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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Форма поиска

Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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Применить Всего найдено 1079. Отображено 100.
24-05-2012 дата публикации

Diagnosis of restenosis in patients undergoing percutaneous coronary intervention

Номер: US20120128660A1
Автор: Kailash Prasad
Принадлежит: UNIVERSITY OF SASKATCHEWAN

The present invention relates to compounds, compositions, methods and/or kits for determining and/or predicting and/or diagnosing and/or treating restenosis in a patient.

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Номер записи: 1
14-06-2012 дата публикации

Detection and quantification of modified proteins

Номер: US20120149883A1
Автор: Junmin Peng, Steven P. Gygi
Принадлежит: Harvard College

The invention provides a method detecting and quantifying proteins by mass spectrophotometric analysis using peptide internal standards and provides a highly sensitive way of detecting protein modifications. In one aspect, the invention provides a method for determining a site of ubiquitination in a polypeptide and for evaluating ubiquitination targets in a population of polypeptides. In this way, a proteome ubiquitination map can be obtained which comprises information relating to the ubiquitination states of a plurality of cellular polypeptides. Maps can be obtained for a variety of different types of cells and cell states. For example, ubiquitination targets in normal and diseased cells can be evaluated. Preferably, the map is stored as data files in a database. Individual ubiquitinated polypeptides identified can be used to generate molecular probes diagnostic of a cell state and/or can serve as targets for agents that modulate one or more cellular processes.

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Номер записи: 2
31-10-2013 дата публикации

Modulation of histone h2b monoubiquitination and treatment of cancer

Номер: US20130287791A1
Автор: C. Wilson Xu, Yasuyo Urasaki
Принадлежит: Individual

Provided are methods and compositions for treatment of cancer. In particular, these methods and compositions may include an inhibitor of a deubiquitinating enzyme. In certain aspects, these methods and compositions may include a modulator of glucose metabolism. Also provided are methods of assaying the glucose content of cells and tissues using detection of uH2B.

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Номер записи: 3
13-03-2014 дата публикации

Method for analyzing glycan structure

Номер: US20140070086A1
Автор: Atsuhiko Toyama, Koji Ueda
Принадлежит: RIKEN Institute of Physical and Chemical Research, Shimadzu Corp

In order to provide an analysis method that is capable of determining a glycan structure with high detection sensitivity, a method of the present invention includes the steps of: carrying out triple quadrupole mass spectrometry at various values of CID energy; creating an energy-resolved profile including yield curves representing relationships between (i) a value of the CID energy and (ii) measured amounts of specific types of product ions; preparing a reference profile, and identifying a glycan structure of a test material by comparing the energy-resolved profile with the reference profile.

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Номер записи: 4
07-01-2016 дата публикации

Proximity Assay for In Situ Detection of Targets

Номер: US20160002701A1
Автор: Farrell Michael, Hong Rui, Jiang Zeyu
Принадлежит:

A proximity detection method is described that utilizes enzymatic biotinylation to detect targets in a sample, particularly formalin fixed paraffin embedded samples using automated staining platforms. One disclosed embodiment comprises contacting the sample with a first conjugate comprising a biotin ligase and a first specific binding moiety that binds proximally to the first target; contacting the sample with a second conjugate comprising a biotin ligase substrate and a second specific binding moiety that binds proximally to the second target; subjecting the sample to conditions that allow biotinylation of the biotin ligase substrate by the biotin ligase when the first target and the second target have a proximal arrangement; and detecting biotinylation of the biotin ligase substrate. The conditions that allow biotinylation of the substrate include addition of biotin and ATP. The method also may comprise contacting the sample with a streptavidin-enzyme conjugate. Signal amplification also can be used. 1. A method for analyzing a sample to determine whether a first target is proximal to a second target , the method comprising:contacting a sample with a first conjugate comprising a biotin ligase and a first specific binding moiety for associating with a first target;contacting the sample with a second conjugate comprising a biotin ligase peptide substrate and a second specific binding moiety for associating with a second target under conditions that allow biotinylation of the peptide substrate; anddetecting the biotin if the first target is proximal to the second target.2. The method according to claim 1 , wherein labeling the first target includes contacting the sample with a first primary antibody specific to the first target and labeling the second target includes contacting the sample with a second primary antibody specific to the second target.3. The method according to claim 2 , wherein the first primary antibody is labeled with a first hapten and labeling the ...

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Номер записи: 5
04-01-2018 дата публикации

SURFACE, ANCHORED FC-BAIT ANTIBODY DISPLAY SYSTEM

Номер: US20180002402A9
Автор: Shaheen Hussam Hisham, Zha Dongxing
Принадлежит: Merck Sharp & Dohme Corp.

The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. A bait complexed with a monovalent antibody fragment can be expressed on the surface of the host cell wherein the fragment may be assayed for antigen binding while full antibody is simultaneously secreted from the host cell. Methods of using the system for identifying antibodies that bind specifically to an antigen of interest are also provided. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of use thereof. 1. An antibody display system comprising:(a) an isolated host cell;(b) a bait comprising a heavy Fc immunoglobulin domain fused to a surface anchor polypeptide;(c) one or more polynucleotides encoding an immunoglobulin light chain variable region; and(d) one or more polynucleotides encoding an immunoglobulin heavy chain variable region.2. The antibody display system of further comprising(i) a non-tethered full antibody comprising said immunoglobulin light and heavy chains; and/or(ii) a monovalent antibody fragment which is complexed with the Fc moiety of the bait.3Pichia. The antibody display system of any one of - wherein the host cell is a cell.4. The antibody display system of any one of -wherein said one or more polynucleotides encoding an immunoglobulin light chain variable region is from a genetically diverse population of immunoglobulin light chain variable regions; and/or,wherein said one or more polynucleotides encoding an immunoglobulin heavy chain variable region is from a genetically diverse population of immunoglobulin heavy chain variable regions.5. The antibody display system of any one of - wherein the host cell comprises a polynucleotide encoding the bait which is operably associated with a regulatable promoter.6. An isolated bait polypeptide comprising an Fc immunoglobulin domain or functional fragment thereof fused claim 1 , optionally by a ...

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Номер записи: 6
07-01-2016 дата публикации

BREAST-CANCER DETERMINATION METHOD

Номер: US20160003828A1
Автор: Kurono Sadamu, Matsuura Nariaki, MATSUURA Shuji, Nakajima Tomoyuki
Принадлежит:

The present invention provides a breast cancer marker selected from the group consisting of fucosylated hornerin, fucosylated Zn-α-2-glycoprotein, fucosylated Ig γ-1 chain C region, and fucosylated desmoplakin; a method for determining breast cancer comprising detecting the breast cancer marker in a sample, and determining on the basis of the results of the results; and a kit to be used for the determination. 1. A breast cancer marker selected from the group consisting of fucosylated hornerin , fucosylated Zn-α-2-glycoprotein , fucosylated Ig γ-1 chain C region , and fucosylated desmoplakin.2. A method for obtaining a data for determining breast cancer , comprising detecting one or more breast cancer markers selected from the group consisting of fucosylated hornerin , fucosylated Zn-α-2-glycoprotein , fucosylated Ig γ-1 chain C region , and fucosylated desmoplakin in a sample.3. The method according to claim 2 , wherein the sample is nipple discharge.4. The method according to claim 2 , wherein the sample is serum claim 2 , plasma claim 2 , or whole blood.5. A method for determining breast cancer claim 2 , comprising:detecting one or more breast cancer markers selected from the group consisting of fucosylated hornerin, fucosylated Zn-α-2-glycoprotein, fucosylated Ig γ-1 chain C region, and fucosylated desmoplakin in a sample, anddetermining on the basis of the results thereof.6. The method according to claim 5 , wherein the sample is nipple discharge.7. The method according to claim 5 , wherein the sample is serum claim 5 , plasma claim 5 , or whole blood.8. A kit for detecting a breast cancer marker for determining breast cancer comprising a reagent for detecting a breast cancer marker selected from the group consisting of fucosylated hornerin claim 5 , fucosylated Zn-α-2-glycoprotein claim 5 , fucosylated Ig γ-1 chain C region claim 5 , and fucosylated desmoplakin.9. The kit according to claim 8 , wherein the reagents for detecting a breast cancer marker comprises ...

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Номер записи: 7
03-01-2019 дата публикации

Determination of glycosylation signature

Номер: US20190004050A1
Автор: Ciaran RICHARDSON, Ivan McConnell, John Lamont, Peter Fitzgerald
Принадлежит: Randox Laboratories Ltd, Randox Teoranta

The present invention also describes use of a patient protein glycosylation profile to identify the presence or absence of a disease in subjects.

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Номер записи: 8
20-01-2022 дата публикации

INFLAMMATION-ENABLING POLYPEPTIDES AND USES THEREOF

Номер: US20220017960A1
Автор: GROS Philippe
Принадлежит:

This present technology relates to the use of inflammation-enabling polypeptides (or their coding sequences) to screen for agents useful for the prevention, treatment and/or alleviations of symptoms associated with an inflammatory disorder, to identify individuals susceptible of developing an exacerbated inflammatory response as well as to determine if a therapeutic regimen is capable of preventing, treating or alleviating the symptoms associated to an inflammatory disorder in an individual. The present technology also provides methods for preventing, treating and/or alleviating the symptoms associated to an inflammatory condition based on the inhibition of expression or activity of the inflammation-enabling targets. 1. A method for assessing the ability of an agent to prevent , treat and/or alleviate the symptoms associated with an inflammatory condition in an individual , said method comprising:a) combining the agent with an inflammatory enabling polypeptide selected from the group consisting of a USP15 polypeptide and a TRIM25 polypeptide;b) measuring a biological activity of the inflammatory enabling polypeptide of step (a) to obtain a test level;c) comparing the test level to a control level, wherein the control level is associated with the biological activity of the inflammatory enabling polypeptide observed during the onset or maintenance of the inflammatory condition; andd) characterizing the agent as (i) useful for the prevention, treatment and/or alleviation of the symptoms associated with the inflammatory condition when the at least one biological activity associated with the test level is lower than the biological activity associated with the control level or (ii) lacking utility for the prevention, treatment and/or alleviation of the symptoms associated with the inflammatory condition when the at least one biological activity associated with the test level is equal to or higher than the biological activity associated with the control level.2. The method ...

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Номер записи: 9
20-01-2022 дата публикации

Materials and Methods for Glycan Profiling

Номер: US20220018832A1
Автор: Okamura Koji, Shibata Mayu, Umezawa Akihiro, Yamada Masao
Принадлежит:

The invention aims to provide a non-destructive method of testing and analysing, amongst other things, cell types such as pluripotent cells, including differentiated cells and MSCs. The invention also aims to provide a non-destructive method to test and analyse the cell conditions, such as aging, activity, multipotency, differentiated directivity and effectiveness. Possibility of analysing cell types or cell conditions without destroying the cells by analyzing glycan profile, which is acquired from glycoconjugates secreted from cells in culture mediums, using microarrays with glycan binding protein, on the surface was discovered. 1. A method for determining the glycan profile of a cell of interest , said method comprising the steps of(a) culturing the cell of interest in a culture medium for a period of time;(b) following said period of time, collecting a sample of supernatant from the culture medium, wherein the supernatant is cell-free;(c) mixing the cell-free supernatant with a fluorescent labelling agent to create a labelled sample solution, wherein the fluorescent labelling agent is capable of fluorescently labelling a glycoconjugate;(d) contacting the labelled sample solution with a plurality of glycan binding proteins under conditions suitable to allow binding of a fluorescently labelled glycoconjugate to bind to at least one of said plurality of glycan binding proteins thereby forming a fluorescent labelled glycoconjugate-glycan binding protein complex, wherein said plurality of glycan binding proteins are immobilized on a solid support;(e) applying an excitation light and measuring the level of intensity of excited fluorescence generated from the fluorescent labelled glycoconjugate-glycan binding protein complex;(f) determining a glycan profile of the cell of interest based on the intensity of excited fluorescence generated.2. The method of wherein the glycoconjugate is a glycoprotein.3. The method of wherein the cell of interest is a human cell.4. The ...

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Номер записи: 10
14-01-2016 дата публикации

METHODS AND COMPOSITIONS FOR TARGETING POLYUBIQUITIN

Номер: US20160009791A1
Автор: KELLEY ROBERT F., Matsumoto Marissa L.
Принадлежит: Genentech, Inc.

Anti-K63-linked polyubiquitin monoclonal antibodies, and methods for using the antibodies, are provided. 134-. (canceled)35. An isolated antibody or antigen-binding fragment thereof that specifically binds to K63-linked polyubiquitin , comprising the HVR-H1 sequence of SEQ ID NO: 5 , the HVR-H2 sequence of SEQ ID NO: 63 , the HVR-H3 sequence of SEQ ID NO: 6 , the HVR-L1 sequence of SEQ ID NO: 3 , the HVR-L2 sequence of SEQ ID NO: 11 , and the HVR-L3 sequence of SEQ ID NO: 4.36. The isolated antibody or antigen binding fragment thereof of claim 35 , which is a mouse claim 35 , chimeric claim 35 , or humanized antibody or antigen binding fragment thereof.37. The isolated antibody or antigen binding fragment thereof of claim 35 , which has an affinity (K) for K63-linked polyubiquitin of less than or equal to 10 nM.38. The isolated antibody or antigen binding fragment thereof of claim 35 , wherein the antibody or antigen binding fragment thereof does not specifically bind to K48-linked polyubiquitin or monoubiquitin.39. The isolated antibody or antigen-binding fragment thereof of claim 35 , wherein the antibody or antigen-binding fragment has at least one property selected from inhibiting degradation of the K63-linked polyubiquitinated protein claim 35 , modulating at least one polyubiquitin-mediated signaling pathway claim 35 , inhibiting at least one polyubiquitin-mediated signaling pathway claim 35 , and stimulating at least one polyubiquitin-mediated signaling pathway.40. A composition comprising the antibody or antigen-binding fragment thereof of and a pharmaceutically acceptable carrier.41. An isolated monoclonal antibody or antigen-binding fragment thereof that binds to the same antigenic determinant on K63-linked polyubiquitin as the antibody or antigen-binding fragment of .42. The isolated monoclonal antibody or antigen binding fragment thereof of claim 41 , which is a mouse claim 41 , chimeric claim 41 , human claim 41 , or humanized antibody or antigen ...

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Номер записи: 11
19-01-2017 дата публикации

SUMOYLATION OF SERCA2A AND CARDIOVASCULAR DISEASE

Номер: US20170014494A1
Автор: Hajjar Roger Joseph, Kho Chang Won, Lee Ah Young
Принадлежит:

Methods for treating cardiovascular disease, and in particular heart failure, are provided comprising administering a therapeutically effective amount of a modulator of SERCA2a post-translation modification such as SUMOylation or acetylation. Also provided are methods of treating cardiovascular disease by inhibiting SERCA2a degradation. Further provided are methods of diagnosing a propensity to develop heart failure comprising determining if a SERCA2a mutant is present or determining the level of expression of SUMO1 in cardiomyocytes. The disclosure also provides methods of screening for therapeutics that modulate the post-translational modification of SERCA2a, such as by modulating post-translational SUMOylation and/or acetylation. 1. A method of treating cardiac dysfunction in a subject comprising administering a therapeutically effective amount of a modulator of SERCA2a post-translational modification to the subject.2. The method according to wherein the cardiac dysfunction is selected from the group consisting of heart failure claim 1 , pressure overload-induced cardiac dysfunction claim 1 , and cardiac dysfunction induced by inhibited calcium decay.3. The method according to wherein the heart failure comprises contractile dysfunction.4. The method according to wherein the heart failure is TAC-induced heart failure.5. The method according to wherein the subject is a human.6. The method according to wherein the modulator modulates SERCA2a post-translational SUMOylation.7. The method according to wherein the modulator is a vector comprising an expressible coding region encoding a protein selected from the group consisting of SERCA2a and SUMO1 claim 6 , and wherein the coding region is operably linked to at least one expression control element.8. The method according to wherein the vector is a recombinant adeno-associated virus.9. The method according to wherein the recombinant adeno-associated virus is rAAV1.10. The method according to wherein the modulator ...

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Номер записи: 12
03-02-2022 дата публикации

ASSAYS FOR DETECTING MODIFIED COMPOUNDS

Номер: US20220034897A1
Автор: Pande Praveen, Rabbani Elazar, Rabbani Joshua, Stavrianopoulos Jannis G.
Принадлежит: ENZO LIFE SCIENCES, INC.

Provided are methods and compositions which are useful for separating, isolating, detecting, and quantifying compounds of interest which have been modified chemically, enzymatically or catalytically from other compounds which have not been so modified. The modifications may take the form of functional groups which are gained, lost or retained by the compounds of interest. 1. A method for detecting or quantifying an enzyme capable of adding a functional group to a compound in a sample , comprising the steps of: (i) a compound which can gain a functional group from an enzyme, the compound comprising at least one non-radioactive signaling moiety;', '(ii) a sample suspected of containing an enzyme that could add a functional group to the compound; and, '(a) providing(b) forming a mixture comprising the compound and the sample; and(c) separating any compound that has gained the functional group; and(d) detecting or quantifying the non-radioactive signaling moiety of any separated compounds having the functional group, thereby detecting or quantifying the enzyme in the sample.2. The method of claim 1 , wherein the enzyme is selected from the group comprising kinases claim 1 , phosphatases claim 1 , sulfatases claim 1 , sufotransferases claim 1 , acetyltransferases claim 1 , deacetylases claim 1 , methylases claim 1 , demethylases claim 1 , carboxylases claim 1 , decarboxylases claim 1 , glycosylases claim 1 , amidases claim 1 , deamidases claim 1 , aminases and deaminases.3. The method of claim 1 , wherein the enzyme is selected from the group comprising kinases claim 1 , phosphatases claim 1 , or sulfotransferases.4. The method of claim 1 , wherein the enzyme is sphingosine kinase 1 or sphingosine kinase 2.5. The method of claim 1 , wherein the enzyme is sphingosine kinase 1.6. The method of claim 1 , wherein the enzyme is sphingosine kinase 2.7. The method of claim 1 , wherein the non-radioactive signaling moiety comprises a fluorescent compound claim 1 , a ...

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Номер записи: 13
21-01-2016 дата публикации

IMPROVED METHOD OF MAPPING GLYCANS OF GLYCOPROTEINS

Номер: US20160018409A1
Автор: Higel Fabian
Принадлежит: HEXAL AG

The use of anthranilic acid (2-AA) to label N-glycans prior to separation using a reversed-phase liquid chromatography (RP-LC) column under acidic conditions using formic acid is described herein. Negatively charged 2-AA offers stronger retention on the reversed phase column than 2-aminobenzamide (2-AB) in RP-LC and allows efficient ionization and detection of 2-AA labeled N-glycans. The acidic conditions used for the RP-LC leads to an efficient separation of acidic 2-AA N-glycans carrying terminal sialylation without the need for an ion-pairing reagent. The method and compositions described herein may be used with RP-nano-LC-MS and a 96-well plate sample preparation, which allows attomolar sensitivity and high throughput.

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Номер записи: 14
18-01-2018 дата публикации

Histone protein ubiquitination as a cancer biomarker

Номер: US20180017575A1
Автор: Deborah Joy Marsh, Michael Anthony HAHN
Принадлежит: Northern Sydney Local Health District

The present invention relates to the identification and use of biomarkers for the diagnosis and prognosis of cancer.

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Номер записи: 15
21-01-2021 дата публикации

Sialylated Glycoproteins

Номер: US20210017563A1
Автор: Bhatnagar Naveen, Lansing Jonathan C., Meccariello Robin, Ortiz Daniel, SARVAIYA Hetal, Washburn Nathaniel J.
Принадлежит:

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described. 1. A method of producing a sialylated IgG antibody preparation wherein at least 60% of the branched glycans on the Fc domain of the IgG antibodies comprise a terminal sialic acid on both the alpha 1 ,3 arm and the alpha 1 ,6 arm , the method comprising:providing an IgG antibody preparation;combining the IgG antibody preparation with a beta 1,4 galactosyltransferase and uridine diphosphate galactose a galactose donor to provide a galactosylation reaction mixture;incubating the galactosylation reaction mixture to allow galactosylation of branched glycans; adding a ST6 beta-galactoside alpha-2,6-sialyltransferase 1 having at least 95% identity to amino acids 95-416 of SEQ ID NO:1 and cytidine-5′-monophospho-N-acetylneuraminic acid (CMP-NANA) an ST6-Gal1 sialyltransferase and a sialic acid donor to the galactosylation reaction mixture to provide a sialylation reaction mixture; andincubating the sialylation reaction mixture for a sufficient period of time to allow least 60% of the branched glycans on the Fc domain of the IgG antibodies in the preparation to be sialylated on both the alpha 1,3 arm and on the alpha 1,6 arm, wherein additional CMP-NANA sialic acid donor is added to the sialylation reaction mixture during the incubation of the sialylation reaction mixture.2. The method of claim 1 , wherein the CMP-NANA sialic acid donor is added periodically.3. The method of or claim 1 , wherein the galactosylation reaction mixture and the sialylation reaction mixture comprise MnCl. This application is a continuation of U.S. application Ser. No. 16/589,634, filed on Oct. 1, 2019, which is a continuation of U.S. application Ser. No. 15/954,146, filed on Apr. 16, 2018, which is a continuation of U.S. application Ser. No. 14/787,403, filed on Oct. 27, 2015 (now abandoned), which is a U.S. national stage application under 35 USC § 371 of International ...

