Abstract: The present invention relates to methods and compositions for monitoring subjects suffering from a renal injury. In particular, the invention relates to using assays that detect Beta-2-glycoprotein 1 as a prognostic biomarker in renal injuries.

Abstract: A real-time portable and rapid detection assay to identify the presence of biologically active toxins such as botulinum toxins. The proteolytic activity of BoNT/A is measured using a peptide cleavage assay, where a fluorescent substrate is cleaved by BoNT/A, resulting in increased fluorescence. This fluorescence can be monitored in real-time using a fluorescence detection instrument, such as a real-time PCR system that has been modified to implement a detection algorithm specific to the identification of the target toxin.

Abstract: Described herein are method(s), kit(s), reagent(s) and the like for determining von Willebrand factor (VWF) activity in a sample in the absence of ristocetin.

Abstract: Biomarkers for pre-Diabetes, Diabetes and/or a Diabetes related conditions, and methods of their use, including the biomarkers in Tables 1 and 2 such as peroxiredoxin-2, complement C1q subcomponent subunit B, sulfhydryl oxidase 1 and apolipoprotein A-IV.

Abstract: The invention generally relates to methods of measuring cleaved von Willebrand factor (VWF) fragments. More specifically, the invention relates to methods of measuring the ability of a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) to cleave VWF in vivo. The invention also relates to methods of using various animal models which demonstrate ADAMTS13 activity similar to that of a human. The invention further relates to methods of measuring the cleavage products of rVWF in mammals, particularly in humans and in human plasma.

Abstract: A method of measuring acidic species generated by degradation of a Fab or Fab? component of a Fab-PEG or a Fab?-PEG is provided. The method involves: a) cleaving PEG and a linker from the Fab-PEG or Fab?-PEG with an enzyme; b) optionally separating the PEG and linker from the Fab or Fab? to provide isolated Fab or Fab?; and c) quantitatively analyzing acidic species associated with the cleaved Fab or Fab? and/or the cleaved PEG.

Abstract: A method of analyzing genetic markers includes binding a set of probes to a segment of single stranded nucleic acids. The segment of single stranded nucleic acids includes a repeat region formed of at least two of a repeat unit. The repeat unit can include at least two nucleic acids. The set of probes includes a first probe complementary to the repeat unit. The method can further include directing the segment through a nanopore device and measuring a signal through the nanopore device. The signal can be indicative of the number of repeat units.

Abstract: Quantitation of analytes, including but not limited to peptides, polypeptides, and proteins, in mass spectrometry using a labeled peptide coupled to a reporter, and a universal reporter.

Abstract: A system for measuring analyte concentrations has porous-walled nanocontainers containing multiple magnetic nanoparticles, the magnetic nanoparticles coated with a selective binder that is analyte-responsive and binds a the analyte, an indicator substance releasable from the selective binder by the analyte, or an indicator substance cleavable by the analyte, apparatus for exposing the nanocontainers to a fluid potentially containing the analyte, and magnetic spectroscopy of Brownian motion sensing apparatus for detecting agglutination of the nanoparticles or binding of analyte to the nanoparticles.

Abstract: Objects of the present invention are to provide a DNA fragment encoding a limulus-derived pro-clotting enzyme, a virus harboring the DNA fragment, a cell harboring the virus, a method of producing the pro-clotting enzyme by use of the cell, and means for assaying an endotoxin or (1?3)-?-D-glucan employing the enzyme, wherein these elements are capable of producing an endotoxin or (1?3)-?-D-glucan assay reagent of satisfactory quality, steadily, at low cost, and on a large scale. In the present invention, for example, a DNA fragment encoding a protein having an amino acid sequence defined by SEQ ID NO: 4 is selected as a nucleic acid fragment encoding a limulus-derived pro-clotting enzyme, and the corresponding recombinant pro-clotting enzyme. Use of the enzyme can provide a high-sensitivity method and kit for detecting (1?3)-?-D-glucan and an endotoxin, utilizing a cascade reaction system in a horseshoe crab lysate.

Abstract: The present invention provides a method for monitoring of profile changes of components in a dynamic system such as a cell-free in vitro protein synthesis system by using liquid chromatography (LC) combined with mass spectrometry (MS). In an additional aspect, this invention provides a method for enhancing the yield and/or reproducibility in a cell-free protein synthesis system by modulating the level and/or activity of a protein component that has regulatory effects on the system.

