Abstract: The present invention relates to a method for detecting proteins on polyacrylamide gels, comprising the steps of sensitizing the proteins with a dye composition comprising an acidic organic dye as a silver ion sensitizing agent, and optionally, a basic organic dye as an auxiliary agent, and washing them, and then silver staining complexes of the protein-dye, and to the dye composition for the same.

Abstract: Described are determination procedures for fibrinogen and/or fibrinogen-derivatives by matrix-independent turbidimetry. In the FIFTA called procedure the fibrinogen activity is preferably determined in an undiluted sample by addition of thrombin and/or albumin, as well as polybrene if appropriate, in PBS and determination of the increase in absorbance at 405 nm. In the FIATA called procedure the fibrinogen-concentration and/or the concentration of fibrinogen-derivatives is preferably determined by addition of vancomycin and determination of the increase in turbidity at 405 nm. It is standardized by usage of plasma standards of known fibrinogen-activity and/or fibrinogen-concentration.

Abstract: A system and method for desorption and ionization of analytes in an ablation medium. In one embodiment, the method includes the steps of preparing a sample having analytes in a medium including at least one component, freezing the sample at a sufficiently low temperature so that at least part of the sample has a phase transition, and irradiating the frozen sample with short-pulse radiation to cause medium ablation and desorption and ionization of the analytes. The method further includes the steps of selecting a resonant vibrational mode of at least one component of the medium and selecting an energy source tuned to emit radiation substantially at the wavelength of the selected resonant vibrational mode. The medium is an electrophoresis medium having polyacrylamide. In one embodiment, the energy source is a laser, where the laser can be a free electron laser tunable to generate short-pulse radiation. Alternatively, the laser can be a solid state laser tunable to generate short-pulse radiation.

Abstract: Amounts of components in a specimen can be analyzed with excellent quantitativity. The analysis includes: measuring an amount of a component to be analyzed in a specimen; measuring an amount of a standard component present originally and homeostatically in the specimen other than the component to be analyzed; determining the amount of the specimen from the amount of the standard component thus measured and a known concentration of the standard component in the specimen; and determining a concentration of the component to be analyzed in the specimen from the amount of the specimen thus determined and the amount of the component to be analyzed thus measured. The quantitative analysis of the present invention allows a component to be analyzed to be measured with high quantitativity as shown in FIG. 1.

Abstract: The invention disclosed is a process for determining the cytoprotective activity of plasma that prevents the destruction of endothelial cells and forestalls the development of a number of diseases such as atherosclerosis, preeclampsia, edema, nephrotic syndrome, and stroke. The present invention includes a method of diagnosing a patient's proclivity to develop a disease having a correlation to a reduction in the concentration of pI 5.6 albumin in the plasma by determining a value indicative of the concentration of the pI 5.6 albumin that is not bound to VLDL (“free pI 5.6 albumin”) in the patient's blood serum. The preferred embodiment of the process utilizes in vitro methods to obtain an indicator of the free pI 5.6 albumin instead of directly measuring the concentration of the free pI 5.6 albumin.

Abstract: With rotating and translating a rotation-translation drum installed in a vacuum chamber of a mass spectrometry interface to constitute a mass spectro-meter, a liquid sample incorporating a dissolved substance to be analyzed in its mass is emitted for the drum from a sample supplying nozzle of the mass spectrometry interface, and the dissolved substance is isolated as a spiral filament on the drum. Then, a laser beam is irradiated onto the filament from a laser source via a laser beam inlet situated at the vacuum chamber and thereby, the dissolved substance is ionized without exposing to the air.

Abstract: A method is provided for joining a microchip device to a capillary tube. The microchip device has a capillary channel opening onto an edge surface of the device. A short hole is drilled into the edge surface, aligned with the capillary channel. The drilling is done with a flat bottom, preferably by a two-step drilling process. Then, the end of the capillary can be inserted into the hole so that its end is substantially flush with the flat bottom of the hole, thereby eliminating dead volume. Testing has shown that this connection provides very little band broadening of samples transported through the capillary channel into the capillary tube. The tip of the capillary tube can be tapered, so that it is suitable for use as an electrospray source for a mass spectrometer.