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Номер записи: 16
16-01-2020 дата публикации

DIAGNOSTIC METHOD OF LIVER CANCER USING ALPHA-FETOPROTEIN DERIVED GLYCOPEPTIDES BY MASS SPECTROMETRY

Номер: US20200018759A1
Автор: Kim Jin Young, Kim Kwang Hoe, Lee Ju Yeon, Yoo Jong Shin
Принадлежит: KOREA BASIC SCIENCE INSTITUTE

The present invention relates to a diagnosis method of liver cancer using mass spectrometry of α-fetoprotein derived glycopeptide. Particularly, according to the AFP glycopeptide analysis method of the invention, fucosylation rate of the glycopeptide having the sequence composed of Val-Asn-Phe-Thr-Glu-Ile-Gln-Lys is analyzed to diagnose liver cancer in the early developmental grade. In particular, fucosylation rate is higher in the liver cancer patients than in other liver disease patients, so that the comparison of the fucosylation rate can be useful to diagnose or distinguish liver cancer from other liver diseases in the early HCC patients. 1. A method for analyzing AFP glycopeptide to provide the information useful for the liver cancer diagnosis which comprises the following steps:1) separating α-fetoprotein (AFP) from the sample obtained from a subject;{'b': '1', '2) obtaining AFP glycopeptide by hydrolyzing the AFP separated in step );'}{'b': '2', '3) analyzing the mass and quantity of the AFP glycopeptide obtained in step );'}4) selecting the glycopeptide containing the sequence composed of Val-Asn-Phe-Thr-Glu-Ile-Gln-Lys_(VNFTEIQK, and having the molecular weight of 2600.1±0.5, 2746.2±0.5, 2891.2±0.5, 3037.2±0.5, 3182.3±0.5, or 3328.3±0.5 Da; and{'b': '4', '5) calculating the rate of fucosylation of the glycopeptide selected in step ) and comparing the calculation result with that of the sample separated from a subject having liver cirrhosis or hepatitis.'}2. The method for analyzing AFP glycopeptide to provide the information useful for the liver cancer diagnosis according to claim 1 , wherein the sample is one or more materials selected from the group consisting of tissue claim 1 , cell claim 1 , cell culture fluid claim 1 , blood claim 1 , and serum.31. The method for analyzing AFP glycopeptide to provide the information useful for the liver cancer diagnosis according to claim 1 , wherein the separation of step ) is performed by using an anti-AFP antibody. ...

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Номер записи: 17
22-01-2015 дата публикации

METHOD FOR DETERMINING THE GLYCOSYLATION OF AN ANTIBODY

Номер: US20150024410A1
Автор: Fink Michel, Jaga Delphine, Martinez Stephane, Mokrane Hamed
Принадлежит: CISBIO BIOASSAYS

The invention relates to a method for detecting the binding of an antibody to an Fc receptor present on the surface of a cell as well as to a method for determining the level of glycosylation of an antibody. The invention also relates to a reagent kit for carrying out these methods. 1. An in vitro method for determining the binding of an antibody to an Fc receptor expressed in cell membranes or intact cells present in a measurement medium , comprising the following steps:(i) direct or indirect labeling of said Fc receptor with the first member of a pair of FRET partners, or else introducing, into the medium, cell membranes or intact cells whose Fc receptors were labeled beforehand directly with the first member of a pair of FRET partners;(ii) adding said antibody, labeled directly or indirectly with the second member of said pair of FRET partners, to the measurement medium;(iii) measuring the FRET signal, the existence of a FRET signal being representative of the binding of the antibody to the Fc receptor.2. The method as claimed in claim 1 , which is repeated with different antibody concentrations and which comprises an additional step (iv) of determining the dissociation constant of the antibody-Fc receptor bond.3. An in vitro method for determining the binding of a competing antibody with an Fc receptor expressed in cell membranes or intact cells present in a measurement medium claim 1 , by competition with a reference antibody claim 1 , comprising the following steps:(i) direct or indirect labeled of said Fc receptor with the first member of a pair of FRET partners, or else introducing, into the medium, cell membranes or intact cells whose Fc receptors were labeled beforehand directly with the first member of a pair of FRET partners;(ii) adding the competing antibody to the measurement medium;(iii) adding a reference antibody, labeled directly or indirectly with the second member of said pair of FRET partners, to the measurement medium;(iv) measuring the FRET ...

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Номер записи: 18
26-01-2017 дата публикации

SURROGATES OF POST-TRANSLATIONALLY MODIFIED PROTEINS AND USES THEREOF

Номер: US20170023589A1
Автор: Chorev Michael, Halperin Jose A.
Принадлежит:

The present invention provides compounds that are surrogates of post-translationally modified proteins and uses thereof. Numerous diseases are associated with post-translationally modified proteins that are difficult to obtain in homogenous form and in quantities needed for immunization and use as convenient standards, calibrators, and/or reference compounds that facilitate the detection and analysis of endogenous post-translationally modified proteins. The surrogate compounds of the invention typically comprise antigenic epitopes (one of which carries a post-translational modification) that are tethered by a flexible and hydrophilic linker. The resulting compound behaves like a surrogate of the post-translationally modified protein because it preserves the character of the included antigens and allows recognition by specific antibodies targeting the individual antigens. The surrogate compounds may be prepared by covalently joining two or more polypeptide epitopes using one or more linkers, wherein at least one of the epitopes comprises a post-translational modification. In one aspect, the surrogate compounds of the invention comprise a C-terminal epitope and a glycated epitope of human CD59. The inventive methods allow quantification of the levels of glycated CD59 in the serum in human subjects, particularly those with diabetes or pre-diabetes. This technological platform of post-translationally modified protein surrogates can be applied to other diseases associated with post-translationally modified proteins (e.g., autoimmune diseases such as multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus). In another aspect, the invention provides antibodies that bind specifically to the compounds of the invention and methods for producing such antibodies. 1140-. (canceled)141. A kit for detecting the presence of a post-translationally modified protein in a biological sample , said kit comprising: a first epitope derived from a non-glycated epitope of ...

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Номер записи: 19
25-01-2018 дата публикации

Methods for Detecting and for Treating Pancreatic Cancer

Номер: US20180024133A1
Автор: Haab Brian B., Reatini Bryan
Принадлежит:

A method of diagnosing pancreatic cancer in a patient by detecting a level of one or more glycoforms of a Lewis antigen and a level of the one or more glycoforms of MUC5AC. The patient diagnosed with pancreatic cancer then may be treated for this disease. Also, a method for detecting a level of a glycan in a sample which includes using a capture reagent to immobilize the glycan on a substrate; exposing the immobilized glycan to a detection reagent; detecting the level of the immobilized glycan; and combining the biological sample with one or more pre-capture enzymes and/or exposing the immobilized glycan to one or more pre-detection enzymes. 1. A method of diagnosing pancreatic cancer in a human patient , said method comprising:obtaining a biological sample from the human patient, wherein the biological sample includes one or more glycoforms of a Lewis antigen and one or more glycoforms of a mucin 5AC (MUC5AC);detecting a level of the one or more glycoforms of the Lewis antigen and a level of the one or more glycoforms of MUC5AC in the biological sample; anddiagnosing the patient with pancreatic cancer when the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC are at a different level than a statistically validated threshold for the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC.2. The method of claim 1 , wherein the one or more glycoforms of the Lewis antigen are sialyl-Lewis A:sialyl-Lewis A (sLeA:sLeA) claim 1 , sialyl-Lewis A:sulfated Lewis A (sLeA:sulfo-LeA) claim 1 , sialyl-Lewis A:sulfated sialyl-Lewis A (sLeA:sulfo-sLeA) claim 1 , sialyl-Lewis A:sialyl-Lewis X (sLeA:sLeX) claim 1 , sialyl-Lewis A:sulfated Lewis X (sLeA:sulfo-LeX) claim 1 , or sialyl-Lewis A:sulfated sialyl-Lewis X (sLeA:sulfo-sLeX).3. The method of claim 1 , wherein the one or more glycoforms of MUC5AC are MUC5AC:type 1 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1) claim 1 , MUC5AC:type1type 2 sialyl-N-acetyl- ...

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Номер записи: 20
24-01-2019 дата публикации

Methods and Compositions for Targeting Polyubiquitin

Номер: US20190023774A1
Автор: KELLEY ROBERT F., Matsumoto Marissa L.
Принадлежит: Genentech, Inc.

Anti-K63-linked polyubiquitin monoclonal antibodies, and methods for using the antibodies, are provided. 134-. (canceled)35. A nucleic acid molecule encoding an antibody or antigen-binding fragment thereof that specifically binds to K63-linked polyubiquitin , comprising:the HVR-H1 sequence of SEQ ID NO: 5; (i) a D at position 1 and an S at position 3,', '(ii) an A or S at position 3,', '(iii) an S at position 3 and an L at position 6, or', '(iv) an F at position 3 and an H at position 6;, 'the HVR-H2 sequence of SEQ ID NO: 60, optionally having one set of substitutions selected from the following sets (i)-(v)the HVR-H3 sequence of SEQ ID NO: 6;the HVR-L1 sequence of SEQ ID NO: 3; (a) if HVR-H2 is SEQ ID NO: 60, then HVR-L2 optionally has a substitution of a T at position 4,', '(b) if HVR-H2 is SEQ ID NO: 60 substituted with a D at position 1 and an S at position 3, then HVR-L2 has a substitution of a A, V, S, or T at position 4 of SEQ ID NO: 8 further wherein if position 4 of SEQ ID NO: 8 is substituted with A then position 2 of SEQ ID NO: 8 is optionally substituted with A;', '(c) if HVR-H2 is SEQ ID NO: 60 substituted with an A at position 3, then HVR-L2 has a substitution of an A, L, S, or N at position 4 of SEQ ID NO: 8, further wherein if position 4 of SEQ ID NO: 8 is substituted with A then position 2 of SEQ ID NO: 8 is optionally substituted with A;', '(d) if HVR-H2 is SEQ ID NO: 60 substituted with an S at position 3, then HVR-L2 optionally has a substitution of a Vat position 4 of SEQ ID NO: 8;', '(e) if HVR-H2 is SEQ ID NO: 60 substituted with an S at position 3 and an L at position 6, then HVR-L2 has a substitution of a V at position 4 of SEQ ID NO: 8; and', '(f) if HVR-H2 is SEQ ID NO: 60 substituted with an F at position 3 and an H at position 6, then HVR-L2 has a substitution of an A at position 2 and an A at position 4 of SEQ ID NO: 8; and, 'the HVR-L2 sequence of SEQ ID NO: 8, whereinthe HVR-L3 sequence of SEQ ID NO: 4.36. The nucleic acid molecule ...

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Номер записи: 21
10-02-2022 дата публикации

CALIBRATORS AND CONTROLS FOR THE DETERMINATION OF PERCENT GLYCATED HEMOGLOBIN IN A PATIENT'S LIQUID TEST SAMPLE

Номер: US20220043009A1
Автор: Hixson Craig
Принадлежит: SIEMENS HEALTHCARE DIAGNOSTICS INC.

Non-limiting embodiments of methodologies for preparing diagnostic assay(s) calibrator(s), calibration material(s), and/or control(s), as well as kits, devices, and method(s) of calibration related thereto. 1. A method for preparing a calibration system comprising at least two calibrators for use in an analyte detection assay instrument , the method comprising the steps of:preparing a first calibrator, the first calibrator having a first concentration of an analyte of interest present in a patient's liquid test sample, wherein the first concentration is higher than an average threshold concentration of the analyte of interest present in the patient's liquid test sample; andpreparing a second calibrator, the second calibrator having a second concentration of the analyte of interest present in the patient's liquid test sample, wherein the second concentration is lower than the average threshold concentration of the analyte of interest present in the patient's liquid test sample.2. The method of claim 1 , wherein the patient's liquid test sample is whole blood.3. The method of claim 2 , wherein the analyte of interest is selected from the group consisting of total hemoglobin claim 2 , glycated hemoglobin claim 2 , and combinations thereof.4. The method of claim 3 , wherein the average threshold concentration of the glycated hemoglobin is from about 4% to about 8% of a concentration the total hemoglobin present in the patient's liquid test sample.5. The method of claim 4 , wherein the first concentration of the glycated hemoglobin is equal to or greater than about 8.1%.6. The method of claim 4 , wherein the second concentration of the glycated hemoglobin is equal to or less than about 3.9%.7. The method of claim 3 , wherein the first concentration of the first calibrator is increased above the average threshold concentration of the glycated hemoglobin by a method selected from the group consisting of glucose addition and reaction claim 3 , removal of heme claim 3 , ...

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Номер записи: 22
10-02-2022 дата публикации

DYE-BASED LIQUID REAGENT VOLUME INDICATOR FOR USE IN ANALYTE DETECTION ASSAYS

Номер: US20220043010A1
Автор: Kauffmann Aaron, Ledden David, Stradinger Jon, Zimmerle Chris
Принадлежит: SIEMENS HEALTHCARE DIAGNOSTICS INC.

Non-limiting embodiments of a modified devices that comprise at least one dye for determining whether results obtained from the conductance of at least one diagnostic assay are biased, as well as kits and methods of use related thereto. 1. A method for detecting and correcting inaccurate concentrations of an analyte of interest present within a patient's liquid test sample , the method comprising the steps of: [ 'at least one dye, wherein the at least one dye produces at least two detectable responses when the at least one dye is mixed with at least one liquid reagent, wherein the at least two detectable responses produced by the at least one dye are measured at different wavelengths; and', 'a reaction chamber for the conductance of one or more diagnostic assays, the reaction chamber further comprising at least one solid reagent zone, wherein at least one of the at least one solid reagent zone comprises, 'a container, wherein the container contains at least one liquid analytical reagent, the container being in fluid communication with the reaction chamber to thereby dispense the at least one liquid analytical reagent into the reaction chamber at a predetermined time;', 'a liquid analytical reagent dispensing apparatus contained within the housing, the apparatus comprising], 'providing a housing for conducting at least one diagnostic assay, wherein the housing comprisesintroducing the liquid test sample into the reaction chamber;introducing the at least one liquid analytical reagent from the dispensing apparatus into the reaction chamber, whereby the at least one liquid analytical reagent mixes with the liquid test sample and the at least one dye to thereby form a reaction mixture in the reaction chamber;measuring a first detectable response in the reaction mixture, wherein the first detectable response is measured at a first wavelength to obtain a first value;measuring a second detectable response in the reaction mixture, wherein the second detectable response is ...

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Номер записи: 23
24-01-2019 дата публикации

METHODS RELATED TO BIOLOGICS

Номер: US20190025325A1
Автор: Robblee John
Принадлежит:

The present disclosure relates to the characterization and production of biologics. 1. A method of manufacturing a drug product , comprising:providing or obtaining a sample of a batch of a test glycoprotein drug substance, wherein the batch of the test glycoprotein drug substance comprises a recombinant antibody composition having a first amino acid sequence with at least 98% identity to SEQ ID NO:1 and a second amino acid sequence with at least 98% identity to SEQ ID NO:2;acquiring a level of sialylated glycans of the test glycoprotein in the sample; andprocessing at least a portion of the batch of the test glycoprotein drug substance as drug product if the acquired level of sialylated glycans is about 10-50% (e.g., about 10%, 15%, 20%, 25% 30%, 35%, 40%, or 50%), ortaking an alternative action if the acquired level of sialylated glycans is not about 10-50%.2. A method of manufacturing drug product , comprising:providing or obtaining a sample of a batch of a test glycoprotein drug substance, wherein the batch of the test glycoprotein drug substance comprises a recombinant antibody composition having a first amino acid sequence with at least 98% identity to SEQ ID NO:1 and a second amino acid sequence with at least 98% identity to SEQ ID NO:2;acquiring a level of sialylated glycans of the test glycoprotein in the sample; andformulating at least a portion of the batch of the test glycoprotein drug substance as drug product if the acquired level of sialylated glycans is about 10-50% (e.g., about 10%, 15%, 20%, 25% 30%, 35%, 40%, or 50%), ortaking an alternative action if the acquired level of sialylated glycans is not about 10-50%.3. A method of manufacturing a drug product , comprising:providing or obtaining a sample of a batch of a test glycoprotein drug substance, wherein the batch of the test glycoprotein drug substance comprises a recombinant antibody composition having a first amino acid sequence with at least 98% identity to SEQ ID NO:1 and a second amino acid ...

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Номер записи: 24
29-01-2015 дата публикации

BIOTINYLATED MHC COMPLEXES AND THEIR USES

Номер: US20150031566A1
Автор: NAPPER Catherine Elizabeth, SCHWABE Nikolai Franz Gregor
Принадлежит:

The invention demonstrates an improved choice of biotinylation peptide to be used in a combination or fusion with an MHC molecule for immobilizing or multimerising such MHC molecules for a variety of purposes. 1. A chimeric peptide comprising an MHC peptide and a biotinylation peptide which either is a natural biotinylation peptide or has a greater than 70% sequence homology to a natural biotinylation peptide , the biotinylation peptide comprising a minimal sequence from a naturally occurring biotinylation domain that can be biotinylated by a corresponding enzyme.2. The chimeric peptide comprising an MHC peptide and a biotinylation peptide wherein the biotinylation peptide has a minimal sequence required for being biotinylated that is longer than 50 amino acids in length the biotinylation peptide comprising a minimal sequence from a naturally occurring biotinylation domain that can be biotinylated by a corresponding enzyme.3. The chimeric peptide of wherein the MHC peptide is a Class I MHC peptide or a Class II MHC peptide.4. The chimeric peptide of wherein the MHC peptide is a Class I MHC or a Class II MHC peptide.5. The chimeric peptide of wherein the biotinylation peptide is biotinylated.6. The chimeric peptide of wherein the biotinylation peptide is located in the chimeric protein after the C-terminal end of the MHC peptide.7. The chimeric peptide of wherein the MHC peptide and the biotinylation peptide are separated by a linker sequence.8Proprionibacterium shermanii. The chimeric peptide of wherein the biotinylation peptide is selected from the group consisting of the biotinylation domain of BCCP and 1.3S subunit of transcarboxilase.9. An MHC peptide complex with the formula (α-β-P) claim 1 , wherein α comprises an α chain of a MHC I or MHC II class molecule claim 1 , β comprises an β chain of a MHC I or MHC II class molecule claim 1 , and P is a peptide antigen bound in the binding groove of the MHC molecule claim 1 , wherein said MHC peptide complex comprises ...

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Номер записи: 25
02-02-2017 дата публикации

USE OF GLYCAN AS BIOMARKERS FOR AUTOIMMUNE DISEASES

Номер: US20170030904A1
Автор: GAO Weina, Huang Hao, JIANG Zhi-Hong, LIU LIANG, MENG Qiong, TONG Tiantian, WANG Jing-Rong, YAU Leefong
Принадлежит:

The present invention discloses a method of determining the presence of autoimmune disease with the use of glycan biomarkers. A method of improving the detection sensitivity of trace glycans from a mixture of glycans and a microfluidic chip therefor are also disclosed. 1. A method of determining the presence of autoimmune disease comprising the steps of:a) generating a glycosylation profile of glycoprotein of a subject;b) identifying a glycan biomarker from said glycosylation profile;c) quantifying relative abundance of said glycan biomarker; andd) determining the presence of said autoimmune disease when relative abundance of said glycan marker exceeds a predetermined threshold value.2. The method of wherein said autoimmune disease is rheumatoid arthritis.3. The method of wherein said glycoprotein is immunoglobulin.4. The method of wherein said glycoprotein is serum IgG.5. The method of wherein said glycosylation profile of glycoprotein is N-glycome of serum IgG.6. The method of wherein said glycan biomarker is sulfated glycan.7. The method of wherein said sulfated glycan comprises a structure set forth in claim 1 , item 10 or in claim 1 , item 12.8. The method of wherein said threshold value is in the range of 70%-90%.9. The method of wherein said threshold value is 80%.10. A microfluidic chip for enriching trace glycan from a mixture of glycans comprisingan enrichment column that attaches to an analytical column, a first section that includes porous graphitized carbon;', 'a second section that connects to said first section and includes titanium dioxide; and', 'a third section that connects to said second section and includes porous graphitized carbon;, 'wherein said enrichment column enriches acidic glycans and includes'}wherein said analytical column includes porous graphitized carbon;wherein said first section and said third section of said enrichment column perform pre-enrichment of glycans to remove non-glycan constituents; said second section of said ...

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Номер записи: 26
04-02-2016 дата публикации

MAMMALIAN PROTEIN CO-RECOGNITION BY BROADLY NEUTRALIZING ANTIBODIES AS MODIFIED IMMUNOGENS FOR RE-ELICITATION

Номер: US20160033532A1
Автор: Guenaga Javier, Kumar Shailendra, Tran Karen, WYATT Richard
Принадлежит:

The present invention relates to using HIV-1 broadly neutralizing antibodies to screen for glycan-dependent or protein-dependent self reactivities inherent in these mutated antibodies, and defining this cross recognition at the molecular level, and utilizing the information to re-elicit trimer-specific and/or N-glycan-dependent or protein-surface-dependent broadly neutralizing antibodies and therapeutic applications thereof. 1. A method of screening for glycan-dependent self reactivities comprising immunoprecipitating a non-human immunodeficiency virus (HIV) protein from a media with a broadly neutralizing antibody.2. The method of wherein the broadly neutralizing antibody is selected from the group consisting of PGT151 and PGT121 and VRC06.3. The method of wherein the media is spent tissue culture (TC) supernatant.4. The method of wherein the broadly neutralizing antibody is PGT151.5. The method of wherein the non-HIV protein is galectin 3 binding protein (gal3BP).65. A method of defining cross recognition comprising determining immuprecipitating putative unmutated ancestral antibodies (UAs) of a broadly neutralizing antibody and the non-HIV protein from any one of - claims 1 , wherein a lack of immunoprecipitation suggests that the germline version of the antibody does not recognize the non-HIV protein. Therefore recognition likely evolved during the affinity maturation process in the germinal center (GC) reaction by somatic hypermutation and breaking of peripheral tolerance to now recognize the human self-protein.7. The method of wherein the broadly neutralizing antibody is PGT151.8. The method of wherein the non-HIV protein is gal3BP.9. A method of eliciting trimer-specific and/or N-glycan-dependent broadly neutralizing antibodies in a patient in need thereof comprising administering the non-HIV protein of to the patient.10. The method of wherein the non-HIV protein is modified.11. The method of wherein the non-HIV protein is arrayed on a particle.12. The method ...

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Номер записи: 27
01-05-2014 дата публикации

Method for analyzing glycan structure

Номер: US20140117225A1
Автор: Atsuhiko Toyama, Koji Ueda
Принадлежит: RIKEN Institute of Physical and Chemical Research, Shimadzu Corp

In order to provide an analysis method that is capable of determining a glycan structure with high detection sensitivity, a method of the present invention includes the steps of: carrying out triple quadrupole mass spectrometry at various values of CID energy; creating an energy-resolved profile including yield curves representing relationships between (i) a value of the CID energy and (ii) measured amounts of specific types of product ions; preparing a reference profile, and identifying a glycan structure of a test material by comparing the energy-resolved profile with the reference profile.

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Номер записи: 28
30-01-2020 дата публикации

Sialylated glycoproteins

Номер: US20200032312A1
Автор: Daniel ORTIZ, Hetal Sarvaiya, Jonathan C. Lansing, Nathaniel J. Washburn, Naveen Bhatnagar, Robin MECCARIELLO
Принадлежит: Momenta Pharmaceuticals Inc

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described.

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Номер записи: 29
05-02-2015 дата публикации

High-Throughput Method For Sialic Acid Quantitation

Номер: US20150038362A1
Автор: Markely Lam Raga Anggara, Prajapati Shashi
Принадлежит:

The present invention relates to a method for specifically measuring sialylation of a biomolecule of interest without the interference of other biomolecules present in the sample. 1. A method of detecting sialylation of a biomolecule of interest in a sample comprising:(a) purifying the biomolecule of interest by contacting the sample with an agent that binds the biomolecule of interest;(b) denaturing the biomolecule of interest by incubating the sample with a surfactant;(c) contacting the denatured sample with an agent capable of removing terminal sialic acid residues from the biomolecule of interest, thereby generating free sialic acid residues;(d) labeling the free sialic acid residues with a detectable label;(e) detecting the labeled sialic acid residues, thereby detecting sialylation of the biomolecule of interest; whereby said method allows for the detection of sialylation of the biomolecule of interest without interference from host cell proteins or impurities.2. The method of claim 1 , further comprising quantifying the level of sialylation of the biomolecule of interest.3. The method of or claim 1 , wherein the sample comprises a cell culture supernatant.4. The method of or claim 1 , wherein the sample comprises a clinical sample.54. The method of any of - claims 1 , wherein the sample is located in a multi-well vessel.6. The method of claim 5 , wherein said multi-well vessel comprises up to 96 wells.7. The method of claim 5 , wherein said multi-well vessel comprises greater than 96 wells.8. The method of claim 5 , wherein said multi-well vessel comprises 384 wells.9. The method of claim 5 , wherein said multi-well vessel comprises 1536 wells.109. The method of any of - claims 1 , wherein the agent that binds the biomolecule of interest is an antibody claims 1 , lectin claims 1 , antigen claims 1 , or receptor that specifically binds the biomolecule of interest.119. The method of any of - claims 1 , wherein the agent that binds the biomolecule of interest is ...

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Номер записи: 30
11-02-2016 дата публикации

ApoIII and the Treatment and Diagnosis of Diabetes

Номер: US20160041191A1
Автор: Berggren Per-Olof
Принадлежит:

The present invention provides methods of identifying candidate compounds for the treatment of type I diabetes comprising contacting pancreatic β cells with an amount of apolipoprotein CIII (“apoCIII”) effective to increase intracellular calcium concentration, in the presence of one or more test compounds, and identifying those test compounds that inhibit an apoCIII-induced increase in intracellular calcium concentration in the pancreatic β cells. The present invention also provides methods for treating patients with type I diabetes comprising administering to the patient an amount effective of an inhibitor of apoCIII to reduce apoCIII-induced increase in intracellular calcium concentration in pancreatic β cells. 1. A method of identifying candidate compounds for the treatment of diabetes comprising contacting pancreatic β cells with an amount of apolipoprotein CIII (“apoCIII”) effective to increase intracellular calcium concentration in the presence of one or more test compounds and identifying those test compounds that inhibit an apoCIII-induced increase in intracellular calcium concentration in the pancreatic β cells.2. The method of wherein the apoCIII comprises sialylated apoCIII.3. The method of wherein the apoCIII is substantially purified.4. The method of further comprising synthesizing the test compounds that inhibit an apoCIII-induced increase in intracellular calcium concentration in the pancreatic β cells.5. A method for treating patients with diabetes comprising administering to a patient with diabetes an amount effective of an inhibitor of apoCIII to reduce apoCIII-induced increase in intracellular calcium concentration in pancreatic β cells claim 1 , wherein the inhibitor is selected from the group consisting of an apoCIII-selective antibody claim 1 , an antisense oligonucleotide directed against the apoCIII mRNA claim 1 , and a small interfering RNA directed against the apoCIII mRNA.7. A method for diagnosing diabetes or a propensity to develop ...

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Номер записи: 31
07-02-2019 дата публикации

SURFACE, ANCHORED FC-BAIT ANTIBODY DISPLAY SYSTEM

Номер: US20190040118A1
Автор: Shaheen Hussam Hisham, Zha Dongxing
Принадлежит: Merck Sharp & Dohme Corp.

The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. A bait complexed with a monovalent antibody fragment can be expressed on the surface of the host cell wherein the fragment may be assayed for antigen binding while full antibody is simultaneously secreted from the host cell. Methods of using the system for identifying antibodies that bind specifically to an antigen of interest are also provided. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of use thereof. 120-. (canceled)21. A method for making an antibody display system comprising:(a) an isolated eukaryotic host cell;(b) a bait comprising a human Fc immunoglobulin domain fused to a surface anchor polypeptide;(c) one or more polynucleotides encoding an immunoglobulin light chain variable region;(d) one or more polynucleotides encoding an immunoglobulin heavy chain variable region;comprising introducing, into said eukaryotic host cell, a polynucleotide encoding said bait, said one or more polynucleotides encoding an immunoglobulin light chain variable region; and said one or more polynucleotides encoding an immunoglobulin heavy chain variable region.22. A method for making an antibody or antigen-binding fragment thereof comprisingintroducing, into an isolated eukaryotic host cell comprising a bait that includes a human Fc immunoglobulin domain or functional fragment thereof fused to a surface anchor polypeptide or functional fragment thereof, one or more polynucleotides encoding an immunoglobulin light chain variable region; and/or one or more polynucleotides encoding an immunoglobulin heavy chain variable region; andculturing the host cell under condition whereby the polynucleotides encoding the immunoglobulin chains are expressed and an antibody or antigen-binding fragment thereof is formed from said chains;wherein said bait is operably associated with a ...

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Номер записи: 32
06-02-2020 дата публикации

CLEAVABLE PROBES FOR ISOTOPE TARGETED GLYCOPROTEOMICS AND METHODS OF USING THE SAME

Номер: US20200041524A1
Автор: Bertozzi Carolyn R., Woo Christina
Принадлежит:

Methods for producing isotopically-labelled peptides are provided. Aspects of the method include: contacting a sample including a metabolically tagged protein with a cleavable probe to produce a probe-protein conjugate; separating the probe-protein conjugate from the sample; digesting the probe-protein conjugate to produce a probe-peptide conjugate; and cleaving a cleavable linker to release an isotopically labelled peptide. The method may further include: identifying a predetermined isotopic pattern in a mass spectrum; determining an amino acid sequence of the isotopically labelled peptide; and identifying the site of protein glycosylation based on the determined amino acid sequence. Also provided are cleavable probes for practicing the subject methods, described by the Formula: A-L-(M-Z) where A is an affinity tag, L is a cleavable linker, M is an isotopic label and Z is a chemoselective tag capable of cross-linking a metabolically tagged protein. Compositions and kits for practicing the subject methods are also provided. 1. A method for producing an isotopically-labelled peptide , the method comprising: [{'br': None, 'A-L-(M-Z)\u2003\u2003 (I)'}, A is an affinity tag', 'L is a cleavable linker;', 'M is an isotopic label; and', 'Z is a chemoselective tag capable of cross-linking the metabolically tagged protein;, 'wherein], 'contacting a sample including a metabolically tagged protein with a cleavable probe under conditions sufficient to produce a probe-protein conjugate, wherein the cleavable probe is described by Formula (I)separating the probe-protein conjugate from the sample;digesting the probe-protein conjugate to produce a probe-peptide conjugate; andcleaving the cleavable linker to release the isotopically labelled peptide.2. The method of claim 1 , wherein the metabolically tagged protein is a metabolically tagged glycosylated protein and the isotopically labelled peptide is an isotopically labelled glycopeptide.3. The method of claim 1 , wherein the ...

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Номер записи: 33
18-02-2021 дата публикации

Plasmonic biosensor

Номер: US20210048435A1
Автор: Alexander BELUSHKIN, Filiz YESILKÖY, Hatice Yanik Altug
Принадлежит: Ecole Polytechnique Federale de Lausanne EPFL

The present invention relates to a plasmonic biosensor system. The system includes a nano-hole array device comprising at least one nano-hole array (NHA) including at least one or a plurality of nano holes (NH), an image sensor (A3) for capturing light provided by a light source (A1) and transmitted through the nano-hole array (NHA), and at least one or a plurality of nano-particles (NP) configured to be received by the nano-holes (NH) of the nano-hole array (NHA).

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Номер записи: 34
16-02-2017 дата публикации

Immunochromatographic analysis device, immunochromatographic analysis method, and immunochromatographic analysis kit

Номер: US20170045510A1
Автор: Aki Miyata, Daisuke Ito, Yuya Kato
Принадлежит: Tanaka Kikinzoku Kogyo KK

An object is to provide an immunochromatographic analysis device capable of measuring various components contained in various analytes such as blood and urine efficiently in a short measurement time without the need for complicated measurement preparation and operations. The above object was achieved by an immunochromatographic analysis device for developing an analyte-containing solution obtained by diluting an analyte containing a detection target with an analyte dilution solution, wherein a sample addition part contains an anionic surfactant.

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Номер записи: 35
16-02-2017 дата публикации

METHOD AND MEANS FOR THE NON-INVASIVE DIAGNOSIS OF TYPE II DIABETES MELLITUS

Номер: US20170045533A1
Автор: FROLOV Andrej, Hoffmann Ralf, LI Yichao, SPILLER Sandro, WELCH Lonnie R.
Принадлежит:

The invention relates to a method and means for the non-invasive diagnosis of type II diabetes mellitus. The glycation state is determined in at least one glycation position of selected plasma proteins. 1. Method for non-invasive diagnosis of diabetes , particularly type II diabetes mellitus , wherein the glycation of human plasma proteins is determined in at least one glycation position selected fromLys 64 in human serum albumin (P02768, SEQ ID No. 31),Lys 73 in human serum albumin (P02768, SEQ ID No. 31),Lys 93 in human serum albumin (P02768, SEQ ID No. 31),Lys 174 in human serum albumin (P02768, SEQ ID No. 31),Lys 181 in human serum albumin (P02768, SEQ ID No. 31),Lys 233 in human serum albumin (P02768, SEQ ID No. 31),Lys 262 in human serum albumin (P02768, SEQ ID No. 31),Lys 359 in human serum albumin (P02768, SEQ ID No. 31),Lys 378 in human serum albumin (P02768, SEQ ID No. 31),Lys 414 in human serum albumin (P02768, SEQ ID No. 31),Lys 525 in human serum albumin (P02768, SEQ ID No. 31),Lys 545 in human serum albumin (P02768, SEQ ID No. 31),Lys 574 in human serum albumin (P02768, SEQ ID No. 31),Lys 41 in the human Ig kappa chain C region (P01834, SEQ ID No. 32),Lys 75 in the human Ig kappa chain C region (P01834, SEQ ID No. 32),Lys 99 in the human Ig kappa chain C region (P01834, SEQ ID No. 32),Lys 163 in the human fibrinogen beta chain (P02675, SEQ ID No. 33),Lys 211 in the human fibrinogen beta chain (P02675, SEQ ID No. 33),Lys 295 in the human fibrinogen beta chain (P02675, SEQ ID No. 33),Lys 1003 of human alpha-2-macroglobulin (P01023, SEQ ID No. 34),Lys 1162 of human alpha-2-macroglobulin (P01023, SEQ ID No. 34),Lys 683 of human serotransferrin (P02787; SEQ ID No. 35),Lys 50 in the human Ig lambda chain C region (P01842; SEQ ID No. 36),Lys 120 of human apolipoprotein A-1 precursor (P02647; SEQ ID No. 37),Lys 131 of human apolipoprotein A-1 precursor (P02647; SEQ ID No. 37),Lys 141 of human haptoglobin (P00738; SEQ ID No. 38),Lys 1325 of human complement C3 ...

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Номер записи: 36
25-02-2016 дата публикации

METHODS FOR IDENTIFYING PROTEINS AND COMPOUNDS THAT MODULATE THE ACTIVITY OF OTUB1

Номер: US20160053298A1
Автор: Dibello Anthony, Wiener Reuven, Wolberger Cynthia
Принадлежит: THE JOHNS HOPKINS UNIVERSITY

The present invention describes that OTUB1 cleavage of K48 poly-ubiquitin is stimulated by a select subset of E2 enzymes, and that this stimulation is regulated by the ubiquitin-charged state of the E2 and free ubiquitin. Structural and biochemical studies of OTUB1 and UBCH5B show that the E2 stimulates binding of the polyubiquitin substrate by contacting the OTUB1 N-terminal ubiquitin-binding helix. Methods for identifying E2 enzymes which stimulate or inhibit cleavage of K48 polyubiquitin, as well as novel target compounds which modulate this interaction are provided. 1. A method for screening compounds which modulate the activity of an E2 peptide on the isopeptidase activity of OTUB1 comprising:a) providing a solution comprising a sufficient amount of K48 UB2 OTUB1 enzyme substrate;b) adding to the solution of a) a known quantity of an E2 enzyme;c) adding to the solution of a) a known quantity of a target compound;d) contacting the solution of c) with a known amount of OTUB1 enzyme for a specified period of time; to allow the enzymatic deubiquination of the K48 UB2 by OTUB1;e) quenching the deubiquination reaction of d);f) separating the resultant reaction products from the reaction of d);g) quantifying the amount of deubiquination of the K48 UB2 and comparing the amount of deubiquination to a control solution without the E2 enzyme and a positive control solution without the target compound; andg) determining whether the amount of deubiquination was increased or decreased relative to the amount of deubiquination in the control solution and whether the amount of deubiquination was increased or decreased relative to the amount of deubiquination in the positive control solution.2. A method for measuring the modulatory activity of an E2 peptide on the isopeptidase activity of OTUB1 comprising:a) providing a solution comprising a sufficient amount of K48 UB2 OTUB1 enzyme substrate;b) adding to the solution of a) a known quantity of an E2 enzyme;c) contacting the ...

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Номер записи: 37
13-02-2020 дата публикации

Immunochromatographic test piece for extracting and measuring sugar chain antigen, which is capable of preventing non-specific reaction

Номер: US20200049703A1
Автор: Daisuke Kato, Shino MURAMATSU, Tomohiro HATTORI
Принадлежит: Denka Seiken Co Ltd

It is intended to provide an immunochromatographic test piece which prevents a non-specific reaction by efficiently and continuously contacting and neutralizing a developing solution containing nitrous acid with a neutralizing reagent in an immunochromatography method of extracting and measuring a sugar chain antigen by nitrous acid extraction on the immunochromatographic test piece. The present invention provides an immunochromatographic test piece for extracting and measuring a sugar chain antigen in a specimen, the immunochromatographic test piece comprising: a sample pad to which a specimen mixed with nitrite or an acid solution is added; a label region comprising a labeled antibody obtained by labeling an antibody against the sugar chain antigen; and a detection region on which the antibody against the sugar chain antigen is immobilized, wherein an antibody-sugar chain antigen-labeled antibody complex is formed in the detection region to measure the sugar chain antigen, and the immunochromatographic test piece having a region impregnated with a neutralizing reagent upstream of the label region, and further having a region impregnated with a solid acid reagent when the specimen mixed with the nitrite is used, or a region impregnated with nitrite when the specimen mixed with the acid solution is used, upstream of the region impregnated with the neutralizing reagent, wherein a material for the region impregnated with the neutralizing reagent is a filter or a glass filter having three properties of being highly absorbable, being highly water-retainable, and being low releasable or continuously releasable, and owing to the high water-absorbable property and the high water-retainable property of the region impregnated with the neutralizing reagent, the acid solution containing the sugar chain antigen is sufficiently neutralized, and owing to the low releasable property or the sustained releasable property of the region impregnated with the neutralizing reagent, a ...

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Номер записи: 38
21-02-2019 дата публикации

ASSAYS FOR B-TYPE NATRIURETIC PEPTIDE ANALOGUES RESISTANT TO PROLYL-SPECIFIC DIPEPTIDYL DEGRADATION

Номер: US20190056386A1
Автор: Buechler Kenneth F., Whittaker Michael A.
Принадлежит: ALERE SAN DIEGO, INC.

The present invention describes compositions and methods for treating cardiovascular disease and myocardial infarction using dipeptidyl peptidase inhibitors. Also provided are methods for increasing natriuretic peptide function by administering one or more analogues of B type natriuretic peptide that provide increased stability in the presence of prolyl-specific dipeptidyl peptidases. 1. A method for the treatment of cardiovascular disease comprisinga) contacting a sample from a subject with a first antibody selected to bind biologically active forms of the natriuretic peptide of interest contacting the sample with a second antibody selected to bind all biologically active and biologically inactive forms of the natriuretic peptide of interest and performing an assay in which the signal depends upon the selected antibody specifically binding to a biologically active form of the natriuretic peptide of interest, but not specifically binding to a biologically inactive form of the natriuretic peptide of interest; andb) administering a therapeutically effective amount of a dipeptidyl peptidase inhibitor to said subject when the presence of said biologically active form of said natriuretic peptide is present, and wherein said administration preserves the presence of said biologically active form of said natriuretic peptide.217.-. (canceled)18. The method of claim 1 , wherein said biologically active natriuretic peptide of interest is BNP-32.19. The method of claim 1 , wherein the first and/or second antibodies are each conjugated to a detectable label.20. The method of claim 19 , wherein the label is a fluorescent or luminescent tag claim 19 , a metal claim 19 , a dye claim 19 , a radionuclide claim 19 , or an enzyme.21. The method of claim 1 , wherein the first and/or second antibodies are immobilized onto a solid support.22. The method of claim 21 , wherein the solid support is a magnetic particle claim 21 , a chromatographic matrix particle claim 21 , the surface of an ...

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Номер записи: 39
01-03-2018 дата публикации

METHOD OF PREPARING SAMPLE FOR ANALYSIS AND ANALYSIS METHOD

Номер: US20180059094A1
Автор: NISHIKAZE Takashi
Принадлежит: SHIMADZU CORPORATION

In the method of preparing a sample for analysis, a reaction is performed that produces different modified product depending on the sialic acid linkage type when a sialic acid is bound to a sugar chain of an analyte. In this reaction, an analyte containing a sugar chain, an amine containing two or more carbon atoms, and a dehydration-condensation agent are used. The sialic acid linkage type can be identified by analyzing the resulting sample with mass spectrometry etc. This method is applicable not only to free sugar chains but also to glycopeptides and glycoproteins. 113-. (canceled)14. A sample preparation method for analyzing a sugar chain contained in an analyte , whereinthe method comprising a first reaction,in the first reaction, an analyte containing a sugar chain, an amine containing two or more carbon atoms or a salt thereof, and a dehydration-condensation agent are reacted such that a modified product is formed from a sialic acid in the sugar chain of the analyte,when the sugar chain has an α2,3-linked sialic acid, a lactone is formed as the modified product by the first reaction, andwhen the sugar chain has an α2,6-linked sialic acid, an amide is formed as the modified product by the first reaction.15. The sample preparation method according to claim 14 , wherein the dehydration-condensation agent includes a carbodiimide.16. The sample preparation method according to claim 14 , wherein the amine is an alkylamine having a branched alkyl group or a salt thereof.17. The sample preparation method according to claim 14 , wherein the amine is a primary alkylamine or a salt thereof.18. The sample preparation method according to claim 14 , wherein the amine is isopropylamine or a salt thereof.19. The sample preparation method according to claim 14 , wherein the first reaction is performed in a state where the analyte is immobilized on a solid-phase carrier.20. The sample preparation method according to claim 14 , further comprising the step of subjecting the ...

