Abstract: A method for diagnostic or prognostic monitoring of premalignant or malignant conditions of human secretory epithelia, particularly colonic epithelia, by determining the extent of expression of a .beta.1-3N-acetylglucosaminyltransferase is described. Associated with the expression of a wide variety of carbohydrate antigens in adenocarcinomas is the induction of .beta.1-3N-acetylglucosaminyltransferase in epithelial cells. This enzyme is not found in normal, healthy adult colonic epithelial cells and thus indicates a novel and potentially sensitive method for screening the disease status of individuals.

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: A method for the determination of glycated protein in a sample characterised in that it comprises treating the sample with a protease and treating the protease-treated sample with a ketoamine oxidase, a product of this reaction being measured is disclosed.

Abstract: A multilayer analytical element for assaying fructosamine, having a liquid-impermeable support, a buffer-containing layer which contains a buffer having pH of 8 to 12 formed on the support, a tetrazolium salt containing layer which is formed on the buffer-containing layer and, optionally, an intermediate layer interposed between the buffer-containing layer and the tetrazolium salt-containing layer to prevent contact of these two layers, which element assays fructosamine in a short time with good accuracy.

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.

Abstract: Disclosed is a method for determining the presence in mammalian serum of antibodies against specific strains of HIV through the use of SP-10 peptides. The method incorporates the use of SP-10 peptides derived from the following HIV strains: IIIB, MN, RF, SC, WMJ-1, WMJ-2, WMJ-3, ARV-2, and LAV-1.

Abstract: O-acylated pyridinium compounds in which the 3-hydroxy moiety derivatized specifically at its aliphatic hydroxyl group by a selected acyl 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: A method for determining whether antiphospholipid antibodies are present in a sample is disclosed. The method comprises contacting the sample with a negatively charged phospholipid and with .beta.-2-glycoprotein-I or a homolog or analog thereof and determining whether any antiphospholipid antibodies have bound to the contacted phospholipid and .beta.-2-glycoprotein-I, wherein detection of binding of antiphospholipid antibodies to the phospholipid and .beta.-2-glycoprotein-I, is indicative that antiphospholipid antibodies are present in the sample.

Abstract: A composition, test device and method of determining the presence or concentration of proteins, such as albumin, in a liquid test sample, such as urine. The test device includes a test pad with a carrier matrix incorporating a indicator reagent composition capable of interacting with proteins to produce a detectable response. The indicator reagent composition includes an indicator dye that is capable of interacting with albumin and undergoing a detectable color transition; a buffer; a hydrophobic polymeric compound having the general structural formula: ##STR1## wherein A is ##STR2## and PO is an oxypropylene unit, EO is an oxyethylene unit, y is a number in the range of 0 to about 20, z is a number in the range of 0 to about 20, the sum y+z is a number in the range of about 2 to about 20, and R.sub.2 and R.sub.3 are selected, independently, from the group consisting of hydrogen, an alkyl group, an aralkyl group, and an aryl group;R.sub.

Abstract: A method and apparatus for determining if a pregnant woman is at significant risk of carrying a fetus with Down syndrome, The method comprises measuring the pregnant woman's maternal blood levels of the free beta subunit of human chorionic gonadotropin. The level of free beta subunit of human chorionic gonadotropin individually or with other markers may be compared to reference data. A computerized apparatus for making the determination preferably using a probability density function generated from a set of reference data by a linear discriminant analysis procedure is disclosed.

Abstract: The present invention relates to a method for detecting fetal Down syndrome (Trisomy 21), trisomy 13, trisomy 18 and other chromosomal anomalies during prenatal screening by analyzing blood samples from a pregnant woman. More particularly the present invention relates to a method for improving detection efficiency in screening for the anomalies by measuring the amount of the free beta human chorionic gonadotropin (HCG) and nicked or fragmented or aberrant forms of free beta (HCG), all of which are referenced throughout this application as free beta (HCG) in blood samples from pregnant women.

Abstract: A method for detecting a glycoprotein using a solid support is disclosed where the glycoprotein is oxidized by periodate, polyacrylic polyhydrazide which is a copolymer having repeating units possessing a hydrazide group and repeating units possessing hydroxyl groups is coupled to the oxidized glycoprotein and a glycoenzyme or radioactive compound containing aldehyde groups or activated ketone groups is coupled to the polyacrylic polyhydrazide which allows for detection of the glycoprotein. The glycoprotein may be directly attached to the solid support or may be bound to an antigen which is immobilized on the solid support.