Abstract: The present description relates to an in vitro method for the diagnosis of endometriosis, an in vitro method for the monitoring of endometriosis and a kit for the diagnosis of endometriosis and/or for the monitoring of endometriosis in a subject.

Abstract: Problem to be Solved An object of the present invention is to provide a separase sensor for visualizing separase activity in a living cell. Another object is to develop a function analysis system of cell division using the separase sensor and further obtain a simple screening method for an anticancer agent. Solution The separase sensor of the present invention has two types of fluorescent proteins different in fluorescence wavelength at both ends of an amino acid sequence containing a separase cleavage site and a localization-targeting sequence targeting to a specific site within a cell. Since timing of activating separase and location of separase in a cell can be specified by use of the sensor, cell division can be analyzed and an anticancer agent can be screened.

Abstract: The invention relates to a polypeptide having chymosin activity which: a. is capable of hydrolysing bovine alpha s1-casein at position F23F24 so as to form ?s1-I CN (f24-199) more rapidly than camel chymosin; and b. has a C/P ratio higher than the C/P ratio of bovine chymosin.

Abstract: The present disclosure provides an automated system and method for protein digestion, recovery, and analysis. The present disclosure also provides a buffer solution for use therein.

Abstract: The present invention provides pregnancy associated plasma protein A2 (PAPP-A2), its nucleotide and amino acid sequences, antisense molecules to the nucleotide sequences which encode PAPP-A2, expression vectors for the production of purified PAPP-A2, antibodies capable of binding specifically to PAPP-A2, hybridization probes or oligonucleotides for the detection of PAPP-A2-encoding nucleotide sequences, genetically engineered host cells for the expression of PAPP-A2, and methods for screening for pathologies in pregnant and non-pregnant patients. Methods for screening for altered focal proliferation states in pregnant and/or non-pregnant patients, which include detecting levels of PAPP-A2, are also described.

Abstract: The present invention provides a diagnostic reagent or assay for assessing the activity of a protease in vivo or in vitro and methods of detecting the presence of a cancerous or precancerous cell. The assays are comprised of two particles linked via an oligopeptide linkage that comprises a consensus sequence specific for the target protease. Cleavage of the sequence by the target protease can be detected visually or using various sensors, and the diagnostic results can be correlated with cancer prognosis.

Abstract: The present invention relates to mass spectrometry methods employing multiple reaction monitoring (MRM) in the field of cancer therapeutics, specifically prostate cancer and melanoma.

Abstract: A method for quantifying metallothionein protein isomers is described herein. Such metallothionein isomer protein quantification is useful for detecting and monitoring disease. As illustrated herein, protein quantification is a more accurate measure of metallothionein induction than is mRNA quantification.

Abstract: Methods for producing an isotope-labeled mammalian, including a human, biomolecule, such as polypeptides and proteins, in a cell-free protein synthesis system. A biomolecule standard is produced having at least one isotope different in abundance than that of the naturally occurring isotopes in the biomolecule. Methods for quantifying biomolecules standards expressed using mammalian cell-free extracts are disclosed. Methods for producing such standards, kits, systems and reagents, relating to the use of isotope-labeled biomolecule as quantification standards in mass spectrometric and nuclear magnetic resonance analysis.

Abstract: Assay methods and systems use enzymatic cleavage resulting from protein-protein interaction to modulate (activate or inactivate) a reporter.

Abstract: The present invention reveals a strong correlation between FGL-2 prothrombinase activity levels and the presence of a malignant proliferative disorder in a subject. Thus, the present invention provides FGL-2 prothrombinase activity as a diagnostic tool for malignancy.

Abstract: Compositions and methods are disclosed that provide a rapid, sensitive, and accurate cell-based assay for enzyme activity, particularly for enzyme activities associated with botulinum toxins. A cell is provided that expresses a construct that includes an anchor region, a cleavage site, and a reporting region having two or more identical reporter peptides. Enzymatic activity at the cleavage site releases the reporter region into the cytosol, where multiple degradation events occur. The observed change in the signal is proportional to the enzymatic activity.