Abstract: A method is disclosed for diagnosing early stage of a disease in which an intact protein found in urine is an indicator of the disease. The method includes assaying urine sample to detect the presence of modified protein using either immunological or non-immunological technique. Methods for preventing and treating the disease are also disclosed.

Abstract: The present invention describes a variety of substituted and unsubstituted carbazolylvinyl dyes and their use for detecting and quantifying poly(amino acids), including peptides, polypeptides and proteins. The labeled proteins or peptides are highly colored, but are also detected by their strong fluorescence enhancement. Poly(amino acids) are detected in solution, in electrophoretic gels, and on solid supports, including blots and dipsticks. The present method of staining is highly sensitive, extremely facile, and relatively non-selective and can be accomplished without the use of organic solvent additives.

Abstract: A sample presentation device, with a surface-bound complex including at least one molecule which chemically modifies a biomolecule, is prepared and exposed to a biomolecule. The molecular weights of the chemically modified biomolecule is then determined by mass spectrometry.

Abstract: The present invention relates to a rapid method for the detection of ischemic states and to a kit for use in such a method. Provided for is a rapid method of testing for the existence of and quantifying ischemia based upon method of detecting and quantifying the existence of an alteration of the serum protein albumin which occurs following an ischemic event; methods for detecting and quantifying this alteration include evaluating and quantifying the cobalt binding capacity of circulating blood, analysis and measurement of the ability of serum albumin to bind exogenous cobalt, detection and measurement of the presence of copper in a purified albumin sample and use of an immunological assay sepcific to the alterated form of serum albumin which occurs following an ischemic event.

Abstract: The present invention teaches a marker useful for detection and measurement of free radical damage. Specifically, the invention takes advantage of alterations which occur to the N-terminus of the albumin molecule, a circulating protein in human blood, in the presence of free radicals. These alterations effect the ability of the N-terminus of the albumin molecule to bind metals. Methods for detecting and quantifying this alteration include evaluating and quantifying the cobalt binding capacity of an albumin containing sample, analysis and measurement of the ability of albumin to bind exogenous cobalt, detection and measurement of the presence of copper in a purified albumin sample and use of an immunological assay specific to the altered form of serum albumin which occurs following free radical damage.

Abstract: The present invention relates to methods and compositions for lipid matrix-assisted chemical ligation and synthesis of membrane polypeptides that are incorporated in a lipid matrix. The invention is exemplified in production of a prefolded membrane polypeptide embedded within a lipid matrix via stepwise chemoselective chemical ligation of unprotected peptide segments, where at least one peptide segment is embedded in a lipid matrix. Any chemoselective reaction chemistry amenable for ligation of unprotected peptide segments can be employed. Suitable lipid matrices include liposomes, micelles, cell membrane patches and optically isotropic cubic lipidic phase matrices. Prefolded synthetic and semi-synthetic membrane polypeptides synthesized according to the methods and compositions of the invention also permit site-specific incorporation of one or more detectable moieties, such as a chromophore, which can be conveniently introduced during synthesis.

Abstract: A method is disclosed for diagnosing early stage of a disease in which an intact protein found in urine is an indicator of the disease. The method includes assaying urine sample to detect the presence of modified protein using either immunological or non-immunological technique. Methods for preventing and treating the disease are also disclosed.