Abstract: The subject invention provides purified polypeptides encoded by naturally-occurring wild-type platelet glycoprotein Ib alpha having a mutation which renders the polypeptide more reactive with von Willebrand factor. Preferably, the mutation is in the hinge region of GP Ib.alpha., such as the substitution of valine for glycine at residue 233. These mutations alter the three-dimensional structure of the mutant polypeptide from a beta bend conformation to an alpha helix formation, and also create an amphipathic region within the mutant polypeptide. DNA encoding the mutant polypeptides, as well as expression systems for the production of the mutant polypeptides, are also provided. Methods and compositions using the mutant polypeptides and DNA oligomers complementary to the mutant polypeptides are further provided.

Abstract: A screening method for fetal Down syndrome using free beta (HCG) as a marker. The maternal blood level of free beta (HCG) is measured and compared to reference data, at a similar gestational age, of the level of free beta (HCG) in pregnant women carrying a fetus with Down syndrome and pregnant women carrying an unaffected fetus. The method may advantageously be utilized during the first and second trimesters of pregnancy. Fragments of free beta and "nicked" free beta may also be utilized as markers.

Abstract: A method for measurement of fructosamines in an aqueous liquid characterized by including the steps of:(a) bringing an aqueous liquid sample into contact with one or more active oxygen producing substances;(b) bringing the product of said step (a) into contact with a reagent producing a detectable change in the presence of hydrogen peroxide derived from the active oxygen, and(c) detecting an occurrence of the detectable change of said step (b).

Abstract: A system for determining the concentration of fructosamine in sera which consists of a first reagent in which a tetrazolium salt which reduces all reactive substances in sera including fructosamine and a second reagent which is responsive to all reactive substance in sera other than fructosamine. The difference in color change as between the two allows the determination of concentration of fructosamine.

Abstract: Physiologically active substances contained in a biological sample not subjected to deproteinization can be converted to their derivatives at a high efficiency without being adversely affected by the protein present in the sample, and accordingly the analysis of said physiologically active substances in the form of said derivatives can be performed at a high precision.

Abstract: The amount of lipid bound sialic acid in a blood plasma or serum sample may be determined by an improved method which may be automated involving the following steps to be performed simultaneously on the sample and a known standard prepared from human or animal blood or tissue; diluting with distilled water; mixing; adding a mixture of a chlorinated lower aklyl alcohol; mixing, diluting with water and then treating by mixing further and centrifuging to yield a substantially clear upper phase; recovering the upper phase and adding to it a protein precipitating agent, mixing the resulting admixture; recovering the resulting precipitate, suspending the precipitate in a hydrolysis agent and determining the amount of lipid bound sialic acid present by comparing the optical density of the sample to the optical density of the standard.

Abstract: A method for diagnosing a patient with an autoimmune disease using monoclonal antibodies to quantitate the amount of 680 kd, 54 kd, 44 kd or 14 kd present, wherein an elevated amount of protein compared to a patient without the disease indicates the presence of autoimmune disease.

Abstract: A method is disclosed for detecting the occurrence of a binding or complex-forming reaction between specific substances by utilizing the binding reaction to modify an electrical circuit, and then measuring a change in the electrical state of the circuit. A diagnostic element useful in such a method includes a layer of a biogenic substance, such as an antigen, coated onto a non-conductive base between a pair of electrical conductors superposed on the base. Antibodies which react with the antigen are treated so that they become bound to particles. The particles having antibody bound thereto are then added to is the antigen lay e base and allowed to react therewith. The particles are thereby bound to the base due to the binding reaction between the antigen and antibody to thereby form aggregates of electrically conductive particles which modify the circuit. The particles are then selectively coated with a conductive substance.

Abstract: A method and kit for the isolation and quantitation of glycated hemoglobin and other glycated proteins, specifically albumin, from a single sample of whole blood. Glycated hemoglobin is calculated from an eluate resulting from the contact of the whole blood hemolysate with a boronated resin. Glycated albumin and other plasma proteins are calculated using immunoturbidimetry, resulting from reaction between a buffered antibody solution and the albumin or other protein in the previous eluate.

Abstract: A method of monitoring collagen degradation, comprising assaying a biological fluid sample which contains a fragment of collagen including lysyl pyridinoline or hydroxylysyl pyridinoline or a substituted form thereof, a method of determining the tissue origin or degraded collagen thereby.

Abstract: A kit for staining proteins and nucleic acids wherein the stain is a suspension of colloidal metal particles and the proteins and nucleic acids are visualized as a colored signal localized at the binding site of the colloidal metal particles to the proteins or nucleic acids or quantitatively determined at this site following art-known spectrophotometric procedures.