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Номер записи: 40
28-02-2019 дата публикации

Sumoylation of Serca2A and Cardiovascular Disease

Номер: US20190060424A1
Автор: Hajjar Roger Joseph, Kho Chang Won, Lee Ah Young
Принадлежит:

Methods for treating cardiovascular disease, and in particular heart failure, are provided comprising administering a therapeutically effective amount of a modulator of SERCA2a post-translation modification such as SUMOylation or acetylation. Also provided are methods of treating cardiovascular disease by inhibiting SERCA2a degradation. Further provided are methods of diagnosing a propensity to develop heart failure comprising determining if a SERCA2a mutant is present or determining the level of expression of SUMO1 in cardiomyocytes. The disclosure also provides methods of screening for therapeutics that modulate the post-translational modification of SERCA2a, such as by modulating post-translational SUMOylation and/or acetylation. 117.-. (canceled)18. A method of diagnosing a subject's propensity to develop heart failure , comprising determining the level of expression of SUMO1 in a cardiomyocyte of the subject and comparing that level to the level of expression of SUMO1 in a cardiomyocyte of a healthy control , wherein reduced expression of SUMO1 relative to the control is indicative of a propensity to develop cardiac failure.19. (canceled)20. A method of diagnosing a patient's disposition towards a cardiovascular disease , comprising (a) obtaining a biological sample from a patient , (b) determining the amino acid sequence of SERCA2a at one or more positions 479-482 and/or one or more positions 584-587 , and (c) diagnosing a disposition towards cardiovascular disease if the amino acid sequence varies from the wild-type sequence of SERCA2a (SEQ ID NO: 2).21. The method of claim 20 , wherein step b is carried out by determining the polynucleotide sequence encoding the amino acid sequence of SERCA2a at the position(s).22. The method of claim 20 , wherein the cardiovascular disease is heart failure.23. A method of diagnosing a subject's propensity to develop heart failure claim 20 , comprising (a) determining the expression level of SERCA2a claim 20 , the level of ...

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Номер записи: 41
10-03-2016 дата публикации

MONOCLONAL ANTIBODY RECOGNIZING SIALYLATED SUGAR CHAINS

Номер: US20160068611A1
Автор: Okuda Tetsuya
Принадлежит: NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY

The purpose of the present invention is to provide a novel monoclonal antibody having high affinity and that strictly recognizes, as a sugar chain epitope, only a “Siaα2,6Galβ1,4GlcNAc (6′-Sialyl-LacNAc): CDw75” sugar chain structure, being a molecular target for diagnosis of the malignancy of tumors. An anti-CDw75 monoclonal antibody is provided that recognizes “CDw75” sugar chain structures but does not recognize similar sugar chain structures indicated by “Galβ1,4GlcNAc”, “Siaα2,3Galβ1,4GlcNAc”, or “Siaα2,6Galβ1,4Glc”, by using a glycolipid antigen bonding a carrier lipid compound “HOCHCH (NH—CO—(CH)—CH)—(CH)—CH(C12L)” developed by the inventors to a “CDw75” sugar chain. The obtained anti-CDw75 monoclonal antibody is an excellent detection drug for B-cell lymphoma, gastric cancer, or colorectal cancer, an excellent diagnostic agent for tumor malignancy, etc., an excellent treatment agent for B-cell lymphoma, gastric cancer, or colorectal cancer, and an excellent prevention/treatment drug for influenza. 1. An anti-CDw75 monoclonal antibody or a fragment thereof , characterized in that recognizes sugar chain structure CDw75 represented by Siaα2 ,6Galβ1 ,4GlcNAc but not a sugar chain structure represented by Galβ1 ,4GlcNAc ,; Siaα2 ,3Galβ1 ,4GlcNAc; or 6′-Sialyllactose (Siaα2 ,6Galβ1 ,4Glc).3. The anti-CDw75 monoclonal antibody or a fragment thereof according to claim 1 , the antibody being produced by hybridoma FR9 (deposit number: NITE BP-01516).4. A composition comprising the anti-CDw75 monoclonal antibody or a fragment thereof according to claim 1 , and a pharmaceutically acceptable carrier.5. The pharmaceutical composition according to claim 4 , the composition being used for inhibition and/or treatment of B-cell lymphoma claim 4 , gastric cancer claim 4 , or colorectal cancer.6. The pharmaceutical composition according to claim 4 , the composition being used for prevention and/or treatment of influenza.7. The composition according to claim 4 , the composition ...

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Номер записи: 42
10-03-2016 дата публикации

Methods of cell culture

Номер: US20160068881A1
Автор: Holly Prentice
Принадлежит: Momenta Pharmaceuticals Inc

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using putrescine, are described.

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Номер записи: 43
10-03-2016 дата публикации

MOLECULAR LABELING METHODS

Номер: US20160069890A1
Автор: Kalyuzhny Alexander, Wu Zhengliang L.
Принадлежит:

The disclosed methods involve deglycosylating a target molecule to remove target carbohydrates to create vacant glycosylation acceptor sites and then using glycosyltransferases to incorporate replacement carbohydrates into those sites. The methods also involve using glycosytransferases to incorporate new carbohydrates into vacant glycosylation acceptor sites without having to perform in vitro deglycosylation. The replacement or new carbohydrates include a click chemistry moiety that reacts to a click chemistry moiety on a label. 1. An in vitro method of labeling a target molecule , comprising:providing a sample containing a target molecule;treating the sample with a glycosidase to remove a target carbohydrate on the target molecule, thereby creating a vacant glycosylation acceptor site on the target molecule, wherein the glycosidase is specific for the target carbohydrate;treating the sample with a glycosyltransferase to incorporate a replacement carbohydrate into the vacant glycosylation acceptor site, wherein the glycosyltransferase has a substrate specificity that matches or overlaps with a substrate specificity of the glycosidase and wherein the replacement carbohydrate includes a click chemistry moiety; andadding a label to the sample, wherein the label includes a click chemistry moiety that reacts to the click chemistry moiety of the replacement carbohydrate such that the label attaches to the replacement carbohydrate.2. The method of wherein the target molecule is a target glycoprotein with a target carbohydrate that can be replaced with a replacement carbohydrate.3. The method of wherein the target carbohydrate is a target carbohydrate selected from the group consisting of sialic acid claim 2 , fucose claim 2 , GlcNAc claim 2 , GalNAc claim 2 , galactose claim 2 , mannose and xylose.4. The method of wherein the glycosidase is a glycosidase selected from the group consisting of sialidase claim 1 , fucosidase claim 1 , hexosaminidase and galactosidase or ...

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Номер записи: 44
08-03-2018 дата публикации

Method of mapping glycans of glycoproteins

Номер: US20180067124A1
Автор: Andreas Seidl, Fabian Higel
Принадлежит: HEXAL AG

The present invention uses anthranilic acid (2-AA) to label N-glycans prior to separation using a reversed-phase liquid chromatography (RP-LC) column under acidic conditions using formic acid. Negatively charged 2-AA offers stronger retention on the reversed phase column than 2-aminobenzamide (2-AB) in RP-LC and allows efficient ionization and detection of 2-AA labeled N-glycans. The acidic conditions used for the RP-LC leads to an efficient separation of acidic 2-AA N-glycans carrying terminal sialylation without the need for an ion-pairing reagent. The present invention may be used with RP-nano-LC-MS and a 96-well plate sample preparation, which allows attomolar sensitivity and high throughput.

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Номер записи: 45
09-03-2017 дата публикации

METHODS AND REAGENTS FOR IMPROVED SELECTION OF BIOLOGICAL MATERIALS

Номер: US20170067886A1
Автор: Ciocci Michael, McGann Pauline, Musick Mike, Russell Thomas R.
Принадлежит:

Methods, apparatus and compositions for separating a desired or undesired population or subpopulation from a biological sample are disclosed herein. The selection procedure is based on ferromagnetic, dense particles in a preferred size range from about 0.8 to about 1.2 microns. Specific binding agents are bound to the particles that recognize and bind to specific molecules on the targeted population or subpopulation, and the particles are mixed with the sample in such a way as to promote movement of the particles relative to the sample, promoting binding to the targeted population or subpopulation without non-specifically binding to non-targeted populations in the sample. Because of the large particle density, the bound population is separated from the fluid sample by gravity. Alternatively, the sample, including the bound, targeted population, is placed in a magnetic field such that the particles separate from the sample by evenly distributing over the vessel wall thus limiting non-specific trapping of the non-targeted population. 161-. (canceled)62. A suspension of particles for selecting a pre-determined target population of cells or cell like material or subpopulation thereof from a fluid sample comprising:a. a plurality of particles having a size range from about 50 nanometers to a diameter of about 2.5 microns, said particles having a density difference sufficiently different from a target population to enable selection by said particle; andb. a binding agent linked to said particles such that said binding agent binds specifically to said target population.63. A suspension of particles as described in claim 62 , wherein said particles are formed from ferromagnetic metal.64. A suspension of particles as described in claim 63 , wherein said ferromagnetic metal is selected from the group consisting of iron claim 63 , cobalt claim 63 , nickel claim 63 , and alloys thereof.65. A suspension of particles as described in claim 63 , wherein said ferromagnetic metal is ...

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Номер записи: 46
17-03-2016 дата публикации

PROBES AND ASSAYS FOR MEASURING E3 LIGASE ACTIVITY

Номер: US20160076074A1
Автор: PARK Sungjin, Statsyuk Alexander V.
Принадлежит:

Provided herein is technology relating to the biological process of protein ubiquitination and particularly, but not exclusively, to compositions and methods for studying protein ubiquitination and developing therapeutics to modulate protein ubiquitination. 1. A composition comprising a ubiquitin C-terminal thioester fluorophore.2. The composition of wherein the ubiquitin C-terminal thioester fluorophore comprises a ubiquitin covalently attached to a fluorophore by a thioester.3. The composition of wherein the ubiquitin C-terminal thioester fluorophore comprises a ubiquitin covalently attached to a linker and a linker covalently attached to the fluorophore.4. The composition of wherein the fluorophore is Fluorescein claim 1 , Rhodamine claim 1 , BODIPY claim 1 , Alexa Fluor 488 claim 1 , Oregon Green 488 claim 1 , or Alexa Fluor 594.5. The composition of wherein the linker is an alkyl claim 3 , cycloalkyl claim 3 , aryl claim 3 , heteroaryl claim 3 , heteroalkyl polymer claim 3 , carbon nanotube claim 3 , quantum dot claim 3 , or nanoparticle.6. The composition of wherein the ubiquitin comprises amino acids 1-76 of the ubiquitin polypeptide (SEQ ID NO: 1).7. The composition of further comprising an E3 ligase.8. The composition of wherein the E3 ligase is NEDD4 claim 7 , NEDD4L claim 7 , ITCH claim 7 , WWP1 claim 7 , WWP2 claim 7 , SMURF1 claim 7 , SMURF2 claim 7 , NEDL1 claim 7 , NEDL2 claim 7 , E6AP claim 7 , HECTD2 claim 7 , KIAA0614 claim 7 , TRIP12 claim 7 , G2E3 claim 7 , EDD claim 7 , HACE1 claim 7 , HECTD1 claim 7 , UBE3B claim 7 , UBE3C claim 7 , KIAA0317 claim 7 , HUWE1 claim 7 , HECTD3 claim 7 , HERC1 claim 7 , HERC2 claim 7 , HERC3 claim 7 , HERC4 claim 7 , HERC5 claim 7 , HERC6 claim 7 , SopA claim 7 , NleL claim 7 , ARIH1 claim 7 , ARIH2 claim 7 , CUL9 claim 7 , ANKIB1 claim 7 , PARK2 claim 7 , RNF144A claim 7 , RNF144B claim 7 , RBCK1 claim 7 , RNF19A claim 7 , RNF19B claim 7 , RNF31 claim 7 , RNF216 claim 7 , RNF14 claim 7 , RNF217 claim 7 , or a NEL ...

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Номер записи: 47
15-03-2018 дата публикации

HYBRID PROTEIN FOR THE IDENTIFICATION OF NEDDYLATED SUBSTRATES

Номер: US20180074072A1
Автор: SANTONICO Elena
Принадлежит:

Disclosed is a recombinant protein composed by the fusion of Glutathione S-transferase (GST), or an esa-histidine peptide (poly-His), or Maltose Binding Protein (MBP), to the Carboxyl-terminus end of the human KHNYN protein, containing residues 597-678 or a region including at least the amino acidic region 630-678. Also disclosed is a second recombinant protein where the Carboxyl-terminus end of the human KHNYN protein, containing residues 627-678 is genetically fused in a tandem construct to the Carboxyl-terminus end of KHNYN including residues 597-678. The tandem construct is N-terminally tagged with Glutathione S-transferase (GST), or an esa-histidine peptide (poly-His), or Maltose Binding Protein (MBP). The potential use of these “Neddylation sensors” also called “Neddylation probes” to isolate mono-, poly-neddylated targets as well as substrates modified by the addition of ubiquitin-NEDD8 mixed chains is considered. 18-. (canceled)9. A protein characterized in that it shows a clear preference for the ubiquitin-like NEDD8 , consisting of KH and NYN Domain Containing Protein (KHNYN) for the isolation and subsequent identification of ubiquitinated and neddylated substrates , such a protein consisting of the nucleotide sequence coding for the carboxyl-terminus end of the KHNYN human as shown in SEQ ID No.: 1 , including aminoacids 597-678 as defined in SEQ ID No.: 2 , and in any case including at least the region spanning amino acids 627-678.10. The protein according to and showing at least 60% sequence identity with SEQ ID No.: 2.11. The protein according to claim 9 , consisting of a tandem repeat of the carboxyl-terminus end of KHNYN for the isolation and subsequent identification of ubiquitinated and neddylated substrates claim 9 , such a protein consisting of the nucleotide sequence coding for the carboxyl-terminus end of the KHNYN human spanning residues 627-678 as shown in SEQ ID No.: 3 (nucleotides) and in SEQ ID No.: 4 (amino acids) genetically fused with ...

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Номер записи: 48
16-03-2017 дата публикации

NOVEL LECTINS AND APPLICATIONS FOR THE DETECTION OF PATHOLOGICAL STATE MARKERS

Номер: US20170074887A1
Автор: ARNAUD Julie, AUDFRAY Aymeric, Imberty Anne
Принадлежит:

Multimenic lectins having a β-propeller architecture, formed from monomer modules of approximately 30 to 60 amino acids, in which the binding sites to the glycans are situated on a given side of the proteins and the O-terminus and N-terminus ends of the peptide chains on the other side of the proteins, characterized in that they are formed from 4 to 7 monomer modules, a single, or a plurality of, or all of the adjacent modules being linked to one another by the linkers linking the N-terminus end of one module to the C-terminus end of the adjacent module. 1. Multimeric lectins with a β-propeller type architecture , formed by monomer modules of about 30 to 60 amino acids , in which the glycan binding sites are located on a same side of the proteins and the C-ter and N-ter ends of the peptide chains on the other side of the proteins , characterised in that they are formed by 4 to 7 monomer modules , a single , or several , or all of the adjacent modules being linked together by linkers linking the N-terminal end of a module to the C-terminal end of the adjacent module.2. The lectins according to claim 1 , characterised in that they include peptide chains with zero claim 1 , one or several mutations in the glycan binding site of one or more monomeric modules.3. The lectins according to claim 2 , characterised in that they are monomeric claim 2 , formed by a single peptide chain with a free C-ter end and a free N-ter end respectively claim 2 , and including linkers linking the C-ter and N-ter ends of the adjacent constituent modules of the lectin.4. The lectins according to claim 3 , characterised in that the linkers claim 3 , being identical or different claim 3 , comprise peptide sequences with a sufficient size to link the C-ter and N-ter ends and advantageously include from 2 to 10 amino acids.5. The lectins according to claim 4 , characterised in that they are multivalent lectins claim 4 , having several glycan binding sites claim 4 , or monovalent lectins having a ...

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Номер записи: 49
14-03-2019 дата публикации

METHODS RELATED TO ADALIMUMAB

Номер: US20190077857A1
Автор: Bosques Carlos J., Collins Brian Edward, Kaundinya Ganesh, Robblee John
Принадлежит:

The present invention relates to the characterization and production of adalimumab. 134.-. (canceled)35. A method , comprising:acquiring, from a test glycoprotein preparation, a value for each of a plurality of adalimumab parameters,wherein the plurality of adalimumab parameters comprises 6 to 22 parameters listed in Table 1, andwherein an adalimumab signature distinguishes adalimumab from a non-adalimumab glycoprotein, and the adalimumab signature consists of the 6 to 22 parameters and corresponding reference criteria in Table 1; andmanufacturing an adalimumab drug product by formulating at least a portion of the test glycoprotein preparation as an adalimumab drug product if all of the values acquired for the parameters in the signature meet the corresponding reference criteria; anddiscarding the test glycoprotein preparation if any one of the values acquired for the parameters in the signature do not meet the corresponding reference criteria;wherein the test glycoprotein preparation comprises a recombinant antibody composition including a first polypeptide having the amino acid sequence of SEQ ID NO:1 and a second polypeptide having the amino acid sequence of SEQ ID NO:2, and wherein the first and second polypeptides together form a recombinant antibody.36. The method of claim 35 , wherein the value for a parameter in the signature is directly acquired by performing an analytical test on the test glycoprotein preparation.37. The method of claim 36 , wherein the value for a parameter in the signature is directly acquired using a method provided in Table 2.38. The method of claim 35 , wherein formulating comprises combining the test glycoprotein preparation with an excipient or buffer.39. The method of claim 35 , wherein the adalimumab drug product is approved under Section 351(k) of the Public Health Service (PHS) Act.40. The method of claim 35 , wherein the adalimumab drug product is not approved under biologics license application (BLA) under Section 351(a) of ...

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Номер записи: 50
14-03-2019 дата публикации

METHODS OF GLYCOPROTEIN ANALYSIS

Номер: US20190079100A1
Автор: Anderson James, Tsao Desiree
Принадлежит:

Methods of assessing biosimilarity of proteins, e.g., therapeutic antibodies, are described. 1. A method of manufacture comprising:producing a batch of test protein drug substance;exposing a sample of the batch of the test protein in a first state to a plurality of stressors to obtain a plurality of test protein in a second state, wherein one or more of the plurality of stressors comprises a condition that alters a higher-order structure of a protein;detecting a signal associated with higher-order structure of the test protein for each of the plurality of test protein in the second state, wherein the detecting comprises use of an NMR method;determining a test protein delta between a signal associated with higher-order structure of the test protein drug product in the first state and the signal associated with higher-order for each of the plurality of test protein drug product in the second state;comparing the determined test protein deltas to corresponding target protein deltas of a target protein drug product approved under a primary approval process to determine whether the test protein deltas and the target protein deltas are tolerable; andprocessing the batch of the test protein drug substance as test protein drug product if the test protein deltas and the target protein deltas are tolerable, ortaking an alternative action if the test protein deltas and the target protein deltas are not tolerable.2. (canceled)3. The method of claim 1 , wherein the first state is a native state and the second state is a non-native state.4. (canceled)5. The method of claim 1 , wherein one or more of the plurality of stressors comprises an NMR shift reagent.6. (canceled)7. The method of claim 1 , wherein the compared test protein deltas and target protein deltas are tolerable if they meet a predetermined value.8. The method of claim 1 , wherein the signal associated with higher-order structure comprises one or more peaks from an NMR spectrum.9. The method of claim 1 , wherein the ...

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Номер записи: 51
14-03-2019 дата публикации

METHODS OF EVALUATING AND MAKING BIOLOGICS

Номер: US20190079101A1
Автор: Bosques Carlos J., Collins Brian Edward, Kaundinya Ganesh, Robblee John
Принадлежит:

Methods of making and evaluating biologic therapeutic products are disclosed. 1147.-. (canceled)148. A method comprising: wherein the test protein is not approved under a biologics license application (BLA) or a supplemental BLA,', 'wherein the plurality of parameters includes two or more determinative parameters,', 'wherein a signature consists of the two or more determinative parameters and values for a target protein for the determinative parameters, and wherein the signature distinguishes the target protein from a plurality of antibody or Fc-containing non-target proteins;, 'acquiring, from a test protein preparation, a value for each of a plurality of parameters,'}manufacturing a protein drug product by formulating at least a portion of the test protein preparation into the protein drug product if all of the values acquired from the test glycoprotein preparation for the determinative parameters in the signature are indistinguishable from the values in the signature; anddiscarding the test protein preparation if any one of the values acquired from the test protein preparation for the determinative parameters in the signature is distinguishable from the values in the signature;wherein the target protein: (i) has an amino acid sequence that is at least 95% identical to the test protein amino acid sequence, or (ii) has an amino acid sequence that differs by less than 10 amino acids from the test protein amino acid sequence, and wherein the target protein is approved under a BLA, a supplemental BLA, or an equivalent thereof.149. The method of claim 148 , wherein at least one value acquired from the test protein preparation for each determinative parameter in the signature is directly acquired or is acquired by performing an analytical analysis on said test protein preparation.150. The method of claim 148 , wherein formulating comprises combining the test protein preparation with a second component.151. The method of claim 148 , wherein a value for a target protein ...