Abstract: The invention discloses a method for detecting and/or quantifying a hapten in a homogeneous phase, comprising the following steps: adding a known quantity of a hapten inhibitor complex to the solution containing the hapten to be detected and/or quantified; adding to the solution a quantity of antibodies corresponding to the quantity of the hapten/inhibitor complex; adding to the solution a type C &bgr;-lactamase having an active site for two substrates in antigenic competition in the said active site, the first substrate being a reporter substrate capable of being transformed into a detectable and/or quantifiable product, preferably by visible UV radiation measurement, the second substrate being the hapten/inhibitor complex acting on the hydrolysis rate of the reporter substrate; detecting and/or quantifying the concentration of the product resulting from the transformation of the reporter substrate, the Km constant of the reporter substrate being at least a hundred times higher than the Km constant of the ha

Abstract: The present invention relates to a method of identifying a molecule of a molecule-substrate complex, wherein the molecule is covalently attached directly to a substrate or indirectly by means of a linking moiety, comprising: (a) bombarding the molecule-substrate complex with energized particles to cleave the molecule from the molecule-substrate complex; and (b) determining the molecular weight of the cleaved molecule by means of mass spectrometry. The inventive method may further comprise irradiating the cleaved molecule with photons.

Abstract: A peritoneal dialysis fluid used for a continuous recirculating peritoneal dialysis wherein the peritoneal dialysis fluid is infused into a peritoneal cavity of a patient and then, typically on a continuous process basis a portion of the dialysis fluid is sequentially drained from the cavity, cleansed through an extracorpreal dialyzer, and reinfused into the cavity. The dialysis fluid contains as an osmotic agent a substance which does not substantially permeate through pores of hollow fiber membrane of an extracorporeal dialyzer, preferably an osmotic agent having a molecular weight of about 20,000 to about 100,000. The supplemented amount of the osmotic agent is reduced or not needed during the dialysis.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: The present invention is drawn to method for removing unconventional transmissible agents (UCTA) from aqueous protein solution. This method involves treating the aqueous protein solution with simultaneous electrostatic adsorption and hydrophobic chromatography. The method of the present invention is particularly suitable for treating calf serum or an albumin solution.

Abstract: The present invention relates to a method of identifying a molecule of a molecule-substrate complex, wherein the molecule is covalently attached directly to a substrate or indirectly by means of a linking moiety, comprising: (a) bombarding the molecule-substrate complex with energized particles to cleave the molecule from the molecule-substrate complex; and (b) determining the molecular weight of the cleaved molecule by means of mass spectrometry. The inventive method may further comprise irradiating the cleaved molecule with photons.

Abstract: A sample presentation device, with a surface-bound complex including at least one molecule which chemically modifies a biomolecule, is prepared and exposed to a biomolecule. The molecular weights of the chemically modified biomolecule is then determined by mass spectrometry.

Abstract: The invention relates to the staining of poly(amino acids), including peptides, polypeptides and proteins in gels and on solid supports, using neutral or anionic complexes of transition metals.

Abstract: Disclosed is an improved method for determining the concentration of a first analyte in a fluid test sample as a function of a second analyte also present in the sample whose concentration in the fluid sample is clinically related to that of the first analyte. The method involves determining the concentration of the first analyte, and, if this concentration is outside of its useful analytical range, dividing this concentration by the normal concentration of the second analyte. This method of ratioing the concentrations of the first and second analyte is advantageous because accuracy is increased with fewer false positive and false negative results being reported.

Abstract: A apparatus for assaying glycated proteins and other analytes in biological samples such as blood, in which a sample is presented to the apparatus, includes an inlet port between moveable between first and second inlets, such that the inlet can be brought into liquid communication with each inlet in turn. The inlet port accommodates a filter or binder. The apparatus also includes a microprocessor operable via a key pad, at least one light emitter and at least one light detector, a display and driver, and an A to D converter, and is operatively connected to a power source.

Abstract: A disposable high density optically readable polydeoxynucleotide array with integral fluorescence excitation and fluorescence emission channels is described. The compact array size allows integration into several types of interventional devices such as catheters, guidewires, needles, trocars and may be used intraoperatively. Highly sensitive monitoring of the metabolic and disease pathways of cells in vivo under varying chemical, genetic and environmental conditions is afforded.