Abstract: A method for diagnosing a patient with an autoimmune disease comprising assaying a blood sample from a patient for increased levels of the carbohydrate moieties of one or more proteins, said increased level of one or more of said proteins indicating the presence of an autoimmune disease.

Abstract: The present invention relates to a highly sensitive diagnostic/prediagnostic test to identify persons at risk of a thrombotic event. Thrombotic events include myocardial infarction, deep venous thrombosis, pulmonary embolism, thromboembolic stroke pulmonary embolism deep venous and cardiovascular disease. The test is based on the early detection of elevated levels of resting platelet surface thrombospondin. The present invention also includes methods of determining the presence of thrombospondin on the surface TSP receptors of resting platelets in a biological sample. An anti-thrombospondin monoclonal antibody which is specific for thrombospondin on resting platelets is also disclosed. A diagnostic test in the form of a test kit for the determination of thrombospondin levels in a patient sample, and also for the prediagnosis of persons at risk for thrombotic events, is also described.

Abstract: The present invention relates to a method for detecting fetal Down syndrome (Trisomy 21), trisomy 13, trisomy 18 and other chromosomal anomalies during prenatal screening by analyzing a dried blood sample from a pregnant woman. More particularly the present invention relates to a method for improving detection efficiency in screening for the anomalies by measuring the amount of the free beta human chorionic gonadotropin (HCG) and nicked or fragmented or aberrant forms of free beta (HCG), all of which are referenced throughout this application as free beta (HCG) in dried blood samples from pregnant women.

Abstract: A method of manufacturing a test device for determining the presence and concentration of proteins, such as albumin or Bence Jones proteins, in a test sample. The test device includes a carrier matrix incorporating a reactant system capable of interacting with proteins to produce a visually or instrumentally detectable and/or measurable response. The carrier matrix of the device can include commonly used bibulous matrices, such as filter paper, or a nonbibulous protein-permeable strip, membrane or layer of a polymerized urethane-containing composition. In addition, a reactant system, including a dual indicator reagent system, such as bromophenol blue, methyl orange and, if necessary, a suitable buffer, is incorporated into the carrier matrix to provide improved color resolution and increased sensitivity to proteins, thereby affording a more accurate and trustworthy protein assay of test samples, such as urine.

Abstract: A method for detecting ischemic states in a patient by contacting a sample of serum, plasma, fluid or tissue with a metal ion capable of binding to metal ion binding sites in the sample to form a mixture, and then detecting the presence of unbound metal ions to determine the occurrence of ischemia.

Abstract: Monoclonal antibodies specific for the glycosylated lysine residue at position 525 in glycoalbumin and a method for producing such antibodies. The monoclonal antibodies are useful as reagents in immunoassays for the specific determination of glycoalbumin in human blood samples which is indicative of the severity of the diabetic condition. The monoclonal antibodies are secreted by hybridomas obtained by fusing a myeloma cell with a lymphocyte that has been taken from an animal, usually a mouse, immunized with a peptide immunogen and which produces antibody to the lysine 525 residue in glycoalbumin. The synthetic peptide immunogen comprises a peptide residue which includes an .epsilon.-amino glucosylated lysine and an adjacent amino acid sequence in which at least one of the amino acid units is in a position corresponding to the peptide sequence of human albumin adjacent to lysine 525, the glycosylated peptide residue being linked to an immunogenic carrier.

Abstract: The present invention relates to a method for an early stage detection of three specific pregnancy-related disorders: preeclampsia, intrauterine growth retardation and preterm delivery. According to the method, an antigen consisting of a specific human-derived placental soluble protein, known as PP-13, is determined by radioimmunoassay or ELISA. In the radioimmunoassay method, the PP-13 is labelled by a radioactive iodine and the bounded iodine is counted and correlated with standard curves. The best results are obtained when the protein to be labelled by radioactive iodine is present at a concentration of above 0.71 mg/ml. In case of the ELISA method, the quantitative determination of PP-13 is carried out with an alkaline phosphatase substrate and measured at an optical density of 405 nm.

Abstract: This invention teaches a process for reducing protein matrix effects in assays for serum fructosamine. Blood or blood derived samples are used, and one adds two reagents, one of which reduces interference caused by non-specific reducing substances, the other of which eliminates turbidity. Incubation follows, and then the pH of the sample is adjusted and color forming reagent is added. In one embodiment, the incubation time is only 1-15 minutes. In another embodiment, the first reagent contains peroxidase.

