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1. 发明申请

US20090028848A1 SEQUENCE BASED ENGINEERING AND OPTIMIZATION OF SINGLE CHAIN ANTIBODIES 有权
标题翻译：基于序列的工程和单链抗体优化
公开(公告)号：US20090028848A1
公开(公告)日：2009-01-29
申请号：US12145749
申请日：2008-06-25
申请人： David Urech , Leonardo Borras
发明人： David Urech , Leonardo Borras
IPC分类号： A61K39/395 , C07K16/18 , A61P43/00 , G06F7/06 , G06F17/30
CPC分类号： C07K16/465 , C07K2317/56 , C07K2317/567 , C07K2317/622 , G06F19/16
摘要： The invention provides methods of using sequence based analysis and rational strategies to modify and improve the structural and biophysical properties of single chain antibodies (scFvs), including stability, solubility, and antigen binding affinity. These methods and strategies can be used individually or in combination. The methods of the present invention also include the use of a database comprising scFv sequences from an experimentally screened scFv library of antibodies that have been selected to have superior solubility and stability. The invention also provides methods of using the properties found for these selected antibodies in a general approach for reshaping scFv antibodies to improve stability and solubility properties of a single chain antibody fragment.
摘要翻译：本发明提供了使用基于序列的分析和合理策略来修饰和改善单链抗体（scFv）的结构和生物物理性质（包括稳定性，溶解度和抗原结合亲和力）的方法。这些方法和策略可以单独使用或组合使用。本发明的方法还包括使用包含已被选择具有优异溶解度和稳定性的抗体的实验筛选的scFv文库的scFv序列的数据库。本发明还提供了在通用方法中使用针对这些所选抗体发现的性质的方法，用于重塑scFv抗体以改善单链抗体片段的稳定性和溶解性质。

2. 发明申请

US20080118512A1 Antibodies binding to the extracellular domain of the receptor tyrosine kinase ALK 有权
标题翻译：结合受体酪氨酸激酶ALK细胞外结构域的抗体
公开(公告)号：US20080118512A1
公开(公告)日：2008-05-22
申请号：US11796549
申请日：2007-04-27
申请人： Adrian Auf Der Maur , Alcide Barberis , Peter Lichtlen
发明人： Adrian Auf Der Maur , Alcide Barberis , Peter Lichtlen
IPC分类号： A61K39/00 , C07K16/00 , C07H21/04 , C12N15/00 , C12N5/04 , C12P21/04 , A61P43/00
CPC分类号： C12N9/12 , A61K39/00 , A61K2039/505 , C07K16/2896 , C07K16/30 , C07K2317/34 , C07K2317/565 , C07K2317/567 , C07K2317/622 , C07K2317/92 , C12Y207/10001
摘要： The present invention concerns an antibody specific for human ALK (Anaplastic Lymphoma Kinase), in particular a scFv, a nucleic acid sequence encoding it, its production and its use as a pharmaceutical or for diagnostic purposes. Said antibody is suitable for the local treatment of tumors, in particular glioblastoma.
摘要翻译：本发明涉及对人ALK（间变型淋巴瘤激酶）特异性的抗体，特别是scFv，编码它的核酸序列，其生产及其作为药物或用于诊断目的的用途。所述抗体适用于局部治疗肿瘤，特别是成胶质细胞瘤。

3. 发明授权

US07258985B2 Intrabodies with defined framework that is stable in a reducing environment and applications thereof 有权
公开(公告)号：US07258985B2
公开(公告)日：2007-08-21
申请号：US09750424
申请日：2000-12-28
申请人： Adrian Auf Der Maur , Alcide Barberis , Dominik Escher
发明人： Adrian Auf Der Maur , Alcide Barberis , