НОВЫЕ ЗОНДЫ НА ОСНОВЕ АКТИВНОСТИ ДЛЯ НЕЙТРОФИЛЬНОЙ ЭЛАСТАЗЫ И ИХ ПРИМЕНЕНИЕ

Группа изобретений относится к соединениям и способам для детектирования активности нейтрофильной эластазы. Раскрыт способ детектирования активности нейтрофильной эластазы в лизате образца ткани, включающий: (1) приготовление лизата из образца ткани, полученного от субъекта; (2) контактирование лизата с соединением-зондом; (3) последующее подвергание по меньшей мере аликвоты лизата, полученного на стадии (2), гель-электрофорезу; (4) измерение детектируемого сигнала. Также раскрыты способ диагностики заболевания, соединения-зонды формулы I, формулы II и формулы IIA, а также композиции для детектирования активности нейтрофильной эластазы и для диагностики заболевания. Группа изобретений обеспечивает возможность анализа лизата тканей для определения активности нейтрофильной эластазы. 7 н. и 33 з.п. ф-лы, 25 ил., 1 табл., 8 пр.

СЕЛЕКТИВНЫЕ ИНГИБИТОРЫ ГЛИКОЗИДАЗЫ И ИХ ПРИМЕНЕНИЕ

Изобретение относится к соединению формулы (I) или его фармацевтически приемлемой соли, где Rи Rнезависимо представляют собой Н или F; Rпредставляет собой ORи Rпредставляет собой Н, или Rпредставляет собой Н и Rпредставляет собой OR; каждый из Rпредставляет собой Н; Rпредставляет собой Н или OR; Rвыбирают из группы, состоящей из Н и C-алкила; Rвыбирают из группы, состоящей из C-алкила, необязательно замещенного 1-3 атомами фтора; и каждый Rнезависимо выбирают из группы, состоящей из Н, C-алкила, или две группы Rсвязаны вместе с атомом азота, к которому они присоединены, с образованием 4-членного кольца, содержащего 3 атома углерода. Соединения формулы (I) предназначены для изготовления лекарственного средства или фармацевтической композиции, обладающих ингибирующей активностью в отношении гликопротеин 2-ацетамидо-2-деокси-β-D-глюкопиранозидазы (O-GlcNAcase). Также изобретение относится к способу селективного ингибирования гликопротеин 2-ацетамидо-2-деокси-β-D-глюкопиранозидазы (O-GlcNAcase ...

ГЕН НОВОЙ СЕРИНОВОЙ ПРОТЕАЗЫ, РОДСТВЕННОЙ DPPIV

Изобретение относится к области биотехнологии и может быть использовано в фармакологии и медицине. Клонирована и определена последовательность ДНК, кодирующая новый вид сериновой протеазы человека (DPRP-2), родственной дипептидилолигопептидазе IV, гомология между которыми на уровне выведенной аминокислотной последовательности определена как 39%. С помощью технологии рекомбинантных ДНК получена рекомбинантная форма DPRP-2 и исследованы кинетические свойства нового фермента (субстратная специфичность, значения Km, ингибиторы активности и т.п.). Осуществление изобретения обеспечивает новые возможности при поиске и создании новых терапевтических средств для лечения широкого круга заболеваний человека. 2 н. и 3 з.п. ф-лы, 4 ил., 6 табл.

СПОСОБ ОБНАРУЖЕНИЯ ГИДРОЛАЗЫ И УСТРОЙСТВО ДЛЯ ЕГО ОСУЩЕСТВЛЕНИЯ

Изобретение относится к биотехнологии, медицинской микробиологии, касается обнаружения гидролитически активного фермента, в частности аспарагиновой протеазы в образце или препарате. Образец или препарат контактирует с твердым носителем, на котором иммобилизован фермент-репортер (т.е. сигналгенерирующий фермент) таким образом, что под действием ферментативно активной гидролазы, если таковая присутствует в образце, этот фермент-репортер отделяется от твердого носителя. После контакта с твердым носителем, образец взаимодействует с индикатором. Индикатор представляет собой любое химическое вещество, которое способно подвергаться детектируемому изменению, обычно изменению окраски под действием фермента-репортера. Детектируемое изменение индикатора указывает на присутствие в образце ферментативно активной гидролазы, что может свидетельствовать о наличии конкретного патогена или патологического состояния, например, кандидоза. Устройство для осуществления способа включает приемник образуемый частично ...

СПОСОБЫ И ФАРМАЦЕВТИЧЕСКИЕ КОМПОЗИЦИИ ДЛЯ ЛЕЧЕНИЯ ЗАБОЛЕВАНИЙ, СВЯЗАННЫХ С ТУБУЛИН КАРБОКСИПЕПТИДАЗАМИ (TCP)

Группа изобретений касается лечения заболеваний, связанных с тубулин карбоксипептидазой (TCP), и скрининга соединений, пригодных для лечения таких заболеваний. Предлагается применение ингибитора экспрессии или активности комплекса вазогибин/SVBP (малый вазогибин-связывающий белок) для лечения заболевания, связанного с циклом детирозинирования/тирозинирования тубулина тубулин карбоксипептидазой, у субъекта, который в этом нуждается. Предлагается также применение фармацевтической композиции, содержащей ингибитор экспрессии или активности комплекса вазогибин/SVBP для лечения указанного выше заболевания. Заболевание, связанное с TCP, представляет собой неврологическое нарушение (в частности, рассеянный склероз, инсульт, боковой амиотрофический склероз, болезнь Паркинсона, болезнь Хантингтона и болезнь Альцгеймера) или сердечно-сосудистое заболевание (в частности, инфаркт миокарда, инфаркт миокарда, индуцирующий сердечно-сосудистую дисфункцию, сердечную недостаточность, кардиомиопатию и дистрофинопатию ...

СПОСОБ ОБНАРУЖЕНИЯ ПРЕДРАСПОЛОЖЕННОСТИ К КАРДИОВАСКУЛЯРНОМУ ЗАБОЛЕВАНИЮ

... 1. Способ обнаружения присутствия, по меньшей мере, одного аллельного варианта у человека, включающий идентификацию в генетическом материале выделенного биологического образца упомянутого человека аллельного варианта, состоящего в замене цитозина тимином в положении 46 относительно транскрипционного источника Фактора XII (46С/Т) в локусе хромосомы 5, ограниченном маркерами D5S400 и D5S408, а присутствие упомянутого аллельного варианта является показателем предрасположенности к кардиоваскулярному заболеванию. 2. Способ по п.1, отличающийся тем, что упомянутый биологический образец представляет собой кровь. 3. Способ по п.1, отличающийся тем, что перед идентификацией проводят полимеразную цепную реакцию (ПЦР) для получения специфического геномного фрагмента. 4. Применение выделенного биологического образца, подозреваемого на наличие аллельного варианта 46С/Т в локусе хромосомы 5, ограниченном маркерами D5S400 и D5S408, для выявления предрасположенности к кардиоваскулярному заболеванию, выражающемуся ...

НОВЫЙ МАРКЕР ДЛЯ ДИАГНОСТИКИ РАКА ПОДЖЕЛУДОЧНОЙ ЖЕЛЕЗЫ

НОВЫЕ ХРОМОГЕННЫЕ СУБСТРАТЫ И ИХ ПРИМЕНЕНИЕ ДЛЯ КОЛИЧЕСТВЕННОГО ОПРЕДЕЛЕНИЯ АКТИВНОСТИ КАРБОКСИПЕПТИДАЗ

... 1. Соединение, соответствующее следующей формуле (I) в которой А представляет собой или или или или R1, R2 представляет собой Н, -СН3, -СН(СН3)2, -ОСН3, -Cl, CF3, -OCF3, -SCH3; R3 представляет собой аминокислотный остаток, который может быть отщеплен карбоксипептидазой А; R4 представляет собой остаток основной аминокислоты. 2. Соединение по п.1, соответствующее следующей формуле (I) в которой А представляет собой или или или или R1, R2 представляет собой Н, -СН3, -СН(СН3)2, -ОСН3, -Cl, -CF3, -OCF3, -SCH3; R3 представляет собой остаток гидрофобной аминокислоты; R4 представляет собой остаток аргинина или лизина. 3. Соединение по п.1, отличающееся тем, что R3 представляет собой остаток одной из следующих аминокислот: тирозин, фенилаланин, аланин, валин, лейцин, изолейцин, фенилглицин. 4. Соединение по п.1, отличающееся тем, что R3 представляет собой остаток фенилаланина. 5. Соединение по п.1, отличающееся тем, что R3 представляет собой остаток фенилаланина или тирозина, a R4 представляет собой ...

СПОСОБ ОПРЕДЕЛЕНИЯ БИОЛОГИЧЕСКОЙ АКТИВНОСТИ ДЕФИБРОТИДА

... 1. Способ определения биологической активности дефибротида, предусматривающий стадии a) приведения в контакт дефибротида, плазмина и субстрата, специфичного для плазмина, который за счет реакции с плазмином обеспечивает измеряемый продукт, и b) измерения количества образовавшегося продукта в последовательных временных точках. 2. Способ по п.1, в котором плазмин является плазмином млекопитающего, предпочтительно плазмином человека. 3. Способ по п.1, в котором субстратом, специфичным для плазмина, является соединение формулы А1-А2-А3-Х, в которой А1 и А2 являются неполярными аминокислотами, А3 является лизином или аргинином и Х является измеряемым продуктом. 4. Способ по п.3, в котором измеряемый продукт Х выбран из пара-нитроанилина и 2-нафтиламина. 5. Способ по п.3, в котором субстратом для плазмина является Н-D-валил-L-лейцил-L-лизин-п-нитроанилин. 6. Способ по п.1, в котором плазмин имеет концентрацию от 0,0064 до 0,050 М.Е./мл, а субстрат для плазмина имеет концентрацию от 2,6 до 3,5 ...

MASP-3, комплемент-св зывающий фермент и его применение

... 1. В значительной степени чистый полипептид протеазы серина 3, ассоциированной с маннан-связывающим лектином (MASP-3), при этом упомянутый полипептид состоит либо из i) аминокислотной последовательности, идентифицируемой как последовательность №5 или ее функционального эквивалента, содержащего аминокислотную последовательность, которая не менее, чем на 85% идентична последовательности №5; либо из ii) аминокислотной последовательности, идентифицируемой как последовательность №1 или ее функционального эквивалента, содержащего аминокислотную последовательность, которая не менее, чем на 85% идентична последовательности №1; либо из iii) аминокислотной последовательности, идентифицируемой как последовательность №2 или ее функционального эквивалента, содержащего аминокислотную последовательность, которая не менее, чем на 50% идентична последовательности №2. 2. Полипептид по п.1, при этом упомянутый полипептид конъюгирован с меткой или токсином. 3. Полипептид по одному из предыдущих пунктов, при ...

АНАЛИЗ РАСЩЕПЛЕНИЯ КЛЕТОЧНОГО VAMP

ANTIDENATURATED AGENT FOR FOOD PASTY PRODUCT

Способ получения индоксильных или тиоиндоксильных сложных эфиров аминокислот или пептидов

СПОСОБ ПОЛУЧЕНИЯ ИНДОКСИЛЬ НЫХ ИЛИ ТИОИНДОКСИЛЬНЫХ СЛОЖНЫХ ЭФИ РОВ АМИНОКИСЛОТ или ПЕПТИДОВ общей формулы I 0-А-В где . - одинаковые или различны и означают водород, гал ид, низпгую . алкильную, а коксильную, арильную, ар кильную, аралкоксильнуго, гидроксильную, карбоксил ную, карбокси (низший) ал ксильную, аралкоксикарбонильную , аралкоксикарбоНШ1 (низший) алкоксипьную , нитро- или низшую ациламиногруппу, или два соседних заместителя означают бензоаннелирован- ный остаток; X - сера или незамещенная иминогруппа или замещенная алкильным, аралкильньм , арильным или ацильным остатком, иминогруп- па; А - моноаминокарбоновая кислота или аланинсодержащий ди- или три-пептидный остаток; В - N-защитная группа в пептидной части (остатке), чающийся тем, что ие общей формулы И имеют указанные значения, ют взаимодействию с галоидани или смешанными ангидридами, вированными сложными эфирами бщей формулы III Н - О - А - В В имеют указанные значения.

Антиденатурирующий агент для пищевого пастообразного продукта

Использование изобретения: в пищевой промышленности при производстве рыбной или мясной пасты. Сущность изобретения: антиденатурирующий агент для съедобного пастообразного продукта содержит экстракт разложения белка, полученный разложением белка протеиназой, и сахариды, используемые в качестве активных компонентов . 4 з.п. ф-лы, 2 табл., 1 ил.

Determination of enzyme activity

Determination of the activity of an enzyme in blood plasma or other biological fluid comprises measuring the enzyme activity or the product of an enzymatic reaction in the presence of a substance (I) that inhibits enzyme processing (i.e. conversion of pro-enzyme to enzyme) and/or protein-protein interaction.

ZUSAMMENSETZUNGEN UND METHODEN DER INHIBITION DER BETA-PROTEINFUNKTION

Verfahren zur Nachweis von Marijuana in einer Haar Probe

VERFAHREN ZUM NACHWEIS EINES GENETISCHEN DEFEKTES, ASSOZIERT MIT THROMBOSE UND/ODER MANGELHAFTER ANTICOAGULANZ-REAKTION AUF AKTIVIERTES PROTEIN C

VERFAHREN ZUR HAARANALYSE

VERFAHREN ZUR BESTIMMUNG DER AFFINITÄT VON FIBRINOGEN UND SEINER DERIVATE GEGENÜBER FILAMENTÄREN FORMEN VON HEFEN, UND VERWENDUNG AUF DEM GEBIET DER BESTIMMUNG VON PATHOGENEN HEFEN INSBESONDERE IN BLUTKULTUR

D-ENZYMZUSAMMENSETZUNGEN UND VERFAHREN ZU DEREN VERWENDUNG

Diagnostikum zum Nachweis der den von Willebrand-Faktorspaltenden Aktivität von ADAMTS13

Verfahren zur Isolation des 20S Proteasoms, und Verfahren zur Identifikation von Inhibitoren des 20S und 26S Proteasoms

ZUSAMMENSETZUNG ZUR ERKENNUNG VON PARADONTALEN KRANKHEITEN.

Chromogene Dibenzoxazepinon- und Dibenzothiazepinon-Enzymsubstrate.

VERWENDUNG VON PROTEIN-KINASE-N-BETA

VERFAHREN ZUR VORBEHANDLUNG VON PROBEN ZUR BESTIMMUNG VON VERZUCKERTEN AMINEN, SOWIE VERFAHREN ZUR BESTIMMUNG VON VERZUCKERTEN AMINEN

METHODE ZUR UNTERSUCHUNG DER AKTIVITÄT VON SPERMIDIN/SPERMIN N1-ACETYLTRANSFERASE (SSAT), WOBEI DAS SUBSTRAT AMANTADIN IST

Novel metalloprotease containing a thrombospondin domain (MPTS protein) is useful to treat aggrecan associated disease including rheumatoid arthritis and osteoarthritis

A new metalloprotease containing a thrombospondin domain (MPTS protein) is useful to treat aggrecan associated disease including rheumatoid arthritis and osteoarthritis Independent claims are also included for the following: (1) a nucleic acid molecule (N1) encoding a metalloprotease containing a thrombospondin domain (MPTS protein) which encodes: (a) a polypeptide comprising the 959, 947, 1690 or 947 amino acid sequences fully defined in the specification, or a polypeptide having at least 35% to one of those sequences; (b) comprises the 2879, 3263, 5338 or 4086 nt sequence fully defined in the specification, or a sequence having at least 75% to one of those sequences; (c) comprises a sequence whose complement hybridizes to (a) or (b) under stringent conditions; and (d) is a fragment comprising the functional domain of (a), (b) or (c); (2) a nucleic acid complementary to N1, preferably an antisense nucleic acid molecule; (3) an expression cassette comprising the one of the above nucleic ...

Immunosorbent assay kit for detecting procollagenase 3 (PC3), useful for diagnosis of breast cancer and arthritis, also new monoclonal antibody specific for PC3

ELISA (enzyme-linked immunosorbent assay) kit for detecting procollagenase 3 (PC3) in body fluids, especially human serum or synovial fluid, or in cell culture supernatant. ELISA (enzyme-linked immunosorbent assay) kit for detecting procollagenase 3(PC3) in body fluids, especially human serum or synovial fluid, or in cell culture supernatant comprises: (1) solid carrier having monoclonal antibodies (MAb) that bind specifically and sensitively to human PC3 bound to it; (2) human recombinant PC3, as standard for quantitative detection of PC3 in samples; (3) buffer for preparing a series of standards of the recombinant PC3; (4) buffer for diluting the test sample; (5) detectably labeled conjugate (C) that binds to PC3; and (6) substrate (S) that allows visualization of (C). Independent claims are also included for the following: (1) MAb that have the same properties as antibodies produced by the hybridoma DSM ACC2572; and (2) the hybridoma DSM ACC2572.

FLUORESZENZPOLARIZATIONSBESTIMMUNGEN UNTER VERWENDUNG VON POLYIONEN

Neue Polynukleotide

VERFAHREN ZUR AUSSETZUNG EINES BAKTERIELLEN ANTIGENS UND DASSELBE VERWENDENDES DIAGNOSTISCHES TESTVERFAHREN.

PEPTID-DERIVATE DES 7-AMINO-4METHYLCUMARINS

VERFAHREN ZUR VERHINDERUNG ODER VERMINDERUNG DER TRÜBUNG IN GETRÄNKEN

VERFAHREN UND VORRICHTUNG ZUR IDENTIFIKATION ODER KARAKTERISIERUNG VON POLYPEPTIDEN

VERFAHREN ZUR ÜBERWACHUNG DER PROTEASOMEINHIBITORARZNEIMITTELEFFEKTE

VERFAHREN UND KULTURMEDIEN ZUM NACHWEIS VON HEFEN UND/ODER SCHIMMELPILZEN IN PROBEN

VERFAHREN ZUR BESTIMMUNG VON LEUCINAMINOPEPTIDASE

VERFAHREN ZUR DETEKTION DER GAMMA-SEKRETASE-AKTIVITÄT

Verfahren zur Identifizierung von Verbindungen zur Therapie von Alterungsprozessen des Herz-Kreislauf Systems

The invention relates to a method for identifying a compound that modifies the activity of the urotensin II receptors by influencing those genes or gene products that are associated with the composition, formation or degradation of the extracellular matrix.

Method of analyzing protein

Alzheimer's disease secretase,app substrates therefor,and uses therefor

Detection of enzymes by detecting binding of substrate recognition molecules to modified substrates

An enzyme detection device (1) for detecting the presence, in a sample, of an enzyme capable of modifying a provided substrate 10. The device (1) comprises a substrate which has a modification region (14) that is sensitive to modification by the enzyme from an unmodified state to a modified state. The device (1) further comprises a substrate recognition molecule 16 (17) which binds the modification region (14) in either the modified or the unmodified state. The enzyme and the substrate recognition molecule 16 (17) competitively bind the modification region (14) when mixed. Cleavage or modification of the substrate by an enzyme is detected as the modification alters binding of the substrate recognition molecule 16 (17) to the substrate. The device further comprises a detectable label (18) coupled to the substrate recognition molecule. Methods of detecting an enzyme using the device are also claimed.

Beta-Secretase enzyme compositions and methods

Disclosed are various forms of an active, isolated b -secretase enzyme in purified and recombinant form. This enzyme is implicated in the production of amyloid plaque components which accumulate in the brains of individuals afflicted with Alzheimer's disease. Recombinant cells that produce this enzyme either alone or in combination with some of its natural substrates ( b -APPwt and b -APPsw) are also disclosed, as are antibodies directed to such proteins. These compositions are useful for use in methods of selecting compounds that modulate b -secretase. Inhibitors of b -secretase are implicated as therapeutics in the treatment of neurodegenerative diseases, such as Alzheimer's disease.

Protease inhibition

A method for inhibiting an ADAM (a disintegrin and metalloprotease) protease, comprising inhibiting binding to an integrin-binding loop of a disintegrin domain in the ADAM protease. Also claimed are ADAM protease inhibitors which inhibit binding to an integrin-binding loop of an ADAM protease; such inhibitors may be cyclic peptides and comprise amino acid sequences TSE, KDE, KDM, SNS, ARQ, MGD, DSD, RGD, NAT, KDK, VRQ, ATD and TDD.

Screening Systems Utilizing RTP801

RTP801 represents a unique gene target for hypoxia-inducible factor 1 (HIF-I). Down regulation of the mTOR pathway activity by hypoxia requires de novo mRNA synthesis and correlates with increased expression of RTP801. The present invention relates to screening systems utilizing RTP801 and/or RTP801 interactors and/or RTP801 biological activity, to drag candidates identified by such screening systems, and to the use of such drug candidates in the treatment of various disorders.

Wound monitoring

Assay

Mass Spectrometric Endopeptidase Assay

URINE ASSAY PROCESS AND KIT

IMMUNOLOGICAL METHODS

Novel compounds

L-LEUCYL-4-HYDROXYANILIDE DERIVATIVES

SUBSTRATE FOR DETERMINING A PROTEINASE

... 1496907 Determination of proteinase BEHRINGWERKE AG 23 Dec 1974 [21 Dec 1973] 55521/74 Heading G1B [Also in Division C3] A method of determining proteinase comprises incubating the proteinase with a substrate com- prising a protein covalently bound to a solid carrier and having a reactive dyestuff covalently bonded to the protein [see Division C3], removing any undergraded substrate by filtration or centrifugation, measuring the absorption of the filtrate or centrifugate at a wavelength or wavelengths determined by the absorption characteristics of the reactive dyestuff, and calculating the activity of the enzyme therefrom.