Abstract: Process for the specific determination of the serum fructosamine content in blood or in samples obtained from blood by reaction with an appropriate color reagent and measurement of the color change thereby brought about, in which before the color reaction sample components with a non-specific reducing action and/or causing turbidity are removed and subsequently the color reagent is added at a pH value of from 10 to 12. The sample components are removed by treatment at approximately neutral pH value with a reagent composition comprising at least one enzymatic oxidation agent, optionally together with peroxidase and/or catalase and/or lipase, as well as with at least one SH group-blocking substance. A kit for the specific determination of the serum fructosamine content in blood or samples obtained from blood, comprises said reagent composition, a rebuffering reagent with a buffer which has an alkaline pH value and a color reagent for the detection of fructosamine.

Abstract: Disclosed are diagnostic reagents for use in detection of Herpes Simplex Virus Type 1 antibodies or Herpes Simplex Virus Type 2 antibodies comprising novel Herpes Simplex Virus envelope glycoproteins gD-1 and gD-2, immunologically active fragments thereof, immunologically synthetic replicas, thereof, and specific polypeptides comprising specific amino acid sequences.

Abstract: A reverse-phase HPLC method is disclosed which allows the separation of DPU and PTH-Trp and, therefore, the correct assignment of tryptophan residues in an amino acid sequence. The method is based on a modification of the conventional HPLC gradient commonly used to elute and separate all PTH amino acids of interest in automated and manual sequencing. In one embodiment, using automated instruments manufactured by Applied Biosystems, gradient modification is achieved by changing the manufacturer-supplied gradient program which controls the operation of the HPLC pumps in the instrument. Using the methods of the present invention, the correct and reliable assignment of tryptophan residues is possible and was reproducible over time, even with small sample sizes.

Abstract: Disclosed herein is a method of labelling a sugar at its reducing end by using 1-(p-methoxyphenyl)-3-methyl-5-pyrazolone, thereby analyzing said sugar with high sensitivity, and to a kit to be used for sugar labelling by this method.

Abstract: This invention provides a primary standard and/or secondary standards for assay for glycated proteins in samples such as blood. The primary standard is composed of a polymer or copolymer of an amino acid, such as lysine, serine or those listed on table 37 (pages 100-110 of the second edition of Organic Chemistry by Robert Morris and Robert Nielson-Boyd) glycated with a known amount of glucose, preferably .sup.14 C glucose or .sup.3 H glucose, and free of unbound glucose. The preferred secondary standards are composed of glycated native protein per se, or a mixture of a glycated native protein and native protein that has been standardized against a primary standard to give the actual glycated protein value. These primary standards and secondary standards may be packaged and sold as a kit that contains a primary standard and/or a secondary standard and the reagents needed to perform the glycated protein assay.

Abstract: Variant somatotropins are described in which the asparagine located in the 95-101 region are replaced by glutamine which somatotropins exhibits enhanced stability.

Abstract: A method for separating HDL from whole blood anticoagulated with EDTA is disclosed. The method includes, as a first step, mixing the anticoagulated whole blood with an aqueous solution composed of magnesium .sup.++ cations and dextran sulfate 500 in such proportions and of such concentration(a) that substantially all of the LDL and VLDL lipoproteins and substantially all of the chylomicrons are precipitated while(b) substantially all of the HDL remains in solution.The method also includes the step of sedimenting the precipitated lipoproteins, the chylomicrons and the red blood cells by centrifuging, which can be low gravity centrifuging, and can include the step of determining the cholesterol, triglyceride, phospholipid or protein content of the supernatant.

Abstract: The invention discloses novel methods of detecting glycated proteins in test samples of blood and methods of detecting chronic hyperglycemia. The test sample is contacted with a binder composition under conditions which allow binding of glycated protein in the test sample to at least a portion of the binder composition. The test sample is then exposed to a glycated compound affixed to a solid support under conditions which allow raction of unbound binder composition from the previous step to at least a portion of the glycated compound on the solid support. The glycated compound affixed to a solid support from the previous step which did not react with the unbound binder composition is then reacted with a detecting agent and the detecting agent is detected.

Abstract: Human tumor associated Thomsen-Friedenreich (TF) antigen is purified from adenocarcinoma conditioned media, adenocarcinoma cell detergent extracts or plural effusion fluid by affinity chromatography using an insolubilized TF-specific monoclonal antibody, MAb 49H.8. The TF antigen is a glycoprotein characterized by a non-cryptic Gal .beta.(1.fwdarw.3) GalNAc epitope, a molecular weight in excess of 1,000,000 daltons, and extractability with perchloric acid, the epitope being sensitive to alkali and periodate but resistant to acid. A heterologous sandwich immunoassay has been developed for human TF antigen using a monoclonal antibody as the catcher and labelled peanut agglutinin as the probe. Since human TF antigen is shed by tumor cells, a positive determination of the TF antigen in a patient sample indicates the presence of cancer.