Abstract: Sensitive methods for identifying compounds having biological activity comprising combining living cells with two fluorescent membrane permeable ionic dyes having the same charge sign, the first of which has an emission spectrum which overlaps the excitation spectrum of the second fluorescent membrane penetrative dye. The fluorescence is then induced by illuminating the dyes at a wavelength corresponding to the excitation spectrum of the first fluorescent dye and emission is then registered at a wavelength corresponding to the emission spectrum of the second fluorescent dye (FRET). The change in the FRET is indicative of a modulation of cell membrane potential by the biologically active compounds.

Abstract: The present invention relates to a method for detection of protein using a counter-dye composition, on polyaciylamide gels, and the counter-dye composition for detection of protein on polyacrylamide gels. More specifically, the present invention relates to a method for detection of protein in a high sensitivity on polyacrylamide gels in a rapid and simple manner, comprising the step of staining the polyacrylamide gels with a counter-dye composition containing an acidic organic dye and a basic organic dye, and the counter-dye composition for detection of protein on polyacrylamide gels.

Abstract: A method for preparing a solventless protein standard is disclosed in which a solventless dye indicator and a predetermined amount of solventless protein are placed in an appropriate receptacle such as the well of a multiwell plate. This results in a solventless protein standard in which the solventless protein and the solventless dye are contained within the same receptacle. When an appropriate solvent is added to the receptacle, the protein and dye react together to produce a color change which is detectable, and which can be used as a standard for a protein assay using that dye. Also claimed is the solventless protein assay standard produced by the process.

Abstract: The invention concerns a method for the determination of an analyte in a sample containing free haemoglobin in which the determination is carried out by an optical measurement and the value measured for the analyte concentration is mathematically corrected. This method is in particular suitable for determining the parameters total protein, iron and albumin in a medical sample e.g. in a serum or plasma sample. The correction of the measured value for the analyte concentration is achieved by the steps (a) measuring the blank value of the sample to be analysed, (b) measuring the blank value of a haemoglobin-free reference sample, (c) measuring the uncorrected value for the analyte concentration and (d) correcting the value obtained in step (c) by correlation with the values obtained in step (a) and (b) in order to obtain the corrected value for the analyte concentration.

Abstract: The present invention provides a process for synthesizing a compound having the structure: ##STR1## wherein R is H, as well as related oligosaccharides useful as a vaccine for inducing antibodies to human breast cancer cells in an adjuvant therapy therefor.

Abstract: A sample presentation device, with a surface-bound complex including at least one molecule which chemically modifies a biomolecule, is prepared and exposed to a biomolecule. The molecular weights of the chemically modified biomolecule is then determined by mass spectrometry.

Abstract: The invention provides a kit for use in assaying protein, which comprises:an alkaline copper solution containing a tartrate; anda solution of Folin reagent.An alkaline copper solution containing tartrate has surprisingly been found to remain stable for several months. This allows a kit to be provided for use in a method of protein assay, in which the alkaline copper solution is provided ready-made up for immediate use in the method.

Abstract: Disclosed is an improvement to a sandwich type immunoassay in which there is immobilized to a solid support an antibody which is specific to an epitope of the analyte whose presence or concentration is being sought and a first labeled antibody which is specific to another epitope of the analyte. The improvement involves providing a second labeled antibody which is specific to the first labeled antibody to thereby form a chain of two or more labeled antibodies which results in amplification of the signal generated upon formation of the sandwich.

Abstract: A method for assessing or determining alcohol consumption rates including using a blood specimen from a human subject to develop an individual blood constituent panel; comparing the individual panel with a reference blood constituent panel to provide categories corresponding to rates of alcohol consumption; and identifying the category of consumption rate. The methods of the invention can be varied through modification of one of several statistical models used therewith to preferentially weigh the analysis and identify one consumption category over another. Multi-variate and similar such statistical techniques correlate comparisons of individual/subject blood and reference panel constituents with recognized consumptions rates.

Abstract: A novel method for monitoring the success or the yield of chemical reactions involving a reactant bound to a solid phase support, or to forecast the success of such reactions, or to quantify the number of deuterium containing groups present in a solid-phase bound sample, using infrared spectroscopy and deuterium-carbon absorbances, along with novel compounds and intermediates useful for carring out the method.