Diagnostic composition

Compositions for estimating protease activity comprise (a) a protein, (b) a dye indicator exhibiting protein error at pH1-2,8 and (c) an acid buffer providing said pH. The composition may be a solution or a bibulous carrier impregnated with a solution, the protein (preferably albumin, haemoglobin, mucin or casein) comprising 0,01-1,0% of the solution and the dye (preferably Orange IV, Metanil yellow, Thymol blue or Bromophenol blue) comprising 0,0005-0,005% of the solution. The acid in the buffer may be hydrochloric, citric or hexamic. Proteases to be estimated include pepsin, uropepsin, trypsin, chymotrypsin, ficin and papain. The composition may alternatively be a powder or tablet containing 1-10% protein, 0,1-1% dye and a buffer, which when dissolved forms a solution as above defined. On adding the composition to a protease-containing liquid, the colour changes as the protein is digested and the colour at a given time or the time to reach a given colour is indicative of the protease ...

DIPEPTIDE DERIVATIVES OF GLYCYLPROLINE

Test for risk of COVID-19 infection

The invention provides a method of determining risk of infection by COVID-19 in a pre-determined subject comprising: determining a reference level of dipeptidyl peptidase 4 (DPP4) in a subject not infected by COVID-19; determining a risk level of DPP4 in a pre-determined subject; and comparing the risk level with the reference level to determine the risk of infection. Preferably DPP4 is measured in serum or plasma from the adult subject and the non-infected subject has a comparable risk to the pre-determined subject as they are of a similar age, ethnic background or diagnosis for dementia, diabetes or immunity. The pre-determined subject is considered at risk if the DPP4 level is at least 100 pg/ml less than the reference level. Also disclosed is a second method of determining risk of infection by COVID-19 in a pre-determined subject comprising; determining a reference level of fragmented platelets in a subject not infected by COVID-19; determining a risk level of fragmented platelets in ...

Genes and proteins, and their uses.

Heterecyclic compounds useful as inhibitors of cysteine protease.

Heterocyclic compounds useful as inhibitors of cysteine protease. A group of compounds of the formula (I) which are inhibitors of cysteine proteases particularly cathepsin K, and are useful in the treatment of excessive bone or cartilege loss.

Proatease inhibitors.

The present invention provides compounds which inhibit proteases, including cathepsin k, pharmaceutical compositions of such compounds, and methods for treating diseases of excessive bone loss or cartilage or matrix degradation, including osteoporosis; gingival disease including gingivitis and periodontitis; arthritis, more specifically, osteoarthritis and rheumatoid arthritis; paget's disease; hypercalcemia of malignancy; and metabolic bone disease, comprising inhibiting said bone loss or excessive cartilage or matrix degradation by administering to a patient in need thereof a compound of the present invention.

Inhibitors of cycteine protease

Genes and proteins and their uses

Genes and proteins, and their use.

Genes and proteins, and their use.

Inhibitors of cysteine protease

Method of inhibiting cathepsin k

Protease inhibitors

Inhibitors of cycteine protease

Genes and proteins, and their use.

Genes and proteins and their uses

Genes and proteins, and their use.

SUBSTRATES AND PROOFS TO THE EFFECTIVENESS OF BETA SECRETASE ONES

PROCEDURE FOR THE TREATMENT OF ILLNESSES IN CONNECTION WITH APOE

PROCEDURE FOR THE ANALYSIS OF PROTEASEN AND THIN DIAPHRAGM FOR THE USE OF THIS PROCEDURE

TUMOR NECROSIS FACTOR RECEPTOR SPLITTING OFF ENZYME, ITS PREPARATIONS AND USES

NUCLEIC ACIDS, WHICH FOR ENDOTHELIASEN CODING, ENDOTHELIASEN, AS WELL AS THEIR USE

SCREENING PROCEDURE FOR SUBSTANCES FOR THE TREATMENT OF DIABETES

PROCEDURE FOR THE REGULATION OF PROTHROMBIN AND REAGENT FOR THE EXECUTION OF THE PROCEDURE

SIGNAL SEQUENCES FOR THE PRODUCTION OF LEU HIRUDIN OVER SECRETION BY E. COLI INTO THE CULTURE MEDIUM

REPORTER CONNECTIONS AND PROCEDURES FOR THE DETERMINATION OF THE LIPIDISIERUNGSSTATUS OF A CELL

CLEANED ACTIVE HCV NS2/3 PROTEASE

PROCEDURE FOR THE REGULATION OF ENZYMES OF THE TYPE SERINPROTEASEN, WITH THAT AMINO ACID RESIDUES ENT ATTITUDE CHROMOGENE SUBSTRATES SPLIT BY THESE ENZYMES

BY MEANS OF BOTULINUMTOXIN A (BONT/A) RECOGNIZABLE PEPTIDSUBSTRATE AND USE

SEPARIN INHIBITORS, PROCEDURES FOR YOUR IDENTIFICATION AND YOUR APPLICATIONS

CATHEPSIN K AND CANCER OF THE BREAST

DIAGNOSTIKUND THERAPY PROCEDURE DISEASES CONNECTED BY WITH PILS

PROCEDURE FOR THE DEMONSTRATION OF A MOLECULAR EVENT IN A CELL OF MEANS FLUORESCENCE MARKER PROTEINS

PROCEDURE AND COMPOSITIONS FOR THE DETERMINATION OF GLYCIERTER PROTEINS

MEDIUM FOR THE PROOF OF CERTAIN MICROBES IN A SAMPLE

NEW AGGRECANASE

INHIBITORI, MONOCLONAL ANTIBODY AGAINST THE BLOOD CLOTTING FACTOR VII THE AKITIVIERENDE PROTEASE

PROCEDURE FOR THE DETERMINATION OF THE EFFECTIVENESS THE PROTEASEN INHIBITORS

LINKED D-PROTEIN AND PROCEDURE FOR THE IDENTIFICATION OF RECEPTOR ACTIVITY MODULATING CONNECTIONS

SOLUBLE NOTCH WAS BASED SUBSTRATES FOR GAMMA SECRETASE, METHODS, COMPOSITIONS AND USES OF IT

PROCEDURE FOR THE PRETREATMENT FROM SAMPLES TO THE REGULATION OF VERZUCKERTEN AMINES, AS WELL AS PROCEDURES FOR THE REGULATION OF VERZUCKERTEN AMINES

VERFAHREN ZUR BESTIMMUNG VON PROTHROMBIN UND REAGENS ZUR DURCHFUEHRUNG DES VERFAHRENS

PROCEDURE AND COMPOSITIONS FOR THE PROOF OF THE ACTIVATION STATUS OF MULTIPLE PROTEINS IN SINGLE CELLS

Releasable nonvolatile mass-label molecules

Releasable tag reagents for use in the detection and analysis of target molecules, particular in mass spectrometric analyses are provided. Also provided are methods of detection that employ releasable tag reagents.

Novel Peptidase Substrates

The present invention relates to the use of a compound of the following formula (I), as an enzyme substrate for the detection of a peptidase activity or as a pH indicator: according to which: Y 1 is a peptide, H or an alkyl; W 1 , W 2 , W 3 and W 4 are independently H, Br, Cl, F, I, alkyl, alkoxy, thiomethyl, perfluoroalkyl, nitro, cyano, carboxyl (including the esters or amides thereof) or any combination thereof; n=0, 1 or 2; U is N or N + R and V is CZ 6 N or N + R or else V is N or N + R and U is CZ 6 ; R is H, alkyl, aralkyl, aryl, alkanoyl or alkylsulfonyl; Z 1 , Z 2 , Z 3 , Z 4 and Z 5 are independently H, Br, Cl, F, I, alkyl, aryl, alkoxy, perfluoroalkyl, nitro, cyano, carboxyl, sulfonyl, including the carboxyl or sulfonyl esters or amides, and salts thereof.

Luminescent live and dead cell assay

A method to detect live and dead cells in a sample with one bioluminogenic reagent is provided.

Detection and quantification of modified proteins

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.

Assay for pcsk9 inhibitors

The present invention provides methods for identifying modulators of PCSK9, for example, using a variety of assay formats. Inhibitors of PCSK9 can be used for example, to treat diseases such as hyperlipidemia and related disorders.

Identification of modulators of serine protease inhibitor kazal and their use as anti-cancer and anti-viral agents

This disclosure describes a relevant etiology of cancer and a novel anti-cancer therapeutic strategy, based on the discovery that a protein named serine protease inhibitor (SPIK/SPINK/PSTI) was up-regulated by hepatitis B and C virus infections consequently suppressing the cell apoptosis. Accordingly, the present disclosure provides, inter alia, an inhibitor of SPIK and/or a technology of suppression of over-expression of SPIK in cells. The inhibitors include: 1) chemical compounds, which can inhibit SPIK transcripts, protein activity, and gene expression, 2) SPIK siRNA (RNAi gene silence or dsRNA of SPIK, 3) DNA anti-sense and anti-SPIK antibody. Further, this disclosure provides methods of using the inhibitor as an anti-cancer agent to re-instate cancer cell apoptosis (e.g., serine protease dependent cell apoptosis).

method circuit and system for detecting a connection request while maintaining a low power mode

Disclosed is a method, circuit and system for communication channel scanning by a video transceiver to determine whether a connection is being requested by another video transceiver. Scanning for connections requests may be performed according to two modes: (1) a first (complete) scanning mode, and (2) a second (partial) scanning mode. The information collected and/or recorded during a scanning sequence in the first scanning mode may be used as part of one or more partial scanning sequences performed in the second scanning mode. A scanning sequence in a first scanning mode may be followed by one or more (e.g. one, two, three etc.) scanning sequences in a second scanning mode. There may be provided a scanning circuit for checking for connection requests on one of a set of channels.

Probe reagent for measurement of proteolytic activity

This invention relates to a probe reagent comprising, in order from the N-terminus to the C-terminus, the amino acid sequences of a fluorescent protein I, a peptide capable of terminating protein degradation (i.e., a degradation-terminating peptide), a spacer peptide, a fluorescent protein II, and a protein to be degraded, wherein the protein to be degraded is a protein degraded by the ubiquitin-proteasome system, and the probe reagent is degraded from the C-terminus, but that the degradation of the probe reagent is terminated at the degradation-terminating peptide, a nucleic acid encoding the probe reagent, and use of the probe reagent or the nucleic acid.

Solid phase-bound elastase-binding assay for the measurement of alpha1-antitrypsin activity

The present invention relates to a method for the measurement of active alpha 1 -proteinase inhibitor (A1PI) in a sample, comprising the steps of binding elastase to a solid support, letting the A1PI contained in the sample bind to the solid phase-bound elastase, and detecting solid phase-bound A1PI with a detection reagent.

Use of a substrate in a method for measuring the activity of proteolytic enzymes

The disclosure relates to the use of a proteolytic enzyme substrate of general formula: Q 1 -Xaa 2 -Xaa 1 -rhodamine 110 -Q 2 , in which Xaa 1 and Xaa 2 are amino acids and Q 2 is Xaa 1 -Xaa 2 -Q 1 , or an amino acid Xaa 3 , or a group comprising an aryl group, or H, so as to be able to inhibit, in a blood sample containing a glycosaminoglycan, the anticoagulant capacity of said glycosaminoglycan. The invention also relates to a method for measuring an enzymatic activity using such a substrate and a kit for implementation thereof.

ASSAYS AND METHODS USING BIOMARKERS

Methods and assays examining expression of one or more biomarkers in a mammalian tissue or cell sample are provided. According to the disclosed methods and assays, detection of the expression of GalNac-T related molecules, such as GalNac-T14 or GalNac-T3, is predictive or indicative that the tissue or cell sample will be sensitive to apoptosis-inducing agents such as Apo2L/TRAIL and anti-DR5 agonist antibodies. Kits and articles of manufacture are also provided. 1. A method for predicting the sensitivity of a mammalian tissue or cell sample to Apo2L/TRAIL , comprising the steps of:obtaining a mammalian tissue or cell sample;examining the tissue or cell sample to detect expression of GalNac-T14, wherein expression of said GalNac-T14 is predictive that said tissue or cell sample is sensitive to apoptosis-inducing activity of Apo2L/TRAIL.2. The method of wherein said expression of GalNac-T14 is examined by detecting expression of GalNac-T14 mRNA.3. The method of wherein said expression of GalNac-T14 is examined by immunohistochemistry.4. The method of further comprising the step of examining expression of DR4 claim 1 , DR5 claim 1 , DcR1 claim 1 , or DcR2 receptors in said tissue or cell sample.5. The method of wherein tissue or cell sample comprises cancer tissue or cells.6. The method of wherein said cancer cells are pancreatic claim 5 , lymphoma claim 5 , non-small cell lung cancer claim 5 , colon cancer claim 5 , colorectal cancer claim 5 , melanoma claim 5 , or chondrosarcoma cells or tissue.7. A method for inducing apoptosis in a mammalian tissue or cell sample claim 5 , comprising the steps of:obtaining a mammalian tissue or cell sample;examining the tissue or cell sample to detect expression of GalNac-T14, and subsequent to detecting expression of said GalNac-T14, exposing said tissue or cell sample to an effective amount of Apo2L/TRAIL.8. The method of wherein said expression of GalNac-T14 is examined by testing for expression of GalNac-T14 mRNA.9. The method ...

BIOMARKERS

The invention relates to a method of diagnosing or monitoring schizophrenia or other psychotic disorder. The biomarkers used are selected from cyclophilin A, cytosalic non-specific dipepditase, Caoctosin-like protein, Glucose-6-phosphate isomerase, uncharacterized protein KIAA0423, myosin 14, myosin 15, nicotinamide phosphoribosyltransferase, pyruvate kinase isozyme R/L, phosphoglyterate mutase 4. 16-. (canceled)7. A method of diagnosing or monitoring schizophrenia or predisposition thereto comprising detecting and/or quantifying , in a sample from a test subject , one or more analyte biomarkers selected from Cyclophilin A , Cytosolic non-specific dipeptidase , Coactosin-like protein , Glucose-6-phosphate isomerase , Uncharacterized protein KIAA0423 , Myosin 14 , Myosin 15 , Nicotinamide phosphoribosyltransferase , Pyruvate kinase isozyme R/L and Phosphoglycerate mutase 4.8. (canceled)9. The method of claim 7 , wherein samples are taken on two or more occasions from the test subject.10. The method of claim 7 , further comprising comparing the level of the one or more analyte biomarkers present in samples taken on two or more occasions.11. The method of claim 7 , further comprising comparing the amount of the one or more analyte biomarkers in said test sample with the amount present in one or more samples taken from said subject prior to commencement of therapy claim 7 , one or more samples taken from said subject at an earlier stage of therapy claim 7 , or both.12. The method of claim 7 , further comprising detecting a change in the amount of the one or more analyte biomarkers in samples taken on two or more occasions.13. The method of claim 7 , further comprising comparing the amount of the one or more analyte biomarkers present in said sample with one or more controls.14. The method of claim 13 , further comprising comparing the amount of the one or more analyte biomarkers in the sample with the amount of the biomarker present in a sample from a normal subject.15. ...

Methods and kits for determining von willebrand factor activity in the absence of ristocetin and for determining the activity of adamts-13 protease

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

Compounds and Methods for FRET Based Measurement of Enzyme Activity

Provided are compositions and methods for Fluorescence/Forster Resonance Energy Transfer (FRET) analysis of cleavage of Von Willebrand Factor (VWF) polypeptides by the metalloprotease ADAMTS13. The polypeptides contain: i) a first fluorescent protein ii) a VWF amino acid sequence and iii) a second fluorescent protein. The VWF amino acid can contain any of a variety of amino acid substitutions. The method involves contacting a sample that contains ADAMTS13 and using FRET based measurements to determine ADAMTS13 cleavage of the recombinant polypeptide.

Urokinase-Type Plasminogen Activator Protein /Plasminogen Activator Inhibitor Type-1 Protein Selected Reaction Monitoring Assay

Specific peptides are provided that are derived from subsequences of the urokinase-type plasminogen activator protein and the plasminogen activator inhibitor type-1 protein along with assays that can measure those peptides directly in complex protein lysate samples, including protein lysates prepared from histologicaly-processed formalin fixed tissue. The presence and amount of those peptides in samples from a subject can be associated with disease, including cancer, in a subject and provide information about the diagnostic stage/grade/status of the disease/cancer. 1. A method for measuring levels of the uPA and PAI-1 proteins in a biological sample , comprising detecting at least one fragment peptide from uPA and at least one fragment peptide from PAI-1 in a protein digest prepared from said biological sample using mass spectrometry; and calculating the levels of the uPA and PAI-1 proteins in said sample , wherein said measured levels of the uPA and PAI-1 proteins are independently selected from a relative level or an absolute quantitative level.2. The method of claim 1 , further comprising the step of fractionating said protein digest prior to detecting said peptides.34-. (canceled)5. The method of claim 1 , wherein said protein digest comprises a protease digest.69-. (canceled)10. The method of claim 1 , wherein the uPA fragment peptide comprises an amino acid sequence as shown in Table 1 claim 1 , and set forth as SEQ ID NO:1 claim 1 , SEQ ID NO:2 claim 1 , SEQ ID NO:3 claim 1 , SEQ ID NO:4 claim 1 , SEQ ID NO:5 claim 1 , SEQ ID NO:6 claim 1 , SEQ ID NO:7 claim 1 , and SEQ ID NO:8.11. (canceled)12. The method of claim 1 , wherein the PAI-1 fragment peptide claim 1 , or peptides claim 1 , comprises an amino acid sequence as shown in Table 2 claim 1 , and set forth as SEQ ID NO:9 claim 1 , SEQ ID NO:10 claim 1 , SEQ ID NO:11 claim 1 , SEQ ID NO:12 claim 1 , SEQ ID NO:13 claim 1 , SEQ ID NO:14 claim 1 , SEQ ID NO:15 claim 1 , SEQ ID NO:16 claim 1 , SEQ ID NO:17 ...

TESTING A MAMMAL FOR PRESENCE, PROGRESSION OR STAGE OF A SHOCK CONDITION

Methods and kits for diagnosis and staging of non-septic shock are presented in which one or more protease activities are measured from a biological sample to so identify and/or stage non-septic shock. Most preferably, at least two protease activities are correlated, however, additional or alternative markers may also be measured. 1. A method of testing a mammal for presence , progression or stage of a shock condition , comprising:obtaining a blood sample from the mammal and removing a cellular fraction from the blood to so obtain a serum or plasma fraction;adding a substrate for at least one of a pancreatic protease, a neutrophil protease, and a matrix metalloprotease to the serum or plasma fraction, and measuring a product formed from the substrate to thereby measure activity of the at least one of the pancreatic protease, the neutrophil protease, and the matrix metalloprotease to obtain a test result; andusing the test result, upon diagnosis of the mammal as having a shock condition, to ascertain at least one of presence, progression, and stage of non-septic hypovolemic shock in the mammal;wherein the step of using the test result comprises a step of correlation with a corresponding test result of a healthy mammal, wherein an increased activity of the at least one of the pancreatic protease, the neutrophil protease, and the matrix metalloprotease as compared to the healthy mammal is indicative of the at least one of the presence, progression, and stage of non-septic hypovolemic shock.2. The method of wherein the pancreatic protease is trypsin.3. The method of wherein the neutrophil protease is a matrix metalloproteinase.4. The method of wherein the matrix metalloproteinase is MMP8 or MMP9.5. The method of further comprising a step of measuring activity of an additional protease.6. The method of wherein the additional protease is selected from the group consisting of thrombin claim 5 , plasmin claim 5 , kallikrein claim 5 , and chymotrypsin.7. A method of testing ...

METHODS AND COMPOSITIONS FOR THE TREATMENT OF SYMPTOMS OF COMPLEX REGIONAL PAIN SYNDROME

A therapeutic composition for the treatment of the symptoms of complex regional pain syndrome and a method for preparing the therapeutic composition is disclosed. The therapeutic composition is a stable pharmaceutical composition comprising one or more digestive and/or pancreatic enzymes. The therapeutic composition may be manufactured by a variety of encapsulation technologies. Delivery of the therapeutic composition may be made orally, through injection, by adherence of a medicated patch or by other methods. Further, a method of using fecal chymotrypsin level as a biomarker for the presence of complex regional pain syndrome, or the likelihood of an individual to develop complex regional pain syndrome is disclosed. 1. A method for treating one or more symptoms associated with Complex Regional Pain Syndrome in a patient diagnosed with Complex Regional Pain Syndrome comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition comprising one or more digestive enzymes.2. The method of wherein the one or more digestive enzymes comprise one or more enzymes selected from the group consisting of proteases claim 1 , amylases claim 1 , celluloses claim 1 , sucrases claim 1 , maltases claim 1 , papaya claim 1 , papain claim 1 , and lipases.3. The method of wherein the one or more digestive enzymes comprise one or more pancreatic enzymes.4. The method of wherein the proteases comprise chymotrypin and trypsin.5. The method of wherein the one or more digestive enzymes are claim 1 , independently claim 1 , derived from an animal source claim 1 , a microbial source claim 1 , or a plant source claim 1 , or are synthetically prepared.6. The method of wherein the animal source is a pig.7. The method of wherein the pharmaceutical composition comprises at least one amylase claim 1 , a mixture of proteases comprising chymotrypsin and trypsin claim 1 , at least one lipase claim 1 , and papain.8. The method of wherein the pharmaceutical ...