Abstract: The elevation of human cellular fibronectin monomers having a variably included Type III repeat in a pregnant woman, as early as the first trimester, has been found to precede the onset of the clinical manifestations of toxemia and correlate with the severity of the disease state. The present invention provides a means of detection for and the monitoring of the toxemias of pregnancy, particularly preeclampsia and eclampsia, by the use a marker, human fibronectin having a variably included Type III repeat sequence, ED1 or ED2.

Abstract: An improved method for carrying out multiple laboratory processes including hybridization, cell growth and chromatographic separations including steps of: providing a laboratory apparatus having a hollow recessed chamber and a removable cover sized to be slidably retained in at least a portion of the hollow recessed chamber for providing a variable volume chamber. Inserting a sandwich including a membrane having DNA fragments bound thereto, and a porous non-reactive filter on either side of the membrane into the chamber and inserting the slidable cover, a distance sufficient to provide a minimum volume chamber for the sandwich. Delivering a pre-hybridization solution to the chamber by utilizing a fluid administration kit in fluid communication with the chamber and pre-hybridizing the DNA fragments at a desired temperature. Removing the pre-hybridization solution from the chamber by the fluid administration kit.

Abstract: For the determination of fructosamine in body fluids a solution containing fructosamine and albumin is used as the standard solution for calibration which standard solution is essentially free of glucose and which has a pH between 5.0 and 6.0 and contains at least 10 mmol/l buffer.

Abstract: The present invention provides a method for extracting lipid bound sialic acid from a sample of blood plasma or serum and determining the amount of sialic acid present in the sample which comprises:(a) adding to the sample a mixture of a lower alkyl alcohol and a chlorinated lower alkyl hydrocarbon so as to form a resulting admixture, the volume ratio of the lower alkyl alcohol to the chlorinated lower alkyl hydrocarbon being in the range from about 70:30 to about 85:15;(b) mixing the resulting admixture for a period of time sufficient to dissolve lipid bound sialic acid present in the sample;(c) treating the admixture so as to form a recoverable, substantially clear lipid bound sialic acid-containing upper phase;(d) separately recovering from the clear upper phase so formed a predetermined volume of the upper phase; and(e) determining the amount of lipid bound sialic acid present in the predetermined volume of the upper phase and thereby the amount of sialic acid present in the sample.

Abstract: A single color determination method of quantization of the amount of fructosamine in a sample such as serum by removing other interfering reducing agents and developing a color with a coloring agent such as tetrazolium salt.

Abstract: A synthetic polypeptide is disclosed that substantially corresponds in sequence to a portion of the amino acid residue sequence of the 90 amino acid residue extra type III domain of human cellular fibronectin from about position 36 to about position 60 from the amino-terminus. The polypeptide contains about 10 to about 25 amino acid residues. Also disclosed are antibodies and methods of using both the polypeptide and antibodies.

Abstract: Detection of solublen Interleukin-2 (IL-2) and Interleukin-2 receptor (IL-2R) levels in the serum, plasma, urine and bile of kidney and liver transplant patients using sensitive immunoassays provides valuable prognostic, monitoring and diagnostic information about impending, acute and chronic or recovering allograft rejection episodes. Additionally, monitoring immune system activation with these sensitive immunoassays can be useful for detecting viral and other infections and for distinguishing cyclosporine toxicity from graft rejection.

Abstract: A new and improved test device and method of determining the presence and concentration of fructosamines in a test sample. The test device includes a test pad comprising a carrier matrix incorporating an indicator reagent composition capable of interacting with fructosamines to produce a detectable or measurable response. The carrier matrix of the device includes a bibulous matrix, such as filter paper, or a nonbibulous matrix, such as a polymeric film. In addition, a new and improved indicator reagent composition comprising a dye, such as tetrazolium violet; a suitable buffer; and an accelerator compound is incorporated into the carrier matrix to provide a test pad demonstrating sufficient color resolution and having sufficient sensitivity to fructosamines in a test sample. Therefore, a fast, more accurate and trustworthy fructosamine assay of a liquid test sample, such as whole blood or serum, is achieved.

Abstract: A method and apparatus for analyzing blood plasma to determine the concentration of its lipoprotein constituents, VLDL, LDL, HDL and proteins includes obtaining the NMR chemical shift spectrum of a sample. Stored reference NMR spectra of the lipoprotein constituents are added together to form a lineshape that best fits the measured blood plasma NMR spectrum, and from this, the concentration of each lipoprotein constituent in the blood plasma is determined.