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1. 发明授权

US08465981B2 Polypeptides, systems, and methods useful for detecting glucose 失效
标题翻译：用于检测葡萄糖的多肽，系统和方法
公开(公告)号：US08465981B2
公开(公告)日：2013-06-18
申请号：US12672403
申请日：2008-08-06
申请人： Sylvia Daunert , Kendrick Turner , Smita Joel , Laura Rowe
发明人： Sylvia Daunert , Kendrick Turner , Smita Joel , Laura Rowe
IPC分类号： G01N33/66 , G01N33/68 , G01N33/50 , G01N21/76 , C07K14/00
CPC分类号： A61B5/14532 , A61B5/1459 , A61B5/412 , Y10T436/13 , Y10T436/144444
摘要： The presently-disclosed subject matter provides biosensors for detecting molecules of interest. The biosensors include a polypeptide capable of selectively-binding glucose, wherein the polypeptide molecule is selected from: an unnatural analogue of wild type glucose binding protein; a fragment of wild type glucose binding protein; and an unnatural analogue fragment of wild type glucose binding protein.
摘要翻译：目前公开的主题提供用于检测感兴趣的分子的生物传感器。生物传感器包括能够选择性结合葡萄糖的多肽，其中多肽分子选自：野生型葡萄糖结合蛋白的非天然类似物; 野生型葡萄糖结合蛋白片段; 和野生型葡萄糖结合蛋白的非天然类似片段。

2. 发明授权

US07625951B2 Stimuli-responsive hydrogel microdomes integrated with genetically engineered proteins for high-throughput screening of pharmaceuticals 有权
标题翻译：与基因工程蛋白整合的刺激响应性水凝胶微团体用于药物的高通量筛选
公开(公告)号：US07625951B2
公开(公告)日：2009-12-01
申请号：US10996068
申请日：2004-11-24
申请人： Sylvia Daunert , Sapna Kamlakar Deo , Jason Douglas Ehrick , Tyler William Browning , Leonidas G. Bachas
发明人： Sylvia Daunert , Sapna Kamlakar Deo , Jason Douglas Ehrick , Tyler William Browning , Leonidas G. Bachas
IPC分类号： B01D21/01 , C09K3/00
CPC分类号： G01N33/84 , G01N33/54373
摘要： A hydrogel microdome that can swell in response to a stimuli or target molecule is formed by polymerizing a mixture comprising a monomer capable of forming a hydrogel with a biopolymer. An array of hydrogel microdomes can be formed on a substrate by microspotting the mixture and polymerizing. The array can be used for high-throughput screening of analytes as well as for use as an actuator and biosensor using the swelling property of the hydrogel.
摘要翻译：可以通过将包含能够形成水凝胶的单体与生物聚合物的混合物聚合来形成可以响应于刺激物或靶分子而溶胀的水凝胶微球。可以通过微滴混合物和聚合在基底上形成水凝胶微阵列。该阵列可用于分析物的高通量筛选，以及用作致动器和使用水凝胶膨胀性质的生物传感器。

3. 发明授权

US07345160B2 Aequorin and obelin mutants with differing wavelengths and bioluminescence 失效
标题翻译：具有不同波长和生物发光的水母发光蛋白和obelin突变体
公开(公告)号：US07345160B2
公开(公告)日：2008-03-18
申请号：US10811138
申请日：2004-03-29
申请人： Sylvia Daunert , Sapna Kamlakar Deo , Emre Dikici , Laura Rowe
发明人： Sylvia Daunert , Sapna Kamlakar Deo , Emre Dikici , Laura Rowe
IPC分类号： C07H21/04 , C12P21/02 , C07K14/435
CPC分类号： C07K14/43595 , G01N33/533
摘要： The invention relates to aequorin and obelin mutants whose emission is shifted with respect to wild type. The shift in emission is accomplished using a combination of mutations of amino acids within aequorin or obelin that affect bioluminescence; use of different types of chromophores, i.e., coelenterazines with variable emission characteristics; and modifications of the photoprotein with fluorophores that will allow for emission of light at longer wavelengths as a result of energy transfer. Additionally, an assay employing aequorin mutants to test for HIV-1 protease inhibitors is disclosed.
摘要翻译：本发明涉及发射相对于野生型移位的水母发光蛋白和obelin突变体。排放的变化是通过使用影响生物发光的水母发光蛋白或obelin内的氨基酸突变的组合完成的。使用不同类型的发色团，即具有可变排放特征的肠溶蛋白酶; 以及通过荧光团修饰光蛋白，这将允许由于能量转移而在较长波长处发出光。另外，公开了使用水母发光蛋白突变体测试HIV-1蛋白酶抑制剂的测定法。