METHODS OF DETECTING PREGNANCY-ASSOCIATED PLASMA PROTEIN-A2 (PAPP-A2)

The present invention provides pregnancy associated plasma protein A2 (PAPP-A2), its nucleotide and amino acid sequences, antisense molecules to the nucleotide sequences which encode PAPP-A2, expression vectors for the production of purified PAPP-A2, antibodies capable of binding specifically to PAPP-A2, hybridization probes or oligonucleotides for the detection of PAPP-A2-encoding nucleotide sequences, genetically engineered host cells for the expression of PAPP-A2, and methods for screening for pathologies in pregnant and non-pregnant patients. Methods for screening for altered focal proliferation states in pregnant and/or non-pregnant patients, which include detecting levels of PAPP-A2, are also described. 1. A method for detecting PAPP-A2 or measuring the level of PAPP-A2 in a sample from an individual , the method comprising the steps of:(a1) determining the presence or amount of a PAPP-A2 polynucleotide in the sample; and/or(a2) determining the presence or amount of a PAPP-A2 polypeptide in the sample; and/or(a3) determining the presence or amount of a PAPP-A2 proteolytic activity in the sample; and(b) analyzing the results of the determining step(s) to ascertain the presence or absence of PAPP-A2 or the level of PAPP-A2 in the sample.2. The method of claim 1 , wherein a PAPP-A2 polynucleotide is determined by contacting the sample with a PAPP-A2 nucleic acid probe.4. The method of claim 1 , wherein a PAPP-A2 polynucleotide is determined claim 1 , and the PAPP-A2 polynucleotide is selected from the group consisting of:(i) a polynucleotide comprising nucleotides 1 to 5376 of SEQ ID NO:1;(ii) a polynucleotide corresponding to the coding sequence of PAPP-A2 as deposited in the Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH (DSMZ) under accession number DSM 13783;(iii) a polynucleotide encoding the amino acid sequence as shown in SEQ ID NO:2; and(iv) a polynucleotide, the complementary strand of which hybridizes under stringent conditions with a ...

Resonance Energy Transfer Assay with Cleavage Sequence and Spacer

A molecular construct comprises a donor label, an acceptor label, a linker peptide disposed between the donor and the acceptor, the linker having a cleavage site sequence, and a spacer between at least one of (a) the donor and the cleavage site sequence and (b) the acceptor and the cleavage site sequence. Preferably, the construct is selected from the group consisting of CFP-(SGLRSRA)-SNAP-25-(SNS)-YFP, and CFP-(SGLRSRA)-synaptobrevin-(SNS)-YFP. In preferred embodiments, the linker peptide is a substrate of a botulinum neurotoxin selected from the group consisting of synaptobrevin (VAMP), syntaxin and SNAP-25, or a fragment thereof that can be recognized and cleaved by the botulinum neurotoxin. Advantageously, the spacer increases the electronic coupling between the donor label and the acceptor label relative to a corresponding construct without the spacer. 1. A method of improving sensitivity of energy transfer between a donor label and an acceptor label , comprising:providing a construct comprising the donor label and the acceptor label physically coupled through a linker;including in the linker a cleavage site sequence; andincluding a spacer in the linker between at least one of the donor and the cleavage site sequence and the acceptor and the cleavage site sequence, whereby electronic coupling between the donor and the acceptor is increased.2. The method of claim 1 , wherein the construct is a protein-based construct claim 1 , and the linker is a peptide sequence.3. The method of claim 1 , wherein the cleavage site sequence comprises a SNARE protein claim 1 , or a fragment or mutein thereof claim 1 , and wherein the spacer comprises at least five amino acids.4. The method of claim 1 , wherein the spacer includes a sequence selected from the group consisting of (GGGGS)n and (EAAAK)n claim 1 , where n is 1-3.5. The method of claim 1 , wherein the spacer increases the electronic coupling between the donor and the acceptor by reducing a tertiary structure distance ...

ANALYSIS OF AMINO ACID COPOLYMER COMPOSITIONS

Methods for analyzing, selecting, characterizing or classifying compositions of a co-polymer, e.g., glatiramer acetate are described. The methods entail analysis of pyro-glutamate in the composition, and, in some methods, comparing the amount of pyro-glutamate present in a composition to a reference standard. 1providing a batch of a composition comprising an amino acid copolymer;measuring the amount of gyro-glutamate (pyro-Glu) in the batch; andselecting the batch if the amount of pyro-Glu in the batch is within a predetermined range,thereby selecting a batch of a composition comprising an amino acid copolymer.. A method of selecting a batch of a composition comprising an amino acid copolymer, the method comprising: This application is a continuation of U.S. patent application Ser. No. 12/408,058, filed Mar. 20, 2009, and claims priority to U.S. Provisional Application Ser. No. 61/045,465, filed Apr. 16, 2008. The entire disclosures of the prior applications are considered part of, and are incorporated by reference in their entireties in, the disclosure of this application.Glatiramer acetate (also known as copolymer-1 and marketed as the active ingredient in COPAXONE® by Teva Pharmaceutical Industries Ltd., Israel) is used in the treatment of the relapsing-remitting form of multiple sclerosis (RRMS). According to the COPAXONE® product label, glatiramer acetate (GA) consists of the acetate salts of synthetic polypeptides, containing four naturally occurring amino acids: L-glutamic acid, L-alanine, L-tyrosine, and L-lysine with a reported average molar fraction of 0.141, 0.427, 0.095, and 0.338, respectively. Chemically, glatiramer acetate is designated L-glutamic acid polymer with L-alanine, L-lysine and L-tyrosine, acetate (salt). Its structural formula is:(Glu,Ala,Lys,Tyr)CHCOOH(CHNO.CHNO.CHNO.CHNO)CHOCAS—147245-92-9The invention is based, at least in part, on the identification and characterization of L-pyroGlutamic Acid (gyro-Glu) as a structural signature of ...

Usp2a peptides and antibodies

The invention relates to novel USP2a peptides and antibodies, as well as nucleic acids related to them. The peptides, antibodies and the nucleic acids are useful for the detection, staging and monitoring of the progression of cancer, as well as for determining or monitoring the efficacy of treatment.

FGL-2 PROTHROMBINASE AS A DIAGNOSTIC TOOL FOR MALIGNANCY

The present: invention reveals a strong correlation between FGL-2 prothrombinase activity levels and the presence of a malignant proliferative disorder in a subject. Thus, the present invention provides FGL-2 prothrombinase activity as a diagnostic tool for malignancy. 1. A method for the diagnosis of a malignant proliferative disorder in a subject , said method comprising the steps of:(a) measuring prothrombinase activity in a sample from said subject;(b) comparing said prothrombinase activity with that of a control value;whereby a sample with prothrombinase activity higher than control is indicative of the presence of a malignant disorder in said subject,2. A method for the prognosis of a malignant proliferative disorder in a subject , said method comprising the steps of:(a) measuring prothrombinase activity in a sample from said subject at different time points, said time points being before, during and after treatment of said disorder;(b) comparing the values of prothrombinase activity obtained for the different time points in step (a);whereby increasing prothrombinase activity over time indicates a poor prognosis or resistance to therapy whereas a decreasing prothrombinase activity over time indicates recovery for said subject.3. The method of claim 1 , wherein said sample comprises FGL-2 expressing cells.4. (canceled)5. The method of claim 1 , wherein said sample comprises blood or peripheral blood mononuclear cells (PBMC).6. The method of claim 1 , wherein said prothrombinase activity comprises FGL-2 prothrombinase activity.7. The method of claim 1 , wherein said malignant proliferative disorder is selected from the group consisting of pancreatic cancer claim 1 , breast cancer claim 1 , squamous cell carcinoma claim 1 , multiple myeloma claim 1 , prostate cancer claim 1 , Langerhans' cell sarcoma claim 1 , thyroid papillary cancer claim 1 , melanoma claim 1 , esophageal cancer claim 1 , endometrial sarcoma claim 1 , mammary gland cancer claim 1 , mediastinal ...

Ubiquitination assay

The present application relates to a method of assaying ubiquitination in a sample by combining ubiquitin together with a substrate in a sample containing UBE1, UbcH3, Skp2-isoform 1, Skp1, Cul1, Rbx1, Cks1, CDK2 and Cyclin E1 under conditions suitable for ubiquitination to take place, exposing the sample to a labelled binding partner which is specific for the ubiquitin, and measuring the amount of ubiquitin bound to the substrate.

NOVEL METHOD FOR CHARACTERIZING AND MULTI-DIMENSIONALLY REPRESENTING THE FOLDING PROCESS OF PROTEINS

The invention relates to a novel method for characterizing and multi-dimensionally representing the folding process of proteins (FIG. ). Said method comprises, in a methodically novel combination, kinetically arranging the hydrodynamic size of the refolding and thus modified protein, associating the proteolytically fragmented intermediates on the basis of the bioinformatic detection patterns, classifying the folding pathway association of the intermediates, characterizing the folding sequences, and multi-dimensionally visualizing the characterized folding process in a computer-aided manner. Said method is characterized in that all intermediates modified during the refolding and thus structurally blocked are identified and digitalized according to the four individual characteristics of said intermediates, namely the hydrodynamic size, the formation time, the folding pathway association, and amount. Said novel method has many applications in the field of research of protein folding and proteopathy, protein engineering, antibody engineering, molecular biology, therapeutic medicine, biotechnology, biotechnological production of protein pharmaceuticals, protein taxonomy, and nanotechnology for developing and producing novel functional protein materials. According to the invention, many and varied products in the form of different assay kits, devices, software, and machines can be produced and used to carry out said method. 1. A method for the characterization and multidimensional visualization of the folding procedure of proteins , wherein all intermediates trapped by chemical modification at various time intervals during the refolding process are identified and digitized by at least 4 individual characteristics , namely its hydrodynamic size , the time of formation , the folding pathway identity , and the quantifiable amount , wherein a folding pathway consists of 4 phases , wherein the method is applicable for the characterization of the folding pathway of all proteins ...

METHOD AND KIT FOR DETECTING THE EARLY ONSET OF RENAL TUBULAR CELL INJURY

A method and kit for detecting the early onset of renal tubular cell injury, utilizing NGAL as an early urinary biomarker. NGAL is a small secreted polypeptide that is protease resistant and consequently readily detected in the urine following renal tubule cell injury. NGAL protein expression is detected predominantly in proximal tubule cells, in a punctate cytoplasmic distribution reminiscent of a secreted protein. The appearance of NGAL in the urine is related to the dose and duration of renal ischemia and nephrotoxemia, and is diagnostic of renal tubule cell injury and renal failure. NGAL detection is also a useful marker for monitoring the nephrotoxic side effects of drugs or other therapeutic agents. 129.-. (canceled)30. A method of diagnosing , monitoring or determining the risk of developing acute renal failure in a human subject , wherein said method discriminates between a subject who does not have acute renal failure and is not at immediate risk of developing acute renal failure , and a subject who may have acute renal failure or is at risk of developing acute renal failure , said method comprising the steps of:1) determining the concentration of human neutrophil gelatinase-associated lipocalin (NGAL) in a sample of bodily fluid from the subject, and2) comparing said concentration with a predetermined cutoff value chosen so that an NGAL concentration below the cutoff value categorizes the subject as not having and not being at immediate risk of developing acute renal failure.3135.-. (canceled)36. The method of claim 30 , wherein the risk of developing acute renal failure is due to ischemic renal injury.3739.-. (canceled)40. The method of claim 30 , wherein the risk of developing acute renal failure is due to the administration of a nephrotoxic agent.41. (canceled)42. The method of claim 30 , wherein the bodily fluid is urine.43. (canceled)44. A method of diagnosing claim 30 , monitoring or determining the likelihood of a renal disorder in a human being ...

SYSTEM AND METHODS OF LOG-SCALE CONCENTRATION GRADIENTS

Devices and methods use an integrated microfluidic system that has the capability of realizing a wide range of accurate dilutions in a logarithmic way through semi-direct dilution of samples inside a chip. The device for dose response analysis is able to contain a first fluid source on a microfluidic chip, wherein the first fluid source comprises a drug, a second fluid source on the microfluidic chip, a mixing area on the microfluidic chip fluidically coupling with the first and the second fluidic source, and a detection area coupling with the mixing area for drug information detection using a detection system. 1. A method of performing quantitative drug information analysis comprising:a) mixing a first fluid and a second fluid to form a third fluid on a microfluidic device;b) receiving the drug information from the third fluid; andc) performing analysis on the drug information.2. The method of claim 1 , wherein the first fluid claim 1 , the second fluid claim 1 , the third fluid or a combination thereof comprises a volume in nanoliter scale.3. The method of claim 1 , wherein the microfluidic device comprises a microfluidic chip.4. The method of claim 1 , wherein the second fluid dilutes the first fluid.5. The method of claim 1 , wherein the first fluid comprises a drug claim 1 , a growth factor claim 1 , a growth stimulant claim 1 , or a combination thereof.6. The method of claim 5 , wherein the drug comprises an inhibitor claim 5 , an enzyme claim 5 , or a chemical having a therapeutical effect.7. The method of claim 5 , wherein the second fluid comprises a buffer solution.8. The method of claim 1 , wherein the drug information comprises dose response data.9. The method of claim 1 , wherein the analysis comprises determining a drug response.10. The method of claim 9 , wherein the drug response is proportional to the log of a drug concentration in the third fluid.11. The method of claim 9 , wherein the drug response is received from a bacterial cell claim 9 , a ...

Method for measuring glycated hemoglobin

It is to provide a method for measuring glycated hemoglobin in a hemoglobin-containing sample, comprising reacting the hemoglobin-containing sample with a proteolytic enzyme in the presence of a surfactant, and then reacting the obtained reaction product with fructosyl peptide oxidase, wherein at least one of the former reaction and the latter reaction is performed in the presence of an isothiazolinone derivative; and measuring the generated hydrogen peroxide. The present invention provides a method for accurately and highly sensitively measuring glycated hemoglobin in a hemoglobin-containing sample without being influenced by hemoglobin.

Analytical method for fab and fab' molecules

A method of measuring acidic species generated by degradation of a Fab or Fab′ component of a Fab-PEG or a Fab′-PEG comprising the steps of: a) cleaving the PEG and linker from the Fab-PEG or Fab′-PEG with an enzyme; b) optionally separating the PEG and linker generated in step a) from the Fab or Fab′, to provide a Fab or Fab′; and c) quantitatively analyzing acidic species associated with the cleaved Fab or Fab′ and/or the cleaved PEG.

Assays for detection of glycosaminoglycans

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

Enzyme detection

An enzyme detection product ( 1 ) for detecting the presence of an enzyme in a sample. The product ( 1 ) comprises: a reaction zone ( 16 ) for receiving the sample; a visualization zone ( 10 ) for presenting a signal in response to the detection of the activity of the enzyme; and a membrane ( 11 ). The membrane ( 11 ) is interposable between the reaction zone ( 16 ) and the visualization zone ( 10 ) and prevents passage from the reaction zone ( 16 ) to the visualization zone ( 10 ) the components having a size greater than a threshold size. The reaction zone ( 16 ) comprises a reactant capable of reacting with the enzyme in order to generate a reaction product having a size less than a threshold size.

NOVEL ULTRASENSITIVE CELL BASED SENSORS AND USES THEREOF

The present invention relates to a novel cell based sensor useful for drug discovery that comprises a cell line with professional regulated exocytosis of secretory granules transfected with a protease as a reporter polypeptide stored in the regulated secretory granules of the cell line with professional regulated exocytosis and having either an endogenous or a heterologous molecule as a modulator of regulated secretory granules exocytosis, such said granule stored protease reporter having at least: a high resistance to conditions already present inside the granules such as low pH and proteolysis by other proteases; enzymatic activity after exocytosis; a highly specific cleavage sequence; a very low level of secretion under unstimulated or basal conditions; and a high signal to background activity in a media compatible with cell culture viability and granule exocytosis for a high throughput robust and sensitive detection. 1. A cell based sensor that comprises:a An hemopoyetic cell line with professional regulated exocytosis of secretory granules.b A protease reporter transfected into the cell line of (a) that is stored into the secretory granules of such cell line,c An endogenous modulator or a transfected heterologous modulator of regulated exocytosis of the secretory granules of the cell line of (a)d A specific substrate for detection of the secreted granule stored protease reporter2. The cell based sensor of wherein the cell line with professional regulated exocytosis of secretory granules is a hemopoyetic cell line and/or their progeny selected from the group consisting of myeloid cell lines claim 1 , comprising monocytes claim 1 , macrophages claim 1 , neutrophils claim 1 , basophils claim 1 , eosinophils claim 1 , mast cells claim 1 , erythrocytes claim 1 , megakaryocytes claim 1 , platelets and dendritic cells or selected from a group consisting of lymphoid cell lines claim 1 , comprising T-cells claim 1 , B-cells and NK-cells.3. The cell based sensor of to ...

Protease for proteomics

Provided herein is technology relating to proteases and proteomics and particularly, but not exclusively, to compositions comprising OmpT, methods of using OmpT, and methods of manufacturing OmpT for proteomics.

Markers of Endothelial Progenitor Cells and Uses Thereof

The present invention provides markers of endothelial progenitor cells (EPCs) and use of those markers and reagents that bind thereto to detect EPC cells or diagnose, prognose, treat or prevent EPC-associated conditions. 1. A method for detecting an endothelial progenitor cell (EPC) comprising determining the level of expression of a nucleic acid or protein set forth in Table 1 , or a nucleic acid or a protein having at least about 70% identity thereto , in , on or secreted from a cell , wherein an increased level of expression of a nucleic acid or protein set forth in Table 1 or a nucleic acid or protein having at least about 70% identity thereto compared to another cell type is indicative of an EPC.2. A method according to claim 1 , wherein the nucleic acid or protein is expressed in claim 1 , on or secreted from EPCs at a level at least 1.5 fold greater or 2 fold greater or 3 fold greater or 4 fold greater or 5 fold greater than in claim 1 , on or secreted by human umbilical cord vascular endothelial cells (HUVECs).3. A method according to claim 2 , wherein the nucleic acid or protein is expressed in claim 2 , on or secreted by non-adherent CD133 EPCs at a level at least 1.5 fold greater or 2 fold greater or 3 fold greater or 4 fold greater or 5 fold greater than in claim 2 , on or secreted by HUVECs.4. A method for detecting an endothelial progenitor cell (EPC) comprising determining the level of expression of a protein that is a cell adhesion molecule or a nucleic acid encoding the protein as set forth in Table 2 claim 2 , or a nucleic acid or protein having at least about 70% identity thereto claim 2 , in claim 2 , on or secreted from a cell claim 2 , wherein an increased level of expression of a nucleic acid or protein set forth in Table 2 or a nucleic acid or protein having at least about 70% identity thereto compared to another cell type is indicative of an EPC.5. A method for detecting an endothelial progenitor cell (EPC) comprising determining the level ...

FUSION PROTEIN COMPRISING SMALL HEAT SHOCK PROTEIN, CAGE PROTEIN FORMED THEREBY, AND NOVEL USE THEREOF

The present invention relates to a fusion protein comprising small heat shock protein, a cage protein formed thereby, and novel use thereof, more particularly, a fusion protein comprising a small heat shock protein, a recognition site of a protease, and a histidine polymer, wherein the recognition site and the histidine polymer are sequentially linked to a carboxyl terminal of the small heat shock protein, a cage protein formed thereby, and novel use thereof. The fusion protein of the present invention, and a cage protein formed by the self-assembly properties of the fusion protein are not cytotoxic, and emits a fluorescence signal of about 20 to about 50 times higher comparing to a single peptide for the conventional molecular imaging, per unit protein. Additionally, cell permeability is very excellent, thereby to be effectively used as a biosensor or a bioactive material carrier. 1. A fusion protein comprising a small heat shock protein (Hsp) , a recognition site of a protease , and a histidine polymer , wherein the recognition site and the histidine polymer are sequentially linked to a carboxyl terminal of the small heat shock protein.2. The fusion protein of claim 1 , wherein the small heat shock protein is selected from the group consisting of a polypeptide represented by the amino acid sequence of SEQ ID NO: 6 claim 1 , a fragment and a variant thereof.3. The fusion protein of claim 1 , wherein the protease is selected from the group consisting of caspase claim 1 , matrix metalloproteinases (MMP) claim 1 , cathepsin S claim 1 , and viral proteases.4. The fusion protein of claim 1 , wherein the recognition site is a polypeptide consisting of amino acid residues of 4 to 20 comprising a cysteine residue.5. The fusion protein of claim 1 , wherein the recognition site is a DEVD motif comprising the amino acid sequence of SEQ ID NO: 8 claim 1 ,6. The fusion protein of claim 5 , wherein the DEVD motif is represented by the amino acid sequence of SEQ ID NO: 7.7. The ...

Measurement Method Using Enzyme

A method for measuring an analyte is described that includes the steps of: i) preparing a reagent (D) in which an enzyme (A) and an enzyme (B) coexist in the absence of the analyte; ii) bringing the analyte into contact with the enzyme (A) and the enzyme (B) so that the enzyme (A) acts on the analyte to produce a product (E), on which the enzyme (B) does not substantially act, from the analyte; iii) producing a product (C) by allowing the enzyme (A) or an enzyme (F) that is different from the enzyme (A) that acts on the analyte to produce a product (C) to act on the analyte and/or the product (E); and iv) detecting the product (C) by the enzyme (B). 1. A method for measuring an analyte using an enzyme (A) that acts on the analyte and an enzyme (B) that specifically detects a product (C) produced from the analyte ,the method comprising steps of:i) preparing a reagent (D) in which the enzyme (A) and the enzyme (B) coexist in the absence of the analyte;ii) bringing the analyte into contact with the reagent (D) so that the enzyme (A) acts on the analyte to produce a product (E), on which the enzyme (B) does not substantially act, from the analyte;iii) producing a product (C) by allowing the enzyme (A) or an enzyme (F) that is different from the enzyme (A) to act on the analyte and/or the product (E); andiv) detecting the product (C) by the enzyme (B).2. The method according to claim 1 , wherein the reagent (D) is a commercially available package that includes the enzyme (A) claim 1 , the enzyme (B) claim 1 , and a container in which the enzyme (A) and the enzyme (B) are packaged claim 1 , or a reagent in part of the commercially available package.3. The method according to claim 1 , wherein the step iii) is performed by further adding the enzyme (A) after the step ii).4. The method according to claim 1 , wherein the step iii) is performed by adding the enzyme (F) after the step ii).5. The method according to claim 1 , wherein the step iii) is performed by activating the ...

