Abstract: Concentrated sugar solutions obtained from polysaccharide enriched biomass by contacting biomass with water and at least one nucleophilic base to produce a polysaccharide enriched biomass comprising a solid fraction and a liquid fraction and then contacting enriched biomass with a dilute mineral acid selected from the group consisting of sulfuric acid, phosphoric acid, hydrochloric acid, nitric acid, or a combination thereof, to produce an intermediate saccharification product, which is contacted with an enzyme consortium to produce a final saccharification product comprising fermentable sugars.

Abstract: A method of detection of cells, microorganisms, or molecules by the use of various combinations of fluorogens and a chromogens which yield fluorophores and chromophores when cleaved by specific enzymes and which can be viewed by UV and visible light. Included is the method of application of a family of compounds producing both insoluble fluorophores and chromophores identified as dual enzyme substrates.

Abstract: The present application relates to procedures for the determination of the activity of the protease which activates factor VII, also known as factor VII activating protease or FSAP. The application also relates to a method of detecting whether an individual has increased or lowered activity in the protease which activates factor VII compared to at least one standard sample, wherein the increased or lowered activity indicates an increased risk for disease or cardiovascular complications.

Abstract: The immunoassay element for quantitatively analyzing an antigen by determining the change in enzymatic activity of an enzyme-labelled antigen or antibody caused by an immunological reaction. The immunoassay element comprises a substrate layer containing a non-diffusible substrate which forms a diffusible material in the presence of the labelling enzyme, and a reagent layer containing a fragmenting enzyme for further fragmenting the diffusible material into a lower molecular weight product. As the non-diffusible substrate, a substrate capable of reacting solely with the labelling enzyme and incapable of reacting the fragmenting enzyme is utilized. When an endo-active glucosidase is used as the labelling enzyme, and an exo-active glucosidase is used the fragmenting enzyme in the reagent layer, the non-diffusible substrate of the substrate layer is preferred to be an endo type selectively reactive substrate, which means a substrate having a reactivity specific to endo-active glucosidase.

Abstract: A sensor is provided including a polymer capable of having an alterable measurable property from the group of luminescence and electrical conductivity, the polymer having an intermediate combination of a recognition element, a tethering element and a property-altering element bound thereto and capable of altering the measurable property, the intermediate combination adapted for subsequent separation from the polymer upon exposure to an agent having an affinity for binding to the recognition element whereupon the separation of the intermediate combination from the polymer results in a detectable change in the alterable measurable property, and, detecting said detectable change in the alterable measurable property.

Abstract: A multiplexed enzyme assay method and substrate set for performing the method are disclosed. The method includes performing a plurality of enzyme reactions in the presence of a plurality of enzymes substrates, under conditions effective to convert an enzyme substrate to a corresponding product, where the product of each substrate and the substrate have different separation characteristics from each other and from the other substrates and their corresponding products. After performing the reactions, which may be carried out in separate or combined reactions, the substrates and products in said reactions are separated in a single separation medium. For each separated product and substrate, a separation characteristic effective to identify that product and substrate and a signal related to the amount of the product and substrate is detected. From this, one can determine the amount of substrate converted to the corresponding product in each of the reactions.

Abstract: A sensor element is provided including a polymer exhibiting a measurable property from the group of luminescence and electrical conductivity, the polymer being complexed with a unit including a recognition element, a tethering element and a property-altering element bound thereto so as to alter the measurable property, the unit being susceptible of subsequent separation from the polymer upon exposure to an agent having an affinity for binding to the recognition element whereupon the separation of the unit from the polymer results in a detectable change in the measurable property.

Abstract: The present invention provides a simple method for the determination of a specific component, e.g. 1,5-anhydroglucitol (1,5-AG) in a sample containing glucose, and a reagent and a reagent kit useful in the method. In one embodiment, a method for the determination of 1,5-AG is provided which comprises contacting the sample with an enzyme system which converts glucose into fructose-1,6-diphosphate and converts 1,5-AG into 1,5-AG-6-phosphate to form 1,5-AG-6-phosphate, dehydrogenating 1,5-AG-6-phosphate in the sample by the action of 1,5-AG-6-phosphate dehydrogenase in the presence of an oxidized coenzyme, and measuring the amount of the reduced coenzyme formed by the dehydrogenation reaction. A reagent and a reagent kit useful in this method are also provided.

