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1. 发明申请

US20090017495A1 ESCHERICHIA COLI-DERIVED VACCINE AND THERAPY AGAINST BOTULISM 有权
标题翻译： ESCHERICHIA派生的疫苗和治疗效果
公开(公告)号：US20090017495A1
公开(公告)日：2009-01-15
申请号：US12040542
申请日：2008-02-29
申请人： Michael Baldwin , Eric A. Johnson , Joseph T. Barbieri , Marite Bradshaw , William H. Tepp , Christina L. Pier
发明人： Michael Baldwin , Eric A. Johnson , Joseph T. Barbieri , Marite Bradshaw , William H. Tepp , Christina L. Pier
IPC分类号： C12N15/09 , C07K14/33
CPC分类号： C12N9/52 , A61K39/0258 , A61K39/08 , A61K2039/523 , C12Y304/24069 , Y02A50/469 , Y02A50/474
摘要： A method of producing botulinum toxin C-terminal receptor binding domain (HCR) is disclosed. The one embodiment, the method comprises the steps of (a) preparing E. coli transformed with an expression vector comprising DNA encoding HCR protein, (b) inducing expression of the HCR protein at a reduced temperature in a culture media, and (c) purifying the HCR protein via extraction, wherein the extraction comprises a clarification by centrifugation and a filtration, wherein the purified HCR protein is at least 10 mg/L of culture medium.
摘要翻译：公开了产生肉毒杆菌毒素C末端受体结合结构域（HCR）的方法。该方法包括以下步骤：（a）用包含编码HCR蛋白的DNA的表达载体转化大肠杆菌，（b）在培养基中降低温度诱导HCR蛋白的表达，和（c）通过提取纯化HCR蛋白质，其中提取物通过离心和过滤进行澄清，其中纯化的HCR蛋白质为至少10mg / L的培养基。

2. 发明授权

US07820411B2 Escherichia coli-derived vaccine and therapy against botulism 有权
标题翻译：大肠杆菌来源的疫苗和针对肉毒中毒的治疗
公开(公告)号：US07820411B2
公开(公告)日：2010-10-26
申请号：US12040542
申请日：2008-02-29
申请人： Michael Baldwin , Marite Bradshaw , William H. Tepp , Eric A. Johnson , Joseph T. Barbieri , Christina L. Pier
发明人： Michael Baldwin , Marite Bradshaw , William H. Tepp , Eric A. Johnson , Joseph T. Barbieri , Christina L. Pier
IPC分类号： C12P21/02
CPC分类号： C12N9/52 , A61K39/0258 , A61K39/08 , A61K2039/523 , C12Y304/24069 , Y02A50/469 , Y02A50/474
摘要： A method of producing botulinum toxin C-terminal receptor binding domain (HCR) is disclosed. The one embodiment, the method comprises the steps of (a) preparing E. coli transformed with an expression vector comprising DNA encoding HCR protein, (b) inducing expression of the HCR protein at a reduced temperature in a culture media, and (c) purifying the HCR protein via extraction, wherein the extraction comprises a clarification by centrifugation and a filtration, wherein the purified HCR protein is at least 10 mg/L of culture medium.
摘要翻译：公开了产生肉毒杆菌毒素C末端受体结合结构域（HCR）的方法。该方法包括以下步骤：（a）用包含编码HCR蛋白的DNA的表达载体转化大肠杆菌，（b）在培养基中降低温度诱导HCR蛋白的表达，和（c）通过提取纯化HCR蛋白质，其中提取物通过离心和过滤进行澄清，其中纯化的HCR蛋白质为至少10mg / L的培养基。