DETECTING INFECTION IN REDUCED PRESSURE WOUND TREATMENT

Provided is a method of detecting infection in a wound caused by an infecting organism at a wound site. Also provided is a system for detecting an infection in a wound at a wound site. Additionally, a porous pad comprising luciferase is provided. 1. A method of detecting infection in a wound caused by an infecting organism at a wound site , the method comprising:applying a reduced pressure to the wound site;withdrawing fluid from the wound site in response to the reduced pressure;collecting the fluid withdrawn from the wound site; andassaying the fluid collected from the wound site for a product of the infection or a component of the infecting organism;whereby the presence of the product or the component indicates the presence of an infection in the wound.2. The method of claim 1 , wherein the infecting organism is a bacterium.3. (canceled)4. The method of claim 1 , wherein the fluid withdrawn is assayed for the product of the infection.5. (canceled)6. (canceled)7. (canceled)8. The method of claim 4 , wherein the product of the infection is a host protein associated with an inflammatory response.9. The method of claim 8 , wherein the host protein is a cytokine claim 8 , a fibronectin fragment claim 8 , a neutrophil protease claim 8 , or a macrophage protease.10. The method of claim 1 , wherein the fluid withdrawn is assayed for the component of an infecting organism.11limulus amebocyte. The method of claim 10 , wherein the component of the infecting organism assayed is lipopolysaccharide (LPS) of Gram negative bacteria claim 10 , which is detected with an assay comprising combining the fluid withdrawn with a lysate or by an assay that primes host neutrophil respiratory burst activity via complement opsonized LPS-IgM immune complexes.12. The method of claim 10 , wherein the component of the infecting organism assayed is lipoteichoic acid (LTA) of Gram positive bacteria claim 10 , which is detected with an assay comprising combining the fluid withdrawn with an ...

LIQUID TISSUE PREPARATION FROM HISTOPATHOLOGICALLY PROCESSED BIOLOGICALLY SAMPLES, TISSUES AND CELLS

The current invention provides a method for directly converting histopathologically processed biological samples, tissues, and cells into a multi-use biomolecule lysate. This method allows for simultaneous extraction, isolation, solublization, and storage of all biomolecules contained within the histopathologically processed biological sample, thereby forming a representative library of said sample. This multi-use biomolecule lysate is dilutable, soluble, capable of being fractionated, and used in any number of subsequent experiments. 1. A method of preparing a multi-use biomolecule lysate , comprising the steps of:(a) heating a composition comprising a histopathologically processed biological sample and a reaction buffer at a temperature and a time sufficient to negatively affect protein cross-linking in said biological sample, and(b) treating the resulting composition with an effective amount of a proteolytic enzyme for a time sufficient to disrupt the tissue and cellular structure of said biological sample.218-. (canceled)19. A method of detecting one or more analytes in a multi-use biomolecule lysate suspected of containing said one or more analytes , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) contacting a multi-use biomolecule lysate according to with an array, wherein said array comprises one or more capture agents of known binding specificity immobilized on a support surface in a positionally distinguishable manner; and'}(b) detecting the binding or absence of binding of one or more analytes in said lysate to said immobilized capture reagents.20. The method according to claim 19 , wherein at least one of said analytes is a protein.21. The method according to claim 20 , wherein said capture reagent is selected from the group consisting of antibodies and antibody fragments claim 20 , single domain antibodies claim 20 , engineered scaffolds claim 20 , peptides claim 20 , nucleic acid aptamers claim 20 , a receptor moiety claim ...

COMPOUNDS AND METHODS FOR TREATING OR PREVENTING DISEASE CONDITIONS ASSOCIATED WITH ALPHA-1-ANTITRYPSIN

Pharmacological chaperone compounds and methods for the treatment of an individual having, or at risk of having, a disease condition associated with alpha-1-antitrypsin by using said compounds are disclosed. In particular, such methods are useful for the treatment or prevention of lung disorders associated with alpha-1-antitrypsin deficiency as well as liver disorders associated with an excess of alpha-1-antitrypsin. Suitable pharmacological chaperones include peptides and low-molecular weight compounds. Also provided is an assay for determining whether a test compound modulates alpha-1-antitrypsin activity. 1. A method of treating an individual having or at risk of having a disease condition associated with α-1-antitrypsin comprising administering to the individual an effective amount of a pharmacological chaperone.3. The method of claim 1 , wherein the pharmacological chaperone is:Ac-Thr-Glu-Ala-Ala-NH2,Ac-Thr-Glu-Ala-Ala-Gly-NH2,Ac-Thr-Ser-Ala-Ala-NH2,Ac-Thr-Glu-Val-Ala-NH2 or combinations of two or more thereof.4. The method of claim 1 , wherein the pharmacological chaperone is Ac-Thr-Glu-Val-Ala-NH2.5. The method of claim 1 , wherein the individual has or is at risk of having one or more disease conditions associated with α-1-antitrypsin selected from cirrhosis claim 1 , chronic obstructive pulmonary disease claim 1 , pneumothorax claim 1 , asthma claim 1 , Wegener's granulomatosis claim 1 , pancreatitis claim 1 , gallstones claim 1 , bronchiectasis claim 1 , pelvic organ prolapse claim 1 , primary sclerosing cholangitis claim 1 , autoimmune hepatitis claim 1 , emphysema and cancer.6. The method of claim 1 , wherein the individual has or is at risk of having a liver disorder associated with alpha-1-antitrypsin.7. The method of claim 1 , wherein the individual has or is at risk of having a lung disorder associated with alpha-1-antitrypsin.8. The method of claim 1 , wherein the individual has an E342 missense mutation in α-1-antitrypsin in at least one allele.9. ...

GREEN TEA POLYPHENOL ALPHA SECRETASE ENHANCERS AND METHODS OF USE

The subject invention concerns materials and methods for treating or preventing a neurodegenerative condition or disease associated with β-amyloid peptide deposition in neural tissue in a person or animal by administering a therapeutically effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein that exhibits α-secretase activity. The subject invention also provides methods to increase α-secretase expression and/or activity in cells by administering polyphenol flavonoids like (−)-epigallocatechin--gallate (EGCG) and epicatechin (EC), two polyphenols derived from green tea and other plants and that can be produced synthetically. Furthermore, there are provided methods to decrease or inhibit the production of Aβor Aβby administering the EGCG and EC compounds, their analogs, metabolites, and prodrugs. 1. A method for:i) treating or preventing a neurodegenerative disease or condition associated with β-amyloid peptide deposition in neural tissue, said method comprising administering to a person or animal in need thereof an effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein having α-secretase enzymatic activity; orii) decreasing or inhibiting deposition of β-amyloid peptide in neural tissue, said method comprising contacting said neural tissue with an effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein having α-secretase enzymatic activity; oriii) decreasing levels of a β-amyloid peptide produced by a cell, said method comprising contacting a cell with an effective amount of a polyphenol, or an analog, isomer, metabolite, or prodrug thereof, that increases expression or activity of a protein having α-secretase enzymatic activity.2. The method according to claim 1 , wherein said polyphenol is epigallocatechin-3-gallate ( ...

PLASMINOGEN AND PLASMIN VARIANTS

The invention relates to variants of plasminogen and plasmin comprising one or more point mutations in the catalytic domain which reduce or prevent autocatylic destruction of the protease activity plasmin. Compositions, uses and methods of using said variants of plasminogen and plasmin are also disclosed. 1. An isolated plasminogen variant or plasmin obtained from it , or an isolated plasmin variant , or a proteolytically active or reversible inactive derivative of any of said plasmins characterized in that:(i) it comprises in its catalytic domain a lysine or arginine at an internal position P and a mutation of at least one amino acid at a position [P+/−n], wherein n is 1, 2, 3, 4 or 5 and wherein the amino acid at position [P+/−n] is not a lysine and not an arginine; and(ii) the presence of the mutation of (i) reduces the extent of autoproteolytic degradation of said plasmin variant compared to the extent of autoproteolytic degradation of wild-type plasmin, such as determined with a chromogenic or biological substrate activity assay.2. The plasminogen variant claim 1 , plasmin variant claim 1 , or plasmin derivative according to comprising in its catalytic domain the mutation of at least two internal amino acids:at positions [P+/−n] and [P+/−n′]; wherein n and n′ are 1, 2, 3, 4 or 5; wherein the amino acids at positions [P+/−n] and [P+/−n′] are not lysine and not arginine; and wherein n and n′ are different from each other if both are either added to or subtracted from P; orat positions [P+/−n] and [P′+/−n]; wherein P′ is the greater value of P and P′; wherein the amino acid at position P′ is a lysine or arginine; wherein n is 1, 2, 3, 4 or 5; wherein the amino acid at position [P′+/−n] is not a lysine and not an arginine; and wherein, if present, positions overlapping between [P+n] and [P′−n] are excluded;and wherein the presence of the mutation of said at least two internal amino acids reduces the extent of autoproteolytic degradation of said plasmin variant ...

COMPOSITION FOR TREATING BLOOD AND SET OF DIAGNOSTIC KIT COMPRISING THE SAME TO DETECT AUTOIMMUNE DISEASE

The present invention relates to a composition for treating blood, a set of a diagnostic kit comprising the same to detect an autoimmune disease, and a method of monitoring an autoimmune disease using the same. 1. A method of treating blood to maximize the difference of expression levels of matrix metalloproteinase (MMP) in blood samples , comprising the step of stimulating blood with the composition comprising one or more blood stimulants selected from the group consisting of LPS (Lipopolysacharide) , PMA (Phorbol 12-myristate 13-acetate) , TNF-α (Tumor necrosis factor-alpha) , IL-1β (interleukin-1β) and GM-CSF (Granulocyte-macrophage colony-stimulating factor).2. A method of quantifying or imaging matrix metalloproteinase (MMP) in blood to diagnose an autoimmune disease , the method comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(i) treating blood using the method of ; and'}(ii) applying the treated blood to a kit coated with a complex comprising a conjugate of fluorophore-peptide-quencher,wherein the peptide is a peptide substrate specifically degraded by matrix metalloproteinase (MMP).3. A method of quantifying matrix metalloproteinase (MMP) in blood so as to diagnose an autoimmune disease , the method comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(i) treating blood using the method of ; and'}(ii) quantifying matrix metalloproteinase (MMP) in the treated blood, using a flow cytometer.4. The method of claim 2 , wherein the autoimmune disease is ostarthritis or rheumatoid arthritis.5. The method of claim 2 , wherein the fluorophore is selected from the group consisting of cyanin claim 2 , fluorescein claim 2 , tetramethylrhodamine claim 2 , alexa and bodipy.6. The method of claim 2 , wherein the quencher is a black hole quencher or a blackberry quencher.7. The method of claim 2 , wherein the complex further comprises a polymer coupled to the peptide.8. The method of claim 7 , wherein the polymer is selected ...

IDENTIFICATION OF MODULATORS OF SERINE PROTEASE INHIBITOR KAZAL AND THEIR USE AS ANTI-CANCER AND ANTI-VIRAL AGENTS

This disclosure describes a relevant etiology of cancer and a novel anti-cancer therapeutic strategy, based on the discovery that a protein named serine protease inhibitor (SPIK/SPINK/PSTI) was up-regulated by hepatitis B and C virus infections consequently suppressing the cell apoptosis. Accordingly, the present disclosure provides, inter alia, an inhibitor of SPIK and/or a technology of suppression of over-expression of SPIK in cells. The inhibitors include: 1) chemical compounds, which can inhibit SPIK transcripts, protein activity, and gene expression, 2) SPIK siRNA (RNAi gene silence or dsRNA of SPIK, 3) DNA anti-sense and anti-SPIK antibody. Further, this disclosure provides methods of using the inhibitor as an anti-cancer agent to re-instate cancer cell apoptosis (e.g., serine protease dependent cell apoptosis). 1. A method for identifying a Serine Protease Inhibitor-Kazal (SPIK) modulatory compound , wherein the modulatory compound modulates SPIK serine protease inhibitory activity , the method comprising:(a) contacting a serine protease with SPIK protein in the presence of a labeled serine protease substrate and measuring a first amount of serine protease activity;(b) contacting a serine protease with SPIK protein in the presence of a labeled serine protease substrate and a test agent and measuring a second amount of serine protease activity;(c) comparing the first amount and second amounts of serine protease activity to identify a compound which modulates SPIK serine protease inhibitory activity, wherein the test agent is selected from the group consisting of small molecules, proteins, nucleic acids, and antibodies.2. The method of wherein the label is fluorescent.3. The method of wherein the label can be detected by FRET.4. The method of wherein the modulation is selected from the group consisting of inhibition and activation.5. The method of claim 1 , wherein a plurality of test agents are screened simultaneously.6. The method of claim 1 , wherein the ...

Engineered conformationally-stabilized proteins

Provided herein are conformationally stabilized ubiquitin proteins and methods for using the same to identify agents that bind to the stabilized ubiquitin protein or that bind to a protein that interacts with or processes the stabilized form of the ubiquitin protein. Also provided herein are methods for screening for conformationally stabilized proteins having increased binding affinity to a binding partner in comparison to the binding affinity of the wildtype form of the protein to the binding partner.

Methods of Screening an Agent for an Activity in an Isolated Eye of a Teleost

The present invention provides methods of screening an agent for an activity in an isolated organ, e.g., eye, from a teleost, e.g., zebrafish. Methods of isolating eyes from zebrafish are provided. Methods of screening an agent for an ocular activity in the isolated eye are provided. Methods of screening an agent for an ocular activity in a model of ocular disease or disorder are provided. Methods of screening an agent for an ocular activity in the isolated eye and for screening the agent for cell death and/or toxic activity in the eye or other organ or tissue are provided. The invention further provides high throughput methods of screening agents for an activity in isolated eyes of zebrafish in multi-well plates. 1. (canceled)2. A method of screening an agent for a protective or therapeutic activity for an ocular disease , the method comprising: contacting a teleost with a first agent that is an inducer of ocular disease , and a second agent that is a protective or therapeutic agent for the disease; contacting the teleost with an enzyme that dissociates the eye from the teleost , wherein the enzyme is a collagenase , a dispase , a trypsin , a chymotrypsin , or a hyaluronidase; isolating the eye of the teleost; and analyzing a response to the second agent in the dissociated eye , wherein the presence of the response is indicative of the protective or therapeutic activity of the agent.3. The method of claim 2 , further comprising collecting the dissociated eye by filtration or density gradient centrifugation.4. The method of claim 2 , wherein the teleost is in a well of a multi-well plate.5. The method of claim 2 , wherein the first and second agents can be administered in either order or together.6. The method of claim 2 , wherein the response is analyzed using a microplate reader claim 2 , a high content imaging system claim 2 , or a microscope.7. The method of claim 2 , wherein the isolated eye is contained on a slide following isolation of the eye from the ...

SCREENING METHOD USING GELATINASE-MEDIATED EphA4 CLEAVAGE REACTION AS AN INDICATOR

The object of the present invention is to provide a method for screening a substance that affects gelatinase-mediated EphA4 processing. The present invention provides a method for screening a substance that affects gelatinase-mediated EphA4 processing, which comprises the steps of: (a) allowing a first biological composition containing gelatinase or a biologically active fragment thereof to be contacted with a second biological composition containing EphA4 in the presence and absence of a candidate substance; (b) measuring the presence or amount of the EphA4 ectodomain and/or endodomain fragment; and (c) selecting the candidate substance as a substance that affects gelatinase-mediated EphA4 processing if the results of the step (b) measured in the presence of the candidate substance are changed in comparison with the results of the step (b) measured in the absence of the candidate substance. 1. A method for screening a substance that affects gelatinase-mediated EphA4 processing , which comprises the steps of:(a) allowing a first biological composition containing gelatinase or a biologically active fragment thereof to be contacted with a second biological composition containing EphA4 in the presence and absence of a candidate substance;(b) measuring the presence or amount of the EphA4 ectodomain and/or endodomain fragment; and(c) selecting the candidate substance as a substance that affects gelatinase-mediated EphA4 processing if the results of the step (b) measured in the presence of the candidate substance are changed in comparison with the results of the step (b) measured in the absence of the candidate substance.2. The method according to claim 1 , wherein the step (c) comprises identifying the candidate substance as a substance that promotes gelatinase-mediated EphA4 processing if the EphA4 endodomain and/or ectodomain fragment measured in the presence of the candidate substance during the step (b) is increased in comparison with the EphA4 endodomain and/or ...

TRUNCATED HER2 SRM/MRM ASSAY

This disclosure provides ten (10) specific peptides, and particular peptide characteristics, from the cell membrane-bound Her2 protein and a diagnostic assay useful for determining the presence and amount of full length and truncated versions of the full-length Her2 protein in cells derived from formalin fixed paraffin embedded tissue. 1. A method for detecting the presence and measuring the level of the Her2 protein and the degree of truncated versions of the Her2 protein in a protein digest prepared from a biological sample , comprising detecting specific peptides from the Her2 protein in a protein digest prepared from said biological sample using mass spectrometry; and calculating the level of these 2 versions of the Her2 protein in said sample wherein said level is an absolute level of both versions of the Her2 protein.2. The method of claim 1 , further comprising the step of fractionating said protein digest prior to detecting said peptides.3. The method of claim 2 , wherein said fractionating step is selected from the group consisting of gel electrophoresis claim 2 , liquid chromatography claim 2 , capillary electrophoresis claim 2 , nano-reversed phase liquid chromatography claim 2 , high performance liquid chromatography claim 2 , or reverse phase high performance liquid chromatography.4. The method of claim 1 , wherein said protein digest of said biological sample is prepared by the Liquid Tissue protocol.5. The method of claim 1 , wherein said protein digest comprises a protease digest.6. The method of claim 5 , wherein said protein digest comprises a trypsin digest.7. The method of claim 1 , wherein mass spectrometry comprises tandem mass spectrometry claim 1 , ion trap mass spectrometry claim 1 , triple quadrupole mass spectrometry claim 1 , MALDI-TOF mass spectrometry claim 1 , MALDI mass spectrometry claim 1 , and/or time of flight mass spectrometry.8. The method of claim 7 , wherein the mode of mass spectrometry used is Selected Reaction Monitoring ( ...

NITRATED CARDIAC TROPONIN I AS A BIOMARKER OF CARDIAC ISCHEMIA

The present invention relates to the identification of a novel biomarker for cardiac ischemia: nitrated cardiac troponin I. The present invention also provides methods for the identification and use of a nitrated cardiac troponin as a biomarker for the diagnosis, prognosis and treatment management of myocardial ischemia, with and without necrosis of heart muscle. 13-. (canceled)4. A method of determining the presence of nitrated cardiac troponin I or fragments thereof in a sample of a subject comprising:a) providing a sample of said subject;b) determining whether nitrated cardiac troponin I is present in said sample.5. The method of claim 4 , which is a method for diagnosis or prognosis of myocardial ischemia and which further comprises comparing a concentration of nitrated cardiac troponin I or fragments thereof in the sample with reference values.6. The method of claim 4 , which is a method for diagnosis or prognosis of myocardial ischemia in a subject and which further comprises:c) determining the ratio of non-nitrated cardiac troponin I or fragments thereof to nitrated cardiac troponin I or fragments thereof present in said sample; andd) correlating said ratios with the reference values in order to diagnose or prognose cardiac ischemia.7. The method of claim 4 , which is a method for diagnosis or prognosis of acute coronary syndrome in a subject and which further comprises comparing a concentration of nitrated cardiac troponin I or fragments thereof with reference values in order to diagnose or prognose acute coronary syndrome.8. The method of claim 4 , which is a method for diagnosis or prognosis of acute coronary syndrome in a subject and which further comprises:c) determining a ratio of non-nitrated cardiac troponin I or fragments thereof to nitrated cardiac troponin I or fragments thereof present in said sample; andd) comparing said ratio with reference values in order to diagnose or prognose acute coronary syndrome.9. The method of claim 4 , which is a ...

CHEMILUMINESCENCE-BASED HAEMOSTASIS ASSAY

The present invention relates to a method for in vitro determining generation of a haemostatis factor such as thrombin and/or plasmin in a test sample using a chemiluminescent substrate specific for said blood clotting factor. Upon cleavage of the substrate, a luminescent signal is generated via aminoluceferin with the aid of a luciferase. The invention also relates to a kit for in vitro determining generation of a haemostasis factor in a test sample, and to novel chemiluminescent substrates for the determination of thrombin and plasmin. 1. A method for in vitro determining generation of a haemostasis factor in a test sample comprising determining the amount of said haemostasis factor generated using a chemiluminescent substrate that is specific for said haemostasis factor , wherein preferably a luminescent molecule is liberated by a haemostasis factor which is subsequently converted to produce a detectable light quant.2. The method according to claim 1 , comprising:providing a reaction mixture comprising a test sample to be tested, a trigger molecule for inducing generation of said haemostasis factor, a chemiluminescent substrate specific for said haemostasis factor; anddetermining the amount of said haemostasis factor generated by measuring a luminescent signal.3. The method according to claim 1 , wherein the chemiluminescent substrate specific for said haemostasis factor comprises a peptide comprising a cleavage site specific for said haemostasis factor coupled to aminoluciferin.4. The method according to claim 1 , wherein the haemostasis factor is selected from the group consisting of serine endopeptidases (EC 3.4.21).5. The method according to claim 1 , wherein the haemostasis factor is selected from the group consisting of thrombin claim 1 , plasmin claim 1 , factor Xa claim 1 , factor IXa claim 1 , factor VIIa claim 1 , factor XIa factor XIIa claim 1 , kallikrein claim 1 , activated Protein C claim 1 , tc-tPA and tc-uPA claim 1 , preferably the haemostasis ...