Abstract: There is presently provided a method of detecting a bio-marker for a genetic disease in an individual. The method involves comprising identifying the presence of a proteolytic peptide in a polypeptide fraction obtained from a cellular extract of cells from the individual. The cells are cells by the disease to be diagnosed, and the proteolytic peptide contains a mutation associated with the genetic disease or directly related to the disease condition.

Abstract: Disclosed herein are novel methods, assays and kits useful for the diagnosis and monitoring of subjects with mucopolysaccharidoses (MPS), The methods, assays and kits are particularly useful for detecting the presence of one or more glycosaminoglycans which correlate to MPS and its severity in a variety of biological samples.

Abstract: The present invention relates to a method for diagnosing, evaluating the prognosis of a cancer patient and evaluating the aggressiveness of a cancer. The methods comprise detecting the phosphorylation status of a Scribble protein, a fragment or variant thereof, wherein the phosphorylation status is indicative of cancer, prognosis and the aggressiveness of a cancer. In one embodiment the cancer is breast cancer.

Abstract: The invention provides a method for analyzing a protamine sample for the presence or amount of at least one nucleotidic impurity. The invention also provides a method for the quantitation of peptides in a sample of protamine comprising four major peptides and at least one related impurity.

Abstract: Objects of the present invention are to provide a DNA fragment encoding a limulus-derived pro-clotting enzyme, a virus harboring the DNA fragment, a cell harboring the virus, a method of producing the pro-clotting enzyme by use of the cell, and means for assaying an endotoxin or (1?3)-?-D-glucan employing the enzyme, wherein these elements are capable of producing an endotoxin or (1?3)-?-D-glucan assay reagent of satisfactory quality, steadily, at low cost, and on a large scale. In the present invention, for example, a DNA fragment encoding a protein having an amino acid sequence defined by SEQ ID NO: 4 is selected as a nucleic acid fragment encoding a limulus-derived pro-clotting enzyme, and the corresponding recombinant pro-clotting enzyme. Use of the enzyme can provide a high-sensitivity method and kit for detecting (1?3)-?-D-glucan and an endotoxin, utilizing a cascade reaction system in a horseshoe crab lysate.

Abstract: The presently disclosed subject matter provides methods using mass spectrometry for direct profiling of N-linked glycans from a biological sample. In addition, the embodiments of the present invention also disclose novel methods, known as targeted analyte detection (TAD), for improving the detection limit of MALDI-MS. These methods take advantage of the carrier effect of the added standard analytes, which occurs due to the generic sigmoidal shape of the calibration curve. The functionality of TAD depends on the relative enhancement of sensitivity over the increase of the standard deviation at the analysis of target analytes with spiking in exogenous concentration. At certain ranges of exogenous concentration, the increment in the sensitivity overcomes the standard deviation, resulting in an improved LOD. Theoretically, exogenous concentrations approximately at 1 LODorig would generate the optimum LOD improvement.

Abstract: Disclosed herein are fluorescent proteins and Split-Fluorescent proteins (SFPs) including Split-Green Fluorescent Proteins, such as tripartite split-GFPs. Nucleic acid molecules encoding the fluorescent proteins and SFPs, as well as methods of using the fluorescent proteins and SFPs, are also disclosed. For example, methods of detecting protein-protein interactions are disclosed herein.

Abstract: The presently-disclosed subject matter provides methods and kits for diagnosing obstructive sleep apnea (OSA) in a subject, such as a human child, wherein a biological sample is provided from the subject and the amount of a Urocortin III peptide is determined from the sample. Further provided are methods for diagnosing OSA in a subject wherein the amount of a Urocortin III peptide and one or more peptides selected from a Uromodulin peptide, an Orosomucoid 1 peptide, and a Kallikrein 1 peptide are determined in a biological sample from a subject.

Abstract: The present invention relates to agonists of Neprilysin and their use in preventing and treating pulmonary vascular remodeling. Also described are diagnostic and screening applications stemming from the inventor's discovery that Neprilysin is expressed at reduced levels in disease tissues.

