Patent classifications
G01N2440/38
Tandem mass tag multiplexed quantitation of post-translational modifications of proteins
Disclosed are methods of quantifying multiple quality attributes, such as post translational modifications, of multiple samples in a single mass spectrometry (MS) run, including contacting two or more samples with a digesting solution under conditions sufficient to digest samples, wherein each sample is digested separately and the digesting solution is a Tris-free buffer solution; contacting each of the two or more digested samples with a specific Tandem Mass Tag (TMT) labeling reagent under conditions sufficient to label peptides within each of the digested samples with the specific TMT labeling reagent; quenching labeling of peptides within each of the two or more digested samples; combining equal volumes of the two or more labeled, digested samples into a single combined sample solution; and analyzing the single combined sample solution by targeted mass spectral analysis, thereby allowing multiple quality attributes of the two or more samples to be quantified in a single mass spectrometry (MS) run.
CIRCULATING TOTAL-NT-PROBNP (GLYCOSYLATED AND UNGLYCOSYLATED NT-PROBNP) AND ITS RATIO WITH NT-PROBNP (UNGLYCOSYLATED NT-PROBNP) IN THE ASSESSMENT OF ATRIAL FIBRILLATION
The present invention relates to a method for diagnosing atrial fibrillation in a subject, said method comprising the steps of a) determining the amount of total NT-proBNP in sample from the subject, b) determining the amount of unglycosylated NT-proBNP in a sample from the subject, c) calculating a score of the amounts determined in steps a) and b), d) comparing the calculated score with a reference score, and e) diagnosing atrial fibrillation in a subject.
Detection and quantification of glycosylated peptides
A method of purification and/or separation of glycopeptides and quantitation of same. The method includes contacting a sample comprising glycopeptides to a hydrophilic enrichment substrate under conditions that permit the glycopeptides to bind to the hydrophilic enrichment substrate. The glycopeptides are eluted from the hydrophilic enrichment substrate with an ammonium formate and acetonitrile (ACN) in water solution to create an enriched glycopeptide sample, which may be subjected to analysis to identify specific glycopeptides.
ANALYTICAL METHOD FOR GLYCOGONJUGATES USING A CAPILLARY-BASED IMMUNOASSAY SYSTEM
The invention provides analytical methods for identifying and quantifying complex glycoconjugate compositions, in particular for the analysis of a glycoconjugate in a sample comprising at least 4 glycoconjugates.
INTEGRATED PROTEOMIC BIOMARKERS FOR THE DETECTION OF AGGRESSIVE PROSTATE CANCER
The present invention provides compositions and methods useful for detecting and treating aggressive prostate cancer. In a specific embodiment, a method for identifying a patient as having aggressive prostate cancer comprises the steps of (a) measuring the concentration of total PSA, free PSA, p2PSA in a serum sample obtained from the patient and calculating phi based on the measured serum concentrations; (b) measuring the concentration of fucosylated PSA (fuc-PSA) in a serum sample obtained from the patient; (c) measuring the concentration in a serum sample obtained from the patient of one or more of the following biomarkers: B7-H3, PLA2G7, GDF-15, IL-6R alpha, SDC1, VCAM-1, s Tie-2, IL-16, CA15-3, MMP-2, and H SP27; and (d) using an algorithm to identify the patient as having aggressive prostate cancer based on a panel of biomarkers comprising phi, fuc-PSA and one or more of the serum concentrations measured in step (c).
Detection and treatment of pregnancy complications comprising determining sialyl Lewis antigens and administering hCG
Disclosed herein is a method of identifying and/or addressing incipient preeclampsia in a patient-subject by the steps of (a) performing a bioassay to determine the level of at least one sialyl Lewis antigen in a said patient-subject at about 25 weeks of pregnancy or earlier; (b) performing a bioassay to determine the level of at least one sialyl Lewis antigen in a pregnant non-preeclampsia one or more subjects at about 30 weeks of pregnancy or later, wherein said at least one sialyl Lewis antigen assay is for a sialyl Lewis antigen assayed in step (a) is and if more than one subject is assayed, averaging said results; and (c) managing said patient-subject for preeclampsia, if said level of at least one sialyl Lewis antigen of step (a) is at or greater than about 20% above the level of such sialyl Lewis antigen assayed in step (b).
A CHEMOENZYMATIC METHOD FOR THE DETECTION OF CELL-CELL PROXIMITY INTERACTION AND ISOLATION OF TUMOR-SPECIFIC ANTIGEN REACTIVE T CELLS FOR IMMUNE THERAPY
The present disclosure provides compositions and methods for monitoring cell to cell (“cell-cell”) interactions in vitro, ex vivo, and in vivo. In some embodiments, the present disclosure provides for the use of these compositions and methods (i) to identify and enrich for tumor-specific antigen (TSA) reactive T cells from tumor infiltrating lymphocytes (TILs) or circulating T cells; (ii) to identify and enrich for T cells that recognize autoantigens in a particular autoimmune disease, and/or (iii) to identify and enrich for antigen specific regulatory T cells that have the potential to be exploited to treat autoimmune disease. In some embodiments, the present disclosure also provides for methods of treating diseases, e.g., cancer with such TSA reactive T cells isolated via the present methods.
COMPOSITIONS AND METHODS FOR IDENTIFYING O-LINKED GLYCOSYLATION SITES IN PROTEINS
The present invention relates to the field of protein post-translational modification. More specifically, the present invention provides compositions and methods useful for identifying O-linked glycosylation sites in proteins. In one embodiment, the present invention provides a method for identifying O-linked glycosylation sites of Tn antigen in proteins comprising the steps of (a) digesting proteins present in a sample into peptides; (b) enriching for Tn-glycopeptides; (c) conjugating Tn-glycopeptides to solid phase; (d) labeling Tn using the glycosyltransferse enzyme C1GalT1 and a labeled uridine diphosphate galactose (UDP-Gal) substrate to produce labeled Tn-glycopeptides; (e) releasing the labeled Tn-glycopeptides from the solid-phase using an endopeptidase that cleaves peptides at the N-terminus of O-linked glycans at serine or threonine residues; and (f) mapping O-linked glycosylation sites of Tn antigen using liquid chromatography-mass spectrometry.
Dye-based liquid reagent volume indicator for use in analyte detection assays
Devices, kits, and methods are disclosed for use in detecting a concentration of an analyte of interest in a patient's liquid test sample. The devices, kits, and methods employ the use of one or more solid reagent zones that includes at least one analytical reagent for detection of an analyte of interest. The solid reagent zone(s) also includes at least one dye for determining whether results obtained from the diagnostic assay for the at least one analyte of interest are biased or inaccurate due to a loss of volume of a liquid reagent during the dispensing of the liquid reagent.
METHODS TO IMPROVE DETECTION OF GLYCOSYLAMINES
The present invention provides methods to improve the sensitivity of detecting glycosylamines released from glycoconjugates, such as glycoproteins or glycopeptides, by enzymatic digestion when labeling them with amine-reactive dyes.