Abstract: Methods and compositions for membrane flow-through assays using substrate and enzyme conjugate is described. A preferred method of the present invention comprises providing E. coli alkaline phosphatase (ECAP) conjugate, with additional color enhancement provided by combining lithium salt with the substrate, calcium and/or magnesium salts with the conjugate, and polyalcohol polymers with the conjugate. Color development is further enhanced by spotting tetrazolium dyes, such as the Nitro Blue tetrazolium series, onto the membrane.

Abstract: The invention provides a method for determining the presence of products of conception in a sample derived from the uterus during a D&C, or a therapeutic or spontaneous abortion, and comprises determining the presence in the sample of a fetal restricted antigen, which is found in products of conception but not found in significant amounts in maternal plasma or serum. Since the fetal restricted antigen is not present in significant quantities in maternal plasma or serum, the methods of this invention are reliable even when the sample is contaminated with maternal blood. One fetal restricted antigen is fetal fibronectin.In one embodiment of this invention, the sample is contacted with an insoluble support to which anti-(fetal restricted antigen) antibody is adhered, and the fetal restricted antigen binding to the support is determined.

Abstract: A method for the detection of the presence of inflammation in a patient by measuring the amount of circulating intercellular adhesion molecule (cICAM-1) in a sample of one or more bodily fluids of the patient and then comparing the amount of cICAM-1 in the sample to standards normal for the bodily fluid or fluids assayed. The amount of cICAM-1 can be measured using anti-ICAM-1 antibodies. Higher than normal amounts of cICAM-1 indicate the presence of inflammation. Also contemplated is a method for the detection of organ transplant or tissue graft rejection.

Abstract: A method and apparatus for automatically and rapidly, retrieving, counting and/or analyzing at least one selected white blood cell population and/or subset thereof of a whole blood sample or portion thereof. A volume of a biological sample containing the white blood cells is prepared and at least one reactant specific or preferential at least to some selected biological cells is introduced thereto and rapidly mixed for a short period of time. The opacity and/or volume parameter of the cells can be modified and the mixture is then counted and analyzed in one or more steps to obtain the desired white blood cell population analysis.The biological sample can be a whole blood sample and the reactant can include or be a lyse or a monoclonal antibody bound to microspheres, which will bind to specific ones of the cells or a combination of lyse and microspheres with antibody bound thereto.

Abstract: Selected polyhydroxyl compounds, comprising monosaccharides, disaccharides and oligosaccharides, as well as certain of their derivatives, are effective in preventing the spontaneous activation of complement in vitro. The effect is enhanced in the presence of an anticoagulant together with a divalent cation chelator. Addition of these compounds makes it possible to store clinical samples at conventional temperatures from -20.degree. to +22.degree. C. for extended periods prior to performing complement protein assays. Effective compounds are selected through a screening protocol which comprises the use of immunoassays for complement fragments together with an algorithm for computing effectiveness. Polyhydroxyl compounds extend similar protection from activation to coagulation proteins.

Abstract: A method and a kit for testing a plant for a suspected antigen. A sample of the plant is taken and rubbed between first and second pads to extract sap from the sample and to collect the sap on the first pad. The sap is transferred to a liquid solution, and that solution is tested for the suspected antigen. This may be done by contacting the solution with a tag antibody and with a carrier having a capture antibody so that a tag antibody-antigen-capture antibody complex forms on the carrier if the suspected antigen is in the liquid solution. The carrier is then tested for the tag antibody. The kit preferably includes the first and second pads, the carrier, containers to hold the liquid solution and the tag antibody, and a device or reagent to test the carrier for the tag antibody.

Abstract: The present invention provides an immunoassay of general applicability. The assay detects an antigen by its inhibition of the reaction between the combining site of two antibodies. One of the antibodies binds antigen, and the second antibody binds to the combining site or idiotype of the first antibody. It is preferable that both the antibody that binds the antigen and the anti-idiotypic antibody are monoclonal antibodies. The assay provides a method for measuring concentrations of single epitopes without requiring purified antigen, and it is particularly useful for measuring the quantity of any nonpurified antigen present in low concentration in a mixture of antigens. The use of monoclonal antibodies as reagents is advantageous in providing an essentially unlimited supply of highly specific and standardized reagents, without the batch-to-batch variation encountered when using conventional polyclonal antibodies.

Abstract: There is provided a stable cholesterol assay composition which comprises an aqueous solution of at least one bile acid or salt thereof being present in an amount of up to about 5 mM; a nonionic surfactant present in an amount of from about 0.15 to about 1.5 percent volume by volume; a buffer in a concentration of from 0 to about 65 mM; and cholesterol oxidase in a concentration of at least about 0.1 KIU/l. Solution pH is from about 5.5 to about 8.5. Addition of cholesterol esterase, phenol, peroxidase and 4-aminoantipyrine provides a total cholesterol chromogen system.

Abstract: A method which assists in the early diagnosis of rejection in a liver transplant recipient comprises measuring an increase in plasma or serum alpha glutathione S-tranferase (.alpha.-GST) from said recipient in the absence of or preceding any change in plasma or serum transaminase. .alpha.-GST is most suitably measured by enzymeimmunoassay, using a solid phase antibody which is monospecific for .alpha.-GST. The monospecific antibody cross-reacts with the .alpha.-GST dimers B.sub.1 B.sub.1, B.sub.1 B.sub.2 and B.sub.2 B.sub.2.