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Номер записи: 52
24-03-2016 дата публикации

CATION EXCHANGER FOR LIQUID CHROMATOGRAPHY, PROCESS FOR PRODUCING SAME, AND USE THEREOF

Номер: US20160084804A1
Автор: MURANAKA Kazuaki, SAKO Miyuki
Принадлежит: TOSOH CORPORATION

To provide a cation exchanger for liquid chromatography having a high retention for an object to be separated or analyzed, a sufficient binding capacity for the object, a high resolution and mechanical strength, and a low operation pressure. 111.-. (canceled)12. A cation exchanger for liquid chromatography , comprising non-porous particles and , immobilized to the surface thereof , a polyacrylic acid into which a dicarboxylic acid compound having an amino group is introduced by an amide bond.13. The cation exchanger for liquid chromatography according to claim 12 , wherein the non-porous particles are an inorganic base matrix of silica claim 12 , zirconia or alumina claim 12 , or an organic base matrix of a crosslinked polysaccharide or a vinyl monomer crosslinked polymer claim 12 , having a volume average particle size of at most 5 μm.14. The cation exchanger for liquid chromatography according to claim 13 , wherein the organic base matrix is a crosslinked polymer of a monofunctional vinyl monomer and a polyfunctional vinyl monomer.15. The cation exchanger for liquid chromatography according to claim 12 , wherein the polyacrylic acid has a weight average molecular weight of at least 5 claim 12 ,000 and at most 20 claim 12 ,000.16. The cation exchanger for liquid chromatography according to claim 12 , wherein the dicarboxylic acid compound having an amino group is aspartic acid or glutamic acid.17. The cation exchanger for liquid chromatography according to claim 12 , which has an ion exchange capacity of from 10 to 50μ-equivalent/mL.18. A process for producing the cation exchanger for liquid chromatography as defined in claim 12 , which comprises reacting a functional group of the non-porous particles with a carboxylic acid in the polyacrylic acid thereby to immobilize the polyacrylic acid onto the surface of the non-porous particles claim 12 , and reacting the immobilized polyacrylic acid with the dicarboxylic acid compound having an amino group.19. A process for ...

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Номер записи: 53
24-03-2016 дата публикации

METHOD FOR QUANTIFYING GLYCATED HEMOGLOBIN

Номер: US20160084850A1
Автор: ENDOH Takashi, SHIGETOH Nobuyuki
Принадлежит:

A method for quantifying glycated hemoglobin (HbA1c) contained in a sample, the method including: (a) a process for mixing the sample with a protease in the presence of a composition of a cationic surfactant and a tetrazolium salt to obtain an aqueous glycated peptide solution containing a glycated peptide; (b) a process for mixing the aqueous glycated peptide solution obtained in process (a) with a fructosyl peptide oxidase to obtain hydrogen peroxide, the aqueous glycated peptide solution here containing the composition; and (c) a process for calculating the concentration of the glycated hemoglobin (HbA1c) on the basis of the amount of hydrogen peroxide solution obtained in process (b). 114-. (canceled)151. A method for quantifying glycated hemoglobin (HbAc) contained in a sample , the method comprising steps of:(a) mixing the sample with a protease in the presence of a composition of a cationic surfactant and a tetrazolium salt to obtain a glycated-peptide aqueous solution containing a glycated peptide;(b) mixing the glycated-peptide aqueous solution obtained in the step (a) with fluctosyl peptide oxidase to obtain hydrogen peroxide; whereinthe glycated-peptide aqueous solution contains the composition; and{'b': '1', 'sub': 1', '2', '3', '4', '1', '2', '3', '4, 'sup': +', '−, '(c) calculating a concentration of the glycated hemoglobin (HbAc) on the basis of an amount of the hydrogen peroxide obtained in the step (b), wherein the cationic surfactant is a quaternary ammonium salt, which is represented by the chemical formula RRRRNX: wherein Ris an alkyl group having a carbon number of not less than 8 and not more than 18; R, Rand Rare lower alkyl groups independently; and X represents halogens.'}16. The method according to claim 15 , whereinthe protease is selected from the group consisting of thermolysin and papain.17. The method according to claim 15 , whereinthe tetrazolium salt is selected from the group consisting of 2-(4-Iodophenyl)-3-(2,4-dinitrophenyl)-5-(2 ...

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Номер записи: 54
28-03-2019 дата публикации

CLEAVABLE PROBES FOR ISOTOPE TARGETED GLYCOPROTEOMICS AND METHODS OF USING THE SAME

Номер: US20190094238A1
Автор: Bertozzi Carolyn R., Woo Christina
Принадлежит:

Methods for producing isotopically-labelled peptides are provided. Aspects of the method include: contacting a sample including a metabolically tagged protein with a cleavable probe to produce a probe-protein conjugate; separating the probe-protein conjugate from the sample; digesting the probe-protein conjugate to produce a probe-peptide conjugate; and cleaving a cleavable linker to release an isotopically labelled peptide. The method may further include: identifying a predetermined isotopic pattern in a mass spectrum; determining an amino acid sequence of the isotopically labelled peptide; and identifying the site of protein glycosylation based on the determined amino acid sequence. Also provided are cleavable probes for practicing the subject methods, described by the Formula: A-L-(M-Z) where A is an affinity tag, L is a cleavable linker, M is an isotopic label and Z is a chemoselective tag capable of cross-linking a metabolically tagged protein. Compositions and kits for practicing the subject methods are also provided. 121.-. (canceled)22. A cleavable probe of Formula (I):{'br': None, 'A-L-(M-Z)\u2003\u2003 (I)'} A is an affinity tag', 'L is a cleavable linker;', 'M is an isotopic label; and', 'Z is a chemoselective tag., 'wherein23. The probe of claim 22 , wherein Z is a chemoselective tag comprising a group selected from an alkyne claim 22 , an azide claim 22 , a phosphine claim 22 , a thiol claim 22 , a maleimide or iodoacetyl claim 22 , an aldehyde claim 22 , a hydrazide and an alkoxyamine.24. The probe of claim 23 , wherein Z comprises an alkyne.25. The probe of claim 22 , wherein A is a biotin moiety.27. The probe of claim 22 , wherein X is —O—Si(R)—O— claim 22 , wherein each R is independently selected from hydrogen claim 22 , an aryl claim 22 , a substituted aryl claim 22 , an alkyl and a substituted alkyl.28. The probe of claim 22 , wherein L is a cleavable silane linker.29. The probe of claim 22 , wherein M comprises two or more bromine atoms.30. The ...

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Номер записи: 55
28-03-2019 дата публикации

METHOD OF DIAGNOSING ARTHRITIS OR OTHER JOINT DEGRADING DISEASE

Номер: US20190094243A1
Автор: FLOWERS Sarah Ann, JIN Chunsheng, KALAMAJSKI Sebastian, Karlsson Niclas
Принадлежит:

The invention provides a method of diagnosing arthritis or other joint degrading disease in a subject which comprises determining whether there is a presence or increase of lubricin having a joint tissue posttranslational modification, in a blood sample from the subject, the presence or increase of the lubricin having the joint tissue posttranslational modification indicating arthritis or other joint degrading disease in the subject. The invention further provides a kit or protocol for detecting arthritis or other joint degrading disease by detecting lubricin, the lubricin comprising a joint tissue posttranslational modification. 1. A method of diagnosing arthritis or other joint degrading disease in a subject , which method comprises determining whether there is a presence or increase of lubricin having a joint tissue posttranslational modification , in a blood sample from said subject , said presence or increase of said lubricin having said joint tissue posttranslational modification indicating arthritis or other joint degrading disease in said subject.2. The method according to claim 1 , said blood sample being selected from at least one of the group consisting of serum and plasma.3. The method according to claim 1 , further comprising discriminating between lubricin having said joint tissue posttranslational modification claim 1 , and lubricin not having said joint tissue posttranslational modification claim 1 , in said blood sample claim 1 , whereby only said presence or increase of said lubricin having said joint tissue posttranslational modification indicates arthritis or other joint degrading disease in said subject.4. The method according to claim 1 , further comprising determining the presence or increase of lubricin having said joint tissue posttranslational modification claim 1 , and which method further comprises not determining the presence or increase of lubricin not having said joint tissue posttranslational modification.5. The method according to ...

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Номер записи: 56
06-04-2017 дата публикации

SUMOylation Assay and Related Reagents

Номер: US20170096698A1
Автор: Bjornsti Mary-Ann, Placzek William J., Whitaker Robert H., Wright Christine M.
Принадлежит: THE UAB RESEARCH FOUNDATION

Provided herein and methods and kits for identifying inhibitors of SUMOylation. The provided methods include contacting a candidate agent with (i) a SUMO-conjugating peptide comprising an N-terminal linker and a detectable tag, and (ii) a SUMO comprising a metal complex, under conditions that allow binding between the SUMO-conjugating peptide and the SUMO, and detecting the detectable tag thereby determining the level of binding between the SUMO-conjugating peptide and SUMO. A reduced level of binding as compared to the level of binding in the absence of the candidate agent indicating the agent inhibits SUMOylation. 1. A method of identifying an inhibitor of SUMOylation comprising:(a) contacting a candidate agent with (i) a SUMO-conjugating peptide comprising an N-terminal linker and a detectable tag, and (ii) a SUMO comprising a metal complex, under conditions that allow binding between the SUMO-conjugating peptide and the SUMO; and(b) detecting the detectable tag thereby determining the level of binding between the SUMO-conjugating peptide and SUMO;a reduced level of binding as compared to the level of binding in the absence of the candidate agent indicating the agent inhibits SUMOylation.2. The method of claim 1 , wherein the detectable tag is a fluorescent molecule.3. The method of claim 1 , wherein the detectable tag is biotin.4. The method of claim 3 , wherein the detecting comprises adding streptavidin labeled with a fluorescent molecule.5. The method of claim 1 , wherein the contacting further comprises a SUMO-conjugating enzyme.6. The method of claim 1 , wherein the SUMO-conjugating enzyme is selected from the group consisting of an E1 enzyme claim 1 , an E2 enzyme claim 1 , an E3 enzyme or a combination thereof.7. The method of claim 1 , wherein the metal complex is a metal chelate or a metal cryptate.8. The method of claim 7 , wherein the metal is a lanthanide metal.9. The method of claim 8 , wherein the lanthanide metal is europium or terbium.10. The ...

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Номер записи: 57
14-04-2016 дата публикации

METHOD FOR THE ANALYSIS OF N-GLYCANS ATTACHED TO IMMUNOGLOBULIN G FROM HUMAN BLOOD PLASMA AND ITS USE

Номер: US20160103137A1
Автор: Lauc Gordan, Pucic Bakovic Maja, Vuckovic Frano
Принадлежит:

The invention discloses a method for the analysis of N-glycans attached to immunoglobulin G (IgG) or IgG N-glycopeptides from human blood plasma in which relative abundance of two or more glycans is determined, out of total six, and for these glycans it is determined that they strongly correlate with age. The glycans have the following structures: 110.-. (canceled)12. The method of claim 11 , further comprising predicting the biological and chronological age of the individual.13. The method of claim 11 , further comprising monitoring efficacy of a method for slowing down the aging process of the individual.14. The method of claim 11 , further comprising monitoring progression of a disease developed as a result of the aging process of the individual.15. The method of claim 14 , wherein the disease is selected from the group consisting of inflammatory diseases including atherosclerosis claim 14 , autoimmune diseases claim 14 , tumours claim 14 , diabetes claim 14 , arthritis claim 14 , osteoporosis claim 14 , and Alzheimer disease. The Invention refers to the method for the analysis of N-glycans attached to immunoglobulin G from human blood plasma for the purpose of: more precise prediction of biological and/or chronological age of a person; possibility to monitor efficacy of methods that slow down the ageing process; possibility to monitor progression of diseases that are developed as a result of the ageing process advancement, like: inflammatory diseases (including atherosclerosis), autoimmune diseases, tumours, diabetes, arthritis, osteoporosis, and Alzheimer disease; as well as evaluation of the overall condition/health of a body.The Invention solves technical problem of precise prediction of a human's biological age by means of quantitative analysis of two or more N-glycans attached to immunoglobulin G (IgG), out of the total of six of them that are most strongly correlated to age.Moreover, the Invention solves the technical problem of possibility to efficiently ...

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Номер записи: 58
08-04-2021 дата публикации

COMPOSITIONS AND METHODS FOR ENRICHMENT AND DETECTION OF UBIQUITIN AND UBIQUITIN CONJUGATES

Номер: US20210102950A1
Автор: Berk Jason, HOCHSTRASSER Mark, ZHANG Mengwen
Принадлежит:

Described herein are compositions and methods for detecting the presence of ubiquitination in a sample. The compositions include synthetic peptides containing a high affinity ubiquitin binding domain and an additional peptide sequence that can be coupled to materials such as resins and dyes. 1. An ubiquitin-binding peptide comprising the sequence of SEQ ID NO: 1 or SEQ ID NO:5 ,{'sub': aa', 'n, 'wherein Z is (X)or L;'}{'sub': 'aa', 'wherein each Xis independently selected from the group consisting of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, selenocysteine, penicillamine, homocysteine, and selenomethionine;'}{'sub': 'aa', 'provided that at least one Xis selected from the group consisting of cysteine, selenocysteine, penicillamine, homocysteine, methionine, and selenomethionine;'}wherein L is a chemical moiety comprising from 1 to 500 atoms selected from the group consisting of C, N, O, S, P, H, F, Cl, and Br; andwherein n is an integer from 1 to 1000.2. The peptide of claim 1 , wherein Z is (X).3. The peptide of claim 2 , wherein n is 6.4. The peptide of claim 3 , wherein at least one Xis cysteine.5. The peptide of claim 3 , wherein at least two Xare independently cysteine.6. The peptide of claim 2 , wherein Z has the sequence of SEQ ID NO: 3.7. The peptide of claim 6 , wherein the peptide has the sequence of SEQ ID NO: 4.8. The peptide of claim 6 , wherein the peptide has the sequence of SEQ ID NO: 6.9. A reaction product of the peptide of and a resin claim 1 , wherein the resin comprises at least one reactive group that can react with a sulfhydryl (SH) or selenohydryl (SeH) functionality.10. The reaction product of claim 9 , wherein the resin comprises agarose.11. The reaction product of claim 9 , wherein the at least one reactive group is an iodoacetyl group.12. A reaction product of the ...

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Номер записи: 59
04-04-2019 дата публикации

Anti-Polyubiquitin Antibodies and Methods of Use

Номер: US20190100578A1
Автор: Dixit Vishva, KELLEY ROBERT F., Matsumoto Marissa L.
Принадлежит: Genentech, Inc.

The invention provides anti-polyubiquitin antibodies and methods of using the same. 145-. (canceled)46. A method of separating K11-linked polyubiquitinated protein from non-K11-linked polyubiquitinated protein in a sample , comprising contacting the sample with an antibody that binds K11-linked polyubiquitin , wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 2 and 57 to 60 , an HVR-L2 sequence selected from SEQ ID NOs: 3 and 61 , an HVR-L3 sequence of SEQ ID NO: 4 , an HVR-H1 sequence selected from SEQ ID NOs: 6 to 11 , an HVR-H2 sequence selected from SEQ ID NOs: 12 to 17 and 67 , and an HVR-H3 sequence selected from SEQ ID NOs: 18 to 23 , 68 and 69.47. The method of claim 46 , wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 2 and 57 to 60 claim 46 , an HVR-L2 sequence selected from SEQ ID NOs: 3 and 61 claim 46 , an HVR-L3 sequence of SEQ ID NO: 4 claim 46 , an HVR-H1 sequence of SEQ ID NO: 11 claim 46 , an HVR-H2 sequence selected from SEQ ID NOs: 17 and 67 claim 46 , and an HVR-H3 sequence selected from SEQ ID NOs: 23 claim 46 , 68 and 69.48. The method of claim 46 , wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 58 and 59 claim 46 , an HVR-L2 sequence of SEQ ID NO: 3 claim 46 , an HVR-L3 sequence of SEQ ID NO: 4 claim 46 , an HVR-H1 sequence of SEQ ID NO: 11 claim 46 , an HVR-H2 sequence of SEQ ID NO: 67 claim 46 , and an HVR-H3 sequence of SEQ ID NO: 23.49. The method of claim 46 , wherein the antibody comprises comprising an HVR-L1 sequence of SEQ ID NO: 59 claim 46 , an HVR-L2 sequence of SEQ ID NO: 3 claim 46 , an HVR-L3 sequence of SEQ ID NO: 4 claim 46 , an HVR-H1 sequence of SEQ ID NO: 11 claim 46 , an HVR-H2 sequence of SEQ ID NO: 67 claim 46 , and an HVR-H3 sequence of SEQ ID NO: 23.50. The method of claim 46 , wherein the antibody comprises a heavy chain variable region comprising a sequence with at least 95% sequence identity to SEQ ID NO: 72 and a light chain ...

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Номер записи: 60
21-04-2016 дата публикации

SIALYLATED GLYCOPROTEINS

Номер: US20160108450A1
Автор: Bhatnagar Naveen, Lansing Jonathan C., Meccariello Robin, Ortiz Daniel, SARVAIYA Hetal, Washburn Nathaniel
Принадлежит:

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described. 1. A method of producing a preparation of glycoproteins comprising Fc regions comprising branched glycans comprising an α1 ,3 arm and an α1 ,6 arm , the preparation comprising (i) a target level of branched glycans having a sialic acid on an α1 ,3 arm and/or (ii) a target level of branched glycans having a sialic acid on an α1 ,6 arm , the method comprising:providing a plurality of glycoproteins comprising Fc regions comprising branched glycans comprising an α1,3 arm and an α1,6 arm; andcontacting the glycoproteins with an ST6 sialyltransferase in the presence of a limited reaction condition, thereby producing a glycoprotein preparation having (i) the target level of branched glycans having a sialic acid on the α1,3 arm and/or (ii) the target level of branched glycans having a sialic acid on an α1,6 arm.2. The method of claim 1 , wherein the limited reaction condition is sufficient for the ST6 sialyltransferase substantially to add a sialic acid to an α1 claim 1 ,3 arm of a branched glycan and not sufficient for the ST6 sialyltransferase substantially to add a sialic acid to an α1 claim 1 ,6 arm of a branched glycan.3. The method of claim 1 , further comprising isolating the glycoprotein preparation.4. The method of claim 3 , further comprising measuring a level of branched glycans comprising a sialic acid on an α1 claim 3 ,3 arm and/or measuring a level of branched glycans having a sialic acid on an α1 claim 3 ,6 arm.5. The method of claim 1 , wherein the target level of branched glycans having a sialic acid on an α1 claim 1 ,3 arm is at least 20% claim 1 , 25% claim 1 , 30% claim 1 , 35% claim 1 , 40% claim 1 , 45% claim 1 , 50% claim 1 , 55% claim 1 , 60% claim 1 , 65% claim 1 , 70% claim 1 , 75% claim 1 , 80% claim 1 , 85% claim 1 , 90% claim 1 , 95% claim 1 , or 100% of sialylated branched glycans.6. The method of claim 1 , wherein the target ...

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Номер записи: 61
20-04-2017 дата публикации

A-FUCOSYLATION DETECTION IN ANTIBODIES

Номер: US20170108510A1
Автор: Jaeger Christiane, Koll Hans, Sondermann Peter, Umana Pablo
Принадлежит: ROCHE GLYCART AG

This invention describes a new analytical method to determine the quantity and distribution of fucose per Fc within an antibody preparation. 1. A method for preparing a homogenously deglycosylated immunoglobulin G antibody or a homogenously deglycosylated Fc fragment thereof , the method comprising(a) providing an antibody preparation, wherein the antibody preparation comprises a glycosylated immunoglobulin G antibody or a glycosylated Fc fragment thereof, and(b) removing heterogeneous saccharide residues from the glycosylated immunoglobulin G antibody or the glycosylated Fc fragment thereof with the enzyme endoglycosidase S and the enzyme endoglycosidase H, thereby preparing a homogenously deglycosylated immunoglobulin G antibody or a homogenously deglycosylated Fc fragment thereof.2. The method of claim 1 , wherein the enzyme endoglycosidase S cleaves complex-type N-linked carbohydrate moieties from the glycosylated immunoglobulin G antibody or the glycosylated Fc fragment thereof claim 1 , and the enzyme endoglycosidase H cleaves hybrid-type N-linked carbohydrate moieties from the glycosylated immunoglobulin G antibody or the glycosylated Fc fragment thereof.3. The method of claim 1 , wherein the method further comprises detecting the presence or absence of fucose residues within the homogenously deglycosylated immunoglobulin G antibody or the homogenously deglycosylated Fc fragment thereof claim 1 , the method comprising(a) removing other heterogeneous residues from the homogenously deglycosylated immunoglobulin G antibody or the homogenously deglycosylated Fc fragment thereof with an enzyme, and(b) analyzing the immunoglobulin G antibody or the Fc fragment thereof for the presence or absence of fucose residues.4. The method of claim 3 , wherein the method further comprises purifying the homogenously deglycosylated immunoglobulin G antibody or the homogenously deglycosylated Fc fragment thereof prior to analyzing the immunoglobulin G antibody or the Fc fragment ...