Abstract: The present invention provides a simple method for the determination of a specific component, e.g. 1,5-anhydroglucitol (1,5-AG) in a sample containing glucose, and a reagent and a reagent kit useful in the method. In one embodiment, a method for the determination of 1,5-AG is provided which comprises contacting the sample with an enzyme system which converts glucose into fructose-1,6-diphosphate and converts 1,5-AG into 1,5-AG-6-phosphate to form 1,5-AG-6-phosphate, dehydrogenating 1,5-AG-6-phosphate in the sample by the action of 1,5-AG-6-phosphate dehydrogenase in the presence of an oxidized coenzyme, and measuring the amount of the reduced coenzyme formed by the dehydrogenation reaction. A reagent and a reagent kit useful in this method are also provided.

Abstract: A chemiluminescent assays for the determination of the presence or amount of a biopolymer in bound assays using 1,2-dioxetanes in connection with AttoPhos™ as chemiluminescent substrates for enzyme-labeled targets or probes is provided. Further disclosed is a kit for conducting a bioassay for the presence or concentration of a biopolymer comprising a) an enzyme complex; b) a 1,2-dioxetane; and c) AttoPhos™.

Abstract: An analytical element for quantitative analysis of glucose, cholesterol, or lactic acid in whole blood, composed of a porous layer which has a void volume of 30 to 80% and containing peroxidase, a leuco dye which gives coloring having an absorption peak in the region of 600 to 700 nm, and an oxidase such as glucose oxidase, cholesterol oxidase, or lactate oxidase, respectively. The porous layer can be placed on a transparent support sheet via an adhesive layer in the form of dots or lines, or a hydrophilic adhesive layer.

Abstract: A method is provided for determining in a medium the presence or amount of an analyte which is capable of binding to a ligand partner to form a ligand complex, which method comprises: (a) contacting the medium with either: (i) a ligand partner conjugated with an enzyme being capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, or (ii) a ligand partner and either a competing analyte or an analog of said analyte, a competing analyte or analyte analog being capable of forming a ligand complex with the ligand partner and the competing analyte or analyte analog being conjugated with an enzyme capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, (b) optionally separating complexed and uncomplexed enzyme conjugates, (c) causing or allowing the reaction or cascade of reactions to occur to produce hydrogen peroxide by contacting the complexed or uncomplexed enzyme conjugate with a corresponding enzyme su

Abstract: A spectrophotometric assay for the detection of acetaminophen in aqueous fluids can be carried out with a dry analytical element. The element comprises a support having thereon one or more reagent layers containing a first enzyme, aryl acylamidase, to cleave the amide bond of acetaminophen to produce p-aminophenol; a second enzyme, ascorbic acid oxidase, to oxidize the p-aminophenol so that it couples to a water soluble coupling agent to form a dye that is read at 670 nm. The assay is precise, accurate on serum and plasma samples, and relatively free from significant interferences. The element also allows measurement over a broad dynamic range.

Abstract: A reagent composition comprising (a) liposomes encapsulating a marker therein, immobilizing a hapten on liposome membranes and having a size in the range of 100 to 500 nm in terms of mean particle size plus twice the standard derivation, and (b) an antibody to the hapten is effective for measuring human complement activity easily and precisely with excellent storage stability.

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: The invention relates to an ammonia elimination reagent for use in an enzymatic assay of a biological substance which produces ammonia as the reaction product. The reagent is an alkaline solution of pH 9 to 11 and contains Sulfolobus-derived thermostable isocitrate dehydrogenase, 2-oxoglutaric acid, a reducing coenzyme, isocitric acid, glutamate dehydrogenase, and a divalent metal salt. The reagent can be stored in solution and has excellent stability under conditions of alkaline pHs.

Abstract: A test kit and method for the highly sensitive detection of specific analytes in a sample is provided. The presence of the analyte in the sample results in a decrease in the concentration of a growth inhibiting substance leading to proliferation of cells in the region of the analyte. The presence or absence of the analyte is determined by detecting the presence of increased numbers of cells. Assay sensitivity is accounted for by the exponential amplification of cell number that occurs during cell proliferation in the presence of analyte.

