Abstract: A leukocyte measurement device is provided with a spread inspection window at a position separated from an opening part along a spreading direction of a blood-derived sample in a carrier, for making the carrier visible from outside. When the blood-derived sample is dropped from the opening part, leukocytes contained in the blood-derived sample are captured by the carrier in the vicinity of a dropping position, while erythrocytes and/or hemoglobin and the like spread through the carrier from the dropping position into a predetermined direction. Therefore, in the leukocyte measurement device, seeing through the spread inspection window the color of the erythrocytes and/or hemoglobin spread from the dropping position can determine whether it has already been used or not, whereby a simple structure can prevent measurers from erroneously reusing it.

Abstract: The invention relates to a method for diagnosing a disease state mediated by pathogenic cells. The method comprises the steps of combining with an ex vivo patient sample a composition comprising a conjugate or complex of the general formula Ab-X, wherein the group Ab comprises a ligand that binds to the pathogenic cells and the group X comprises an imaging agent, and detecting the pathogenic cells that express a receptor for the ligand using flow cytometry.

Abstract: The present invention provides the amino acid and polynucleotide sequences of the transcobalamin receptor, as well as modulators of the transcobalamin receptor. Accordingly, the present invention provides compositions and methods for the treatment and prevention of diseases and disorders associated with cobalamin deficiency, including compositions and methods that promote cobalamin uptake. In addition, the present invention provides compositions and methods for the detection, treatment, and prevention of diseases associated with deregulated cell growth, including, e.g., cancer and autoimmune disorders, including compositions and methods that inhibit cobalamin uptake.

Abstract: Disclosed are immunoassay methods for the diagnosis/prognosis of diseases and disease susceptibility traits associated with gene mutations that cause protein truncation or allelic loss. The levels of one or more targeted wild-type proteins expressed by a subject gene or genes are immunologically quantitated in biological samples. Results indicating that a targeted wild-type protein is not present in an assayed sample, or that approximately 50% of the normal amount of such a wild-type protein is present in an assayed sample are considered to be positive for a mutation in one or both alleles of a subject gene, and correlated with the disease or the disease susceptibility trait associated with that mutation or mutations. Normal cells, particularly normal peripheral blood lymphocytes, are preferred biological samples.

Abstract: Disclosed are immunoassay methods for the diagnosis/prognosis of diseases and disease susceptibility traits associated with gene mutations that cause protein truncation or allelic loss. The levels of one or more targeted wild-type proteins expressed by a subject gene or genes are immunologically quantitated in biological samples. Results indicating that a targeted wild-type protein is not present in an assayed sample, or that approximately 50% of the normal amount of such a wild-type protein is present in an assayed sample are considered to be positive for a mutation in one or both alleles of a subject gene, and correlated with the disease or the disease susceptibility trait associated with that mutation or mutations. Normal cells, particularly normal peripheral blood lymphocytes, are preferred biological samples.

Abstract: An isolated nucleotide sequence or fragment thereof encoding the porcine intrinsic factor, wherein the porcine intrinsic factor comprises an amino acid sequence having at least 85% amino acid sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 6, and SEQ ID NO: 9. The invention also encompasses an isolated nucleic acid sequence or fragment thereof comprising, or complementary to, a nucleotide sequence having at least 85% nucleotide sequence identity to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 4, and SEQ ID NO: 7. The porcine intrinsic factor can be use is an assay to determine the quantity of vitamin B12 in a biological sample.

Abstract: Provided are methods and compositions for assaying for ubiquitin agents that are enzymatic components of ubiquitin-mediated proteolysis and, more particularly, methods and compositions for assaying for agents that modulate the activity of such ubiquitin agents.

Abstract: An assay method for determining transcobalamin saturation wherein a transcobalamin containing liquid sample is contacted with a porous substrate with immobilized thereon a transcobalamin immobilizing ligand and with a reporter-labelled transcobalamin binding partner and wherein signals from reporter labels which become immobilized on said substrate are detected, characterised in that one of same ligand or said binding partner comprises a first ligand or binding partner capable of specific binding to holo transcobalamin and a second ligand or binding partner capable of binding to apo transcobalamin or to holo and apo transcobalamin.