3. 发明申请

US20090182123A1 Method of Preparing Botulinum Neurotoxin Type E Light Chain 有权
标题翻译：肉毒杆菌神经毒素E型轻链的制备方法
公开(公告)号：US20090182123A1
公开(公告)日：2009-07-16
申请号：US12273157
申请日：2008-11-18
申请人： Eric A. Johnson , Marite Bradshaw , Michael Baldwin , Joseph T. Barbieri
发明人： Eric A. Johnson , Marite Bradshaw , Michael Baldwin , Joseph T. Barbieri
IPC分类号： C07K14/33
CPC分类号： C07K14/33
摘要： The present invention provides a preparation of botulinum toxin light chain type A or E, wherein the preparation is both catalytically active and soluble. Preferably, the preparation consists essentially of amino acid residues 1 through 425 of the botulinum toxin light chain type A. A method of screening inhibitors is also provided, wherein the method comprises exposing a test inhibitor to the preparation of botulinum toxin light chain type A and evaluating the biological activity of the preparation. In another embodiment, a method of providing a catalytically active, soluble preparation of botulinum toxin light chain, type A is provided, wherein the method comprises obtaining an expression vector comprising a DNA sequence encoding amino acid residues 1-425 and expressing a polypeptide.
摘要翻译：本发明提供了A型或E型轻链型肉毒杆菌毒素的制备方法，其中所述制剂既具有催化活性，又具有可溶性。优选地，制剂基本上由A型肉毒毒素轻链的氨基酸残基1至425组成。还提供了筛选抑制剂的方法，其中所述方法包括将测试抑制剂暴露于A型轻链A型肉毒杆菌毒素的制备中，评估制剂的生物活性。在另一个实施方案中，提供了提供A型肉毒毒素轻链的催化活性可溶性制剂的方法，其中所述方法包括获得包含编码氨基酸残基1-425的DNA序列并表达多肽的表达载体。

4. 发明授权

US07465457B2 Method for preparing botulinum neurotoxin type A light chain 有权
标题翻译：制备肉毒杆菌神经毒素A型轻链的方法
公开(公告)号：US07465457B2
公开(公告)日：2008-12-16
申请号：US11404289
申请日：2006-04-14
申请人： Eric A. Johnson , Marite Bradshaw , Michael Baldwin , Joseph T. Barbieri
发明人： Eric A. Johnson , Marite Bradshaw , Michael Baldwin , Joseph T. Barbieri
IPC分类号： A61K39/08 , C12P21/02
CPC分类号： C07K14/33
摘要： The present invention provides a preparation of botulinum toxin light chain type A or E, wherein the preparation is both catalytically active and soluble. Preferably, the preparation consists essentially of amino acid residues 1 through 425 of the botulinum toxin light chain type A. A method of screening inhibitors is also provided, wherein the method comprises exposing a test inhibitor to the preparation of botulinum toxin light chain type A and evaluating the biological activity of the preparation. In another embodiment, a method of providing a catalytically active, soluble preparation of botulinum toxin light chain, type A is provided, wherein the method comprises obtaining an expression vector comprising a DNA sequence encoding amino acid residues 1-425 and expressing a polypeptide.
摘要翻译：本发明提供了A型或E型轻链型肉毒杆菌毒素的制备方法，其中所述制剂既具有催化活性，又具有可溶性。优选地，制剂基本上由A型肉毒毒素轻链的氨基酸残基1至425组成。还提供了筛选抑制剂的方法，其中所述方法包括将测试抑制剂暴露于A型轻链A型肉毒杆菌毒素的制备评估制剂的生物活性。在另一个实施方案中，提供了提供A型肉毒毒素轻链的催化活性可溶性制剂的方法，其中所述方法包括获得包含编码氨基酸残基1-425的DNA序列并表达多肽的表达载体。

5. 发明授权

US07833524B2 Method of preparing botulinum neurotoxin type E light chain 有权
标题翻译：制备肉毒杆菌神经毒素E型轻链的方法
公开(公告)号：US07833524B2
公开(公告)日：2010-11-16
申请号：US12273157
申请日：2008-11-18
申请人： Eric A. Johnson , Marite Bradshaw , Michael Baldwin , Joseph T. Barbieri
发明人： Eric A. Johnson , Marite Bradshaw , Michael Baldwin , Joseph T. Barbieri
IPC分类号： C12P1/04
CPC分类号： C07K14/33
摘要： The present invention provides a preparation of botulinum toxin light chain type A or E, wherein the preparation is both catalytically active and soluble. Preferably, the preparation consists essentially of amino acid residues 1 through 425 of the botulinum toxin light chain type A. A method of screening inhibitors is also provided, wherein the method comprises exposing a test inhibitor to the preparation of botulinum toxin light chain type A and evaluating the biological activity of the preparation. In another embodiment, a method of providing a catalytically active, soluble preparation of botulinum toxin light chain, type A is provided, wherein the method comprises obtaining an expression vector comprising a DNA sequence encoding amino acid residues 1-425 and expressing a polypeptide.
摘要翻译：本发明提供了A型或E型轻链型肉毒杆菌毒素的制备方法，其中所述制剂既具有催化活性，又具有可溶性。优选地，制剂基本上由A型肉毒毒素轻链的氨基酸残基1至425组成。还提供了筛选抑制剂的方法，其中所述方法包括将测试抑制剂暴露于A型轻链A型肉毒杆菌毒素的制备评估制剂的生物活性。在另一个实施方案中，提供了提供A型肉毒毒素轻链的催化活性可溶性制剂的方法，其中所述方法包括获得包含编码氨基酸残基1-425的DNA序列并表达多肽的表达载体。