4. 发明申请

US20050059097A1 Quantitative binding assays using green fluorescent protein as a label 审中-公开
标题翻译：使用绿色荧光蛋白作为标签的定量结合测定
公开(公告)号：US20050059097A1
公开(公告)日：2005-03-17
申请号：US10765063
申请日：2004-01-28
申请人： Sylvia Daunert , Jennifer Lewis , Emily Hernandez
发明人： Sylvia Daunert , Jennifer Lewis , Emily Hernandez
IPC分类号： C12Q1/68 , G01N33/53 , G01N33/533 , G01N33/543 , G01N33/58 , C07K16/46
CPC分类号： G01N33/543 , G01N33/533 , G01N33/582
摘要： A heterogeneous binding assay for an analyte in a fluid sample is developed, which uses a green fluorescent protein (GFP) label. A ligand-GFP conjugate has a specific binding affinity for an anti-ligand immobilized on a support. The anti-ligand also has a specific binding affinity for the analyte. Competition between the analyte and ligand-GFP conjugate for binding sites on the anti-ligand permits an assay for an unknown amount of the analyte. Preferred specific binding pairs for use in the assay are biotin:avidin, and a selected antibody and its antigen. A preferred assay employing an antibody and its antigen is illustrated for a fusion protein containing GFP and an antigenic determinant. Picomolar amounts of analyte can be detected. The mutant of GFP that contains a six-histidine tail to facilitate purification on an immobilized metal affinity column is chemically modified to incorporate biotin moieties. The resulting conjugates retain the fluorescence characteristics of the unmodified protein and are used along with avidin-coated magnetic beads in the development of the assay.
摘要翻译：开发了流体样品中分析物的异质结合测定法，其使用绿色荧光蛋白（GFP）标记。配体-GFP缀合物对固定在载体上的抗配体具有特异性结合亲和力。抗配体还对分析物具有特异性结合亲和力。分析物与配体-GFP缀合物之间在抗配体上的结合位点之间的竞争允许测定未知量的分析物。用于测定中的优选的特异性结合对是生物素：抗生物素蛋白和选择的抗体及其抗原。对于含有GFP和抗原决定簇的融合蛋白说明了使用抗体及其抗原的优选测定法。可以检测到皮摩尔量的分析物。包含六组氨酸尾部以促进固定化金属亲和柱上的纯化的GFP突变体被化学修饰以掺入生物素部分。所得到的缀合物保留未修饰蛋白质的荧光特性，并在测定开发中与抗生物素蛋白包被的磁珠一起使用。