Abstract: Disclosed are immunoassay methods for the diagnosis/prognosis of diseases and disease susceptibility traits associated with gene mutations that cause protein truncation or allelic loss. The levels of one or more targeted wild-type proteins expressed by a subject gene or genes are immunologically quantitated in biological samples. Results indicating that a targeted wild-type protein is not present in an assayed sample, or that approximately 50% of the normal amount of such a wild-type protein is present in an assayed sample are considered to be positive for a mutation in one or both alleles of a subject gene, and correlated with the disease or the disease susceptibility trait associated with that mutation or mutations. Normal cells, particularly normal peripheral blood lymphocytes, are preferred biological samples.

Abstract: Processes for removing endogenous Vitamin B12 and folate are provided. The albumin preparations produced by the process are suitable for use as reference control samples in assays for determining these analytes in a test sample. Kits and methods related to the albumin preparations are also provided.

Abstract: The present invention relates to folic acid-polysaccharide complexs and method of preparation thereof, more particularly relates to folic acid-Dextran complexs, method of preparation thereof, pharmaceutical compositions having said complex as active component and uses of said composition in therapy of tumors. The folic acid-polysaccharide complexs of the present invention have general formula of: (X)n—Y, wherein X is identical or different, and is selected from folic acid, derivatives of folic acid and other substances that can enter into cell via the pathway of folic acid receptor; Y is polysaccharide; n?1.

Abstract: The present invention relates to fluorescent cobalamins and uses of these compounds. More particularly, this invention relates to fluorescent cobalamins that comprise a fluorescent, phosphorescent, luminescent or light-producing compound covalently linked to cobalamin. These fluorescent cobalamins can be used to as diagnostic and prognostic markers (a) to distinguish cancer cells and tissues from healthy cells and tissues, including identifying lymph nodes containing cancer cells, and (b) to determine if an individual will respond positively to chemotherapy using cobalamin-therapeutic bioconjugates.

Abstract: This invention relates to binding protein assays. In particular, this invention relates to binding protein assays for B12 and folate in serum or plasma. More specifically, this invention provides a sequential assay that uses a combination of specific binding proteins, and anti-binding protein antibodies to measure B12 and folate in serum or plasma.

Abstract: Vitamin B.sub.12 receptor modulating agents capable of modulating cell surface receptors by affecting the cell surface receptor trafficking pathway are disclosed. The vitamin B.sub.12 receptor modulating agents are comprised of a covalently bound rerouting moiety and targeting moiety linked by a water-solublizing linker.

Abstract: Methods for detecting schizophrenia or depression based on modifications of the contribution of the D.sub.4 receptor to phospholipid methylation levels are described herein. Individuals with schizophrenia or depression have a deficiency in phospholipid methylation activity compared with normal individuals. Methods for screening therapeutic processes or agents for use in treatment of schizophrenia or related neuropsychiatric disorders are also described.

Abstract: Methods and compositions are provided for specific binding assays in which specific binding reagents are immobilized on a solid phase. Immobilization is facilitated by covalently coupling specific binding assay reagents such as polypeptide receptors or analytes with water soluble polymers. Such water soluble polymers, for example star polymers such as dendrimers, provide production advantages of lot-to-lot uniformity and homogeneity, and can enhance sensitivity due to low non-specific binding to the solid phase.

Abstract: A composition is described which is useful in the assay of folate from a hemolysate prepared from red blood cells using non-radiolabeled assay techniques such as fluorometric, colorimetric, enzymatic or chemiluminescent assay methods. Methods to ready hemolysates for the assay of folate are also described.

Abstract: An IgM class monoclonal antibody which specifically reacts with mucus glycoproteins produced by human gastric gland-type mucous cells is provided. By performing immunohistochemical staining using a labeled derivative of the monoclonal antibody, human gastric gland-type mucous cells as well as mucus secreted by these cells can be specifically stained. The monoclonal antibody can also be used for the analysis of gastric gland-type mucous cell-derived mucus glycoproteins in human body fluids as well as for the examination or diagnosis of cancer.