Abstract: The present disclosure provides peptide amino acid sequencing and identification methods and kits for performing such methods. For example, single-molecule detection of fluorophore-labeled peptides is disclosed using multiple rounds of standard Edman degradation or using digestion by chemicals or enzymes. Different fluorophores covalently attached to each of a specific type of amino acid side chain of a peptide provide for the derivation of the peptide's encoded amino acid sequence following image alignments of multiple Edman cycles or following digestion by chemicals or enzymes. The amino acid sequence of a peptide and/or the identity of the peptide can be determined by bioinformatic analysis based on the encoded amino acid sequence. The present disclosure further provides peptide derivatization and immobilization strategies to enable the sequencing and identification of a single peptide or a plurality of peptides.

Abstract: The present invention relates to a method of obtaining peptides from procaryotic and/or eucaryotic cells.

Abstract: Embodiments described herein relate to compositions, methods and uses for alpha-1 antitrypsin (AAT) or derivatives or analogs or peptides or mutants thereof for treating a subject exposed to radiation. In certain embodiments, AAT or derivatives thereof or analogs thereof can be used to modulate adverse effects of radiation therapy on subjects undergoing treatment for cancer. In other embodiments, compositions disclosed herein can be used for treating a subject having been exposed to non-therapeutic or accidental radiation. Some embodiments reported herein concern compositions and methods for enhancing radiotherapy. Certain embodiments relate to methods and compositions for preventing or reducing radiation exposure-induced cellular damage in a subject.

Abstract: The present invention relates to a method aiding in the assessment of cancer. It discloses the use of the human fibroblast activation protein (FAP/seprase) as a universal marker of different cancer types. Seprase aids in the assessment of pulmonary or lung cancer (LC) or of colon cancer, e.g., of non-small cell lung carcinoma (NSCLC) or colorectal cancer (CRC), but also likely of other specific types of cancer. Such specific cancer types are, e.g., esophagus, head and neck cancer, stomach cancer, bile duct cancer, pancreas cancer, kidney cancer, cervix cancer, ovary cancer, breast cancer, bladder cancer, endometrium cancer or prostate cancer. Furthermore, it especially relates to a method for assessing cancer from a liquid sample, derived from an individual by measuring seprase in said sample. Measurement of seprase can, e.g., be used in the early detection of cancer or in the surveillance of patients who undergo surgery.

Abstract: The invention provides a novel class of cyanine dyes that are functionalized with sulfonic acid groups and a linker moiety that facilitates their conjugation to other species and substituent groups which increase the water-solubility, and optimize the optical properties of the dyes. Also provided are conjugates of the dyes, methods of using the dyes and their conjugates and kits including the dyes and their conjugates.

Abstract: The current disclosure provides for specific peptides, and derived ionization characteristics of the peptides, from the KRT5, KRT7, NapsinA, TTF1, TP63, and/or MUC1 proteins that are particularly advantageous for quantifying the KRT5, KRT7, NapsinA, TTF1, TP63, and/or MUC1 proteins directly in biological samples that have been fixed in formalin by the method of Selected Reaction Monitoring (SRM) mass spectrometry, or what can also be termed as Multiple Reaction Monitoring (MRM) mass spectrometry. Such biological samples are chemically preserved and fixed wherein said biological sample is selected from tissues and cells treated with formaldehyde containing agents/fixatives including formalin-fixed tissue/cells, formalin-fixed/paraffin embedded (FFPE) tissue/cells, FFPE tissue blocks and cells from those blocks, and tissue culture cells that have been formalin fixed and or paraffin embedded.

Abstract: Methods for assessing the condition of keratinized structures, including hair, in particular methods to determine the condition of keratinized structures in relation to suitability for analysis of analytes of interest in a test sample, are presented. The methods comprise contacting the keratinized structure with a non-proteolytic reducing agent and an optional proteolytic agent. The methods further include inspection of the hair sample, or measurement of free protein eluted from the keratinized structure, after reduction and optional proteolysis to determine condition prior to analyte identification and quantitation by known techniques such as immunoassays.

Abstract: The present invention provides a method for screening a therapeutic and/or prophylactic agent for Alzheimer's disease using at least one index selected from the group consisting of the levels of A? oligomers, BiP, cleaved caspase 4, PRDX4 and ROS in nerve cells or the like whose differentiation has been induced from iPS cells derived from somatic cells of a patient with Alzheimer's disease.