Abstract: Compositions and processes to alleviate oxygen toxicity are disclosed based on the use of nitroxides in association with physiologically compatible macromolecules. In particular, hemoglobin-based red cell substitutes are described featuring stable nitroxide free radicals for use in cell-free hemoglobin solutions, encapsulated hemoglobin solutions, stabilized hemoglobin solutions, polymerized hemoglobin solutions, conjugated hemoglobin solutions, nitroxide-labelled albumin, and nitroxide-labelled immunoglobulin. Formulations are described herein that interact with free radicals, acting as antioxidant enzyme-mimics, which preserve nitroxides in their active form in vivo. Applications are described including blood substitutes, radioprotective agents, imaging agents, agents to protect against ischemia and reperfusion injury, and in vivo enzyme mimics.

Abstract: A pyridinoline composition in which pyridinoline is derivatized specifically at its aliphatic hydroxyl group by a selected chemical group is disclosed. In various embodiments, the composition may be used as a standard for HPLC or immunoassay of pyridinoline, a pyridinoline immunogen for producing anti-pyridinoline antibodies, and a solid-phase reagent for use in an immunoassay kit. Also disclosed are methods for making and using the composition.

Abstract: The determination of human serum albumin (HSA) in urine is carried out by contacting the urine with a reagent system containing a buffer and a protein error indicator. Other proteins normally found in urine can compete with the HSA for interaction with the protein error indicator thereby affecting the sensitivity of the test for HSA. The present invention involves the addition of certain polymeric competitive inhibitors to the reagent system which inhibit the interaction of the urinary proteins other than HSA with the protein error indicator to a greater extent than the interaction with HSA is inhibited thereby increasing the reagent's sensitivity for HSA. Suitable polymeric inhibitors include poly(vinyl alkyl esters), poly(alkyl acrylates), poly(vinyl alkyl carbonates) and poly(vinyl alkyl ketones).

Abstract: Levuglandin (LG) derivatives are used as antigens for raising antibodies useful in diagnostic assays. The antibodies produced by LG-carrier protein adducts can be used to detect adducts of LGE.sub.2 with human low density lipoprotein (LDL). LGE.sub.2 -protein adduct immunoreactivity may be generated during in vitro free-radical oxidation of LDL. An enzyme-linked immunosorbent assay for detecting adducts of LGE.sub.2 with human LDL is also described.

Abstract: The present invention describes the use of a variety of merocyanine dyes and substituted merocyanine dyes for detecting and quantifying poly(amino acids), including peptides, polypeptides and proteins. The labeled proteins or peptides are highly colored, but are also detected by their strong fluorescence enhancement. Poly(amino acids) are detected in solution, in electrophoretic gels, and on solid supports, including blots and dipsticks. The present method of staining is highly sensitive, extremely facile, and relatively non-selective.

Abstract: A method, composition, and kit for isolating biomolecules from a biological sample wherein the sample is mixed with a soluble, linear polymer, such as polyvinylpyrrolidone, to form a precipitate. The biomolecule of interest is found in the precipitate or is isolated from the supernatant.

Abstract: An improved method of testing individuals for diabetes, even if they have levels of interfering substances (e.g., uric acid, bilirubin, and glutathione) that would otherwise interfere with such testing, is disclosed. The individual's protein-bound glucose level and glucose level are compared to the analogous values for a reference population to enable the risk of that individual's having diabetes to be assessed. The substances that would otherwise tend to interfere with the assay for protein-bound glucose are removed before the assay, desirably by precipitating the protein-bound glucose using uranyl acetate, which desirably leaves substantially all of the interfering substances in the supernatant, then separating the precipitate from the supernatant, redissolving the precipitate, and conducting the colorimetric assay on the resulting solution. An improved colorimetric test for protein-bound glucose using viologens as the colorimetric electron acceptors is also disclosed.