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Номер записи: 62
09-06-2022 дата публикации

SUMO INHIBITOR COMPOUNDS AND USES THEREOF

Номер: US20220177488A1
Автор: JUDD Ted Charles, Kubota Miles, Ouyang Xiaohu, TASKER Andrew S.
Принадлежит:

The present invention relates to compounds and compositions capable of acting as inhibitors of small ubiquitin-like modifier (SUMO) family of proteins. The compounds and compositions may be used in the treatment of cancer. There are disclosed, inter alia, methods of inhibiting an E1 enzyme, and compounds useful for inhibiting an E1 enzyme. 2. The compound of wherein ring A is selected from dihydropyran claim 1 , dihydrothienyl claim 1 , dihydrofuryl claim 1 , oxo-dihydrofuryl claim 1 , pyrrolinyl claim 1 , dihydrothiazolyl claim 1 , dihydro-oxazolyl claim 1 , dihydro-isothiazolyl claim 1 , dihydro-isoxazolyl claim 1 , midazolinyl and pyrazolinyl claim 1 , wherein ring A is optionally substituted with one or more substituent groups.3. The compound of wherein ring A is selected from thienyl claim 1 , furanyl claim 1 , pyrrolyl claim 1 , thiazolyl claim 1 , oxazolyl claim 1 , midazolyl claim 1 , pyrazolyl claim 1 , isoxazolyl claim 1 , triazolyl and isothiazolyl claim 1 , wherein ring A is optionally substituted with one or more substituent groups.6. The compound of wherein ring A is selected from phenyl claim 1 , pyridyl claim 1 , pyrazinyl claim 1 , pyrimidinyl claim 1 , pyridazinyl claim 1 , and triazinyl claim 1 , wherein ring A is optionally substituted with one or more substituent groups.8. The compound of wherein ring A is selected from phenyl or thienyl claim 1 , wherein ring A is optionally substituted with one or more substituent groups.9. The compound of any one of - wherein Ris selected from substituted or unsubstituted phenyl claim 1 , substituted or unsubstituted cycloalkyl and substituted or unsubstituted 5- or 6-membered heteroaryl.10. The compound of any one of - wherein Ris selected from substituted or unsubstituted phenyl claim 1 , substituted or unsubstituted cyclopentyl claim 1 , substituted or unsubstituted cyclohexyl and substituted or unsubstituted pyridyl.11. The compound of any one of - wherein E is selected from —C(═O)CH═CH claim 1 , —C(═O)- ...

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Номер записи: 63
09-06-2022 дата публикации

LABELLED COMPOUNDS AND METHODS FOR MASS SPECTROMETRY-BASED QUANTIFICATION

Номер: US20220178942A1
Автор: MÜLLER Susanne Leslie, Reiter Lukas
Принадлежит: BiognoSYS AG

Methods for peptide and/or protein quantification by mass spectrometry using labeled peptides, wherein multiple labels lead to distinct fragments for the labeled peptides and their unlabeled variant, thus facilitating data analysis and enhancing the potential for quantification. Methods for selecting the label and label position are further given, as well as sets of labeled peptides resulting from or for use in the above-mentioned methods. The methods and substances are especially useful for data-independent or multiplexed parallel reaction monitoring proteomics applications involving peptide quantification. 115.-. (canceled)16. A method for selecting the label and label position of at least one suitable reference peptide for use in a method for the absolute or relative quantitative analysis of at least one of proteins or peptides , in each case with or without post translational modification(s) , using a mass spectrometry method comprising:a first step where unlabeled proteins from an endogenous mixture are digested and subsequently digestion products thereof selected,a second step where said digestion products are fragmented, anda third step where a combined fragment spectrum is acquired comprising b-ions as well as y-ions of said digestion products,wherein at least one reference peptide with or without post translational modification(s) is added to said mixture before or after digestion or both, is fragmented, acquired, and stored in said combined fragment spectrum comprising also b-ions and y-ions of said digestion products,wherein said at least one reference peptide is added in a known concentration in case of absolute quantification or in always the same concentration in a series of experiments for relative quantitative analysis, 'one isotopically labeled amino acid forming its very C-terminus or being one of the four terminal amino acids at the C-terminus, and additionally one further isotopically labeled amino acid forming its very N-terminus, or being one ...

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Номер записи: 64
09-04-2020 дата публикации

METHOD FOR PREPARING ANALYTICAL SAMPLE, ANALYSIS METHOD, AND KIT FOR PREPARING ANALYTICAL SAMPLE

Номер: US20200109159A1
Автор: NISHIKAZE Takashi
Принадлежит: SHIMADZU CORPORATION

A method for preparing an analytical sample for analysis of a glycan contained in a sample includes: performing an amidation reaction that amidates a lactone structure included in the glycan through contacting the sample with a reaction solution that is basic; adding an acidic solution to the reaction solution after the reaction solution is subjected to the amidation reaction; and purifying the sample contained in the reaction solution after the acidic solution is added to the reaction solution by using a carrier for hydrophilic interaction chromatography. 1. A method for preparing an analytical sample for analysis of a glycan contained in a sample , the method comprising:performing an amidation reaction that amidates a lactone structure included in the glycan through contacting the sample with a reaction solution that is basic;adding an acidic solution to the reaction solution after the amidation reaction; andpurifying the sample contained in the reaction solution by using a carrier for hydrophilic interaction chromatography after the acidic solution is added to the reaction solution.2. The method for preparing an analytical sample according to claim 1 , whereinpH of the reaction solution after the acidic solution is added to the reaction solution is 10 or lower.3. The method for preparing an analytical sample according to claim 1 , the method further comprising:performing a lactonization reaction that lactonizes at least a part of sialic acids included in the glycans before the amidation reaction.4. The method for preparing an analytical sample according to claim 2 , the method further comprising:performing a lactonization reaction that lactonizes at least a part of sialic acids included in the glycans before the amidation reaction.5. The method for preparing an analytical sample according to claim 3 , whereinthe lactonization reaction is performed through contacting the sample with a lactonization reaction solution containing a dehydration condensation agent.6. ...

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Номер записи: 65
03-05-2018 дата публикации

ANTIBODIES SPECIFIC TO GLYCOSYLATED PD-L1 AND METHODS OF USE THEREOF

Номер: US20180118830A1
Автор: CHUNG Ezra M., Hung Mien-chie, Kim Yong-Soo, Li Chia-Wei, LIM Seung-Oe, YOO Stephen S.
Принадлежит:

Antibodies that bind specifically to glycosylated PD-L1 relative to unglycosylated PD-L1 are provided. Antibodies that recognize specific epitopes of glycosylated PD-L1 protein and can block the binding of PD-L1 to PD-1 are provided. In some aspects, PD-L1 polypeptides comprising glycosylated amino acid residues at amino and carboxy terminal positions of the PD-L1 extracellular domain are also provided. Methods for making and using such antibodies and polypeptides (e.g., for the treatment of cancer) are also provided. 1. An isolated antibody which selectively binds to glycosylated PD-L1 relative to unglycosylated PD-L1 and inhibits binding of glycosylated PD-L1 to PD-1.2. The isolated antibody of which is recombinantly engineered claim 1 , chimeric claim 1 , humanized or fully human.3. The isolated antibody of or claim 1 , wherein the antibody selectively binds to amino acid residues within regions of the PD-L1 protein comprising amino acid positions 35 claim 1 , 192 claim 1 , 200 and/or 219 in SEQ ID NO: 1 relative to unglycosylated PD-L1.4. The isolated antibody of any one of to claim 1 , wherein the antibody binds to glycosylated PD-L1 with a Kthat is less than half of the Kexhibited by the antibody binding to unglycosylated PD-L1.5. The isolated antibody of claim 4 , wherein the antibody binds to glycosylated PD-L1 with a Kat least 10 times less than the Kexhibited by the antibody binding to unglycosylated PD-L1.6. The isolated antibody of any one of to claim 4 , wherein the antibody binds glycosylated PD-L1 with an affinity of from 5-20 nM.7. The isolated antibody of any one of to claim 4 , wherein the antibody claim 4 , which is directly or indirectly detectable by a fluorescent label claim 4 , preferentially binds to cells expressing glycosylated PD-L1 with a measured fluorescence intensity (MFI) that is 2-fold to 10-fold higher than the MFI exhibited by the antibody binding to cells expressing unglycosylated PD-L1 in a cell flow cytometry assay.8. The ...

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Номер записи: 66
04-05-2017 дата публикации

Enzymatically responsive magnetic particles and their use

Номер: US20170119910A1
Автор: Bing Xu, Jie Zhou, Xuewen DU
Принадлежит: BRANDEIS UNIVERSITY

The invention relates to an enzymatically responsive product that includes an amino acid residue conjugated to a magnetic particle, wherein the amino acid residue is phosphorylated or sulfated or comprises an ester-moiety linked via peptide bond. Compositions containing the enzymatically responsive product, and the use thereof for separating distinct types of mammalian cells (e.g., cancer cells from normal cells), for treating a cancerous condition, and imaging cancer cells are also disclosed.

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Номер записи: 67
04-05-2017 дата публикации

Sandwich assay using labeled lectin and kit therefor

Номер: US20170122940A1
Автор: Takatoshi KAYA, Tomonori Kaneko
Принадлежит: KONICA MINOLTA INC

The present invention provides a sandwich assay for quantifying a glycoprotein, which is a substance to be detected, in a sample using a labeled lectin, wherein the effect attributed to a contaminant, namely noise on the quantified value of the substance to be detected, is suppressed by introduction of a simple treatment. The sandwich assay includes a treatment for inhibiting the binding of the labeled lectin to a sugar chain carried by the contaminant non-specifically adsorbed to the measurement region, which contaminant is contained in the sample and which sugar chain is the same as that of the substance to be detected.

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Номер записи: 68
25-08-2022 дата публикации

METHODS FOR THE RAPID PREPARATION OF LABELED GLYCOSYLAMINES AND FOR THE ANALYSIS OF GLYCOSYLATED BIOMOLECULES PRODUCING THE SAME

Номер: US20220268779A1
Автор: Brousmiche Darryl W., Koza Stephan M., Lauber Matthew A.
Принадлежит: WATERS TECHNOLOGIES CORPORATION

Methods of analyzing glycosylated biomolecules include the steps of producing a deglycosylation mixture of biomolecules deglycosylated by natural or synthetic enzymatic or chemical techniques; providing a reagent solution having a labeling reagent in a polar aprotic, non-nucleophilic organic solvent; and mixing the deglycosylation mixture with the reagent solution in an excess of labeling reagent to produce derivatized glycosylamines. The method steps can be carried out purposefully without depletion of protein matter. A quenching solution can be added to the reaction mixture so that the pH of the reaction mixture is shifted to above 10. The yield of derivatized glycosylamines can be in an amount of about 80 to about 100 mole percent of the reaction mixture with minimal overlabeling, less than 0.2 mole percent. The derivizated glycosylamines can be separated from the reaction mixture and detected by chromatographic detection, fluorescence detection, mass spectrometry (“MS”), or Ultra Violet (“UV”) detection and/or a combination thereof. 118-. (canceled)19. A method of rapid derivatization of glycosylamines comprising the steps of:providing a biological sample comprising a glycoprotein;contacting the glycoprotein with an enzyme to produce a deglycosylation mixture; andmixing (a) a reagent solution comprising a labeling reagent selected from an N-hydroxysuccinimide ester reagent and an N-hydroxysuccinimide carbamate reagent combined with a polar aprotic, non-nucleophilic organic solvent, (b) the deglycosylation mixture, and (c) a buffer solution to produce a reaction mixture, the reaction mixture being formed under conditions in which protein matter has not been depleted from the deglycosylation mixture, the reaction mixture comprising a molar excess of labeling reagent, the reaction mixture having the labeling reagent, released glycosylamines, proteinaceous amines comprising lysine residues, and derivatized glycosylamines wherein the derivatized glycosylamines are ...

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Номер записи: 69
25-04-2019 дата публикации

METHOD FOR ANALYZING SIALYL SUGAR CHAIN

Номер: US20190120793A1
Автор: NISHIKAZE Takashi
Принадлежит: SHIMADZU CORPORATION

Provided is a method for efficiently analyzing the structure of a sialylated glycan, including the linkage type of a sialic acid linked to a terminal of the glycan. After the sialic acid residues in a sample (glycan) to be analyzed are modified in a linkage-type-specific way (S), a mass spectrum for the sample is obtained by mass spectrometry (S). Based on the masses of the modified glycans estimated from the m/z values of the peaks observed on the mass spectrum, all possible monosaccharide compositions are exhaustively estimated under specific conditions including the kinds of monosaccharides and the range of the number of occurrences of each monosaccharide (S). Monosaccharide composition candidates listed for each peak are narrowed down by using the mass difference between different modifications of a sialic acid residue generated by the linkage-type-specific modification (S). Specifically, peaks whose mass differences correspond to the mass difference between different modifications of the sialic acid residue generated by the linkage-type-specific modification are located and assumed as a cluster of peaks originating from the same glycan including linkage isomers. For each peak belonging to this cluster, the corresponding monosaccharide composition candidates are narrowed down by excluding candidates that do not satisfy specific conditions, such as the presence or absence of a modified sialic acid residue and the number of occurrences of the modified sialic acid residue. 1. A sialylated glycan analysis method using mass spectrometry for analyzing a structure of a sialylated glycan to which a sialic acid is linked , the method comprising:a) a modification step in which a linkage-type-specific modification is performed on a sialic acid residue included in a sialylated glycan to be analyzed;b) an analysis execution step in which the sialylated glycan after the modification by the modification step is subjected to a mass spectrometric analysis to obtain mass spectrum ...

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Номер записи: 70
14-05-2015 дата публикации

Method for Determining Ubiqutin Chain Length

Номер: US20150132779A1
Автор: Kaiho Ai, Saeki Yasushi, TANAKA Keiji, Tsuchiya Hikaru
Принадлежит:

Protein ubiquitylation, an essential post-translational modification, regulates almost every cellular process including protein degradation, protein trafficking, signal transduction, and DNA damage response in eukaryotic cells. The diverse functions of ubiquitylation are thought to be mediated by distinct chain topologies resulting from eight different ubiquitin linkages, chain lengths, and complexities. Currently, ubiquitin linkages are generally thought to be a critical determinant of ubiquitin signaling. However, ubiquitin chain lengths, another key element of ubiquitin signaling, have not been well documented especially in vivo situation during past three decades from the discovery of ubiquitin. The reason of this was simply because no method has been available for determination of ubiquitin chain length in endogenous ubiquitylated substrates. In the present invention, a practical technique for determining the actual length of substrate-attached polyubiquitin chains from biological samples is established. Using the method, the mean length of substrate-attached polyubiquitin chains was determined and the robustness of ubiquitin chain length regulation in cells is investigated. The following is a summary of findings in this invention: 1. A method for determining ubiquitin chain length was developed and this method was named ‘ubiquitin protection from trypsinization’ (Ub-ProT). 2. Using Ub-ProT, it was determined that the mean length of substrate-attached ubiquitin chains is in the dimer to decamer range. 3. By quantitative proteomics, it was found that the mean lengths of five major types of ubiquitin chains can be divided into two groups. 4. Proteasome-inhibition did not alter the mean length of substrate-attached polyubiquitin chains, indicating that cells have a robust system for regulating ubiquitin chain length. 1. A polypeptide comprising ubiquitin binding domains , wherein said ubiquitin binding domains are linked to each other via a linker amino acid ...

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Номер записи: 71
01-09-2022 дата публикации

ANTIBODIES SPECIFIC TO GLYCOSYLATED PD-L1 AND METHODS OF USE THEREOF

Номер: US20220276252A9
Автор: CHUNG Ezra M., Hung Mien-chie, Kim Yong-Soo, Li Chia-Wei, LIM Seung-Oe, YOO Stephen S.
Принадлежит:

Antibodies that selectively bind to glycosylated PD-1 relative to unglycosylated PD-1 are provided. In some aspects, PD-1 polypeptides comprising glycosylated amino acid positions are also provided. Methods for making and using such antibodies and polypeptides (e.g., for the treatment of cancer) are also provided. 1. An isolated chimeric , humanized , or fully human monoclonal antibody which selectively binds to glycosylated PD-1 relative to unglycosylated PD-1.2. The isolated antibody of claim 1 , wherein the antibody selectively binds to PD-1 glycosylated at positions N49 claim 1 , N58 claim 1 , N74 claim 1 , N116 claim 1 , or any combination thereof claim 1 , relative to unglycosylated PD-1.3. (canceled)4. The isolated antibody of claim 1 , wherein the antibody claim 1 , when it is directly or indirectly detectable by a fluorescent label claim 1 , preferentially binds to cells expressing glycosylated PD-1 with a measured fluorescence intensity (MFI) that is 10-fold to 50-fold higher than the MFI exhibited by the antibody binding to cells expressing unglycosylated PD-1 in a cell flow cytometry assay.5. (canceled)6. The isolated antibody of claim 1 , wherein said antibody binds to an epitope of glycosylated PD-1 comprising amino acids 34 to 44 claim 1 , 49 to 59 claim 1 , and 104 to 124 of SEQ ID NO: 1 and contacts amino acids at positions 36 claim 1 , 38 claim 1 , 51 claim 1 , 53 claim 1 , 55 claim 1 , 109 claim 1 , 115 claim 1 , 118 and 121 of SEQ ID NO: 1 or to an epitope of glycosylated PD-L1 comprising amino acids 91 to 107 and 123 to 134 of SEQ ID NO: 1 and contacts amino acids at positions 95 claim 1 , 103 claim 1 , 127 and 131 of SEQ ID NO: 1.7. The isolated antibody of claim 1 , wherein said antibody competes or cross competes for specific binding to glycosylated PD-1 with MAb STM418 or MAb STM432.8. The isolated antibody of claim 1 , wherein the VH domain of said antibody has an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ...

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Номер записи: 72
02-05-2019 дата публикации

Methods and kits for the diagnosis and risk stratification of patients with ischemia

Номер: US20190128900A1
Автор: Judit CUBEDO RÀFOLS, Lina Badimon Maestro, Teresa PADRÓ CAPMANY
Принадлежит: Consejo Superior de Investigaciones Cientificas CSIC, Fundacio Institut de Recerca de lHospital de La Santa Creu i Sant Pau

The invention relates to methods for the diagnosis of ischemia or ischemic tissue damage, methods for predicting the progression of ischemia in a patient having suffered an ischemic event, for determining the prognosis of a patient having suffered an ischemic event and for determining the risk that a patient suffering from stable coronary disease suffers a recurrent ischemic event based on the detection of the levels of glycosylated Apo J. The invention relates as well to a method for the determination of glycosylated Apo J in a sample.

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Номер записи: 73
23-04-2020 дата публикации

METHODS FOR LIQUID CHROMATOGRAPHY CALIBRATION FOR RAPID LABELED N-GLYCANS

Номер: US20200124574A1
Автор: Brousmiche Darryl W., Lauber Matthew A., Morris Michael F.
Принадлежит:

Methods are provided for making rapid labeled dextran ladders and other calibrants useful in liquid chromatography. The methodologies include a two-step process comprising a reductive amination step of providing a reducing glycan and reacting it with a compound having a primary amine to produce an intermediate compound. The intermediate compound is then rapidly tagged with a rapid tagging reagent to produce the rapid labeled dextran ladder. 1. A method of making a calibrant useful in liquid chromatography , comprising the steps of:providing a reducing glycan having an aldehyde group;reacting the aldehyde group of the reducing glycan with a compound having a primary amine to produce an intermediary compound;mixing the intermediary compound with the rapid tagging reagent, wherein the intermediary compound is rapidly labeled with the rapid tagging reagent; andproducing a rapid labeled dextran ladder having substantially identical optical properties as a rapid labeled N-glycan wherein the rapid label N-glycan is produced by rapid tagging of a N-glycan with the rapid tagging reagent.2. (canceled)3. A method of generating a glucose unit useful in glycan database investigations , comprising the steps of:preparing a plurality of N-glycans from a biological sample;reacting the N-glycans with a rapid tagging reagent to produce a plurality of rapid labeled N-glycans;providing a reducing glycan;reacting the reducing glycan with a compound having a primary amine to produce an intermediary compound;mixing the intermediary compound with the rapid tagging reagent, wherein the intermediary compound is rapidly labeled with the rapid tagging reagent to produce a rapid labeled dextran ladder having substantially identical optical properties as the rapid labeled N-glycan;calibrating a hydrophilic interaction chromatography separation of the rapid labeled N-glycans with the rapid labeled dextran to provide peak retention times for the rapid labeled N-glycans; andconverting the peak ...

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Номер записи: 74
19-05-2016 дата публикации

METHODS AND COMPOSITIONS RELATING TO NEURODEGENERATIVE DISEASES

Номер: US20160139151A1
Автор: Liang Hui-Chung, Pike Ian Hugo, Ward Malcolm Andrew
Принадлежит: Electrophoretics Limited

The present invention provides a method for diagnosing or assessing a neurodegenerative disease in a test subject, comprising: (i) providing a protein-containing sample that has been obtained from the test subject; (ii) determining the concentration, amount or degree of expression of at least one specific protein isoform and/or glycoform derived from a protein biomarker selected from the group consisting of: clusterin precursor; apolipoprotein A-IV precursor; apolipoprotein C-III precursor; transthyretin; galectin 7; complement C4 precursor; alpha-2-macroglobulin precursor; Ig alpha-1 chain C; histone 2B; Ig lambda chain C region; fibrinogen gamma chain precursor; complement factor H; inter-alpha-trypsin heavy chain H4 precursor; complement C3 precursor; gamma or beta actin; haptoglobin precursor; and the serum albumin precursor, or a fragment thereof; (iii) comparing said concentration, amount or degree determined in (ii) with a reference from a control subject with a specific neurodegenerative disease, dementia or stage of disease, or from a control subject that does not have a neurogenerative disease or dementia; and (iv) based on the level of the at least one specific protein isoform and/or glycoform of the protein biomarker in the test subject relative to the reference, making a diagnosis or assessment as to the presence of and/or stage of neurodegenerative disease or dementia of the test subject. Also provided are related products and systems for use in such a method. 144-. (canceled)45. A method for diagnosing or assessing a neurodegenerative disease or neurodegenerative dementia in a test subject , comprising:(i) providing a protein-containing sample that has been obtained from the test subject;(ii) determining the concentration, amount or degree of expression of at least one specific protein isoform and/or glycoform derived from a protein biomarker selected from the group consisting of: clusterin precursor; apolipoprotein A-IV precursor; apolipoprotein C- ...

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Номер записи: 75
07-08-2014 дата публикации

ApoIII and the Treatment and Diagnosis of Diabetes

Номер: US20140220039A1
Автор: Per-Olof Berggren
Принадлежит: BIOCRINE AB

The present invention provides methods of identifying candidate compounds for the treatment of type I diabetes comprising contacting pancreatic β cells with an amount of apolipoprotein CIII (“apoCIII”) effective to increase intracellular calcium concentration, in the presence of one or more test compounds, and identifying those test compounds that inhibit an apoCIII-induced increase in intracellular calcium concentration in the pancreatic β cells. The present invention also provides methods for treating patients with type I diabetes comprising administering to the patient an amount effective of an inhibitor of apoCIII to reduce apoCIII-induced increase in intracellular calcium concentration in pancreatic β cells.

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Номер записи: 76
18-05-2017 дата публикации

COMPOSITIONS AND METHODS FOR TREATING CARDIOVASCULAR DISEASE AND MYOCARDIAL INFARCTION WITH DIPEPTIDYL PEPTIDASE INHIBITORS OR B TYPE NATRIURETIC PEPTIDE ANALOGUES RESISTANT TO PROLYL-SPECIFIC DIPEPTIDYL DEGRADATION

Номер: US20170138932A1
Автор: Whittaker Michael A.
Принадлежит: ALERE SAN DIEGO, INC.

The present invention describes compositions and methods for treating cardiovascular disease and myocardial infarction using dipeptidyl peptidase inhibitors. Also provided are methods for increasing natriuretic peptide function by administering one or more analogues of B type natriuretic peptide that provide increased stability in the presence of prolyl-specific dipeptidyl peptidases. 117-. (canceled)18. A method for improving the accuracy of an assay for a natriuretic peptide of interest , comprising:selecting one or more antibodies that specifically bind to a biologically active form of the natriuretic peptide of interest, but that do not specifically bind to a biologically inactive form of the natriuretic peptide of interest, andperforming an assay in which the signal depends upon the selected antibody specifically binding to a biologically active form of the natriuretic peptide of interest, but not specifically binding to a biologically inactive form of the natriuretic peptide of interest,thus generating an assay more specific for the biologically active form of the natriuretic peptide of interest,wherein the difference in biological activity between the active and inactive forms of the natriuretic peptide of interest is due to the glycosylation state of the natriuretic peptide.19. The method of claim 18 , wherein the step of selecting one or more antibodies comprises selecting antibodies claim 18 , that claim 18 , when used in an assay claim 18 , detect a biologically active form of the natriuretic peptide of interest and exhibits a signal at least a factor of 5 greater claim 18 , at least a factor of 10 greater claim 18 , at least a factor of 20 greater claim 18 , at least a factor of 50 greater claim 18 , or at least a factor of 100 greater claim 18 , than the signal exhibited from detection of a biologically inactive form of the natriuretic peptide of interest.20. The method of claim 18 , wherein the step of selecting one or more antibodies comprises ...

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Номер записи: 77
18-05-2017 дата публикации

ApoIII and the Treatment and Diagnosis of Diabetes

Номер: US20170138968A1
Автор: Per-Olof Berggren
Принадлежит: BIOCRINE AB

The present invention provides methods of identifying candidate compounds for the treatment of type I diabetes comprising contacting pancreatic β cells with an amount of apolipoprotein CIII (“apoCIII”) effective to increase intracellular calcium concentration, in the presence of one or more test compounds, and identifying those test compounds that inhibit an apoCIII-induced increase in intracellular calcium concentration in the pancreatic β cells. The present invention also provides methods for treating patients with type I diabetes comprising administering to the patient an amount effective of an inhibitor of apoCIII to reduce apoCIII-induced increase in intracellular calcium concentration in pancreatic β cells.

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Номер записи: 78
28-05-2015 дата публикации

METHODS RELATED TO BEVACIZUMAB

Номер: US20150147317A1
Автор: Bosques Carlos J., Collins Brian Edward, Kaundinya Ganesh, Robblee John
Принадлежит:

The present invention relates to the characterization and production of bevacizumab. 128-. (canceled)29. A method of manufacturing a bevacizumab drug product , comprising:providing or obtaining a test glycoprotein preparation;acquiring at least one value for a bevacizumab parameter listed in Table 1 for the test glycoprotein preparation; andprocessing at least a portion of the test glycoprotein preparation as bevacizumab drug product if the at least one value for the test glycoprotein preparation meets a reference criterion shown in Table 1 for the parameter,thereby manufacturing a bevacizumab drug product.30. The method of claim 29 , comprising:acquiring values for any combination of two or more bevacizumab parameters listed in Table 1; andprocessing at least a portion of the test glycoprotein preparation as bevacizumab drug product if the values for the any combination of two or more bevacizumab parameters for the test glycoprotein preparation meet the corresponding reference criterion shown in Table 1 for the parameters.31. The method of claim 30 , wherein the any combination of two or more bevacizumab parameters comprises:2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, all, or a plurality of the bevacizumab parameters listed in Table 1; orany two or more of parameter numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and/or 15 shown in Table 1.32. The method of claim 29 , wherein the test glycoprotein preparation comprises a recombinant antibody composition having a first amino acid sequence with at least 85% identity to SEQ ID NO:1 (e.g. claim 29 , 90 claim 29 , 95 claim 29 , 98 claim 29 , or 100% identity to SEQ ID NO:1) and a second amino acid sequence with at least 85% identity to SEQ ID NO:2 (e.g. claim 29 , 90 claim 29 , 95 claim 29 , 98 claim 29 , or 100% identity to SEQ ID NO:2).33. The method of claim 32 , wherein the first and second amino acid sequences form a recombinant antibody.34. The method of claim 29 , wherein the test glycoprotein ...

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Номер записи: 79
28-05-2015 дата публикации

METHOD FOR DETECTING CANCER, AND ANTIBODY CAPABLE OF RECOGNIZING PANCREATIC RIBONUCLEASE 1

Номер: US20150147762A1
Автор: Nakatani Daisuke
Принадлежит: TOSOH CORPORATION

Pancreatic cancer can be detected using a monoclonal antibody which binds to pancreatic RNase 1 when a site in pancreatic RNase 1 capable of being modified with an N-glycan chain is not linked to a glycan chain, but which does not bind to pancreatic RNase 1 when an N-glycan chain is linked to the site. Also provided is a monoclonal antibody which can bind to pancreatic RNase 1 simultaneously with the binding of the aforementioned antibody to pancreatic RNase 1, and determining the ratio of A to B using the antibodies, wherein A represents the amount of the site located in the pancreatic RNase 1 capable of being modified with an N-glycan chain, wherein an N-glycan chain is linked or not linked to the site; and B represents the amount of the site located in the pancreatic RNase 1 capable of being modified with an N-glycan chain. 1. A method for detecting cancer , characterized by measuring the amount of a putative N-glycosylation site of a glycoprotein linked with an N-glycan chain or not linked with an N-glycan chain.2. The method according to claim 1 , wherein the cancer is pancreatic cancer.3. The method according to claim 1 , wherein the glycoprotein is pancreatic ribonuclease 1.4. The method according to claim 2 , wherein the amount of the site linked with the N-glycan chain is increased in pancreatic cancer patients in comparison with healthy individuals.5. A method for detecting cancer claim 2 , comprising: determining the ratio A/B for A and B indicated below:A: amount of putative N-glycosylation site of a glycoprotein linked with an N-glycan chain or not linked with an N-glycan chain; and,B: amount of putative N-glycosylation site of the glycoprotein.6. The method according to claim 5 , wherein the cancer is pancreatic cancer.7. The method according to claim 5 , wherein the glycoprotein is pancreatic ribonuclease 1.8. The method according to claim 6 , wherein A represents the amount of the site not linked with an N-glycan chain claim 6 , and the value of A/B ...

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Номер записи: 80
28-05-2015 дата публикации

SURFACE ANCHORED LIGHT CHAIN BAIT ANTIBODY DISPLAY SYSTEM

Номер: US20150148246A1
Автор: Shaheen Hussam Hisham, Zha Donxing
Принадлежит:

The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. Embodiments of the invention provide a system in which a bait complexed with a monovalent antibody fragment can be captured prior to secretion in a host cell by virtue of surface displaying an antibody light chain and utilizing the covalent interaction of this light chain with the heavy chain of an antibody molecule that is co-expressed in the same host. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of using the system for identifying antibodies that bind specifically to an antigen of interest. 1. An antibody display system comprising:(a) an isolated host cell;(b) a bait comprising a light immunoglobulin chain or functional fragment thereof fused to a surface anchor polypeptide or functional fragment thereof;(c) one or more polynucleotides encoding an immunoglobulin light chain variable region; and(d) one or more polynucleotides encoding an immunoglobulin heavy chain variable region.2. The antibody display system of further comprising:a non-tethered full antibody or antigen-binding fragment thereof comprising said immunoglobulin light and heavy chains encoded by said polynucleotides; and/or(ii) a monovalent antibody fragment which is complexed with an immunoglobulin heavy chain which is complexed with the immunoglobulin light chain of the bait.3PichiaSaccharomyces cerevisiae. The antibody display system of wherein the host cell is a or cell.4. The antibody display system of claim 1 ,wherein said one or more polynucleotides encoding an immunoglobulin light chain is from a genetically diverse population of immunoglobulin light chains; and/or,wherein said one or more polynucleotides encoding an immunoglobulin heavy chain is from a genetically diverse population of immunoglobulin heavy chains.5. The antibody display system of claim 1 , wherein the host cell comprises ...

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Номер записи: 81
26-05-2016 дата публикации

Method of monitoring cellular trafficking of peptides

Номер: US20160146786A1
Автор: Cunningham Paula, Heinrich Tatjana, Hoffmann Katrin, Hopkins Richard, Milech Nadia, Watt Paul
Принадлежит:

This disclosure provides a method of isolating peptides having cell-penetrating function, wherein the peptides are detected as biotinylated molecules only following their translocation through the cell membrane. The disclosure also provides methods for validating the cell-penetrating function of the peptides, or that may be employed in their own right to isolate such peptides, wherein the peptides are detectable by virtue of their ability to transport a detectable cargo into the cytoplasm, such as a cargo toxin or a fragment of a green fluorescent protein (GFP) that is required for complementation of a functional GFP. The disclosure also provides non-canonical peptides having cell-penetrating function that differ structurally from known CPPs such as TAT, VP22, transportan and penetratin, and that are capable of translocating cell membranes and escaping the endosome. The disclosed peptides have utility in transporting cargo therapeutics and diagnostics into cells. 1. A method of determining or identifying a peptide capable of translocating a membrane of a cell , the method comprising the steps:(i) contacting host cells expressing a biotin ligase with a plurality of non-biotinylated members, wherein the members comprise scaffolds displaying fusion proteins, each of the fusion proteins comprising a candidate peptide moiety and a biotin ligase substrate domain, and wherein said contacting is for a time and under conditions sufficient for at least the displayed fusion proteins of members to enter the host cells;(ii) incubating the host cells for a time and under conditions such that the biotin ligase substrate domain of the at least fusion proteins that have translocated a membrane of the host cell are enzymatically biotinylated by the expressed biotin ligase; and (a) detecting the presence of a biotinylated fusion protein in a host cell or cell lysate or extract thereof, wherein the presence of a biotinylated fusion protein indicates that the candidate peptide moiety ...

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Номер записи: 82
09-05-2019 дата публикации

SATURATION BINDING RATIOMETRIC ASSAY

Номер: US20190137522A1
Автор: MANNEH Victor, Svarovsky Sergei A
Принадлежит:

Methods, devices, and reagents are described for performing ratiometric assays for hemoglobin A1c. The methods involve a direct ratio determination between Hb A1c and normalized total hemoglobin utilizing a saturating amount of hemoglobin so that Hb A1c binds proportionately to a substrate. In some applications, the assay utilizes LOCI or other proximity label for signal generation and/or labeled magnetic beads. The methods can be configured as homogeneous or heterogeneous assays. 1. A method for determining the fraction of a glycated hemoglobin in a sample , comprising a) a sample containing total hemoglobin in sufficient volume whereby binding of total hemoglobin on said solid phase surface is saturated; and', 'b) second binding member specific for hemoglobin A1c of said total hemoglobin and labeled with a second label;, 'contacting optionally labeled particles comprising a solid phase substrate bearing first binding members specific for total hemoglobin with'}detecting at least one signal from said second label from specific binding member bound to said particles, wherein the level of said at least one signal is indicative of the fraction of hemoglobin A1c relative to total hemoglobin.2. The method of claim 1 , wherein said solid phase substrate is labeled with a first label claim 1 , and said method further comprises detecting the distinguishable signal from first label as an indication of the number of particles present.3. The method of claim 1 , wherein the binding member specific for total hemoglobin binds within beta chain amino acids 7-25 and preferentially binds to HbA rather than to HbF.4. The method of claim 1 , wherein said method is performed as a homogenous assay.5. A method for determining the fraction of Hb A1c in a sample containing hemoglobin claim 1 , comprising a) a sample containing total hemoglobin in sufficient volume and concentration whereby binding of total hemoglobin on said particles is saturated; and', 'b) second specific binding member ...

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Номер записи: 83
24-05-2018 дата публикации

SURFACE ANCHORED LIGHT CHAIN BAIT ANTIBODY DISPLAY SYSTEM

Номер: US20180142233A1
Автор: Shaheen Hussam Hisham, Zha Dongxing
Принадлежит: Merck Sharp & Dohme Corp.

The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. Embodiments of the invention provide a system in which a bait complexed with a monovalent antibody fragment can be captured prior to secretion in a host cell by virtue of surface displaying an antibody light chain and utilizing the covalent interaction of this light chain with the heavy chain of an antibody molecule that is co-expressed in the same host. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of using the system for identifying antibodies that bind specifically to an antigen of interest. 15-. (canceled)6: An isolated bait polypeptide comprising a light immunoglobulin chain or functional fragment thereof fused , optionally by a peptide linker , to a surface anchor polypeptide or a functional fragment thereof which bait polypeptide is optionally amino-terminally fused to a signal peptide.7: The polypeptide of wherein the surface anchor polypeptide is SED-1 or a functional fragment thereof.8: The polypeptide of wherein the light immunoglobulin chain or functional fragment thereof comprises a kappa or lambda constant immunoglobulin domain.9: An isolated polynucleotide encoding the polypeptide of .10: A vector comprising the polynucleotide of .11: An isolated host cell comprising the polypeptide of or a polynucleotide encoding said polypeptide.12: The host cell of further comprising one or more polynucleotides encoding an immunoglobulin light chain; and/or one or more polynucleotides encoding an immunoglobulin heavy chain.13PichiaSaccharomyces cerevisiae: The host cell of which is a or cell.14: The host cell of wherein the bait polypeptide is located on the surface of the cell membrane.15: A composition comprising the host cell of claim 12 , further comprising a non-tethered full antibody or antigen-binding fragment thereof comprising said immunoglobulin light ...

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Номер записи: 84
04-06-2015 дата публикации

Methods related to panitumumab

Номер: US20150152184A1
Автор: Brian Edward Collins, Carlos J. Bosques, Ganesh Kaundinya, John Robblee
Принадлежит: Momenta Pharmaceuticals Inc

The present invention relates to the characterization and production of panitumumab.

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Номер записи: 85
24-05-2018 дата публикации

METHODS OF IDENTIFYING SENP1 INHIBITORS

Номер: US20180143196A1
Автор: Chen Chi-Hong, Chen Yuan, Namanha Andrew
Принадлежит:

Provided herein are methods of detecting binding of an SENP1 polypeptide to a compound and methods for screening for inhibitors of SENP1. Further provided are aqueous compositions comprising SENP1 polypeptides and NMR apparatuses comprising the compositions for NMR analysis. 118.-. (canceled)19. A method of screening for an inhibitor of SENP1 comprising contacting a composition comprising an SENP1 polypeptide with a test compound and detecting whether the test compound binds the SENP1 polypeptide or fragment thereof by nuclear magnetic resonance.20. A method of identifying an SENP1 inhibitor , the method comprising:combining an SENP1 polypeptide, a SUMO protein, and a test compound in a reaction vessel;allowing the SENP1 polypeptide, SUMO protein and test compound to form a SENP1-SUMO-compound complex; anddetecting the SENP1-SUMO-compound complex thereby identifying the compound as a SENP1 inhibitor. This application is a divisional of U.S. patent application Ser. No. 14/247,153, filed Apr. 7, 2014, now U.S. Pat. No. 9,791,447, issued Oct. 17, 2017, which claims the benefit of U.S. Provisional Patent Applications 61/809,208, filed Apr. 5, 2013, and 61/813,832, filed Apr. 19, 2013, each of which is incorporated herein by reference in its entirety and for all purposes.This invention was made with government support under NM Grant Nos. R01GM074748, R01GM086171 and R01GM102538. The government has certain rights in the invention.The Sequence Listing written in file 48440-521C01US_ST25.TXT, created on Oct. 13, 2017, 22,281 bytes, machine format IBM-PC, MS-Windows operating system, is hereby incorporated by reference.Post-translational modifications with the small ubiquitin-like modifiers (SUMO) are initiated and removed by the activities of SUMO-specific proteases (SENPs). Unlike ubiquitylation, which has one modifier (i.e., ubiquitin) and one dominant role, namely protein degradation, SUMOylation involves three modifiers (SUMO-1, -2, and -3) and affects diverse cellular ...

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Номер записи: 86
24-05-2018 дата публикации

METHOD FOR SCREENING DIABETIC NEPHROPATHY IN A SUBJECT

Номер: US20180143207A1
Автор: CHANG CHIA CHU, CHANG CHUNG HO
Принадлежит:

A urine biomarker is provided, and more particularly, a screening method using the urine biomarker to screen a subject having a high risk of developing diabetic nephropathy is also provided. The urine biomarker of glycosylated uromodulin can be used to detect and predict diabetic patients having a high risk of developing diabetic nephropathy more accurate and earlier than general detection methods of PCR or ACR tests, thereby the patient being screened can be treated earlier and thus prevent deterioration of the progression of chronic kidney disease, and therefore dramatically decrease the risk of death. 1. A method for evaluating a subject having a high risk of developing a diabetic nephropathy , comprising the steps of:providing a urine sample from the subject; anddetecting the presence of glycated uromodulin in the urine sample.2. The method according to claim 1 , wherein the subject is a diabetic patient.3. The method according to claim 2 , wherein the age of the subject is less than 65 years old.4. The method according to claim 1 , wherein the urine sample is obtained from the supernatant fraction of the urine sample centrifugation.5. The method according to claim 4 , wherein the urine sample is centrifuged at about 16 claim 4 ,000×g to about 120 claim 4 ,000×g.6. The method according to claim 1 , wherein the level of glycated uromodulin is determined by Western blotting claim 1 , mass spectrometry claim 1 , immunoassay claim 1 , or chromatography.7. The method according to claim 1 , wherein the level of glycated uromodulin in the urine sample is 9 claim 1 ,000 a.u. or more.8. The method according to claim 7 , wherein an albumin-creatinine ratio (ACR) or a protein-creatinine ratio (PCR) of the urine sample is further detected. This application claims priority to Taiwan Patent Application No. 105138384 filed on 23 Nov. 2016, all disclosures of which are incorporated herein by reference in its entirety.The present invention relates generally to a urinary ...