5. 发明授权

US08389263B2 Spores for the stabilization and on-site application of bacterial whole-cell biosensing systems 失效
标题翻译：用于稳定和现场应用细菌全细胞生物传感系统的孢子
公开(公告)号：US08389263B2
公开(公告)日：2013-03-05
申请号：US12676302
申请日：2008-09-10
申请人： Sylvia Daunert , Sapna K. Deo , Patrizia Pasini , Amol Date
发明人： Sylvia Daunert , Sapna K. Deo , Patrizia Pasini , Amol Date
IPC分类号： C12N1/00 , C12N1/12 , C12N1/20 , C12N3/00
CPC分类号： C12N3/00 , C12Q1/02
摘要： The presently-disclosed subject matter is directed to biosensors comprising spore-forming bacterial cells and/or spores generated therefrom, a recognition unit within each spore-forming cell for binding an analyte of interest, and a reporter molecule within each spore-forming cell for detecting binding of the analyte of interest, wherein the reporter molecule generates a detectable signal upon binding of the analyte by the recognition element. The presently-disclosed subject matter further provides methods of using the biosensors and systems and kits including the biosensors.
摘要翻译：目前公开的主题涉及包含由其产生的形成孢子的细菌细胞和/或孢子的生物传感器，用于结合感兴趣分析物的每个孢子形成细胞内的识别单元和每个孢子形成细胞内的报道分子检测感兴趣的分析物的结合，其中报告分子在通过识别元件结合分析物时产生可检测的信号。目前公开的主题还提供使用生物传感器和包括生物传感器的系统和试剂盒的方法。

6. 发明申请

US20100304379A1 SPORES FOR THE STABILIZATION AND ON-SITE APPLICATION OF BACTERIAL WHOLE-CELL BIOSENSING SYSTEMS 失效
标题翻译：用于稳定和现场应用细菌生物传感系统的体育活动
公开(公告)号：US20100304379A1
公开(公告)日：2010-12-02
申请号：US12676302
申请日：2008-09-10
申请人： Sylvia Daunert , Sapna K. Deo , Patrizia Pasini , Amol Date
发明人： Sylvia Daunert , Sapna K. Deo , Patrizia Pasini , Amol Date
IPC分类号： G01N33/53 , C12N1/20 , C12N1/21 , C12N11/00 , C12Q1/02 , C12Q1/68
CPC分类号： C12N3/00 , C12Q1/02
摘要： The presently-disclosed subject matter is directed to biosensors comprising spore-forming bacterial cells and/or spores generated therefrom, a recognition unit within each spore-forming cell for binding an analyte of interest, and a reporter molecule within each spore-forming cell for detecting binding of the analyte of interest, wherein the reporter molecule generates a detectable signal upon binding of the analyte by the recognition element. The presently-disclosed subject matter further provides methods of using the biosensors and systems and kits including the biosensors.
摘要翻译：目前公开的主题涉及包含由其产生的形成孢子的细菌细胞和/或孢子的生物传感器，用于结合感兴趣分析物的每个孢子形成细胞内的识别单元和每个孢子形成细胞内的报道分子检测感兴趣的分析物的结合，其中报告分子在通过识别元件结合分析物时产生可检测的信号。目前公开的主题还提供使用生物传感器和包括生物传感器的系统和试剂盒的方法。

