSV-2gpt, a transformant of E. coli MC 1000 strain transformed with the plasmid pSV2-H1G1 and a transformant of mouse myeloma cells J558L or NS-1 transformed with the plasmid pSV2-H1G1 are also provided.

Abstract: A method is disclosed for conducting an assay for an analyte. The method comprises causing a specific binding pair member in a first aqueous medium to become bound to a first bibulous member by contacting a portion of the first bibulous member with the medium. The first bibulous member is in liquid receiving relationship with an absorbent member. The contacting is carried out under conditions wherein the first medium traverses the first bibulous member and at least a portion of the absorbent member by capillary action. The method further comprises causing the first bibulous member to come into liquid receiving relationship with a second bibulous member. A reagent in a second aqueous medium is absorbed by and preferably becomes a non-diffusively bound to the second bibulous member in relation to the presence of analyte in the first medium.