Abstract: This invention is directed to the novel assay methods utilizing nucleophilic polysubstituted aryl acridinium ester conjugates as the tracers. Conjugates prepared by covalent coupling of novel nucleophilic polysubstituted aryl acridinium esters with biological compounds including small organic molecules such as Vitamin B12, folate, cortisol, estradiol, and thromboxane B2, were found useful in the development of highly sensitive assays for the analytes of diagnostic interest.

Abstract: Immunoassays for vitamin B.sub.12 using novel monoclonal antibodies to the intrinsic factor:vitamin B.sub.12 complex and to the vitamin B.sub.12 binding site on intrinsic factor.

Abstract: An improved method for performing immunoassays whereby specific binding proteins for vitamin B12, folate and other target analytes are utilized with antibodies with different specificities for the binding proteins. Antibodies bridge the specific binding protein directly or indirectly to a capturable material.

Abstract: A method for purifying an aqueous intrinsic factor solution which contains R-protein is disclosed. The method involves adding to the intrinsic factor solution an amount of colloidal silica to disperse lipid emulsion, an amount of cobinamide sufficient to bind substantially all of the R-protein in the solution and an amount of an intrinsic factor affinity resin sufficient to bind the intrinsic factor in the solution, washing the bound cobinamide and the R-protein from the resin, eluting the intrinsic factor from the resin, and dialyzing the eluted intrinsic factor. The purified intrinsic factor possesses less than 0.004 percent cross reactivity with cobinamides, and at least 95 percent of the proteins in the purified material can bind cobalamins. A conjugate of microparticles and the purified intrinsic factor is also disclosed, as is a kit for conducting an assay for cobalamins which includes a conjugate of microparticles and purified intrinsic factor.

Abstract: Denaturing agents used to pretreat test samples for immunoassays are neutralized and deactivated by the addition of a treatment agent which is a combination of a buffering agent in acid form and an agent for converting the sulfhydryl groups of the denaturing agents to a non-active form such as disulfides or alkylthio groups. The treatment agent serves this function without having sufficient denaturing activity by itself to interfere with subsequent steps of the assay procedure. Also disclosed is a protocol for a vitamin B.sub.12 assay which involves a sequential rather than competitive binding of the B.sub.12 with excess intrinsic factor followed by the binding of excess enzyme-labeled B.sub.12 to any remaining intrinsic and immobilizing such intrinsic factor on a solid phase. Various problems associated with B.sub.12 assays are avoided by this technique.

Abstract: The current invention is a method, kit and reagents for detecting cobalamins in a sample. The method involves introducing into the sample a first conjugate of a latex or latex-like solid phase linked to affinity purified intrinsic factor with a first linking group at least three angstroms long. The first conjugate and the bound cobalamins in the sample are then exposed to a second conjugate of a cobalamin linked to a detectable enzyme to produce second conjugate bound to first conjugate and unbound second conjugate. The enzyme activity associated either with the solid phase first conjugate or the unbound second conjugate is then detected. By linking affinity purified intrinsic factor to a latex or latex-like solid phase with a first linking group at least three angstroms long, an exzyme assay can be performed which detects levels of cobalamins in patient samples within or below normal ranges found in such samples.

Abstract: A method is provided for combining normally interacting reagents in a liquid single reagent and preventing complex formation using a surfactant and then reversing the inhibition by adding a cyclodextrin. The method finds particular use in diagnostic immunoassays. Reagents facilitating the invention are also provided.

Abstract: Compositions and methods are disclosed for multiple simultaneous assays of different analytes using radioactive labeled antibodies to the analytes, at least one portion of the assay being an immunoradiometric assay in which there is employed a metal isotope label, e.g., .sup.57 Co, attached to an antibody to the analyte through a chelator, e.g., ethylenediaminetetraacetic acid. Multiple simultaneous immunoradiometric assays can be performed by this method, as can multiple simultaneous assays in which one portion of the assay is an immunoradiometric assay and another portion or portions involve one or more other radioassay techniques.

Abstract: Serum samples for vitamin B.sub.12 analysis can be pretreated using a combination of peroxy acid and dithiothreitol. The pretreatment makes vitamin B.sub.12 bound to serum binding proteins available for analysis by any of a variety of currently available assay techniques. The pretreatment finds particular use in an assay using solid phase intrinsic factor as a specific binding protein.