Abstract: Disclosed herein is a cell containing a modified peptide. More specifically, the N-terminal amino acid residue of the peptide is modified by the addition of an aryl ketone group which, when contacted with an appropriate substrate, and exposed to light having a wavelength of about 330 nm or greater, results in the covalent bonding of the peptide to the substrate by a C--H insertion dominant mechanism. In preferred, embodiments, the aryl ketone is a benzophenone moiety. The peptide can be designed to specifically bind to a protein of interest in the cell. The cell is then contacted with light having a wavelength of greater than about 330 nm to bind the peptide covalently to the binding site on the intracellular protein of interest. In this way, the modified peptide can be used to specifically and irreversibly block a binding site on an intracellular protein of interest.

Abstract: A chemiluminescent assay method, compositions, kits and chemiluminescent acridan compounds are described which use a two-step chemiluminescent reaction process. The reaction involves an acridan compound, preferably a derivative of an N-alkylacridan-9-carboxylic acid, which undergoes a reaction with a peroxide compound, a peroxidase enzyme and an enhancer under conditions of time, temperature and pH which permit the accumulation of an intermediate compound, which is subsequently induced to produce a burst of light by raising the pH. The result is generation of very high intensity light from the reaction. The peroxidase enzyme is present alone or linked to a member of a specific binding pair in an immunoassay, DNA probe assay or other assay where the hydrolytic enzyme is bound to a reporter molecule. The method is particularly amenable to automated assays because of the separation of the incubation and light generating steps.

Abstract: A chemiluminescent assay method, compositions and kits are described which use a protected phenolic enhancer compound which is deprotected by a hydrolytic enzyme and then enhances a chemiluminescent reaction. The reaction involves an acridan compound, preferably a derivative of an N-alkylacridan-9-carboxylic acid, which is activated to produce light by a peroxide compound and a peroxidase enzyme in the presence of the deprotected enhancer. The result is enhanced generation of light from the reaction. The hydrolytic enzyme is present alone or linked to a member of a specific binding pair in an immunoassay, DNA probe assay or other assay where the hydrolytic enzyme is bound to a reporter molecule.

Abstract: An assay for photometrically detecting and/or quantitating the presence of an analyte in a sample in which the signal generated by a label associated with the analyte is photometrically detected in the presence of a suspended solid support.

Abstract: The subject invention provides an antibody which specifically binds to the product of a reaction between a labeling substance and a substrate. The subject invention also provides a method of making an immunogen used to produce the antibody of the subject invention. The invention further provides methods of using the subject antibody for detecting an antigen of interest in a sample, for example detecting a protein comprising an amino acid sequence of interest and detecting a nucleic acid molecule comprising a nucleic acid sequence of interest.

Abstract: Disclosed is a method for the quantitative determination of D-3-hydroxybutyric acid and acetoacetic acid, which comprises reacting a biological sample containing D-3-hydroxybutyric acid and acetoacetic acid, with a reagent comprising: (1) a D-3-hydroxybutyrate dehydrogenase, (2) A.sub.1 and (3) B.sub.1, the components (1), (2) and (3) participating in the following cycling reaction: ##STR1## thereby effecting the enzymatic cycling reaction, and measuring a change in the amount of A.sub.2 formed or the amount of B.sub.1 consumed. Also disclosed is an analytical reagent comprising the components (1), (2) and (3) for use in the above method. The method and the analytical reagent ensure rapidness and accuracy in the determination of D-3-hydroxybutyric acid and acetoacetic acid, even with the use of a small quantity of a biological sample, so that they are very useful in application fields, such as clinical diagnosis and food testing.

Abstract: A chromogenic assay for determination of blood coagulation Factor IX (christmas factor) utilizes Factor Xa formed by the conversion of Factor X by activated Factor IX and Factor VIII to cleave a chromogenic substrate. The sample is combined into a mixture with Factor XIa in the presence of Factor IIa (thrombin), incubated, combined with Factor X and Factor VIII, and incubated. A thrombin inhibitor, and an indicator agent which reacts with Factor Xa, are added and the resulting signal measured and correlated to the level of Factor IX in the sample.

Abstract: An immunoassay element for quantitatively analyzing a ligand by determining the change in enzymatic activity caused by a reaction between the ligand, a linked product of the ligand and a high molecular weight compound and an enzyme-labelled antibody. The immunoassay element comprises a substrate layer containing a non-diffusible substrate which forms a diffusible material in the presence of the enzyme, and a reagent layer containing a fragmenting enzyme for further fragmenting the non-difussible material. Also provided is a process for quantitatively analyzing a ligand contained in a sample by the use of the immunoassay element.