Abstract: A novel chromogenic substrate to peroxidase enzymes is provided which is comprised of a mixture of an adduct formed from a hydrozone and a dienophile, and an aromatic nucleophile. The mixture undergos an oxidative coupling in the presence of peroxidase enzymes and peroxides forming a purple indamine dye. The mixture is also stable and unaffected by oxygen of the air or by hydrogen peroxide.

Abstract: Methods for detecting the propensity for an individual to be affected by a polyomavirus are disclosed. The methods include an assay wherein a biological specimen from a female is contacted with at least one probe capable of determining whether the female has been exposed to a polyomavirus. A method for prophylactically treating the female is also described.

Abstract: A process and medium are disclosed for the lyophilization of cells, specifically platelets, and cell-like matter, which comprises the use of solutions including monosaccharide hexoses and pentoses, and biocompatible amphipathic polymers to permit the reconstitution of transfusably useful cells, specifically platelets, and cell-like matter.

Abstract: The use of pyridoxal-5'-phosphate as an in vitro anticoagulant agent which retains the platelet activity of stored whole blood or stored plasma for more than about six hours is disclosed.

Abstract: Preservation of leukocytes by suspending in storage medium containing modified fluid gelatin, plasma, and a non-toxic buffer. Preferred additives include heterocyclic bases which occur in nucleic acids, or nucleosides or nucleotides containing the same.

Abstract: Biologically competent non-pasteurized albumin wherein virus present in the source fluid has been inactivated with one or more of a class of compounds exemplified by glycyrrhizin, glycyrrhizinic acid or glycyrrhetinic acid glycoside, and analogous triterpenes, e.g. carbenoxolone and cicloxolone and their derivatives, and blood substitutes comprising such albumin and hemoglobin are disclosed.

Abstract: The new hybridoma cell lines RF-HBs-1, RF-HBs-2 and RF-HBs-4 each secrete a nonoclonal antibody to hepatitis B surface antigen. The production of the antibodies may be carried out in vitro by culturing one of the cell lines or in vivo by establishing one of the cell lines as an ascites tumour in a mouse and isolating antibodies from the ascites fluid or from the serum. The antibodies have therapeutic, preventative and diagnostic uses in respect of hepatitis B virus infections and can be used to purify hepatitis B surface antigen. The relative specificities of the three monoclonal antibodies make them particularly useful in radiometric assay techniques employing specific combinations of the antibodies in solid phase.

Abstract: Methods are provided for preparing just before administration unit doses of therapeutic solutions which contain redox active unstable and/or diffusable metabolites such as a ketoacid, a sulfhydryl-contining amino acid, or carbon dioxide. The method involves preparing and storing an aqueous solution of stable components which may or may not contain carbon dioxide. A dry powder comprised of unstable components is also prepared and stored separately. These separate component compositions are packaged in, for example, individual chambers of a common scaled container which is so constructed as to permit the opening, by externally applied manual means or the like, of a passageway between such chambers at the time when usage is contemplated. Thus, a fresh solution in desired full dosage form is preparable just befor administration. Improved container structures for practice of this method are also provided.

Abstract: Diagnosis of insulin-dependent (Type I) diabetes mellitus (IDDM) by contacting a blood sample from a patient with an immunoreagent comprising epitopes of two or more of glutamic acid decarboxylase (GAD) and the pancreatic islet cell antigens referred to as ICA512 and ICA12. Binding of antibodies present in the blood sample with one or more of such epitopes correlates with IDDM or a potential for developing IDDM. In clinical testing, about 80 percent of sera from newly diagnosed IDDM patients react positively with at least one epitope in a GAD/ICA512 panel. Reactivity with the GAD/ICA512/ICA12 panel is between about 80 and 90 percent. The method is useful in screening patients for pre-IDDM, for distinguishing IDDM from Type II diabetes, and for monitoring therapy.

Abstract: The present invention relates to an in vitro method to determine the safety of modified hemoglobin blood substitutes for human subjects prior to their clinical usages, wherein the method is based on complement activation reaction from adding modified hemoglobin blood substitutes to a human plasma sample and comprises the steps of: a) obtaining at least one plasma sample from at least one human subject by i) taking a blood sample and immediately centrifugating; and ii) separating the centrifuged blood sample of step i) and retaining the supernatant plasma; b) mixing the plasma of step ii) with the modified hemoglobin blood substitutes or control-ringer in a weight/volume ratio of about 4:1; c) incubating for a time sufficient to allow for a complement activation reaction to occur; d) adding the product of step c) to an appropriate volume of saline in an EDTA tube; and e) analyzing the degree of complement activation by analysis of the product of step d); thereby determining the safety of the modified hemoglobi

Abstract: A reagent for proteolytic enzyme assays has the general formula ##STR1## where RCO-- is an enzyme reactive acyl, such as an amino acid, peptide or substituted amino acid or peptide. The reagent may be hydrolysed by proteolytic enzymes and developed to form a distinctive color. The reagent may be formed by reacting RCOOH with N-hydroxysuccinimide to form the acyl N-hydroxysuccinimide ester. The ester may then be reacted to form the reagent.