Abstract: A method and composition for isolating biomolecules from a biological sample wherein the sample is mixed with a soluble, linear polymer, such as polyvinylpyrrolidone, to form a precipitate. The biomolecule of interest is found in the precipitate or is isolated from the supernatant.

Abstract: A method for preparing a solventless protein standard is disclosed in which a solventless dye indicator and a predetermined amount of solventless protein are placed in an appropriate receptacle such as the well of a multiwell plate. This results in a solventless protein standard in which the solventless protein and the solventless dye are contained within the same receptacle. When an appropriate solvent is added to the receptacle, the protein and dye react together to produce a color change which is detectable, and which can be used as a standard for a protein assay using that dye. Also claimed is the solventless protein assay standard produced by the process.

Abstract: Processes are provided for pretreating body fluid compositions and subsequently analyzing the pretreated body fluid compositions for analytes of interest. Processes for pretreating the compositions include providing size exclusion gel having a molecular weight fractionation range or a molecular weight exclusion such that the size exclusion gel is capable of excluding or fractionating the analytes of interest, and then causing the composition to contact the size exclusion gel in order to separate the analytes from low molecular weight composition components which interfere with the separation and analysis of the analytes of interest. Processes for analyzing pretreated compositions include electrophoretic methods such as capillary zone electrophoresis which involve the separation and detection of analytes of interest.

Abstract: A modification of the standard BCA protocol is provided that allows sensitive and reliable protein determinations in a matter of seconds utilizing a microwave oven to irradiate the samples. Methods of determining protein concentrations in a sample are disclosed comprising the steps of combining the sample with a BCA assay reagent in a sample container, placing the sample container into a microwave oven, irradiating the sample and measuring an absorbance value at 562 nm for the sample. Protein concentrations are then determined by comparing the absorbance value with a known value based on a calibration curve. In preferred embodiments, the calibration curve is generated by placing one or more standards in the microwave oven and irradiating the standards. Preferably, the step of irradiating the sample is carried out for a duration of less than about 60 seconds and most preferably, for a duration of about 20 seconds.

Abstract: The detection of protein is accomplished using novel composition and method involving phenolsulfonephthalein protein error indicator, buffer and an aliphatic ether-polycarbonate present in an amount equal to or less than ten percent by weight.

Abstract: A modification of the standard BCA protocol is provided that allows sensitive and reliable protein determinations in a matter of seconds utilizing a microwave oven to irradiate the samples. Methods of determining protein concentrations in a sample are disclosed comprising the steps of combining the sample with a BCA assay reagent in a sample container, placing the sample container into a microwave oven, irradiating the sample and measuring an absorbance value at 562 nm for the sample. Protein concentrations are then determined by comparing the absorbance value with a known value based on a calibration curve. In preferred embodiments, the calibration curve is generated by placing one or more standards in the microwave oven and irradiating the standards. Preferably, the step of irradiating the sample is carried out for a duration of less than about 60 seconds and most preferably, for a duration of about 20 seconds.

Abstract: Disclosed is an improvement to the assay for protein in urine involving the use of a molybdate or tungstate salt and an indicator dye which forms a complex with molybdate or tungstate whose absorption band is shifted in the presence of protein. The improvement involves the use of an ionizable phosphate containing compound characterized by the formula: ##STR1## to reduce background interference caused by constituents normally present in urine.

Abstract: It has been found that aluminum, which is considered as to cause various diseases such as dialysis dementia, bone diseases, hypochromic anemia and Alzheimer's disease, would be eluted into a conventional albumin preparation preserved in a hard glass container, which increases the aluminum concentration of the preparation. According to the present invention, it has been found that the substitution of the conventional hard glass container with a soft glass container dealkalized by, for example, treatment with sulfurous acid gas or ammonium sulfate solution for preserving an albumin preparation can make it possible to prevent the elution of aluminum from the container, thus giving a highly safe albumin preparation which can be maintained with an extremely low aluminum concentration over a long period of time.