Abstract: The present invention is directed to a method of quantitative assay for a vitamin B.sub.12 -containing substance which comprises culturing marine methanol-utilizing bacteria having B.sub.12 auxotrophy in a medium for quantitative assay of the bacteria containing pyrroloquinoline-quinone and a polyoxyethylene sorbitan fatty acid ester and, quantitatively determining vitamin B.sub.12 in the vitamin B.sub.12 -containing substance as a function of the degree of growth thereof and, to a reagent for the B.sub.12 -containing substance comprising a kit for combination of an ampule or vial having sealed therein a dry viable bacteria composition of a marine methanol-utilizing bacteria for inoculation and an ampule or vial having sealed therein a medium for the bacteria containing pyrroloquinoline-quinone and a polyoxyethylene sorbitan fatty acid ester, or in addition thereto, further an ampule or vial having sealed therein a standard dilution of vitamin B.sub.12 having a serial concentration.

Abstract: A method for enzyme immunoassay includes contacting under binding conditions a liquid suspected of containing an analyte, an antianalyte affixed to a solid support and a tracer having an enzyme conjugated thereto. A bound fraction is separated from the liquid and incubated in a second liquid with a masked ligand. The masked ligand is converted by the enzyme on the bound fraction to give free lignad which binds to an antiligand. A signal system, such as a signal enzyme and substrate therefor, or a label-loaded vesicle and vesicle lysing agent, is added to generate a signal used to detect or measure the analyte in the liquid. The invention includes a kit of materials useful in performing the assay of the invention.

Abstract: A biological sample containing an analyte, partly free and partly bound to natural binders, is incubated with a labelled specific binder for the analyte and a derivative of the analyte which does not react with the natural binders. Part of the labelled specific binder binds to the analyte derivative, the proportion depending on the free analyte concentration in the sample. The proportion is measured and used to determine the free analyte concentration. For example, the analyte may be thyroxine or cortisol, the labelled specific binder may be an antibody thereto, and the analyte derivative may be in the form of a solid matrix. The labelled specific binder should have an affinity for the analyte, expressed as a dissociation constant of the complex of the two, approximately equal to the concentration of free analyte in the sample.

Abstract: A serum folate assay wherein the serum folate is stabilized with dimercaptosuccinic acid or thioctic acid.

Abstract: The present invention provides a method for detection of protozoan parasites in blood or other specimen from their mammalian hosts. The method comprises nucleic acid hybridization of repetitive nuclear DNA fragments of the parasites. Hybridization probes have been prepared for this purpose by cloning the repetitive nuclear elements that are species-specific in appropriate vectors. The sensitivity of these probes has been increased by further sub-cloning to make them capable of cascade hybridization. The assay is highly specific and sensitive for detection of disease-causing protozoan parasites of the commonly occurring Trypanosomatidae of the genus Leishmania and genus Trypanosoma, as well as for malaria-causing protozoan and other parasitic microorganims of mammals.

Abstract: An assay for Vitamin B.sub.12 deficiency by determining for a blood sample that essentially all of the Vitamin B.sub.12 is carried by a combination of TCI and TCIII and/or essentially no Vitamin B.sub.12 is carried by TCII. TCII may be conveniently separated from a combination of TCI and TCIII by precipitating TCII with silica. Vitamin B.sub.12 deficiency may be determined prior to the onset of anemia and/or nerve damage by determining a decrease in Vitamin B.sub.12 carried by TCII.

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: This invention relates to a highly-sensitive mini-iodinated radioactive hormone tracer which has both initial low iodine-to-hormone molar ratios and low specific activity. This invention also relates to methods for preparing and using such tracers in a radio-immunoassay which takes much less time than used heretofore and to a method of preparing said tracers.

Abstract: A continuous hybridoma cell line which secretes recoverable quantities of monoclonal antibodies having specificity against Vitamin B.sub.6, which antibodies are useful in a method for detecting the presence of vitamin B.sub.6 in an animal sample.