7. 发明授权

US07659078B1 Method and kit for determination of prostacyclin in plasma 失效
标题翻译：血浆前列环素测定方法及试剂盒
公开(公告)号：US07659078B1
公开(公告)日：2010-02-09
申请号：US10620806
申请日：2003-07-17
申请人： Sylvia Daunert , Michael Poon , Urvee Desai , Sapna K. Deo
发明人： Sylvia Daunert , Michael Poon , Urvee Desai , Sapna K. Deo
IPC分类号： G01N33/53 , G01N33/537 , G01N33/543 , G01N33/88 , C12Q1/66 , C07K16/18 , C07K16/26
CPC分类号： G01N33/88 , G01N2800/12 , Y10S435/971 , Y10S435/975 , Y10S530/807
摘要： A solid-phase immunoassay for 6-keto-Prostaglandin F1α, the stable hydrolysis product of prostacyclin (Prostaglandin I2) is disclosed. Prostacyclin, a potent vasodilator with anti-platelet and anti-proliferative properties is an effective treatment for primary pulmonary hypertension and pulmonary arterial hypertension associated with scleroderma and scleroderma-like syndrome. Levels of 6-keto-Prostaglandin F1α can be directly correlated with levels of prostacyclin. Therefore, 6-keto-Prostaglandin F1α has become the indicator of choice to measure prostacyclin levels. The single step immunoassay for 6-keto-Prostaglandin F1α uses the bioluminescent protein, aequorin as a label. Analyte-label conjugates were constructed by linking the carboxyl group of 6-keto-Prostaglandin F1α and lysine residues of aequorin by chemical conjugation methods. The binding properties of 6-keto-Prostaglandin F1α towards its antibody and the bioluminescent properties of aequorin are retained in the conjugate. The concentration of 6-keto-Prostaglandin F1α after extraction from plasma shows good correlation with the concentration of 6-keto-Prostaglandin F1α obtained without prior extraction of the same plasma sample. The assay allows the measurement of 6-keto-Prostaglandin F1α directly in plasma without any pre-treatment of the samples, which results in a much simpler method with a faster assay time.
摘要翻译：公开了前列环素（前列腺素I2）的稳定水解产物6-酮 - 前列腺素F1alpha的固相免疫测定。前列环素，具有抗血小板和抗增殖特性的强效血管扩张剂是与硬皮病和硬皮病样综合征相关的原发性肺动脉高压和肺动脉高压的有效治疗方法。 6-keto-Prostaglandin F1alpha的水平可以与前列环素的水平直接相关。因此，6-keto-Prostaglandin F1alpha已经成为衡量前列环素水平的首选指标。 6-keto-Prostaglandin F1alpha的单步免疫测定使用生物发光蛋白，水母发光蛋白作为标记。通过化学共轭方法连接6-酮基前列腺素F1al的羧基和水母发光蛋白的赖氨酸残基，构建了分析标记物缀合物。 6-酮前列腺素F1alpha对其抗体的结合性质和水母发光蛋白的生物发光性质保留在缀合物中。从血浆中提取6-keto-Prostaglandin F1alpha的浓度与未经提取相同血浆样品的6-酮 - 前列腺素F1al的浓度呈现良好的相关性。该测定允许直接在血浆中测量6-酮基前列腺素F1alpha，而没有对样品进行任何预处理，这导致以更快的测定时间简单的方法。

8. 发明申请

US20050282225A1 Quantitative binding assays using green fluorescent protein as a label 审中-公开
标题翻译：使用绿色荧光蛋白作为标签的定量结合测定
公开(公告)号：US20050282225A1
公开(公告)日：2005-12-22
申请号：US11197429
申请日：2005-08-05
申请人： Sylvia Daunert , Jennifer Lewis , Emily Hernandez
发明人： Sylvia Daunert , Jennifer Lewis , Emily Hernandez
IPC分类号： C12Q1/68 , G01N33/53 , G01N33/533 , G01N33/543 , G01N33/58
CPC分类号： G01N33/543 , G01N33/533 , G01N33/582
摘要： A heterogeneous binding assay for an analyte in a fluid sample is developed, which uses a green fluorescent protein (GFP) label. A ligand-GFP conjugate has a specific binding affinity for an anti-ligand immobilized on a support. The anti-ligand also has a specific binding affinity for the analyte. Competition between the analyte and ligand-GFP conjugate for binding sites on the anti-ligand permits an assay for an unknown amount of the analyte. Preferred specific binding pairs for use in the assay are biotin:avidin, and a selected antibody and its antigen. A preferred assay employing an antibody and its antigen is illustrated for a fusion protein containing GFP and an antigenic determinant. Picomolar amounts of analyte can be detected. The mutant of GFP that contains a six-histidine tail to facilitate purification on an immobilized metal affinity column is chemically modified to incorporate biotin moieties. The resulting conjugates retain the fluorescence characteristics of the urunodified protein and are used along with avidin-coated magnetic beads in the development of the assay.
摘要翻译：开发了流体样品中分析物的异质结合测定法，其使用绿色荧光蛋白（GFP）标记。配体-GFP缀合物对固定在载体上的抗配体具有特异性结合亲和力。抗配体还对分析物具有特异性结合亲和力。分析物与配体-GFP缀合物之间在抗配体上的结合位点之间的竞争允许测定未知量的分析物。用于测定中的优选的特异性结合对是生物素：抗生物素蛋白和选择的抗体及其抗原。对于含有GFP和抗原决定簇的融合蛋白说明了使用抗体及其抗原的优选测定法。可以检测到皮摩尔量的分析物。包含六组氨酸尾部以促进固定化金属亲和柱上的纯化的GFP突变体被化学修饰以掺入生物素部分。所得到的缀合物保留尿素化蛋白质的荧光特性，并与抗生物素蛋白包被的磁珠一起用于测定的开发。