ANTIMICROBIAL AGENTS

The present invention relates to a fusion protein composed of an enzyme having the activity of degrading the cell wall of Gram-negative bacteria and/or Gram-positive bacteria and at least two peptide stretches fused to the enzyme at the N- or C-terminus, wherein the peptide stretches are distinct and selected from the group of synthetic amphiphatic peptide, synthetic cationic peptide, synthetic polycationic peptide, synthetic hydrophobic peptide, or naturally occurring antimicrobial peptide, like sushi peptide and defensin. Moreover, the present invention relates to nucleic acid molecules encoding said fusion protein, vectors comprising said nucleic acid molecules and host cells comprising either said nucleic acid molecules or said vectors. In addition, the present invention relates to said fusion protein for use as a medicament, diagnostic means, cosmetic substance, a disinfectant or a food additive. The present invention also relates to the treatment or prevention of Gram-positive and/or Gram-negative bacterial contamination of foodstuff, of food processing equipment, of food processing plants, of surfaces coming into contact with foodstuff, of medical devices, of surfaces in hospitals and surgeries. 1. A fusion protein comprising an enzyme having the activity of degrading the cell wall of Gram-negative bacteria and/or Gram-positive bacteria and at least two peptide stretches fused to the enzyme at the N- or C-terminus , wherein the peptide stretches are distinct and selected from the group of synthetic amphiphatic peptide , synthetic cationic peptide , synthetic polycationic peptide , synthetic hydrophobic peptide , or naturally occurring antimicrobial peptide , like sushi peptide and defensin.2. The fusion protein according to claim 1 , wherein the enzyme is an endolysin claim 1 , autolysin or bacteriocin.3. The fusion protein according to claim 1 , wherein the synthetic peptide or naturally occurring peptide has a length of 6 to 39 amino acid residues.4. The ...

Quantitation of GCP Activity in Biological Samples

Provided herein are methods for quantitating glutamate carboxypeptidase II activity in a biological sample, such as a skin biopsy sample, and for determining whether an agent is inhibiting GCP in a subject. 1. A method quantitating glutamate carboxypeptidase II (GCPII) activity in a biological sample that contains endogenous N-acetyl-aspartate glutamate (NAAG) , the method comprising the steps of:(a) reducing the amount of the endogenous NAAG in the biological sample;(b) contacting the biological sample with exogenous NAAG; and(c) detecting an exogenous NAAG cleavage product in the biological sample.2. The method of claim 1 , wherein the biological sample comprises a skin sample claim 1 , optionally claim 1 , wherein the skin sample is a skin biopsy collected from a subject.3. (canceled)4. The method of claim 2 , wherein the subject had been administered a GCPII inhibitor before the skin biopsy was collected.5. The method of claim 1 , wherein the step of reducing the amount of endogenous NAAG in the biological sample comprises the steps of:(i) homogenizing the biological sample in a buffer to create a homogenate;(ii) centrifuging the homogenate to create a sample pellet that contains the GCPII and a supernatant that contains the endogenous NAAG; and(iii) removing the supernatant from the sample.6. The method of claim 5 , wherein the sonication is used to homogenize the biological sample.7. The method of claim 1 , wherein the exogenous NAAG comprises a detectable label that is detected in step (c) as an indication of the presence of the exogenous NAAG cleavage product.8. The method of claim 7 , wherein the detectable label is a radioactive claim 7 , fluorescent claim 7 , enzymatic claim 7 , colorimetric or chemiluminescent label.9. (canceled)10. The method of claim 1 , wherein the exogenous NAAG cleavage product is glutamate or N-acetyl-aspartate (NAA).11. (canceled)12. The method of claim 1 , wherein the exogenous NAAG cleavage product is separated from remaining ...

METHOD FOR THE DETERMINATION OF BOTULINUM NEUROTOXIN BIOLOGICAL ACTIVITY

The present invention relates to means and methods for determining neurotoxin activity. Specifically, it relates to a polypeptide having caspase activity comprising a large subunit and a small subunit wherein the caspase further comprises a neurotoxin cleavage site which upon cleavage activates the caspase activity. Also encompassed are polynucleotides encoding the polypeptides as well as vectors or host cells comprising the polynucleotides. The present invention further relates to a method for determining neurotoxin activity in a sample based on the polypeptide of the invention as well as the use of the polypeptide for determining neurotoxin activity in a sample, in general. 117-. (canceled)18. A polypeptide which exhibits caspase activity comprising a caspase large subunit and a caspase small subunit and a neurotoxin cleavage site which upon cleavage activates the caspase activity.19. The polypeptide of claim 18 , wherein the neurotoxin cleavage site is located between the C-terminal proximal region of the caspase large subunit and the N-terminal proximal region of the caspase small subunit.20. The polypeptide of claim 19 , wherein(i) the C-terminal proximal region of the caspase large subunit consists the amino acids corresponding to amino acids C170 to D175, L168 to E173, Q161 to D175, or Q161 to E173 of the large subunit of caspase 3 and the N-terminal proximal region of the caspase small subunit consists of the amino acids corresponding to amino acids S176 to V190 or S176 to C184 of the small subunit of caspase 3;(ii) the C-terminal proximal region of the caspase large subunit consists the amino acids corresponding to amino acids V174 to D179, II172 to V177, Q165 to D179, or Q165 to V177 of the large subunit of caspase 6 and the N-terminal proximal region of the caspase small subunit consists of the amino acids corresponding to amino acids S180 to A194 or S180 to V188 of the small subunit of caspase 6;(iii) the C-terminal proximal region of the caspase large ...

DETECTION OF PROTEASE AND PROTEASE ACTIVITY USING A SINGLE NANOCRESCENT SERS PROBE

This invention pertains to the in vitro detection of proteases using a single peptide-conjugate nanocrescent surface enhanced Raman scattering (SERS) probes with at least nanomolar sensitivity. The probe enables detection of proteolytic activity in extremely small volume and at low concentration. In certain embodiments the probes comprise an indicator for the detection of an active protease, where the indicator comprises a nanocrescent attached to a peptide, where said peptide comprises a recognition site for the protease and a Raman tag attached to the peptide. 1. An indicator for the detection of an active protease , said indicator comprising:a nanocrescent attached to a peptide, wherein said peptide comprises a recognition site for said protease; and wherein said nanocrescent comprises a shell without a core.233-. (canceled)34. A method of detecting or quantifying the presence , amount , or activity of at least one protease in a sample , said method comprising:contacting said sample an indicator comprising a nanocrescent attached to a peptide, wherein said peptide comprises a recognition site for said protease; andmonitoring differences in spectral characteristics of detected surface-Raman scattering spectra, the differences being indicators of the presence, amount or activity of protease present in the sample.35. The method of claim 34 , wherein said indicator further comprises a Raman label attached to said peptide.36. The method of claim 34 , wherein said monitoring comprises monitoring surface enhanced Ramen scattering (SERS).37. The method of claim 34 , wherein said sample comprises material selected from the group consisting of sample selected from the group consisting of whole blood claim 34 , plasma claim 34 , serum claim 34 , synovial fluid claim 34 , cerebrospinal fluid claim 34 , bronchial lavage claim 34 , ascites fluid claim 34 , seminal fluid claim 34 , bone marrow aspirate claim 34 , pleural effusion claim 34 , urine claim 34 , and tumor cells or ...

FITNESS ASSAY AND ASSOCIATED METHODS

The present invention provides an assay for determining the biochemical fitness of a biochemical species in a mutant replicating biological entity relative to its predecessor. The present invention further provides a continuous fluorogenic assay for measuring the anti-HIV protease activity of protease inhibitor. The present invention also provides a method of administering a therapeutic compound that reduces the chances of the emergence of drug resistance in therapy. The present invention also provides a compound of formula (I) or a pharmaceutically acceptable salt, a prodrug, a composition, or an ester thereof, wherein A is a group of formulas (A), (B), (C) or (D); R, R, R, Ror Ris H, or an optionally substituted and/or heteroatom-bearing alkyl, alkenyl, alkynyl, or cyclic group; Y and/or Z are CH, O, S, SO, SO, amino, amides, carbamates, ureas, or thiocarbonyl derivatives thereof, optionally substituted with an alkyl, alkenyl, or alkynyl group; n is from 1 to 5; X is a bond, an optionally substituted methylene or ethylene, an amino, O or S; Q is C(O), C(S), or SO; m is from 0 to 6; Ris OH, ═O (keto), NH, or alkylamino, including esters, amides, and salts thereof; and W is C(O), C(S), S(O), or SO. Optionally, Rand R, together with the N—W bond of formula (I), comprise a macrocyclic ring. 1. An assay for determining the biochemical fitness of a biochemical target of a mutant replicating biological entity relative to its predecessor , comprising:obtaining said predecessor,determining the biochemical vitality of said biochemical target of said predecessor in the presence of a compound capable of inhibiting said biochemical target of said predecessor,determining the biochemical vitality of said biochemical target of said mutant replicating biological entity in the presence of said compound, andcomparing the biochemical vitality of said biochemical target of said mutant replicating biological entity relative to the biochemical vitality of said biochemical target of said ...

METHOD FOR IDENTIFICATION OF PROTEASE ACTIVITY INHIBITORS AND ASSAYING THE PRESENCE OF PROTEASE ACTIVITY

A system for the identification of proteases and protease inhibitors is provided. The system has at least two components. The first component is a reporter construct with at least one binding site, a transcriptional promoter, an inducible promoter region, and at least one reporter gene, all functionally connected for expression of the reporter gene(s) in functional coordination with a transcriptional activation agent. The second component is a transcriptional activation agent comprising a nucleic acid binding domain, at least one protease substrate domain, and at least one transcriptional activation domain for an inducible promoter. The system allows detection and evaluation of agents affecting protease activity directed to the protease substrate domain. The system also allows for the detection of the presence of proteases in environmental samples. 1. A system for the assessment of protease activity , comprising:at least two molecules, wherein one molecule is a reporter construct incorporated into a mammalian cell DNA and a second molecule is a transcriptional activation agent;wherein the reporter construct comprises at least one binding site capable of being bound by a nucleic acid binding domain of the transcriptional activation agent, an inducible promoter region, and at least one reporter gene, all functionally connected for expression of the reporter gene(s), and in functional coordination with the transcriptional activation agent upon binding of the transcriptional activation agent; andwherein the transcriptional activation agent comprises the nucleic acid binding domain, at least one cleavable protease substrate domain, and at least one transcriptional activation domain for an inducible promoter, said cleavable protease substrate domain capable of being cleaved by an exogenous protease, wherein said protease is expressed outside the nucleus.2. The system of claim 1 , further comprising at least one protease or protease candidate claim 1 , wherein the protease ...

METHOD AND REAGENTS FOR TREATING HEPATIC FIBROSIS AND INFLAMMATION

The invention relates to methods for identifying an anti-fibrotic or anti-inflammatory agent comprising determining cathepsin S expression in activated hepatic stellate cells which have been exposed to a test compound and comparing expression to non-exposed hepatic stellate cells. The invention also relates to methods for treating a disorder characterised or caused by hepatic fibrosis or inflammation, comprising administering a cathepsin S inhibitor to a subject. 1. A method of identifying a hepatic anti-fibrotic agent or hepatic anti-inflammatory agent , the method comprising:a) determining a first expression level of Cat S in a first activated hepatic stellate cell;b) exposing a second activated hepatic stellate cell to a test compound;c) determining a second expression level of Cat S in said second activated hepatic stellate cell;d) comparing the first expression level and the second expression levelwhereby the first expression level which is greater than the second expression level indicates that the test compound is a hepatic anti-fibrotic agent or hepatic anti-inflammatory agent.2. The method of further comprising the step of exposing the first and second hepatic stellate cells to a cytokine prior to determining the first and second expression levels.3. The method of wherein the first hepatic stellate cell and the second hepatic stellate cell are provided in vitro.4. The method of wherein the first hepatic stellate cell and the second hepatic stellate cell are a HSC-T6 cell.5. The method of wherein the cytokine is IFN-γ.6. A kit comprising:a) a hepatic stellate cell; andb) a reagent for detecting the expression level of Cat S.7. The kit according to wherein the reagent for detecting the expression level of Cat S is a nucleic acid complementary to a portion of the Cat S gene.8. The kit according to wherein the reagent for detecting the expression level of Cat S is an anti-cathepsin S antibody.9. The kit according to wherein the antibody is a monoclonal antibody ...

METHODS AND COMPOUNDS FOR INCREASING SENSITIVITY OF BOTULINUM ASSAYS

Apparatus, systems and methods can provide improved detection of botulinum neurotoxins. In one aspect an isoquinolynyl compound can be used to enhance the sensitivity of both Förster resonance energy transfer (FRET) and non-FRET cell-based assays. In another aspect, non-FRET assays and constructs utilize a reporter that is not coupled with the second fluorophore in a manner that produces significant FRET. In that subject matter an environment cell can include an enzyme that facilitates degradation of the reporter significantly faster after the cleavage than before the cleavage, and presence of the Botulinum toxin correlates with reduction of the signal from a baseline signal. Where the environment is a cell, the cell can advantageously express both the construct that includes the reporter, and an enzyme that facilitates the degradation. 1. A method of detecting presence of a Botulinum toxin , comprising:providing an assay with a cell having (a) a molecule with cleavage site that interacts with a portion of the Botulinum toxin and (b) a reporter that reports a signal responsive to cleavage of the molecule at the cleavage site induced by Botulinum toxin; andincubating the cell in a culture media that contains a compound effective to increase sensitivity of the assay by at least 0.5 log;treating the cell with an amount of the Botulinum toxin, then sensing the signal, and correlating the signal with presence or absence of the Botulinum toxin.2. The method of claim 1 , wherein the compound is selected from (a) an isoquinolynyl and (b) a protein kinase inhibitor.3. The method of claim 1 , wherein the compound comprises 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine dichloride (H7).4. The method of claim 1 , wherein the culture media comprises B275. The method of claim 1 , wherein osmolarity of the media is optimized for Botox®.6. The method of claim 1 , wherein osmolarity of the media is optimized for Dysport®/Reloxin®.7. The method of claim 1 , wherein osmolarity of the ...

MONITORING OF WOUNDS BY MEASUREMENT OF PROTEASE AND PROTEASE INHIBITOR LEVELS IN WOUND FLUIDS

A diagnostic test apparatus for determining a ratio of: (a) at least one endogenous protease enzyme inhibitor, to (b) at least one endogenous protease enzyme, in a sample of a wound fluid. Suitably the protease enzyme is a neutrophil elastase and the protease enzyme inhibitor is alpha-1-antitrypsin. Also provided are wound treatment systems comprising an apparatus according to the invention and a wound dressing comprising an oxidized cellulose. Also provided are methods of diagnosis, prognosis and treatment of wounds using the apparatus and systems of the invention. 1. A diagnostic test apparatus for determining a ratio of: (a) at least one endogenous protease enzyme inhibitor , to (b) at least one endogenous protease enzyme , in a sample of a wound fluid , the diagnostic test apparatus comprising:an indicator moiety that is immobilized or inhibited by a chemical moiety, wherein said chemical moiety comprises an exogenous peptide substrate for a protease enzyme, and said exogenous peptide substrate is cleavable by the protease enzyme to release or activate said indicator moiety.2. The diagnostic test apparatus according to claim 1 , wherein said step of determining comprises determining whether said ratio falls within a predetermined range.3. The diagnostic test apparatus according to claim 1 , wherein the said at least one protease enzyme comprises one or more proteases selected from the group consisting of neutrophil elastase claim 1 , matrix metalloproteinases ( e.g. MMP-9 claim 1 , MMP-8 claim 1 , MMP-1 claim 1 , MMP-12) claim 1 , proteinase 3 claim 1 , plasmin claim 1 , low molecular weight gelatinases and latent or active elastases claim 1 , interleukin converting enzymes and tumor necrosis factor (TNFa) converting enzymes.4. The diagnostic test apparatus according to claim 1 , wherein the said at least one protease comprises one or more proteases selected from the group consisting of neutrophil elastase and matrix metalloproteinases.5. The diagnostic test ...

Mutant protease biosensors with enhanced detection characteristics

A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

METHOD AND APPARATUS FOR FRACTIONATION-BASED CHEMICAL ANALYSES

A method () for analyzing chemicals includes fractionating a complex sample into at least two sample portions that each includes potions of two polypeptides though in different concentration ratios, digesting and performing LC/MS on each of the sample portions ( and associating precursor ions observed via LC/MS with their corresponding polypeptide in response to LC/MS provided intensity data (). A set of precursor ions that has substantially similar intensity ratios in both sample portions is determined to be associated with the same polypeptide. 114-. (canceled)15. An apparatus for performing sample analysis comprising:a chromatography module;a mass-spectrometry module in communication with said chromatography module; anda control unit in communication with said chromatography module and said mass spectrometry module, said control unit including at least one processor and a memory for storing a plurality of instructions executed by said processor, said plurality of instructions causing said processor to perform a method comprising:providing a first sample portion and a second sample portion, wherein said first sample portion and said second sample portion are included in a plurality of samples portions produced by fractionating a complex sample, wherein said complex sample includes at least a first compound and a second compound, wherein said first sample portion and said second sample portion each include portions of the first compound and the second compound though in different concentration ratios;performing analysis of the first sample portion to observe intensities of precursor ions associated with the first and second compounds in the first sample portion, wherein said analysis of the first sample portion includes performing a chromatographic separation using the chromatography module and performing mass analysis using the mass-spectrometry module;performing analysis of the second sample portion to observe intensities of precursor ions associated with the ...

METHOD FOR SCREENING INHIBITORY SUBSTANCES TO INFLAMMATORY SKIN-AGING

Disclosed is a method for screening a substance inhibiting inflammatory skin aging, comprising: treating an immune cell with an inflammatory stimulant; treating a skin cell with a candidate substance; treating the skin cell treated with the candidate substance, with the immune cell treated with the inflammatory stimulant; and measuring the concentration of matrix metalloproteinase-1 (MMP-1) in the skin cell treated with the immune cell. 1. A method for screening a substance inhibiting inflammatory skin aging , comprising:treating an immune cell with an inflammatory stimulant;treating a skin cell with a candidate substance;treating the skin cell treated with the candidate substance, with the immune cell treated with the inflammatory stimulant; andmeasuring a concentration of matrix metalloproteinase-1 (MMP-1) in the skin cell treated with the immune cell.2. The method for screening a substance inhibiting inflammatory skin aging according to claim 1 , wherein the immune cell comprises a THP-1 cell (human acute monocytic leukemia cell).3. The method for screening a substance inhibiting inflammatory skin aging according to claim 1 , wherein the skin cell comprises a normal human fibroblast (NHF).4. The method for screening a substance inhibiting inflammatory skin aging according to claim 1 , wherein the inflammatory stimulant comprises a lipopolysaccharide (LPS) and interferon-γ (INF-γ).5Escherichia coli.. The method for screening a substance inhibiting inflammatory skin aging according to claim 4 , wherein the LPS comprises an LPS derived from6. The method for screening a substance inhibiting inflammatory skin aging according to claim 4 , wherein a concentration of the LPS is 0.001-20 μM and a concentration of the INF-γ is 1-2000 UNITs. The present disclosure relates to a method for screening a substance inhibiting inflammatory skin aging.In general, chronic skin inflammation is a major cause of wrinkles since it reduces elasticity of skin. Once inflammation occurs in ...

EVALUATION OF COPOLYMER DIETHYLAMIDE

Methods of analyzing glatiramer acetate (GA) or a polymeric precursor thereof are provided. The methods can include determining a level of one or more diethylamide-modified amino acids in a sample comprising GA or a polymeric precursor thereof, and selecting at least a portion of the sample based on the assessment of the one or more diethylamide-modified amino acids in the sample. 1. (canceled)2. A method of manufacturing glatiramer acetate drug product , the method comprising:preparing an amino acid copolymer of L-glutamic acid, L-alanine, L-Lysine, and L-tyrosine;measuring, in at least one sample of the copolymer, the levels of at least two, three, or four individual diethylamide-modified amino acids selected from the group consisting of: diethylamide-modified alanine, diethylamide-modified lysine, diethylamide-modified glutamic acid, and diethylamide-modified tyrosine; andprocessing the copolymer to produce glatiramer acetate drug product if the measured levels of at least one of the at least two, three, or four measured individual diethylamide-modified amino acids meets commercial release specification (i), (ii), (iii), or (iv):(i) the level of diethylamide-modified alanine in the sample is 59.5-76.1% of the total diethylamide-modified amino acids in the sample on a mol percent basis;(ii) the level of diethylamide-modified lysine detected in the sample is 11.3-17.3% of the total diethylamide-modified amino acids in the sample on a mol percent basis;(iii) the level of diethylamide-modified glutamic acid detected in the sample is 9.9-15.0% of the total diethylamide-modified amino acids in the sample on a mol percent basis; or(iv) the level of diethylamide-modified tyrosine detected in the sample is 4.8-7.2% of the total diethylamide-modified amino acids in the sample on a mol percent basis,thereby manufacturing glatiramer acetate drug product.3. The method of wherein:the step of preparing comprises co-polymerizing N-carboxy anhydrides of L-alanine, benzyl- ...