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: An improved competitive protein binding assay comprising; incubating an analyte in the presence of labeled analyte and a protein binder suitable for competitive binding activity by the analyte and the labeled analyte to provide a mixture having free analyte, free labeled analyte, bound analyte and bound labeled analyte, substantially separating the bound analyte and the bound labeled analyte from the free analyte and free labeled analyte to form a first fraction containing substantially the bound analyte and the bound labeled analyte and a second fraction containing substantially the free analyte and the free labeled analyte by causing a predetermined level of the mixture to flow through a bed of an organo-silane coupled to silica gel, and detecting the labeled analyte of at least one of the first and second fractions to determine the concentration of the analyte by comparison to a reference.

Abstract: Vitamin B.sub.12 (cobalamin) and/or folate (folic acid) assay techniques can be simplified, by eliminating the step of heating or boiling the to-be-tested sample prior to its assay. This is accomplished by utilizing compositions and procedures:(1) which substantially completely liberate all vitamin B.sub.12 and/or folate from endogenous binding protein without heating or boiling;(2) which substantially completely destroy all endogenous binding protein which is present in the to-be-assayed sample, and which may bind natural and radioactive vitamin B.sub.12 or folate both due to its liberation from vitamin B.sub.12 or folate in the original sample or due to its unbound presence in the original sample;(3) which substantially completely inhibit or block any undestroyed endogenous binding protein; and(4) which substantially completely inhibit or destroy any intrinsic factor-blocking antibodies which may be present in the to-be-assayed sample.

Abstract: Vitamin-containing multicomponent compositions are assayed to determine the extent of conversion of the vitamin to one or more analogues thereof due to interaction between the vitamin and the other components of the composition, employing a radioactive labeled form of the vitamin. The assay is useful for quality control in the manufacture of multivitamin and multivitamin-mineral formulations and vitamin-supplemented foods intended for human or animal ingestion.

Abstract: A method of assaying for vitamin B12 achieves denaturation of serum binding proteins which normally bind the vitamin B12 by treatment with alkali at ambient temperature. The assay is characterized by the use, in the alkali denaturation step, of a dithiopolyol and cyanide, and by the use, during the intrinsic factor assay step, of a vitamin B12 analogue such as cobinamide to bind with any remaining serum proteins. The invention also includes a kit, in which the dithiopolyol is provided in admixture with the alkali.

Abstract: A serum sample is heated at an alkaline pH to release folate and vitamin B.sub.12 from endogenous binders. A simultaneous radioassay for folate and vitamin B.sub.12 is effected by contacting the sample with binder for folate, binder for vitamin B.sub.12, folate labeled with one radioactive isotope and vitamin B.sub.12 labeled with another radioactive isotope, followed by separation of bound and free portions, and determination of the radioactivity of at least one of the portions. The amounts of folate and vitamin B.sub.12 present in the sample may be determined from standard curves.

Abstract: Competitive protein binding radioassay for sera (or cell) vitamin B.sub.12 and/or serum folate (target components, or analytes) or other target components in liquid ambient utilizing a highly alkaline (pH 12-14) environment and reducing agent for denaturing separating the target component(s) from serum without boiling, consistent with other requirements of such assays, then reducing pH to an 8-10 range for effecting the competitive protein binding after which protein bound and unbound groups of radioactively tagged replicates of the target component(s) can be separated and detected to determine content of the target component(s) in the original serum (or cell), i.e. original endogenous analyte(s).

Abstract: Improvement in a competitive protein binding assay for N-5-methyltetrahydrofolic acid wherein any folate standard used is N-5-methyltetrahydrofolic acid complexed with folate binder protein. The protein of the complex is destroyed prior to the addition of immobilized folate binder protein. The immobilized folate binder protein participates in competitive binding with unknown folate and labeled folate reagents.

Abstract: The performance of double receptor, specific binding assays is improved by use of a receptor complex having the structureA.sub.BL (BL).sub.n A.sub.1wherein BL is a binding ligand, A.sub.BL is a receptor specific for binding ligand, A.sub.1 is a receptor, BL is covalently bonded to A.sub.1 and A.sub.BL is reversibly bonded to BL. Generally A.sub.BL is absorbed onto an insoluble surface and A.sub.1 is an antibody to the substance being assayed. The complex has particular utility in coated tube and rechargeable radioimmunoassay systems.