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Номер записи: 87
25-05-2017 дата публикации

AGENTS AND METHODS FOR DETERMINING COLORECTAL CANCER STATUS

Номер: US20170146537A1
Автор: Yeung Trevor Ming-Yee
Принадлежит: OXFORD UNIVERSITY INNOVATION LIMITED

The invention provides a method of determining the colorectal cancer status of subject, comprising applying to a colon and/or rectal surface an agent which is able to distinguish between a) mature glycosylated forms of MUC2 and b) incomplete or aberrant glycosylated forms of MUC2, and determining the extent to which the agent binds. 1. A method of determining the colorectal cancer status of subject , comprising applying to a colon and/or rectal surface an agent which is able to distinguish between a) mature glycosylated forms of Mucin 2 (MUC2) and b) incomplete or aberrant glycosylated forms of MUC2 , and determining the extent to which the agent binds.2. An agent which is able to distinguish between a) mature glycosylated forms of MUC2 and b) incomplete or aberrant glycosylated forms of MUC2.3. (canceled)4. A method of treating colorectal cancer or dysplasia in a subject comprising:i) topically applying to the colon and/or rectal region of a subject a labeled agent capable of binding predominantly to either mature glycosylated forms of MUC2 associated with healthy tissue or to incomplete or aberrant glycosylated forms of MUC2 associated with dysplasia or cancer;ii) visualizing the colorectal region to which the agent was applied and observing where the agent has bound; andiii) excising or treating tissue associated with the incomplete or aberrant glycosylated forms of MUC2.5. A kit for identifying cancerous or dysplastic colorectal tissue , or for determining the colorectal cancer status , in a subject comprising i) a labeled agent which binds predominantly to either mature glycosylated forms of MUC2 or to incomplete or aberrant glycosylated forms of MUC2; and ii) instructions to apply the labeled agent topically to the surface of the colon and/or rectal region of the subject.6. The method claim 1 , wherein the agent binds only claim 1 , or predominantly claim 1 , to mature glycosylated forms of MUC2; or wherein the agent binds only claim 1 , or predominantly claim ...

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Номер записи: 88
02-06-2016 дата публикации

METHOD FOR DETERMINING DEPRESSION, KIT FOR ANALYZING SEROTONIN TRANSPORTER, AND KIT FOR ANALYZING UBIQUITINATED SEROTONIN TRANSPORTER IN BLOOD

Номер: US20160154014A1
Автор: Mouri Akihiro, Nabeshima Toshitaka
Принадлежит: MEIJO UNIVERSITY

[Problem] To provide: a method for utilizing a novel marker, including a method for determining depression; and a kit for analyzing an ubiquitinated serotonin transporter. 14.-. (canceled)5. A kit for analyzing a serotonin transporter , comprising a proteasome inhibitor and an anti-serotonin transporter antibody , and being used for the analysis of the amount of the untreated serotonin transporter and the amount of the inhibitor-treated serotonin transporter in the collected blood sample.6. The kit for analyzing a serotonin transporter of claim 5 , which is used for the diagnosis of depression.7. A method for determining depression claim 5 , comprising a step of analyzing the proportion of the ubiquitinated serotonin transporter in a blood sample collected from a subject.8. The method for determining depression of claim 7 , which further includes a step of comparing the proportion of the ubiquitinated serotonin transporter obtained from the blood sample collected from the subject and the proportion of the ubiquitinated serotonin transporter as the control obtained from at least one healthy subject.9. The method for determining depression of claim 8 , which determines that the subject has depression when the proportion of the ubiquitinated serotonin transporter obtained from the blood sample collected from the subject is at a lower level than that in the control.10. A kit for analyzing an ubiquitinated serotonin transporter in the blood claim 8 , comprising an ubiquitinated protein collector and an anti-serotonin transporter antibody claim 8 , and being used for the analysis of the proportion of the ubiquitinated serotonin transporter in the collected blood sample.11. The kit for analyzing an ubiquitinated serotonin transporter in the blood of claim 10 , which is used for the diagnosis of depression. The present invention relates to a method for utilizing a novel marker, a kit for analyzing a serotonin transporter, and a kit for analyzing an ubiquitinated serotonin ...

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Номер записи: 89
31-05-2018 дата публикации

METHODS AND KITS FOR CANCER ANTIGEN AND HEPARAN SULFATE IMAGING

Номер: US20180149658A1
Автор: Person Anthony D., Wu Zhengliang L.
Принадлежит:

An in vitro method of imaging a cancer antigen includes providing a cell sample including a cancer antigen selected from the group consisting of a Tn antigen, a T antigen, a sialylated-Tn antigen, and a sialylated-T antigen, treating the sample with a glycosyltransferase to incorporate a carbohydrate with a click chemistry moiety into the cancer antigen, adding a label to the sample that includes a click chemistry moiety that reacts to the click chemistry moiety of the carbohydrate such that the label attaches to the carbohydrate to form a labeled cancer antigen, and imaging the sample with a camera. 1. An in vitro method of imaging a cancer antigen , the method comprising:providing a sample comprising a cancer antigen selected from the group consisting of a Tn antigen, a T antigen, a sialylated-Tn antigen, and a sialylated-T antigen;treating the sample with a glycosyltransferase to incorporate a carbohydrate into the cancer antigen, wherein the carbohydrate includes a click chemistry moiety;adding a label to the sample, wherein the label includes a click chemistry moiety that reacts to the click chemistry moiety of the carbohydrate such that the label attaches to the carbohydrate to form a labeled cancer antigen; andimaging the sample with a camera.23613132616264. The method of claim 1 , wherein the glycosyltransferase is selected from the group consisting of BGNT claim 1 , GCNT claim 1 , STGal claim 1 , STGal claim 1 , STGalNAc claim 1 , STGalNAc claim 1 , and STGalNAc.3. The method of claim 1 , further comprising treating the sample with sialidase prior to treating the sample with the glycosyltransferase to expose a glycosyltransferase recognition site.4. The method of claim 1 , wherein the label is a fluorescent label claim 1 , a colorimetric label claim 1 , a biotin linked to a fluorescent label claim 1 , or a biotin linked to a colorimetric label claim 1 , and wherein the carbohydrate includes a click chemistry moiety selected from one of an azido group or an ...

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Номер записи: 90
11-06-2015 дата публикации

METHODS RELATED TO ADALIMUMAB

Номер: US20150158943A1
Автор: Bosques Carlos J., Collins Brian Edward, Kaundinya Ganesh, Robblee John
Принадлежит:

Production and bioanalytical characterization of determinative parameters of adalimumab is presented. 134-. (canceled)35. A method of manufacturing an adalimumab drug product , comprising:providing or obtaining a test glycoprotein preparation;acquiring at least one value for an adalimumab parameter listed in Table 1 for the test glycoprotein preparation; andprocessing at least a portion of the test glycoprotein preparation as adalimumab drug product if the at least one value for the test glycoprotein preparation meets a reference criterion shown in Table 1 for the parameter,thereby manufacturing an adalimumab drug product.36. The method of claim 35 , comprising:acquiring values for any combination of two or more adalimumab parameters listed in Table 1; andprocessing at least a portion of the test glycoprotein preparation as adalimumab drug product if the values for the any combination of two or more adalimumab parameters for the test glycoprotein preparation meet the corresponding reference criterion shown in Table 1 for the parameters.37. The method of claim 36 , wherein the any combination of two or more adalimumab parameters comprises:2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, all, or a plurality of the adalimumab parameters listed in Table 1; orany two or more of parameter numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and/or 22 shown in Table 1.38. The method of claim 35 , wherein the test glycoprotein preparation comprises a recombinant antibody composition having a first amino acid sequence with at least 85% identity to SEQ ID NO:1 (e.g. claim 35 , 90 claim 35 , 95 claim 35 , 98 claim 35 , or 100% identity to SEQ ID NO:1) and a second amino acid sequence with at least 85% identity to SEQ ID NO:2 (e.g. claim 35 , 90 claim 35 , 95 claim 35 , 98 claim 35 , or 100% identity to SEQ ID NO:2).39. The method of claim 38 , wherein the first and second amino acid sequences form a recombinant antibody. ...

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Номер записи: 91
11-06-2015 дата публикации

Identification and Quantification of Intact Glycopeptides in Complex Samples

Номер: US20150160233A1
Автор: Mechref Yehia S., Tang Haixu
Принадлежит:

Illustrative embodiments of methods and apparatus for identifying one or more intact glycopeptides in a sample are disclosed. According to one illustrative embodiment, a method may comprise receiving data representing a plurality of mass spectra obtained from mass spectrometry of the sample, scoring data representing each of the plurality of mass spectra against data associated with target glycopeptides, and identifying one or more intact glycopeptides in the sample based at least in part on the scoring of the data representing each of the plurality of mass spectra. 1. A method of identifying one or more intact glycopeptides in a sample , the method comprising:receiving data representing a plurality of mass spectra obtained from mass spectrometry of the sample;scoring data representing each of the plurality of mass spectra against data associated with target glycopeptides;identifying one or more intact glycopeptides in the sample based at least in part on the scoring of the data representing each of the plurality of mass spectra;scoring data representing each of the plurality of mass spectra against data associated with decoy glycopeptides; andestimating a false detection rate (FDR) based at least in part on the scoring of the data representing each of the plurality of mass spectra against the data associated with target glycopeptides and the scoring of the data representing each of the plurality of mass spectra against the data associated with decoy glycopeptides, wherein the FDR comprises a ratio of a number of decoy ETD scores that exceed a threshold to a total number of ETD scores that exceed the threshold.2. The method of claim 1 , wherein each of the one or more intact glycopeptides comprises one or more glycans attached to a peptide.3. The method of claim 2 , wherein identifying one or more intact glycopeptides in the sample comprises identifying claim 2 , for each of the one or more intact glycopeptides claim 2 , one or more glycosylation sites at which the ...

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Номер записи: 92
28-08-2014 дата публикации

Composition for diagnosis of lung cancer and diagnosis kit for lung cancer

Номер: US20140242608A1
Автор: Je Yoel Cho, Jung Mo AHN
Принадлежит: SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION

The present invention provides serum paraoxonase 1 protein as a biomarker useful for early diagnosis of lung cancer.

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Номер записи: 93
16-06-2016 дата публикации

METHOD FOR DIAGNOSING CANCER THROUGH DETECTION OF DEGLYCOSYLATION OF GLYCOPROTEIN

Номер: US20160168618A1
Автор: Jin Jonghwa, Kim Hyunsoo, Kim Kyunggon, Kim Youngsoo, YOON Jung-Hwan
Принадлежит: SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION

Provided is a method for diagnosing cancer through a difference with a control group in view of the ratio of a deglycosylated peptide fragment and a non-glycosylated peptide fragment in a protein including an N-linked glycosylation motif. Further provided is a method for diagnosing cancer through the detection of the glycosylation ratio in the protein according to the subject matter enables the diagnosis of cancer with high specificity and sensitivity using at least one existing marker, and can be useful in discovering new markers for diagnosing cancer. 1. A method of diagnosing cancer in a subject or a sample in need thereof comprising steps of:providing a sample from the subject comprising proteins having a N-linked glycosylation motif;de-glycosylating the proteins comprised in the sample;fragmenting the de-glycosylated proteins;determining in the fragmented proteins the amount of the de-glycosylated peptide at the N-linked motif and the amount of the non-glycosylated peptide which does not contain the N-linked motif and the ratio therebetween; anddiagnosing the subject or the sample as cancer or susceptible to cancer if the ratio is changed in the subject or in the sample compared to that of a control.2. The method of claim 1 , wherein the N-linked glycosylation motif is represented by an amino acid sequence of AsnXxxSer (SEQ ID NO:3) claim 1 , AsnXxxThr (SEQ ID NO:4) or AsnXxxCys (SEQ ID NO:5).3. The method of claim 2 , wherein a residue of the de-glycosylation at the motif is Asn and a peptide fragment AspXxxSer (SEQ ID NO:6) claim 2 , AspXxxThr (SEQ ID NO:7) or AspXxxCys (SEQ ID NO:8) is generated by the de-glycosylation.4. The method of claim 1 , wherein the de-glycosylation is performed by using a PNGase-F.5. The method of claim 1 , wherein the fragmentation is performed by treating the sample with at least one of a trypsin claim 1 , a lysine-C claim 1 , an arginine-C or an aspartic acid N.6. The method of claim 1 , wherein the cancer is selected from the ...

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Номер записи: 94
15-06-2017 дата публикации

IN VITRO PROCESS FOR THE QUANTIFICATION OF CARBOXYMETHYL AND CARBOXYETHYL LEVEL OF ALBUMIN IN A SAMPLE

Номер: US20170168068A1
Автор: Kulkarni Mahesh Jagdishrao
Принадлежит:

An in vitro method for the identification and quantification of glycated human serum albumin to assess the extent of diabetic complications in diseased individuals. Further, a diagnostic kit for identifying the extent of diabetes in a diseased individual by estimating glycated serum albumin levels in such individuals. 1. An in vitro process for the identification , quantification or identification and quantification of carboxymethyl and carboxyethyl level of albumin in a sample comprising;(a) subjecting the sample to mass spectrometry to generate fragment ions;(b) identifying the specificity of fragment ions obtained in step (a) to advanced glycation end-product (AGE) modified glycation sites selected from carboxymethyl and carboxyethyl modified amino acid residues in human serum albumin by comparing said fragment ions with signature ions specific to modified carboxymethyl and carboxyethyl glycated sites selected from the group consisting of K36, K88, K160, K161, K183, K375, K438, K490 and K549 in Seq Id No. 1; and(c) further quantifying the levels of carboxymethyl and carboxyethyl modified peptide content wherein AGE modified glycation sites are situated at lysine (K) residues.2. The process as claimed in claim 1 , wherein AGE modified glycated sites are identified and quantified in human serum albumin represented by amino acid sequence having at least 80% similarity with Seq Id No. 1.3. The process as claimed in claim 1 , wherein AGE modified glycated sites are preferably selected from the group consisting of K549 claim 1 , K438 claim 1 , K490 claim 1 , K88 claim 1 , and K375.4. A diagnostic kit for the identification claim 1 , quantification or identification and quantification of carboxymethyl and carboxyethyl level of albumin in a sample withdrawn from a subject claim 1 , said kit comprising;a sterilized device to draw out serum from said subject which serum is to be subjected to fragmentation by mass spectrometry methods;a chart, figure or representation of ...

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Номер записи: 95
30-05-2019 дата публикации

ASSAYS FOR DETECTING MODIFIED COMPOUNDS

Номер: US20190162733A1
Автор: Pande Praveen, Rabbani Elazar, Rabbani Joshua, Stavrianopoulos Jannis G.
Принадлежит: Enzo Life Sciences, Inc. c/o Enzo Biochem

Provided are methods and compositions which are useful for separating, isolating, detecting, and quantifying compounds of interest which have been modified chemically, enzymatically or catalytically from other compounds which have not been so modified. The modifications may take the form of functional groups which are gained, lost or retained by the compounds of interest. 1. A method for detecting or quantifying an activity in a sample that causes a compound to gain a functional group , the method comprising the steps of: (i) a compound which can gain a functional group, the compound comprising at least one non-radioactive signaling moiety;', '(ii) a chemical source of the functional group which can be gained by the compound; and', '(iii) a sample;, '(a) providing(b) forming a mixture comprising the compound, the source of the functional group and the sample, and incubating the mixture under conditions suitable for the compound to gain the functional group if the activity is present in the sample; and(c) capturing any compound that has gained the functional group and separating said compound which has gained the functional group from any compound which has not gained the functional group; and(d) detecting or quantifying the non-radioactive signaling moiety of any captured separated compounds having the functional group, thereby detecting or quantifying the activity in the sample.2. The method of claim 1 , wherein the capturing and separating step comprises capturing any compound that has gained the functional group on a matrix.3. The method of claim 2 , wherein the matrix comprises a metal ion.4. The method of claim 3 , wherein the metal ion is Fe claim 3 , Ga claim 3 , Zn claim 3 , Zn claim 3 , or Mn.5. The method of claim 1 , wherein the compound comprises a macromolecule or a small biological compound.6. The method of claim 5 , wherein the macromolecule or small biological compound comprises a nucleic acid claim 5 , a protein claim 5 , a sugar claim 5 , a ...

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Номер записи: 96
25-06-2015 дата публикации

Diagnosis and treatment of alzheimer's disease

Номер: US20150177261A1
Автор: Adnan Halim, Erik Portelius, Göran Larson, Gunnar Brinkmalm, Henrik Zetterberg, Jonas Nilsson, Kaj Blennow
Принадлежит: Individual

Methods are provided for the prevention, treatment and diagnosis of Alzheimer's disease, based on the glycosylation pattern of amyloid-beta peptides in body fluids and tissues.

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Номер записи: 97
28-05-2020 дата публикации

Detection and Treatment of Pregnancy Complications Comprising Determining Sialyl Lewis Antigens and Administering hCG

Номер: US20200166527A1
Автор: Jeschke Udo, KALKUNTE Satyan, Sharma Surendra
Принадлежит:

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 silalyl Lewis antigen assayed in step (b). 1. A method of identifying and/or addressing incipient preeclampsia in a patient-subject by the steps of 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 silalyl Lewis antigen assayed in step (b).2. The method of claim 1 , wherein step (a) is performed at about 20 weeks of pregnancy or earlier.3. The method of claim 2 , wherein step (a) is performed at about 10 weeks of pregnancy or earlier.4. A method of identifying and/or addressing incipient preeclampsia in a patient-subject by the steps of by the steps ...

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Номер записи: 98
08-07-2021 дата публикации

GLYCOPEPTIDE ANALYZER

Номер: US20210208117A1
Автор: MURASE Masaki, NISHIKAZE Takashi
Принадлежит: SHIMADZU CORPORATION

A glycopeptide analyzer that performs a structural analysis on glycoforms of a glycoprotein, including: a spectrum creator creating an MS/MS spectrum for each elution time based on data acquired by an LC/MS analysis of a sample containing glycopeptides originating from a target glycoprotein; a peptide mass calculator selecting a glycopeptide-related spectrum from a plurality of MS/MS spectra and calculating the mass of a peptide from the selected spectrum; a similarity determiner determining a similarity between the glycopeptide-related spectrum and each of the other MS/MS spectra; an elution-time range estimator estimating an elution-time range based on a distribution of the frequency of occurrence of an MS/MS spectrum for which a high level of similarity has been determined on a time axis; and a glycan composition estimator selecting an ion peak corresponding to a mass equal to or greater than a peptide mass and estimating a glycan composition based on the peak. 1. A glycopeptide analyzer configured to perform a structural analysis on glycoforms of a glycoprotein using an analyzing device including a liquid chromatograph coupled with a mass spectrometer capable of MS/MS analysis , the glycopeptide analyzer comprising:a spectrum creator configured to create an MS/MS spectrum for each elution time, based on data acquired by analyzing a sample containing glycopeptides originating from a target glycoprotein by the analyzing device;a peptide mass calculator configured to select a glycopeptide-related spectrum which is likely to have originated from a glycopeptide, from a plurality of MS/MS spectra created by the spectrum creator for different elution times, and to calculate a mass of a peptide in the selected glycopeptide based on the glycopeptide-related spectrum;a spectrum similarity determiner configured to determine a similarity between the glycopeptide-related spectrum and each of the other MS/MS spectra or each MS/MS spectrum corresponding to a specific range of ...

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Номер записи: 99
08-07-2021 дата публикации

GLYCAN ANALYSIS OF PROTEINS AND CELLS

Номер: US20210208156A1
Автор: Angel Peggi M., Drake Richard R., Haab Brian, MEHTA Anand
Принадлежит:

The present invention provides methods and compositions for glycan analysis of complex solutions, including proteins and cells in a biological sample. The method includes the preparation of substrates for the capture of proteins and cells for multiplexed analysis. Cells and proteins may be captured by antibody arrays, culture, or direct deposition. The invention further relates to the use of protein and cell glycan analysis in the diagnosis and screening of disease states and disease progression. 1. A method for glycan analysis of at least one sample , the method comprising the steps of:providing a substrate having a surface spotted with a plurality of antibodies;incubating the substrate in a blocking solution;incubating the substrate in at least one sample;spraying the substrate with an enzymatic releasing solution; andscanning the substrate by mass spectrometry to detect and identify the presence of glycans.2. The method of claim 1 , wherein the at least one sample comprises at least one protein solution.3. The method of claim 1 , wherein the at least one sample comprises at least one population of cells.4. The method of claim 3 , wherein the at least one population of cells is incubated in a fixing and rinsing agent prior to the step of spraying the substrate with an enzymatic releasing solution.5. The method of claim 4 , wherein the fixing and rinsing agent is selected from the group consisting of: formalin claim 4 , Carnoy's solution claim 4 , paraformaldehyde claim 4 , an ethanol-based fixative claim 4 , and a polyethylene glycol-based fixative.6. The method of claim 1 , wherein the substrate is a glass or plastic microscope slide or multiwell plate.7. The method of claim 1 , wherein the blocking solution is a serum.8. The method of claim 7 , wherein the serum is 1% BSA in PBS and detergent.9. The method of claim 1 , wherein the blocking solution is removed with a wash step comprising 3×PBS baths and 1× water bath.10. The method of claim 1 , wherein the at ...

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Номер записи: 100