GLUTAMYL AMINOPEPTIDASE AS A MARKER OF RENAL DAMAGE

The present invention relates to a method and a kit for the diagnosis and/or prognosis of kidney injury comprising analysing a sample obtained from a patient and determining the activity of at least one aminopeptidase selected from aspartyl aminopeptidase, glutamyl aminopeptidase, alanyl aminopeptidase and leucyl-cystinyl aminopeptidase. 1. (canceled)2. A method for obtaining useful data in the diagnosis and/or prognosis of kidney injury comprising determining in an isolated sample the glutamyl aminopeptidase activity.3. The method according to claim 2 , where the aminopeptidase activity is determined by fluorometric analysis.4. A method for the diagnosis and/or prognosis of kidney injury comprising determining in an isolated sample the glutamyl aminopeptidase activity and comparing said activity with a control value claim 2 , where the alteration of said activity is indicative of kidney injury.5. The method according to claim 4 , where the kidney injury is an acute kidney injury.6. The method according to claim 4 , where the sample is urine.7. The method according to claim 4 , where the aminopeptidase activity is determined by fluorometric analysis.8. The method according to claim 4 , where the substrate used is glutamyl-β-naphthylamide.9. The method according to claim 4 , where the increase in the glutamyl aminopeptidase activity claim 4 , with respect to control value is indicative of kidney injury.10. The method according to claim 4 , where the increase in the glutamyl aminopeptidase activity with respect to control value is indicative of early kidney injury.11. The method according to claim 4 , where the patient suffers a disease selected from the group consisting of hyperthyroidism claim 4 , hypertension claim 4 , diabetes claim 4 , transplant claim 4 , and secondary effects of drugs that cause kidney injury.12. A kit for the diagnosis and/or prognosis of kidney injury according to the method of claim 4 , comprising the reagents necessary to determine the ...

WOUND PROGNOSIS

Methods and devices for determining the healing status of wounds, in particular of chronic wounds are provided. Also provided are kits comprising the diagnostic devices, and methods of wound diagnosis and treatment using the diagnostic devices and methods. A diagnostic apparatus (device) for monitoring wounds that exude a wound fluid and determining whether said wounds would be responsive to treatment with wound therapy such as oxidized cellulose therapy is also provided along with kits comprising the diagnostic apparatus and a wound dressing. Furthermore, methods of prognosing and treating wounds that exude a wound fluid are also provided. 1. A method of wound prognosis comprising the steps of:determining if the level of human neutrophil elastase (hNE) in a sample of wound fluid from a wound exceeds a first predetermined threshold;determining if the level of at least one matrix metalloproteinase (MMP) in a sample of wound fluid from said wound exceeds a second predetermined threshold; and assigning said wound to a non-healing category if either said level of hNE or said level of MMP exceeds said first or second threshold, respectively.2. A method according to claim 1 , wherein the matrix metalloproteinases determined are selected from MMP-1 claim 1 , MMP-2 claim 1 , MMP-8 claim 1 , MMP-9 claim 1 , MMP-12 claim 1 , MMP-13 claim 1 , combinations thereof claim 1 , and total MMP.3. A method according to claim 2 , wherein the matrix metalloproteinases determined comprise MMP-1 and MMP-9.4. A method according to claim 1 , wherein said level of MMP is a weighted total of individual levels of a plurality of different MMPs.5. A method according to claim 1 , wherein said steps of determining are performed on a sample of wound fluid that has been removed from the body.6. A method according to claim 1 , wherein one or both of said steps of determining comprise allowing the analyte protease(s) to cleave a peptide substrate claim 1 , whereby said level corresponds to an activity ...

DESTRUCTIBLE SURFACTANTS AND USES THEREOF

The present invention provides methods for enhancing chemical reactions of molecules, e.g., biomolecules, with destructible surfactants. The chemical reactions may involve and/or be associate with analysis, e.g., solubilizing, separating, purifying and/or characterizing the molecules. In one aspect, the anionic surfactants of the present invention may be selectively broken up at relatively low pH. The resulting breakdown products of the surfactants may be removed from the molecule/sample with relative ease. The invention has applicability in a variety of analytical techniques. 2. The method of claim 1 , wherein the molecule is a biomolecule.3. The method of claim 2 , further comprising analyzing the biomolecule following the chemical reaction.4. The method of claim 2 , wherein the biomolecule is contained in a biological sample.5. The method of claim 4 , wherein the biological sample is selected from the group consisting of inclusion bodies claim 4 , biological fluids claim 4 , biological tissues claim 4 , biological matrices claim 4 , embedded tissue samples claim 4 , and cell culture supernatants.6. The method of claim 2 , wherein the biomolecule is selected from the group consisting of a protein and a peptide.7. The method of claim 6 , wherein the biomolecule is selected from the group consisting of a lipophilic protein claim 6 , a receptor claim 6 , a proteolytic protein claim 6 , and a membrane-bound protein.8. The method of claim 3 , wherein the analysis is selected from the group consisting of solid phase extraction claim 3 , solid phase micro extraction claim 3 , electrophoresis claim 3 , mass spectrometry claim 3 , liquid chromatography claim 3 , liquid-liquid extraction claim 3 , membrane extraction claim 3 , soxhlet extraction claim 3 , precipitation claim 3 , clarification claim 3 , electrochemical detection claim 3 , staining claim 3 , elemental analysis claim 3 , Edmund degradation claim 3 , nuclear magnetic resonance claim 3 , infrared analysis claim ...

Di- and Poly-Ubiquitin Deubiquitinase Substrates and Uses Thereof

Methods for detection of the activity of proteolytic enzymes, particularly isopeptidases, are disclosed. 1. A diubiquitin for measuring the isopeptidase activity of a deubiquitinase comprising:A) a first ubiquitin molecule operably linked to at least one first energy transfer pair member; andB) a second ubiquitin molecule operably linked to at least one second energy transfer pair member;wherein said first ubiquitin molecule is operably linked to said second ubiquitin molecule by an isopeptide bond from the C-terminus of said first ubiquitin molecule to the side chain of a lysine residue of said second ubiquitin molecule,wherein said first and second energy transfer pair members are individually either a fluorescent group or a quenching group.2. The diubiquitin of claim 1 , wherein said first and second ubiquitin molecules have at least 95% identity with SEQ ID NO: 1.3. The diubiquitin of claim 1 , wherein said first energy transfer pair member is a fluorescent group and said second energy transfer pair member is a quenching group.4. The diubiquitin of claim 1 , wherein said second energy transfer pair member is a fluorescent group and said first energy transfer pair member is a quenching group.5. The diubiquitin of claim 1 , wherein said first and second ubiquitin molecules comprise a cysteine residue.6. The diubiquitin of claim 5 , wherein said first and second energy transfer pair members are attached to said first and second ubiquitin molecules at said cysteine residues.7. The diubiquitin of claim 5 , wherein said cysteine residue is introduced at any position between residues 8 and 70.8. The diubiquitin of claim 7 , wherein said cysteine residues is introduced at position 11 claim 7 , 20 claim 7 , 31 claim 7 , 34 claim 7 , 48 claim 7 , 57 claim 7 , or 63.9. The diubiquitin of claim 1 , wherein said isopeptide bond is between the C-terminus of said first ubiquitin molecule and the lysine at position 6 of said second ubiquitin molecule.10. The diubiquitin of ...

MAST CELL MARKERS AND PREVENTION, DIAGNOSIS, AND THERAPY FOR CHRONIC PELVIC PAIN SYNDROME

The present invention provides compositions and methods for detection, diagnosis, treatment and/or prevention of chronic pelvic pain syndrome. In particular, the present invention provides biomarkers of chronic pelvic pain syndrome (e.g., mast cell markers (e.g., tryptase)), and/or inhibition of mast cell function (e.g. inhibition of MCP-1 and/or MIP-1α) to treat or prevent chronic pelvic pain syndrome. 1. A method for treating and/or preventing chronic pelvic pain syndrome in a subject , comprising administering therapeutic composition comprising a therapeutically effective amount of an inhibitor of mast cell function to said subject.2. The method of claim 1 , wherein said inhibitor of mast cell function comprises a combination of a mast cell stabilizer and a histamine receptor antagonist.3. The method of claim 2 , wherein said therapeutic composition further comprising one or more inhibitors of MCP-1 and/or MIP-1α.4. The method of claim 2 , wherein said histamine receptor antagonist comprises a histamine receptor 1 antagonist and/or histamine receptor 2 antagonist.5. A method for treating and/or preventing chronic pelvic pain syndrome in a subject claim 2 , comprising co-administering a mast cell stabilizer and a histamine receptor antagonist.6. The method of claim 5 , further comprising administering one or more inhibitors of MCP-1 or MIP-1α.7. A composition for the treatment or prevention of CPPS claim 5 , comprising:a) a mast cell stabilizer; andb) a histamine receptor antagonist.8. The composition of claim 7 , further comprising an inhibitor of a CPPS biomarker.9. The composition for the treatment or prevention of CPPS of claim 7 , wherein said histamine receptor antagonist comprises a histamine receptor 1 antagonist and/or a a histamine receptor 2 antagonist.10. The composition for the treatment or prevention of CPPS of claim 7 , comprising inhibitors of MIP-1α and/or MCP-1.11. A method for detecting chronic pelvic pain syndrome in a subject claim 7 , ...

FLUORESCENCE LIFETIME EPIGENETICS ASSAYS

The invention is concerned with methods of assaying the activity of enzymes based on measurement of fluorescence lifetime (FLT). In particular, the invention relates to assays for enzymes which are capable of modifying the structure of peptide substrates, including for example enzymes catalysing methylation, demethylation, acetylation, deacetylation or deimination of peptide substrates. 1. A method of assaying the activity of a modifying enzyme in a test sample , comprising:(a) contacting the test sample with a fluorescent-modulated enzyme substrate comprising a linker molecule conjugated to a fluorescent moiety and a fluorescence lifetime modulator moiety configured to modulate the fluorescence lifetime of the fluorescent moiety, wherein the substrate is modified by the action of the modifying enzyme to form a modified substrate, the linker molecule of said modified substrate either being rendered susceptible to cleavage by a second enzyme or protected from cleavage by a second enzyme between the fluorescent moiety and the fluorescence lifetime modulator moiety as a result of the modification, wherein cleavage of the substrate or the modified substrate by the second enzyme separates a portion of the substrate containing the fluorescence lifetime modulator moiety from a portion of the substrate containing the fluorescent moiety; and(b) detecting formation of the modified substrate by detecting changes in the fluorescence lifetime of the fluorescent moiety as a result of the action of the second enzyme on the modified substrate and/or the substrate, wherein formation of the modified substrate provides an indication of the activity of the modifying enzyme.2. The method according to wherein the linker molecule of the fluorescent-modulated enzyme substrate is a peptide linker and the second enzyme is a protease which cleaves either the peptide linker or a modified peptide linker formed by action of the modifying enzyme on the peptide linker to separate a portion of the ...

Methods of Measuring Protein and/or Fat Digestibility and Uses Thereof

The present invention provides methods for continuous measurement of protein and/or triglyceride digestibility during a meal. Stable isotope of N-spirulina protein and H-phenylalanine was added to the nutritional supplement. Protein digestibility is calculated by measuring the ratio [N]PHE to [H]PHE in plasma and the nutrition. In another embodiment, [1,1,1-C]tripalmitin and [2,2-H]palmitic acid were added to the meal. The ratio between [1-C]palmitic acid/[2,2-H]palmitic acid will represent the percentage digestion of triglycerides by lipase from the pancreas. 1. A method for determining reduced protein digestibility in an individual in need of such determination , comprising the step of:administering a meal comprising an isotope labeled protein and a free amino acid tracer to said individual and to a control subject; andmeasuring the amount of isotope labeled amino acid formed after digestion of said isotope labeled protein and measuring the amount of free amino acid tracer in a biological sample of said individual and said control subject to determine a ratio of isotope labeled amino acid to free amino acid tracer for said individual and for said control subject; wherein a lower ratio of labeled amino acid to free amino acid tracer in said individual compared to the ratio of labeled amino acid to free amino acid tracer from said control subject indicates that said individual has a reduced capacity to digest protein.2. The method of claim 1 , wherein said free amino acid tracer is H-labeled amino acid claim 1 , N-labeled amino acid or C-labeled amino acid.3. The method of claim 2 , wherein said labeled amino acid is H-phenylalanine claim 2 , H-lysine claim 2 , H-leucine claim 2 , N-phenylalanine claim 2 , N-lysine claim 2 , N-leucine claim 2 , C-phenylalanine claim 2 , C-lysine claim 2 , or C-leucine.4. The method of claim 1 , wherein said isotope labeled protein is N-labeled spirulina protein claim 1 , N-labeled milk protein claim 1 , N-labeled soy protein claim 1 ...

Tailored cyclodepsipeptides as potent non-covalent serine protease inhibitors

The present invention pertains to an improved chemical synthesis method for Ahp-cyclodepsipeptides which allows straight forward and easy synthesis of tailor-made Ahp-cyclodepsipeptides. The invention further provides Ahp-cyclodepsipeptides for use as HTRA protease inhibitors and their medical use.

CARTRIDGE FOR ENDOTOXIN DETECTION

The present invention is directed to methods, compositions and devices useful for the detection and/or quantification of a microbial contaminant, including an endotoxin. In embodiments, cartridges are provided that include dried compositions that are useful in absorbance-based assays and in combination with portable readers/devices. 1. A cartridge for determining the presence and/or amount of a microbial contaminant in a sample , the cartridge comprising:a. a housing including an optical sample well, a fluid inlet port and a conduit fluidly connecting the fluid inlet port and the optical sample well;b. a pump mechanism associated with the housing and fluidly connected to the fluid inlet port and the conduit; andc. a dried composition including a hemocyte lysate dried on the optical sample well.2. The cartridge of claim 1 , wherein the housing includes four optical sample wells claim 1 , each comprising the dried composition including the hemocyte lysate and further including a chromogenic substrate dried on the optical sample wells claim 1 , and wherein two of the four optical sample wells further include an agent representative of the microbial contaminant dried on the optical sample wells.3. The cartridge of claim 1 , wherein the housing includes a liquid impermeable membrane fluidly connected to the optical sample well.4. The cartridge of claim 1 , wherein the housing includes a top section and a bottom section claim 1 , mechanically connected to each other.5. The cartridge of claim 1 , wherein the pump mechanism is a two-position syringe claim 1 , wherein a first position creates a vacuum to introduce the sample into the fluid inlet port and the conduit claim 1 , and a second position provides transport from the conduit to the optical sample well.6limulus. The cartridge of claim 1 , wherein the hemocyte lysate is amoebocyte lysate.7. The cartridge of claim 2 , wherein the agent representative of the microbial contaminant is a bacterial endotoxin.8. A cartridge ...

BLUE COLLAGENASE ASSAY

A method of measuring soluble or insoluble cell or tissue-associated collagenase activity. The substrate includes native (fibrillar) collagen fragments that were stained with Coomassie Brilliant Blue R-250. Incubation with collagenase can be observed in real-time by the generation of digested smaller fragments. The degraded blue fragments are obtained by filtration through class fibers, onto which intact collagen fibrils are retained. The filtrate containing the blue collagen fragments is incubated with a detergent in order to extract the blue dye, and the mixture is centrifuged in order to separate the dye in the supernatant from the pellet, which contains de-stained collagen fragments and other insoluble materials contained in the test samples (such as bacterial cells or tissues). The amount of dye extracted is quantified by measuring the amount of dye extracted from these fragments, i.e., the absorbance at 600 nm using a spectrophotometer or an ELISA reader. 1. An assay for measuring soluble or insoluble cell or tissue-associated collagenase activity , comprising:obtaining dry collagen fibrils suitable for dying;staining said collagen fibrils with blue dye at saturation level;suspending said blue collagen fibrils in a collagenase substrate buffer;incubating said blue collagen fibril suspension with a test sample at 37° C. on a rotator, wherein collagenase activity results in production of blue collagen particulates that can be readily observed in a resulting mixture;filtering said mixture to retain said blue collagen fibrils, wherein said hlu collagen particulates are digested and collected in a resulting filtrate;extracting said blue dye from said filtrate to determine an amount of said digested blue collagen particulates; andmeasuring absorbance of said digested blue collagen particulates.2. An assay as in claim 1 , wherein the step of measuring absorbance is performed using a spectrophotometer at an optimal wave length of 600 nm.3. An assay as in claim 1 , ...

COMPOSITIONS AND METHODS FOR DIAGNOSIS OF SHOCK

Methods and kits for diagnosis and staging of shock, and especially non-septic shock are presented in which protease activities and/or volatile compounds are measured from a biological sample to so identify and/or stage shock. 1. (canceled)2. A method for detecting putrescine , cadaverine , or a combination thereof , in a human having septic shock or non-septic shock , the method comprising:(i) obtaining a breath sample from the human;(ii) applying the sample to a solid carrier or a liquid carrier;(iii) detecting putrescine, cadaverine, or a combination thereof, in the solid carrier or the liquid carrier by infrared spectroscopy, ultraviolet spectroscopy, or visible spectroscopy; thereby detecting putrescine, cadaverine, or the combination thereof in the human having septic shock or non-septic shock.3. A method for detecting a volatile organic amine compound in a human having a shock condition , the method comprising:(i) obtaining a breath sample from the human;(ii) applying the sample to a solid carrier or a liquid carrier;(iii) detecting the volatile organic amine compound in the solid carrier or the liquid carrier; thereby detecting the organic amine compound in the human having the shock condition.4. The method of claim 3 , wherein the volatile organic amine compound comprises putrescine.5. The method of claim 3 , wherein the volatile organic amine compound comprises cadaverine.6. The method of claim 3 , wherein the volatile organic amine compound comprises putrescine and cadaverine.7. The method of claim 3 , wherein the shock condition is septic shock.8. The method of claim 3 , wherein the shock condition is non-septic shock.9. The method of claim 3 , further comprising quantifying the amount of volatile organic amine compound in the solid carrier or the liquid carrier.10. The method of claim 3 , wherein step (i) comprises obtaining and concentrating the breath sample.11. The method of claim 3 , wherein step (iii) comprises detecting the volatile organic amine ...

KIT FOR DETECTING VIRUS

The present invention relates to a kit for detecting a virus, a composition for detecting a virus and a method for detecting a virus. According to the present invention, viruses may be detected with high efficiency at low cost within a short period of time. 1. A kit for detecting a virus , comprising:a biomolecule that specifically reacts with the surface protein of a virus; anda probe that reacts with the virus activated by the biomolecule.2. The kit of claim 1 , wherein the virus is influenza virus claim 1 , coronavirus or paramyxovirus.3. The kit of claim 1 , wherein the surface protein of a virus is hemagglutinin (HA) claim 1 , spike protein or F protein.4. The kit of claim 1 , wherein the biomolecule that specifically reacts with the surface protein of a virus is an enzyme.5. The kit of claim 4 , wherein the enzyme includes one or more selected from the group consisting of furin claim 4 , trypsin claim 4 , serine claim 4 , endoprotease and carboxypeptidase.6. The kit of claim 1 , wherein the probe is a micelle claim 1 , a polymersome claim 1 , a colloidsome claim 1 , a vesicle claim 1 , a liposome or a droplet.7. The kit of claim 1 , wherein the probe is selected from the group consisting of an amphiphilic particle comprising a marker; organic and inorganic particles; an antibody; an aptamer; and combinations thereof.8. The kit of claim 7 , wherein the marker is selected from the group consisting of a self-quenched dye claim 7 , a fluorescent dye claim 7 , an electrochemiluminescent material or combinations thereof.9. The kit of claim 7 , wherein the amphiphilic particle includes a hydrophilic polymer and a hydrophobic polymer.10. The kit of claim 9 , wherein the hydrophilic polymer includes one or more selected from the group consisting of polyalkyleneglycol (PAG) claim 9 , polyacrylic acid (PAA) claim 9 , polyacrylonitrile (PAN) claim 9 , polyethyleneoxide (PEO) claim 9 , polyvinylacetate (PVAc) claim 9 , polyvinylalcohol (PVA) claim 9 , polyvinylpyrrolidone ...

Enzymatic sample purification

An enzymatic purification method involves the introduction of a sample comprising a target analyte and amino acids into a porous matrix of a reaction chamber. The reaction chamber includes first pores and second pores. The first pores contain polypeptide synthesis enzymes that react with the amino acids to form polypeptides. First pores having a first size to be accessible by amino acids but inaccessible by the subsequently formed polypeptides. The second pores have a second size greater than the first size, are in contact with the first pores and form a series extending from within the reaction chamber to a waste chamber. The formed polypeptides are migrated through the series of second pores to the waste chamber. The target analyte of the sample is extracted from the reaction chamber.

METHOD FOR EVALUATING METABOLIZABLE ENERGY IN GOOSE DIET BY USING A SIMULATIVE DIGESTION GROSS ENERGY TECHNIQUE

Embodiments of the present disclosure belongs to the technical field of animal feed and provides a method for rapidly evaluating metabolizable energy of goose diet by using a technique of simulative digestion gross energy. By using the technical means combining the biological method and the simulative digestion gross energy technique, metabolizable energy of goose feed can be evaluated quickly. Based on the “stomach-small intestine” two-step enzymatic methods, it is the first time to establish a regression equation between the metabolizable energy change and fiber level in the cecum to rectify the value of simulative digestion gross energy in the cecal microbial digestion phase, making the simulative digestion gross energy technique more reasonable in the assessment of metabolizable energy in geese. Results show that the use of simulative digestion gross energy technique to assess the metabolizable energy of goose feed value is highly feasible. 1. A method for rapidly evaluating metabolizable energy of a goose diet by using a simulative digestion gross energy technique , comprising the following steps: a digestive enzyme in the simulated gastric fluid is pepsin, and the concentration of the pepsin 1475 U/ml;', 'a digestive enzyme in the simulated intestinal fluid is selected from trypsin, chymotrypsin, and amylase; and every 2000 ml of the simulated intestinal fluid contains 0.1900 g of trypsin, 0.0526 g of chymotrypsin and 4.45 ml of amylase solution;', 'a digestion temperature both in the gastric phase and the in intestinal phase is in a range of 40.5-41.5° C.; and', 'a digestion time in the gastric phase is 4-5 hours, and a digestion time in the intestinal phase is in a range of 12-16 hours; and, 'measuring and calculating, for the diet, a value of simulative digestion gross energy of one or more fiber levels by using a simulative digestion gross energy technique, wherein the calculation using the simulative digestion gross energy technique utilizes parameters ...

Clostridium Histolyticum Enzymes and Methods for the Use Thereof

The invention relates to recombinant nucleic acid and polypeptides encoding collagenase I and collagenase II, methods for the preparation thereof and methods for the use thereof. The invention also encompasses methods related to releasing a composition comprising collagenase prior to therapeutic administration. 1. A recombinant nucleic acid molecule comprising a polynucleotide having a sequence of SEQ ID NO: 1 or the complement of SEQ ID NO: 1.2. The recombinant nucleic acid of claim 1 , further comprising a heterologous regulatory sequence operably linked to the polynucleotide.3. The recombinant nucleic acid of claim 2 , wherein the regulatory sequence is a promoter.4. A recombinant nucleic acid molecule comprising a polynucleotide having a sequence of SEQ ID NO: 2 or the complement of SEQ ID NO: 2.5. The recombinant nucleic acid of further comprising a heterologous promoter operatively linked to the polynucleotide.6. A recombinant polypeptide encoded by the nucleic acid of .7. A recombinant polypeptide encoded by the nucleic acid of .8. (canceled)9. (canceled)10. A vector comprising the nucleic acid of .11. The vector of claim 10 , wherein the vector is a plasmid.12. A vector comprising the nucleic acid of .13. The vector of claim 12 , wherein the vector is a plasmid.14. A recombinant host cell comprising the vector or plasmid of .15. The recombinant host cell of claim 14 , wherein the host cell is selected from the group consisting of a bacterial cell claim 14 , a fungal cell claim 14 , an insect cell claim 14 , a plant cell and a mammalian cell.16E. coli, Streptomyces, Pseudomonas, Serratia marcescens, Salmonella typhimuriumlactococcus lactis.. The recombinant host cell of claim 15 , wherein the host cell is selected from the group consisting of claim 15 , a yeast cell claim 15 , plant cells claim 15 , thymocytes claim 15 , Chinese hamster ovary cell (CHO) claim 15 , COS cell claim 15 , and17. (canceled)18. (canceled)19. A method of producing collagenase I ...

CO-TRANSLATIONAL ACTIVATION OF A TRANSCRIPTION FACTOR BY PORTEOLYTIC CLEAVAGE AND METHODS OF USE

A method for measuring expression of autoregulatory molecules within living cells is provided. An autoregulatory molecule and marker construct is expressed in vivo, where the marker is cleaved from the construct during translation. The method comprises the expression of a construct having an autoregulatory molecule bound to a measurable expression marker by a cleavable linker. The cleavable linker is the substrate of a protease, which acts on its substrate in vivo during translation. Cleavage during translation, allows the autoregulatory molecule to fold normally as it would in its native form. The measurable marker is released and available for detection upon cleavage by the protease. As a result, the concentration of the measurable marker is directly related to the level of expression of the autoregulatory molecule. 1. A method for measuring expression of an autoregulatory molecule , comprising:expressing a construct in a cell, wherein the construct comprises the autoregulatory molecule, a measurable marker, and a cleavable substrate in a single molecule;expressing a protease capable of cleaving the cleavable substrate in the cell, wherein the protease cleaves the cleavable substrate during translation allowing the autoregulatory molecule to fold into a functional molecule; andevaluating the cell for the presence of the measurable substrate.2E. coli. The method of claim 1 , wherein the cell is an cell.3. The method of claim 1 , wherein the autoregulatory molecule is CI.4. The method of claim 1 , wherein the measurable marker is selected from the group consisting of fluorescent peptides claim 1 , colorimetric compounds claim 1 , chemiluminescent peptides claim 1 , and combinations thereof.5. The method of claim 5 , where the fluorescent peptide is selected from the group consisting of yellow fluorescent protein (YFP) claim 5 , blue fluorescent protein (BFP) claim 5 , green fluorescent protein (GFP) claim 5 , red fluorescent protein (RFP) and fluorescing mutants ...

N-end rule protease activity indication methods and uses thereof

A cell based assay for detection for protease activity is disclosed. In the assay a cell is engineered to express a protease substrate with at least one label, preferably on its C-terminus. Cleavage of the substrate by the protease that recognizes it results in a C-terminal fragment and a N-terminal fragment, where the fragment having the label is subject to ubiquitin proteasome degradation. The assay measures the disappearance of the label due to degradation of the fragment to which it is attached. A cell free assay is also described for detection of protease activity. In the cell free assay, the protease substrate is expressed in a solution that includes the elements of the ubiquitin proteasome pathway for degradation of the fragment. The assay measures the disappearance of the label attached to the fragment that results from cleavage by the protease.

Cellular Libraries of Peptide Sequences (CLiPS) and Methods of Using the Same

The present invention provides compositions including peptide display scaffolds that present at least one candidate peptide and at least one detectable moiety in at least one of the N-terminal and C-terminal candidate peptide presenting domains that when expressed in a cell are accessible at a surface of the cell outermembrane. In addition, the present invention also provides kits and methods for screening a library of cells presenting the candidate peptides in peptide display scaffolds to identify a ligand for an enzyme. 146-. (canceled)47. A peptide display scaffold comprising a fusion protein comprising the formula:{'br': None, 'sub': 1', '1', '2, '[D-C]-TM-[D],'}wherein TM is a circularly permuted transmembrane protein;{'sub': '1', 'wherein Ccomprises a candidate peptide;'}{'sub': '1', 'wherein Dis a first detectable moiety and is a heterologous peptide or polypeptide;'}{'sub': '2', 'wherein Dis a second detectable moiety and is a heterologous peptide or polypeptide;'}{'sub': 1', '2, 'wherein Dand Dare not the same; and'}{'sub': 1', '2', '1, 'wherein D, D, and C, when the peptide display scaffold is expressed in a host cell, are exposed at an extracellular surface of the host cell outer membrane.'}48. The peptide display scaffold of claim 47 , wherein the fusion protein comprises at least one linker claim 47 , wherein the linker is between Cand TM or between Dand TM.49. The peptide display scaffold of claim 47 , wherein detectable moieties Dand Dare affinity tags.50. The peptide display scaffold of claim 47 , wherein when Dprovides a detectable Dsignal claim 47 , Ddoes not provide a detectable Dsignal above a background level of a detectable Dsignal.51. The peptide display scaffold of claim 47 , wherein the Ccomprises a member of a library of candidate peptide substrates for an enzyme.52. The peptide display scaffold of claim 47 , wherein the fusion protein comprises a linker between the Cand the TM and a linker between the Dand the TM.53. The peptide display ...

ELECTROCHEMICAL DETECTION OF PROTEASES USING AC VOLTAMMETRY ON NANOELECTRODE ARRAYS

An electrochemical method for measuring the activity of enzymes using nanoelectrode arrays fabricated with vertically aligned carbon nanofibers. Short peptide substrates specific to disease-related enzymes are covalently attached to the exposed nanofiber tips. A redox moiety, such as ferrocene, can be linked at the distal end of the nanofibers. Contact of the arrays with a biological sample containing one or more target enzymes results in cleavage of the peptides and changes the redox signal of the redox moiety indicating the presence of the target enzymes. 1. A nanoelectrode array comprising a substrate having a plurality of carbon nanofibers extending vertically therefrom , said array being capable of detecting the presence of one or more enzymes present within a biological sample.2. The array according to claim 1 , wherein said nanofibers comprise a proximal end that is attached to said substrate and a distal end that is spaced from said substrate.3. The array according to claim 2 , wherein said nanofibers are encapsulated claim 2 , except for at least one of said distal ends claim 2 , within a matrix of an insulative material.4. The array according to claim 3 , wherein said insulative material comprises silicon dioxide.5. The array according to claim 2 , wherein said distal end of at least one of said nanofibers comprises a peptide or peptide residue covalently attached thereto.6. The array according to claim 5 , wherein said peptide or peptide residue is capable of being cleaved by a protease enzyme that is expressed by a cancer-causing cell.7. The array according to claim 6 , wherein said peptide is a tetrapeptide selected from the group consisting of Ala-Ala-Asn-Leu (SEQ ID NO:1) claim 6 , Leu-Arg-Phe-Gly (SEQ ID NO:2) claim 6 , and Pro-Leu-Ser-Leu (SEQ ID NO:3).8. The array according to claim 5 , wherein said peptide or peptide residue is a ferrocenyl peptide or peptide residue.9. The array according to claim 5 , wherein said peptide or peptide residue is ...

NOVEL DIAGNOSTIC AND THERAPEUTIC TARGET IN INFLAMMATORY AND/OR CARDIOVASCULAR DISEASES

Methods for diagnosing inflammatory and/or cardiovascular diseases by assaying for Fibroblast Activation Protein (FAP) expression in a body fluid is provided as well as therapeutic means based thereon. 124.-. (canceled)26. The method of claim 25 , wherein the cardiovascular disease or condition is atherosclerosis claim 25 , stroke claim 25 , acute coronary syndrome claim 25 , heart attack claim 25 , chronic liver disease claim 25 , cerebral venous thrombosis claim 25 , coronary artery disease claim 25 , deep venous thrombosis or pulmonary embolism.27. The method of claim 26 , wherein the acute coronary syndrome is myocardial infarction.28. The method of claim 25 , wherein the disease is thrombosis.29. The method of claim 25 , wherein the condition is vulnerable atherosclerosis plaques or atherothrombosis.30. The method of claim 25 , wherein said capture antibody is a monoclonal or polyclonal antibody.31. The method of claim 30 , wherein said monoclonal antibody is a mouse anti-human FAP antibody.32. A diagnostic composition for determining an amount of Fibroblast Activation Protein (FAP) in a sample of blood taken from a patient suffering from or at risk of suffering from a cardiovascular disease or condition claim 30 , comprising an antibody raised against a catalytic domain of FAP and/or a monoclonal antibody raised against FAP.33. The diagnostic composition of claim 32 , wherein the composition comprises the antibody raised against a catalytic domain of FAP and the monoclonal antibody raised against FAP claim 32 , wherein the antibody raised against a catalytic domain of FAP is a capture antibody and the monoclonal antibody raised against FAP is a detection antibody for detecting the capture antibody.34. The diagnostic composition of claim 33 , wherein the capture antibody is a mouse anti-human FAP antibody.35. The diagnostic composition of claim 33 , wherein the capture antibody is a monoclonal antibody.36. The diagnostic composition of claim 34 , wherein the ...

BIOANALYTICAL ANALYSIS OF SITE-SPECIFIC ANTIBODY DRUG CONJUGATES

Methods to rapidly and accurately detect, characterize, measure, and quantify site-specific antibody drug conjugates, that may be present in pre-clinical animal biological samples, or human biological samples, including plasma/serum and tissue samples. 124-. (canceled)25. A method of evaluating an antibody drug conjugate (ADC) , wherein the ADC is suspended in whole blood , serum , plasma , or tissue of a mammal selected from a human , a cynomolgus monkey , a rat , and a mouse , comprising: a cysteine amino acid residue,', 'a selenocysteine amino acid residue,', 'a glutamine amino acid residue,', 'a non-naturally occurring amino acid residue, and', 'a sugar-modified glycan residue,, 'a. digesting an ADC comprising at least one drug moiety linked to an antibody at a recombinantly-engineered site selected fromwith IdeS protease that cleaves the ADC, to form a digested ADC composition comprising at least one peptide fragment that is not linked to the at least one drug moiety, and at least one peptide fragment that is linked to the at least one drug moiety; and,b. analyzing the digested ADC composition by at least one of RP-LC, RP-LC/MS, and LC-MS/MS to detect at least one peptide fragment that is not linked to the at least one drug moiety.26. The method of claim 25 , wherein the antibody is selected from an IgG antibody claim 25 , an antibody fragment claim 25 , a human or humanized antibody claim 25 , a glycosylated or phosphorylated antibody claim 25 , and a cysteine-engineered antibody.27. The method of claim 25 , wherein the antibody portion of the ADC is an antibody which binds to one or more tumor-associated antigens or cell-surface receptors selected from (1)-(53):(1) BMPR1B (bone morphogenetic protein receptor-type IB);(2) E16 (LAT1, SLC7A5);(3) STEAP1 (six transmembrane epithelial antigen of prostate);(4) MUC16 (0772P, CA125);(5) MPF (MPF, MSLN, SMR, megakaryocyte potentiating factor, mesothelin);(6) Napi2b (NAPI-3B, NPTIIb, SLC34A2, solute carrier family 34 ( ...

COMPOSITIONS AND METHODS FOR IMPROVING SENSITIVITY IN CELL BASED ASSAYS

Compositions and methods are provided that reduce the time required for detection of botulinum neurotoxins in cell-based assays. In one aspect an isoquinolynyl compound can be used for this purpose. In such cell-based assays the cell can include an enzyme that facilitates degradation of the reporter significantly faster after the cleavage than before the cleavage, and presence of the Botulinum toxin correlates with reduction of the signal from a baseline signal. The cell can advantageously express both the construct that includes the reporter, and an enzyme that facilitates the degradation. 1. A method of detecting the presence of a Botulinum toxin of a specified Botulinum toxin serotype , comprising:providing an assay with a cell having (a) a molecule with a cleavage sequence that is cleaved by a light chain of said Botulinum toxin and (b) a reporter that reports a signal responsive to cleavage of said molecule at said cleavage sequence induced by the light chain of said Botulinum toxin; andincubating the cell in a culture media that contains a compound effective to decrease time for performance of the assay by at least 66% relative to a corresponding assay performed in the absence of the compound;treating the cell with said Botulinum toxin, then detecting the signal, and correlating the signal with the presence or absence of said Botulinum toxin,wherein said compound is an isoquinolynyl compound that inhibits protein kinase C (PKC), and wherein said cleavage sequence comprises a SNARE protein, SNARE motif, or SNARE mutein.2. The method of claim 1 , wherein said compound comprises 1-(5-isoquinolinyl sulfonyl)-2-methylpiperazine dichloride (H7).3. The method of claim 1 , wherein said reporter is an artificial construct expressed in the cell.4. The method of claim 1 , wherein said molecule includes a first fluorophore and the cleavage sequence.5. The method of claim 4 , wherein said first fluorophore is selected from the group consisting of Yellow Fluorescent Protein ...

DEVICE FOR THE DIAGNOSIS OF INFLAMMATORY TISSUES IN DENTAL APPLICATIONS

The document proposes a diagnostic chewing gum for identifying the presence of inflammatory tissues in the mouth, in particular in or adjacent to the mandible, the maxilla, an implant or the teeth of a user, comprising a base material or particles () embedded and/or attached to said base material; an element (-), like e.g. a releasable flavor molecule, attached to said base material and/or said particles, for the generation of a change in the chewing gum directly detectable by the user; wherein the element (-) generates the change upon direct or indirect contact with a marker (), e.g. a proteolytic enzyme, which is released by inflammatory tissue in response to bacterial mediators. 1. A diagnostic chewing gum for identifying the presence of inflammatory tissues in the mouth , comprisinga base material or particles embedded, attached, or both, to said base material;an element attached to said base material or said particles or both for the generation of a change in the chewing gum directly detectable by the user;wherein the element generates the change upon direct or indirect contact with a marker which is released by inflammatory tissue in response to bacterial mediators.2. The chewing gum according to claim 1 , wherein the marker inducing the change is a proteolytic enzyme released by inflammatory tissue.3. The chewing gum according to claim 1 , wherein the marker inducing the change is a matrix metalloproteinase.4. The chewing gum according to claim 1 , wherein the element is a molecule or molecular assembly which claim 1 , upon direct or indirect contact with the marker undergoes a color change perceivable by the naked eye of the user claim 1 , and which is embedded or attached to the base material or to particles embedded or attached to said base material.5. The chewing gum according to claim 1 , wherein the element is a flavor molecule releasably attached to the base material or to particles embedded or attached to said base material.6. The chewing gum ...

QUANTITATIVE DETECTION METHOD FOR SNAKE VENOM THROMBIN-LIKE ENZYME (SVTLE)

The present invention relates to the technical field of chemical analysis and quantitative detection, in particular to a quantitative detection method for snake venom thrombin-like enzyme (SVTLE) from . The quantitative detection method for the SVTLE includes the following steps of taking a reference substance of marker peptide for the SVTLE from with an amino acid sequence of LDSPVSNSAHIAPLSLPSSAPSVGSVCR, and preparing a series of reference solutions with different concentrations; adding the reference solutions in test solutions respectively for enzymolysis, and then taking a supernatant after enzymolysis as a series of solutions to be detected; and adding the solutions to be detected in a liquid chromatogram-mass spectrometer, and then selecting a qualitative ion pair and a quantitative ion pair to detect contents of marker peptide in the solutions to be detected. 1. A quantitative detection method for snake venom thrombin-like enzyme (SVTLE) , comprising the following specific steps of:{'i': 'Agkistrodon halys pallas', '(1) taking a reference substance of a marker peptide for SVTLE from with an amino acid sequence of LDSPVSNSAHIAPLSLPSSAPSVGSVCR, and dissolving and diluting the reference substance to prepare a series of reference solutions with different concentrations;'}(2) taking a to-be-detected sample, and dissolving the to-be-detected sample to prepare a test solution;(3) taking the series of reference solutions with different concentrations in step (1), adding the test solutions in step (2) respectively for uniform mixing, conducting enzymolysis with trypsin, and then taking supernatants after enzymolysis as a series of solutions to be detected;(4) injecting the solutions to be detected in step (3) into a liquid chromatogram-mass spectrometer respectively, conducting multiple-reaction monitoring by adopting an electrospray positive ion mode, and conducting detection with tricharged 935.8→602.3 as a qualitative ion pair and tricharged 935.8→861.4 as a ...

BIOMARKER FOR SELECTING A SUBJECT FOR APPLICATION OF AN ANTI-C-MET ANTIBODY

A method of selecting a subject for administration of an anti-c-Met antibody, comprising (i) determining the presence or the amount of a ubiquitin peptidase, (ii) determining the presence or expression level of a ubiquitin peptidase coding gene, or (iii) measuring the activity of a ubiquitin peptidase, in a biological sample; as well as related methods. 1. A method of selecting a subject for administration of an anti-c-Met antibody , comprising(i) determining the presence or the amount of a ubiquitin peptidase or a ubiquitin peptidase-coding gene, or measuring the activity of a ubiquitin peptidase, in a biological sample from a subject; and(ii) selecting the subject for administration of an anti-c-Met antibody if the amount of ubiquitin peptidase or ubiquitin peptidase-coding gene, or activity level of ubiquitin peptidase, in the sample is absent or at a low level as compared to a reference sample in which the anti-c-Met antibody has no effect or has a resistance to the anti-c-Met antibody, or the activity level has a score measured by immunohistochemical staining of “−”, “0”, or “+1.”2. The method according to claim 1 , wherein the ubiquitin peptidase is ubiquitin specific peptidase 8 (USPS).3. The method according to claim 1 , wherein the anti-c-Met antibody is an antibody or an antigen-binding fragment thereof comprising:{'sup': rd', 'th', 'st', 'th, 'at least one heavy chain complementarity determining region (CDR) selected from the group consisting of (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 4; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 2, or an amino acid sequence comprising 8-19 consecutive amino acids of SEQ ID NO: 2 including the 3to 10positions of SEQ ID NO: 2; and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 6, SEQ ID NO: 85, or an amino acid sequence comprising 6-13 consecutive amino acids of SEQ ID NO: 85 including the 1to 6positions of SEQ ID NO: 85, or a heavy chain variable region ...

GUT FLORA-DERIVED EXTRACELLULAR VESICLES, AND METHOD FOR SEARCHING FOR A DISEASE MODEL, VACCINE, AND CANDIDATE DRUG AND FOR DIAGNOSIS USING THE SAME

The present application relates to a composition including gut flora-derived extracellular vesicles, and to an animal disease model using same. In addition, the present application relates to a method for using the gut flora-derived extracellular vesicles to efficiently search for a candidate drug which may prevent or treat diseases that occur due to gut flora-derived extracellular vesicles, and to a vaccine which can efficiently prevent or treat infections caused by gut flora or diseases that occur due to gut flora-derived extracellular vesicles. Further, the development of diagnostic technology to discover, using the gut flora-derived extracellular vesicles of the present application, the etiology of diseases that occur due to gut flora-derived extracellular vesicles, can be achieved. 16.-. (canceled)7. A method of assessing a subject's susceptibility to one or more diseases , the method comprising:providing a database listing pre-identified pathogenic microorganisms and one or more substances of the pre-identified pathogenic microorganisms;providing a sample comprising gut flora isolated from a subject;processing the sample to obtain a concentrated population of extracellular vesicles contained in the gut flora;processing at least part of the concentrated population to determine if the concentrated population contains at least one substance of a first one of the pre-identified pathogenic microorganisms listed in the database; andonce determined that at least one substance is of the first pre-identified pathogenic microorganism, further determining that the subject is potentially susceptible to one or more diseases that are associated with the first pre-identified pathogenic microorganism.8. The method of claim 7 , wherein the at least one substance comprises at least one genetic substance claim 7 , wherein the database further comprises sequences of genetic substances of at least part of the pre-identified pathogenic microorganisms claim 7 , sequencing at least ...