Abstract: An assay method, compositions and test kits using a hydroxyaryl cyclic diacylhydrazide is described. A hydrogen peroxide and peroxidase enzyme. The preferred compositions incorporate enhancer compounds and a chelating agent which suppresses light production prior to addition of a peroxidase enzyme. The assay method can test for a peroxidase enzyme, a peroxide or can be used in immunoassays and probe assays.

Abstract: A method and apparatus for determining the enzyme or enzymes in the human body which metabolize a particular drug. Microsomes are obtained from each of several donors. The microsome for one donor is reacted with a test drug and the quantity of metabolites produced is determined. The microsomes are similarly each reacted with the test drug and the quantity of metabolites produced for each microsome is determined to generate drug metabolism data. The drug metabolism data obtained is compared to reference data which indicates the activity of a select number of major enzymes in each of the donors. The reference data for each enzyme is separately tabulated. The enzyme responsible for the metabolism of the test drug is identified when the metabolism data correlates with the tabulated reference data for that enzyme.

Abstract: Subject matter of the invention is a new method of detecting a substance with hydrolase activity in a sample, characterized in that the sample is brought into contact with an indicator enzyme, which is covalently bound to an insoluble carrier material and can be rendered soluble by the hydrolase activity, in that the cleaved off indicator is separated and in that its enzymatic activity is determined and test means for its implementation.

Abstract: An assay for photometrically detecting and/or quantitating the presence of an analyte in a sample in which the signal generated by a label associated with the analyte is photometrically detected in the presence of a suspended solid support.

Abstract: A liposome immunoassay comprising the steps of reacting an analyte antigen, a liposome bearing antibody comprising a first antibody to the analyte antigen and a liposome encapsulating a marker therein linked to the antibody, and a second antibody to the analyte antigen to form an antigen-antibody complex, releasing the marker from the liposome in an amount depending on an amount of the analyte antigen in the presence of a complement, and measuring the released marker to determine the analyte antigen, characterized in using a third antibody capable of binding directly or indirectly to the second antibody and having an ability to activate the complement; andA kit for liposome immunoassay comprising at least one of; (1) an liposome bearing antibody comprising a first antibody to an analyte antigen and liposome encapsulating a marker therein linked to the antibody; (2) a second antibody to the analyte antigen; (3) a third antibody capable of binding directly or indirectly to the second antibody and having an abil

Abstract: Microcapsule-reagents are prepared by previously reacting at least a part of an antigen or antibody attached to microcapsules encapsulating a labeling substance with an antibody or antigen which is specifically reactive therewith, and then the reagent thus prepared is reacted with a sample containing an antigen or antibody in the presence of a complement, whereby highly sensitive immunoassay of the antigen or antibody in the sample can be realized even when the antigen or antibody attached to the microcapsules has a lowered activity or has only low reactivity with the antibody or antigen in the sample.

Abstract: A method of binding an antibody with protein A cells is provided which includes a sequence of incubation and dilution steps to produce a preselected amount and concentration of antibody with a preselected distribution of antibody among the protein A cells. In addition, a method is provided for preparing an antibody entrapped porous matrix and apparatus which includes a specific porous matrix with a preselected position of antibody bound bacterium cells therein along with means for drawing fluids through the porous medium and means for facilitating the deposition of fluids onto the surface of the porous matrix. The apparatus and method is useful for testing for the level of progesterone in animal body fluids, such as milk, plasma, serum, whole blood and saliva.

Abstract: A small enzymically inactive peptide fragment of an enzyme (e.g. ribonuclease S-peptide) is used as the label and conjugated with the complementary fragment (S-protein) to form an enzyme which catalyses a primary reaction whose product is, or leads to, an essential coenzyme or prosthetic group for a second enzyme which catalyses a secondary reaction leading to a detectable result indicating the presence of analyte. Also disclosed are novel synthetic substrates for the primary reaction. Substrates for ribonuclease S conjugate enzyme are of the formula R-X where R is pyrimidine 3'-phosphate moiety and X is a leaving group linked to R through the 3'-phosphate group and leads to said coenzyme or prosthetic group, e.g. via riboflavin, thiamine, pyridoxal, pyridoxine or pyridoxine phosphate.