6. 发明授权

US10011823B2 Engineered botulinum neurotoxin 有权
公开(公告)号：US10011823B2
公开(公告)日：2018-07-03
申请号：US13264078
申请日：2010-04-13
申请人： Joseph T. Barbieri , Sheng Chen
发明人： Joseph T. Barbieri , Sheng Chen
IPC分类号： C12N9/52 , C07K14/33 , A61P35/00 , A61P11/00 , A61P29/00 , A61P37/00 , A61K39/08 , A61P11/06 , A61K38/00
CPC分类号： C12N9/52 , A61K38/00
摘要： The present invention provides a novel modified BoNT/E catalytic domain and methods of use thereof. In one embodiment, the light chain residue 224, or a residue corresponding to residue 224, of the modified BoNT/E catalytic domain has been altered to be aspartic acid or glutamic acid. The modified catalytic domain cleaves SNAP23 but does not cleave SNAP29 or SNAP47, providing novel methods of treating diseases including without limitation, asthma, CF, chronic obstructive pulmonary, gastric acid efflux and inflammation, immune disorders with a cytokine component or cancers with a cytokine component.

7. 发明授权

US5599665A Pseudomonas aeruginosa nucleic acids encoding exoenzyme S activity and use thereof in detecting pseudomonas aeruginosa infection 失效
标题翻译：编码外切酶S活性的铜绿假单胞菌核酸及其在检测铜绿假单胞菌感染中的应用
公开(公告)号：US5599665A
公开(公告)日：1997-02-04
申请号：US171299
申请日：1993-12-21
申请人： Joseph T. Barbieri , Dara W. Frank , Scott M. Kulich
发明人： Joseph T. Barbieri , Dara W. Frank , Scott M. Kulich
IPC分类号： A61K38/00 , A61K39/00 , C07K14/21 , C12N9/10 , C12Q1/68 , C12N15/85
CPC分类号： C07K14/21 , C12N9/1077 , C12Q1/689 , A61K38/00 , A61K39/00
摘要： A genetic construct containing a coding region for exoenzyme S activity from Pseudomonas aeruginosa is disclosed. A essentially pure protein preparation of the 49 kDa form of exoenzyme S is also disclosed. The protein product of the genetic construct may be used to modify the RAS protein function in mammalian carcinomas, used as a vaccine, or used to diagnose Pseudomonas aeruginosa infection.
摘要翻译：公开了一种含有来自绿脓假单胞菌的外切酶S活性的编码区的遗传构建体。还公开了49KDa形式的外切酶S的基本上纯的蛋白质制剂。遗传构建体的蛋白质产物可用于修饰用作疫苗的哺乳动物癌症中的RAS蛋白质功能，或用于诊断铜绿假单胞菌感染。