ASSAYS FOR DETECTION OF GLYCOSAMINOGLYCANS

Disclosed herein are novel methods, assays and kits useful for the diagnosis and monitoring of subjects with mucopolysaccharidoses (MPS). The methods, assays and kits are particularly useful for detecting the presence of one or more glycosaminoglycans which correlate to MPS and its severity in a variety of biological samples. 1. A method for determining the concentration of one or more glycosaminoglycans in a sample comprising: (a) combining a serine protease , a labeled substrate for the serine protease , an inhibitor of the serine protease , and a sample suspected of comprising one or more glycosaminoglycans in an assay buffer solution comprising NaCl , under conditions and for a time suitable for cleavage of the labeled substrate by the serine protease to produce a detectable signal; (b) detecting the detectable signal; and (c) comparing the amount of detectable signal with a standard to determine the concentration of the one or more glycosaminoglycans in the sample;wherein the serine protease is a serine protease of the clotting cascade;wherein the inhibitor of the serine protease is selected from the group consisting of heparin cofactor II and antithrombin III;wherein the one or more glycosaminoglycans are selected from the group consisting of dermatan sulfate (DS) and heparan sulfate (HS); andwherein the sensitivity of the method is modulated by the concentration of NaCl in the assay buffer solution.2. A method according to wherein the serine protease is selected from the group consisting of the serine proteases shown in and .3. A method according to wherein the labeled substrate is a chromogenic or fluorogenic substrate.4. A method according to wherein the serine protease is thrombin and the labeled substrate is a chromogenic thrombin substrate.5. A method according to wherein the sample is treated to inactivate all but one of the glycosaminoglycans in the sample.6. A method according to wherein the sample is treated with chondroitinase B and the active ...

QUANTIFICATION OF MISFOLDED TNFR2:FC

The present invention is directed to methods for determining the relative amount of wrongly disulphide bridged TNFR2:Fc in a sample of TNFR2:Fc, a fusion protein which is used in a variety of therapeutic applications. In addition, the invention pertains to a method for purifying TNFR2:Fc using said method for determining the percentage of wrongly disulphide bridged TNFR2:Fc, and to TNFR2:Fc compositions obtained thereby. 1. A method for determining Cys-Cysdisulfide bridged TNFR2:Fc in a sample comprising Cys-Cys/Cys-Cysdisulfide bridged TNFR2:Fc and Cys-Cysdisulfide bridged TNFR2:Fc , wherein the method comprises the steps of:{'sub': 78', '88', '74', '88', '78', '96, '(f) providing a sample comprising a mixture of Cys-Cysdisulfide bridged TNFR2:Fc and Cys-Cys/Cys-Cysdisulfide bridged TNFR2: Fc;'}(g) denaturing and alkylating the sample of step (a);(h) subjecting the sample resulting from step (b) to tryptic digestion;{'sub': 78', '88, '(i) subjecting the sample resulting from step (c) to HPLC, thereby separating fragments indicative of Cys-Cysdisulfide bridged TNFR2:Fc; and'}{'sub': 78', '88', '74', '78', '88', '96, '(j) conducting a peak integration for the peak indicative of Cys-Cysdisulfide bridged TNFR2:Fc and for a peak not affected by disulfide bridging of Cys, Cys, Cysand Cys, as obtained from step (d);'}wherein the amino acid sequence of the TNFR2 part of TNFR2:Fc has at least 97% identity to the amino acids 1-235 of the amino acid sequence of SEQ ID NO: 1.2. The method of claim 1 , wherein the amino acid sequence of the TNFR2:Fc applied to step (a) has at least 97% identity to the amino acid sequence of SEQ ID NO: 3 (etanercept).3. The method of claim 1 , wherein the peak not affected by disulfide bridging of Cys claim 1 , Cys claim 1 , Cysand Cysis not affected by disulfide bridging at all and indicative of the total TNFR:Fc in the sample.4. The method of claim 3 , wherein the fragments indicative of Cys-Cysdisulfide bridged TNFR2:Fc comprise the amino ...

METHOD FOR IN-GEL VISUAL DETECTION OF BIOANALYTES

The present invention relates to a method for in-gel visual detection and quantitative detection of proteins in activity based protein profiling (ABPP) using horseradish peroxidase mimic Fe-TAML complex of ligand as a catalytic probe. The invention further relates to kit comprising compounds of formula (I) and method for the detection of bioanalytes using kit comprising compounds of formula (I). 2. The method as claimed in claim 1 , wherein said method is conducted by catalytic signal amplification in activity based protein profiling (ABPP).3. The method as claimed in claim 1 , wherein claim 1 , the compound of formula I is a horseradish peroxidase mimic Fe-TAML complex of a ligand.4. The method as claimed in claim 1 , wherein claim 1 , the method further comprises:a) treating the mixture of bioanalyte with an inhibitor probe that has an organoazide handle so as to obtain an azide labelled bioanalyte;b) subjecting the mixture of step (a) to click reaction with an alkyne-tagged biuret-Fe-TAML;{'sub': 2', '2, 'c) running the mixture of step (b) on a polyacrylamide gel followed by probing with HOand 3,3′,5,5′-Tetramethylbenzidine (or related probes) and'}d) observing blue colored band for the bioanalyte.5. The method as claimed in claim 4 , wherein claim 4 , the organoazide is maleimide-azide.6. The method as claimed in claim 4 , wherein claim 4 , the bioanalyte is selected from the group proteins claim 4 , enzymes claim 4 , antibodies claim 4 , or nucleic acids.8. The kit as claimed in claim 7 , wherein said bio-analyte is selected from azide labeled Bovine Serum Albumin(BSA-N) or a serine protease.10. The gel as claimed in claim 9 , wherein claim 9 , the compound is biuret-Fe-TAML complex.11. The gel as claimed in claim 9 , wherein claim 9 , the gel is a polyacrylamide gel.12. The gel as claimed in claim 10 , wherein claim 10 , the gel is a polyacrylamide gel.13. The gel as claimed in claim 9 , wherein claim 9 , the bioanalyte is selected from proteins claim 9 , ...

Translocation of a polymer through a nanopore

Embodiments of the present disclosure are directed to methods, systems and devices, for analyzing the molecules. For example, in some embodiments, a system is provided which includes a first volume of conducting fluid, a second volume of conducting fluid, an orifice in communication with said first and second volumes of fluid, and means for applying an electric potential difference between said first and second volumes of fluid. In some such embodiments, a conjugate product is provided which comprises charged polymers each having attached thereto at least one first molecule for analysis, where the product carries a predetermined charge greater than the charge on the first molecule, and upon dissolving a product in the first volume of fluid, the product is directed into the orifice.

BRET SENSOR MOLECULES FOR DETECTING HYDROLASES

The present invention relates to bioluminescence resonance energy transfer sensor molecules having the structure R-L-R—B or B—R-L-R, wherein Ris a bioluminescent protein, L is a linking element, Ris a non-protein acceptor domain and B is a blocking group, and wherein Rbound to B comprises a hydrolysable bond which produces a change in BRET when hydrolysed. The invention also discloses a method of detecting a hydrolase by contacting a sample with a molecule B—R, then contacting with a compound R-L or L-Runder conditions to cause attaching of Rto L, and detecting a change in the BRET ratio. Specifically exemplified sensors comprise luciferase and fluorescein diacetate, which is hydrolysed by an esterase. The invention also discloses luciferase enzymes derived from RLuc8 by removing cysteine residues. 2. The sensor molecule of claim 1 , wherein the blocking group stabilises the acceptor domain in a low-fluorescent or non-fluorescent state.3. The sensor molecule of or claim 1 , wherein the blocking group comprises a phosphate containing moiety claim 1 , sugar containing moiety claim 1 , amino acid containing moiety claim 1 , nucleotide claim 1 , nucleoside claim 1 , ester or ether.4. The sensor molecule of any one of to claim 1 , wherein the linking element comprises an alkyl chain claim 1 , glycol claim 1 , ether claim 1 , polyether claim 1 , polyamide claim 1 , polyester claim 1 , peptide claim 1 , polypeptide claim 1 , amino acid or polynucleotide.5. The sensor molecule of claim 4 , wherein the linking element comprises a polypeptide.6. The sensor molecule of claim 5 , wherein R-L or L-Rare a single polypeptide.7. The sensor molecule of or claim 5 , wherein the linking element comprises a cysteine residue and/or a lysine residue.8. The sensor molecule of claim 7 , wherein Ris attached to the linking element via the cysteine residue.9. The sensor molecule of any one of to claim 7 , wherein Ris selected from an Alexa Fluor dye claim 7 , Bodipy dye claim 7 , Cy dye ...

LIQUID PROCESSING SYSTEM AND LIQUID PROCESSING METHOD

A liquid processing system to process liquid biological material includes: a trunk provided turnable on an axis, set within a predetermined work space; a first arm provided to the trunk and having at least three degrees of freedom or higher degrees of freedom; a second arm provided to the trunk and having at least three degrees of freedom or higher degrees of freedom; a driving mechanism configured to drive each of the trunk, the first arm, and the second arm; and physiochemical equipment situated within the work space and within the range of movement of at least one of the first and the second arm. The driving mechanism is operated by teaching playback based on the positions and shapes of the physiochemical equipment, and the biological material is processed using the physiochemical equipment. 1. A liquid processing system to process liquid biological material , the system comprising:a trunk provided turnable on an axis, set within a predetermined work space;a first arm provided to the trunk and having at least three degrees of freedom or higher degrees of freedom;a second arm provided to the trunk and having at least three degrees of freedom or higher degrees of freedom;a driving mechanism configured to drive each of the trunk, the first arm, and the second arm; andphysiochemical equipment situated within the work space and within the range of movement of at least one of the first and the second arm;wherein the driving mechanism is operated by teaching playback based on the positions and shapes of the physiochemical equipment, and the biological material is processed using the physiochemical equipment.2. The liquid processing system according to ;wherein the first arm includes a first robot hand to handle the physiochemical equipment;and wherein the second arm includes a second robot hand to handle the physiochemical equipment;and wherein a plurality of physiochemical equipment to perform processing different from one another is situated within a range of movement ...

DETERMINING THE CONDITION OF A WOUND

A product for monitoring the condition of the wound comprising a biologically inert matrix which absorbs wound exudate and one or more reagents on or in the matrix for measuring one or more markers comprised within the wound exudate. A change in the one or more reagents caused by the one or more markers comprised within the wound exudate provides a visual indication of an alteration in the condition of the wound. Companion wound dressings, kits and methods are also provided. 2. The product according to wherein the one or more reagents comprise a complete test unit integrated on or in the matrix.3. The product according to wherein the one or more reagents form a discrete reaction zone on or within the matrix.4. The product according to wherein the alteration is a deterioration.5. The product according to wherein the matrix is able to absorb and retain a volume of wound exudate sufficient for further analysis of the wound exudate.6. The product according to wherein the matrix has the capacity to absorb a volume of at least 0.2 ml wound exudate.7. The product according to wherein the matrix is dimensioned to facilitate positioning between a wound dressing and the wound.8. The product according to wherein the matrix comprises:(i) a first matrix portion comprising one or more reagents on or in the matrix portion for measuring one or more markers comprised within the wound exudate; and(ii) a second matrix portion which is able to absorb and retain a volume of wound exudate sufficient for further analysis of the wound exudate.9. The product according to wherein:(a) the two matrix portions are laminated together; or(b) the two matrix portions are separate, independent components.10. The product according to wherein the surface of the first matrix portion that is not in contact with the second matrix portion is coated or surrounded by a transparent protective layer.11. The product according to wherein the matrix does not measurably alter the condition of the exudate or its ...

Labile Linkers for Biomarker Detection

Disclosed herein are methods and compositions for electronic detection and/or quantification of enzymes or enzymatic activity in a sample using a pore system. 1. A method of detecting the presence or absence of a target molecule suspected to be present in a sample , comprising:contacting the sample with a fusion molecule comprising a cleavable linker, wherein said cleavable linker is specifically cleaved in the presence of said target molecule;loading said sample into a device comprising a nanopore, wherein said nanopore separates an interior space of the device into two volumes;configuring the device to pass a polymer scaffold through said nanopore, wherein a first portion of said fusion molecule is bound to said polymer scaffold, wherein a second portion of said fusion molecule is bound to a payload molecule, and wherein the device comprises a sensor configured to identify objects passing through the nanopore; anddetermining with the sensor whether the cleavable linker has been cleaved, thereby detecting the presence or absence of the target molecule in said sample.2. The method of claim 1 , wherein contacting the sample with said fusion molecule is performed prior to loading said sample into said device.3. The method of claim 1 , wherein loading said sample into said device is performed prior to contacting the sample with said fusion molecule.4. The method of claim 1 , wherein said fusion molecule comprises a polymer scaffold binding domain.5. The method of claim 4 , further comprising contacting the sample with a polymer scaffold.6. The method of claim 4 , further comprising binding said polymer scaffold to said polymer scaffold binding domain.7. The method of claim 6 , wherein said polymer scaffold is bound to said polymer scaffold binding domain via a covalent bond claim 6 , a hydrogen bond claim 6 , an ionic bond claim 6 , a van der Waals force claim 6 , a hydrophobic interaction claim 6 , a cation-pi interaction claim 6 , a planar stacking interaction claim ...

MULTIPLEX ENZYME ASSAY USING MASS SPECTROMETER-BASED FLOW CYTOMETER

The present invention generally relates to methods for the detection of enzymes using elemental analysis. 1. A method for determining protease activity in a biological fluid , the method comprisingattaching a coded bead to a first amino acid of a peptide substrate to form an immobilized peptide substrate, the peptide substrate comprising the first amino acid and a last amino acid and being a substrate for a protease enzyme:attaching an element tag to the last amino acid of the peptide substrate to form an immobilized tagged peptide substrate:incubating the immobilized, tagged peptide substrate with a biological fluid:detecting the element tag and the coded bead in the biological fluid by elemental analysis: anddetermining protease activity within the biological fluid based on detecting the element tag.2. The method of claim 1 , wherein detecting the element tag and the coded bead comprises using mass cytometry.3. The method of or claim 1 , wherein the peptide substrate comprises two or more peptide substrates claim 1 , wherein the two or more peptide substrates are substrates for two or more different protease enzymes.4. The method of any one of to claim 1 , wherein detecting the element tag and the coded bead comprises quantifying the elemental tags using mass cytometry.5. The method of any one of to claim 1 , further comprising separating the immobilized peptide substrate from the biological fluid to form an analyte solution and detecting the element tag.6. The method of claim 5 , wherein the element tag is detected by mass cytoinetry.7. The method of any one of to claim 5 , wherein the coded bead comprises a unique element or a unique combination of elements for the coded bead.8. The method of any one of to claim 5 , wherein the first amino acid of the peptide substrate is a C-terminal amino acid.9. The method of any one of to claim 5 , wherein the first amino acid of the peptide substrate is a N-terminal amino acid.10. The method of any one of to claim 5 , ...

STABLE VAMP REPORTER ASSAY

The present invention provides a polypeptide comprising an N-terminal polypeptide domain having luciferase activity and a C-terminal polypeptide domain having VAMP1, VAMP2 or VAMP3 activity where VAMP stands for vesicle-associated membrane protein. Corresponding nucleic acid molecules, expression vectors and genetically modified cells are also provided. The invention also provides methods and uses of the same. 1. A polypeptide comprising an N-terminal polypeptide domain having luciferase activity and a C-terminal polypeptide domain having VAMP1 , VAMP2 or VAMP3 activity.2. The polypeptide of claim 1 , wherein the polypeptide domain having VAMP2 activity comprises the amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 2 claim 1 , or a conservative amino acid sequence variant thereof.3. The polypeptide of claim 1 , wherein the polypeptide domain having VAMP1 activity comprises the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 1 claim 1 , or a conservative amino acid sequence variant thereof.4. The polypeptide of claim 1 , wherein the polypeptide domain having VAMP3 activity comprises the amino acid sequence of SEQ ID NO:9 or SEQ ID NO: 3 claim 1 , or a conservative amino acid sequence variant thereof.5. The polypeptide of claim 1 , wherein the polypeptide domain having luciferase activity comprises the amino acid sequence of SEQ ID NO: 4 or a conservative amino acid sequence variant thereof.6. A nucleic acid molecule encoding the polypeptide of .7. An expression vector comprising the nucleic acid molecule of .8. A genetically modified cell comprising the nucleic acid molecule of claim 6 , or an expression vector comprising the nucleic acid molecule of .9. The genetically modified cell of claim 8 , wherein the cell is selected from the group consisting of a SiMa neuroblastoma cell claim 8 , a LANS neuroblastoma cell claim 8 , a NG108 neuroblastoma cell claim 8 , an immortalised neuron and a primary neuron.1013.-. (canceled)14. A method of detecting neurotoxin ...

GLYCATED HEMOGLOBIN MEASUREMENT

Described herein are devices, systems, and methods used to measure glycated hemoglobin. 1. A slide comprising: a first film layer comprising a cross-linked gel, wherein said cross-linked gel comprises a detection agent, a fructosyl oxidase, an interference prevention agent, and a peroxidase;', 'a second film layer comprising a first gel; and', 'a third film layer comprising a lysing agent, a denaturing agent and a protease., 'a stack of film layers comprising, from bottom to top,'}2. The slide of claim 1 , wherein said lysing agent is selected from the groups consisting of octylphenol ethoxylate (TRITON X-100) claim 1 , TWEEN (TWEEN 20) claim 1 , sodium dodecyl sulfate (SDS) claim 1 , cetyltrimethylammonium bromide (CTAB) claim 1 , tetradecyltrimethylammonium bromide (TTAB) claim 1 , polyoxyethylene lauryl ethers (POEs) and NONIDET P-40 (NP-40).3. The slide of claim 1 , wherein said denaturing agent is one or more of sodium nitrite or N-lauroylsarcosine (NLS).4. The slide of claim 1 , wherein said protease is a metalloproteinase claim 1 , an endoprotease or an exoprotease.5. The slide of claim 1 , wherein said detection agent is selected from the group consisting of N-carboxymethylaminocarbonyl)-4 claim 1 ,4′-bis(dimethylamino)-diphenylamine sodium (DA-64) claim 1 , N claim 1 ,N claim 1 ,N′N′ claim 1 ,N″ claim 1 ,N″-hexa(3-sulfopropyl)-4 claim 1 ,4′ claim 1 ,4″-triamino-triphenylmethane hexasodium salt (TPM-PS) claim 1 , 10-(carboxymethylaminocarbonyl)-3 claim 1 ,7-bis(dimethylamino)-phenothiazine sodium (DA-67) claim 1 , and 2-(3 claim 1 ,5-dimethoxy-4-hydroxyphenol)-4 claim 1 ,5-bis-(4-dimethylamino phenyl) imidazole6. The slide of claim 1 , wherein said second film layer further comprises a reflective material portion.7. The slide of claim 6 , wherein said reflective material portion comprises titanium.8. The slide of claim 1 , wherein said third film layer further comprises a layer with particles having a diameter of about 25 μm.9. The slide of claim 1 , wherein ...

SUPRAMOLECULAR NANOBEACON IMAGING AGENTS AS PROTEASE SENSORS

Disclosed herein are novel nanobeacon imaging agents having the following formula: 2. The imaging agent of claim 1 , wherein D is chemically linked either directly or indirectly to a separate amino acid of the enzymatically cleavable oligopeptide.3. The imaging agent of claim 1 , wherein L is an enzymatically cleavable peptide having between 4 and 20 amino acids.4. The imaging agent of claim 1 , wherein L is an enzymatically cleavable peptide having the amino acid sequence GFLG (SEQ ID NO: 1).5. The imaging agent of claim 1 , wherein PEP is a fragment of the HIV Tat protein.6. The imaging agent of claim 1 , wherein A is a lysine chemically linked directly to PEP.7. The imaging agent of claim 1 , wherein Q is chemically linked directly to at least one separate side chain moiety of an amino acid of PEP.8. The imaging agent of claim 2 , wherein D comprises a fluorescent dye.9. The imaging agent of claim 7 , wherein Q is a non-fluorescent quencher.11. A method of identifying a cell or a population of cells in vivo expressing a protease of interest comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) contacting the cell or a population of cells expressing a protease of interest with the imaging agent of , which is selectively cleavable by a protease of interest;'}b) allowing the imaging agent to be selectively cleaved the protease of interest in the cell or population of cells; andc) detecting the presence of the fluorescent imaging agent after being cleaved by the protease of interest in the cell or population of cells.12. The method of claim 11 , wherein the cell or population of cells is a tumor cell.13. A method of diagnosing a disease in a patient comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) administering to a patient suspected of having said disease, an imaging agent which is selectively cleavable by a protease of interest, the cleavage of which indicates the presence of the disease, wherein said imaging agent is an imaging agent of ...

COMPOSITIONS AND METHODS RELATING TO FUSION PROTEIN BIOMARKERS

The present invention provides fusion proteins as biomarkers specific for chromosomal translocation-based conditions (e.g., cancer), related methods for detecting fusion protein biomarkers associated with chromosomal translocation-based conditions, related methods for quantifying amount of fusion protein expression, and related methods for diagnosing chromosomal translocation-based conditions through detection of such fusion protein biomarkers. Such fusion protein biomarkers and related methods additionally find use in research settings. 1. A method for detecting the presence of one or more fusion proteins within a biological sample , comprising: a biological sample, wherein said biological sample is a blood sample and/or a tissue sample,', wherein said labeled peptides within each distinct group comprise peptides having one isotopically labeled amino acid residue and peptides having two isotopically labeled amino acid residues,', 'wherein said amino acid sequence of said labeled peptides within each distinct group is identical with an amino acid sequence of a particular fusion protein peptide fragment generated through digestion of said particular fusion protein with a protease,, 'one or more distinct groups of labeled peptides, wherein each distinct group of labeled peptides is specific for a particular fusion protein to be detected, wherein the labeled peptides within each distinct group have labeled peptides having an amino acid sequence that spans the fusion junction of said particular fusion protein encoded by a fusion of a first nucleic acid to a second nucleic acid,'}, 'one or more proteases known to generate upon digestion of said two or more fusion proteins to be detected particular fusion protein peptide fragments,, 'a) providingb) digesting said biological sample with said one or more proteases resulting in generation of biological sample-based peptide fragments,c) combining said one or more distinct groups of labeled peptides with said generated ...