Abstract: The current invention is directed to a method for determining amino acid sequence mutations in a produced polypeptide, comprising the following steps of a) providing a sample of a produced polypeptide, b) incubating the polypeptide in the sample with a protease, c) performing a two dimensional analysis using reversed phase chromatography coupled with a high resolution mass spectroscopy (FT-ICR/FT-orbitrap) and MS/MS analysis of the amino acid sequence fragments of the peptides, d) data evaluation by comparing the LC-MS data sets obtained for the samples side by side with the data set of a reference sample, by searching for differences in the signal intensities at given retention times and by evaluation of differential signals with respect to amino acid sequence mutations. The reference sample for data evaluation (d) can be either a well characterized standard or one of the samples to be analyzed.

Abstract: The present application relates to extracellular vesicles (EVs) derived from gram-positive bacteria. In detail, the present application provides animal models of disease using extracellular vesicles derived from gram-positive bacteria, provides a method for screening an active candidate substance which is capable of preventing or treating diseases through the animal models of disease, provides vaccines for preventing or treating diseases caused by extracellular vesicles derived from gram-positive bacteria, and provides a method for diagnosing the causative factors of diseases caused by gram-positive bacteria using extracellular vesicles.

Abstract: Methods for liquid-liquid extraction, as can be effected using polymeric reverse micelles, such methods as can be used in conjunction with various mass spectrometric techniques.

Abstract: A novel reagent for clarification of an emulsion containing a salt and first and second surfactants is used in a method of clarifying emulsions which may contain inorganic, organic or biological particles to be measured. The reagent may be proved as kit of parts for in situ preparation thereof.

Abstract: Methods and systems for using fibrosis biomarkers associated with a prolonged period of physical inactivity are provided. Also provided is a method of reducing the effect of prolonged physical inactivity on the development of fibrosis in a subject who is experiencing or is expected to experience prolonged physical inactivity in the near future by administering a therapeutically effective amount of a leucine metabolite (e.g., ?-hydroxy-?-methylbutyric acid (HMB)) to the subject.

Abstract: Sirt5, a mitochondrial Sirtuin, has been identified herein as an efficient demalonylase and desuccinylase. Disclosed herein are assays to identify Sirt5 modulators based on this robust enzymatic activity. Sirt5-specific modulators can be used study the biological function of Sirt5 and to target Sirt5 activities in treating human diseases.

Abstract: In one form, a fructosyl peptidyl oxidase derived from a budding yeast Phaeosphaeria nodorum for assaying a glycated protein in a sample is provided. The fructosyl peptidyl oxidase has higher activity toward fructosyl valine as well as fructosyl valyl histidine, and may be useful in assaying HbA1c with higher sensitivity and specificity. Still, other forms include unique methods, techniques, systems and devices involving a fructosyl peptidyl oxidase.

Abstract: The invention provides methods and compositions for simultaneously detecting the activation state of a plurality of proteins in single cells using flow cytometry. The invention further provides methods and compositions of screening for bioactive agents capable of coordinately modulating the activity of a plurality of proteins in single cells. The methods and compositions can be used to determine the protein activation profile of a cell for predicting or diagnosing a disease state, and for monitoring treatment of a disease state.

Abstract: A biological tissue processing substrate for fixing proteins in a biological tissue or degradation products of the proteins, the substrate comprising: a porous body that forms a contact surface with the biological tissue, the porous body holding in pores an enzyme for obtaining the proteins or the degradation products of the proteins from the biological tissue, wherein the proteins or the degradation products obtained by the action of the enzyme are brought into contact with a member consisting of a metal.

Abstract: The present invention provides a method of pretreating a sample containing a glycated amine as an analyte, thereby enabling highly reliable measurement of a glycated amine. A glycated amino acid in the sample is degraded by causing a fructosyl amino acid oxidase (FAOD) to act thereon, and thereafter, a FAOD further is caused to act on the glycated amine as the analyte in the sample to cause a redox reaction. The amount of the glycated amine is determined by measuring the redox reaction. The substrate specificity of the FAOD caused to act on the glycated amino acid may be either the same as or different from that of the FAOD caused to act on the glycated amine. When using the same FAOD, a FAOD is caused to act on the glycated amino acid to degrade it, and thereafter, the sample is treated with a protease to inactivate the FAOD and also to degrade the glycated amine.