8. 发明申请

US20120039941A1 ENGINEERED BOTULINUM NEUROTOXIN 有权
标题翻译：工程BOTULINUM神经毒素
公开(公告)号：US20120039941A1
公开(公告)日：2012-02-16
申请号：US13264078
申请日：2010-04-13
申请人： Joseph T. Barbieri , Sheng Chen
发明人： Joseph T. Barbieri , Sheng Chen
IPC分类号： C12N9/52 , C07K14/33 , A61P35/00 , A61P11/00 , A61P29/00 , A61P37/00 , A61K39/08 , A61P11/06
CPC分类号： C12N9/52 , A61K38/00
摘要： The present invention provides a novel modified BoNT/E catalytic domain and methods of use thereof. In one embodiment, the light chain residue 224, or a residue corresponding to residue 224, of the modified BoNT/E catalytic domain has been altered to be aspartic acid or glutamic acid. The modified catalytic domain cleaves SNAP23 but does not cleave SNAP29 or SNAP47, providing novel methods of treating diseases including without limitation, asthma, CF, chronic obstructive pulmonary, gastric acid efflux and inflammation, immune disorders with a cytokine component or cancers with a cytokine component.
摘要翻译：本发明提供了一种新的改进的BoNT / E催化结构域及其使用方法。在一个实施方案中，改性的BoNT / E催化结构域的轻链残基224或对应于残基224的残基已被改变为天冬氨酸或谷氨酸。修饰的催化结构域切割SNAP23但不切割SNAP29或SNAP47，提供治疗疾病的新方法，包括但不限于哮喘，CF，慢性阻塞性肺，胃酸流出和炎症，具有细胞因子成分的免疫疾病或具有细胞因子成分的癌症。

9. 发明授权

US5986080A Cloned nucleotide pyrophosphohydrolase and uses thereof 失效
标题翻译：克隆核苷酸焦磷酸水解酶及其用途
公开(公告)号：US5986080A
公开(公告)日：1999-11-16
申请号：US954333
申请日：1997-10-17
申请人： Ikuko Masuda , Joseph T. Barbieri , Arthur L. Haas , Brian D. Halligan , Daniel J. McCarty , Lawrence M. Ryan
发明人： Ikuko Masuda , Joseph T. Barbieri , Arthur L. Haas , Brian D. Halligan , Daniel J. McCarty , Lawrence M. Ryan
IPC分类号： C12N9/16 , C07N21/04 , C12N1/20 , C12N9/14 , C12N15/00
CPC分类号： C12N9/16
摘要： We have cloned and sequenced the cDNA encoding the 61 kD active fragment of a unique porcine chondrocyte nucleotide pyrophosphohydrolase (NTPPHase) from a porcine chondrocyte library. Degenerate oligonucleotides, corresponding to the N-terminal amino acid sequence of this peptide were hybridized to porcine chondrocyte cDNA and used to amplify DNA encoding the N-terminal sequence of 61 kD with the polymerase chain reaction (PCR). The PCR products were then used as probes to clone the entire open reading-frame for the 61 kD fragment from a porcine chondrocyte cDNA library. The length of the cloned cDNA was 2509 bp. Translation of the open-reading-frame predicts the 61 kD fragment to be a 459 amino acid protein. BLAST and FASTA analysis confirmed that this amino acid sequence was unique and did not possess high homology to any known proteins in the non-redundant protein data base. Limited homology (17%) between the 61 kD fragment and several prokaryotic and eukaryotic ATP pyrophosphate-lyase (adenylate cyclase) was detected. Northern blot analysis of porcine chondrocyte RNA showed that the DNA encoding the 61 kD fragment hybridized to a 4.3 kbp RNA transcript. Human chondrocyte RNA also hybridized to this porcine DNA probe. Coupled in vitro transcription translation of an expression vector containing the DNA insert in frame showed the expression of a 61 kD protein.
摘要翻译：我们已经从猪软骨细胞库克隆并测序了编码独特猪软骨细胞核苷酸焦磷酸水解酶（NTPPHase）的61kD活性片段的cDNA。对应于该肽的N末端氨基酸序列的简并寡核苷酸与猪软骨细胞cDNA杂交，并用聚合酶链反应（PCR）扩增编码61kD N末端序列的DNA。然后将PCR产物用作探针，以克隆来自猪软骨细胞cDNA文库的61kD片段的整个开放阅读框。克隆的cDNA的长度为2509bp。开放阅读框的翻译将61 kD片段预测为459个氨基酸的蛋白质。 BLAST和FASTA分析证实，该氨基酸序列是独特的，并且与非冗余蛋白质数据库中的任何已知蛋白质不具有高同源性。检测到61kD片段与几种原核和真核ATP焦磷酸裂解酶（腺苷酸环化酶）之间的有限同源性（17％）。猪软骨细胞RNA的Northern印迹分析表明，编码61kD片段的DNA与4.3kbp RNA转录物杂交。人类软骨细胞RNA也与猪DNA探针杂交。在框架中包含DNA插入片段的表达载体的体外转录翻译显示61kD蛋白的表达。

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