9. 发明申请

US20110229415A1 SEMI-SYNTHETIC ANTIBODIES AS RECOGNITION ELEMENTS 审中-公开
标题翻译：作为识别元素的半合成抗体
公开(公告)号：US20110229415A1
公开(公告)日：2011-09-22
申请号：US12672397
申请日：2008-08-06
申请人： Sylvia Daunert , Leonidas G. Bachas , Boyd Haley , Smita Joel
发明人： Sylvia Daunert , Leonidas G. Bachas , Boyd Haley , Smita Joel
IPC分类号： A61K49/00 , C07K16/00 , G01N33/53 , A61B5/1459
CPC分类号： C12Q1/6876
摘要： The presently-disclosed subject matter is directed to biosensors for detecting molecules of interest, and systems and methods for using same. The biosensors include an antibody and a probe covalently-linked to the antibody. The antibody has an antigen-binding site that selectively binds the molecule of interest and a purine-binding site, which is at a location distinct from that of the antigen-binding site. The probe includes a purine molecule, which is covalently bound at the purine-binding site to the antibody, and a label linked to the purine molecule. Upon binding of the molecule of interest to the biosensor antigen-binding site, the biosensor undergoes a conformational change, which detectably alters a signal of the label such that the molecule of interest can be detected.
摘要翻译：目前公开的主题涉及用于检测目标分子的生物传感器，以及用于其的系统和方法。生物传感器包括与抗体共价连接的抗体和探针。抗体具有选择性结合感兴趣分子的抗原结合位点和位于与抗原结合位点不同的位置的嘌呤结合位点。探针包括嘌呤结合位点与抗体共价结合的嘌呤分子和与嘌呤分子连接的标记。当感兴趣的分子与生物传感器抗原结合位点结合时，生物传感器经历构象变化，其可检测地改变标记的信号，使得可以检测到感兴趣的分子。

10. 发明申请

US20110117661A1 POLYPEPTIDES, SYSTEMS, AND METHODS USEFUL FOR DETECTING GLUCOSE 失效
标题翻译：用于检测葡萄糖的多肽，系统和方法
公开(公告)号：US20110117661A1
公开(公告)日：2011-05-19
申请号：US12672403
申请日：2008-08-06
申请人： Sylvia Daunert , Kendrick Turner , Smita Joel , Laura Rowe
发明人： Sylvia Daunert , Kendrick Turner , Smita Joel , Laura Rowe
IPC分类号： G01N33/50 , C07K14/00
CPC分类号： A61B5/14532 , A61B5/1459 , A61B5/412 , Y10T436/13 , Y10T436/144444
摘要： The presently-disclosed subject matter provides biosensors for detecting molecules of interest. The biosensors comprise a polypeptide capable of selectively-binding glucose, wherein the polypeptide molecule is selected from: an unnatural analogue of wild type glucose binding protein; a fragment of wild type glucose binding protein; and an unnatural analogue fragment of wild type glucose binding protein.
摘要翻译：目前公开的主题提供用于检测感兴趣的分子的生物传感器。生物传感器包含能够选择性结合葡萄糖的多肽，其中多肽分子选自：野生型葡萄糖结合蛋白的非天然类似物; 野生型葡萄糖结合蛋白片段; 和野生型葡萄糖结合蛋白的非天然类似片段。

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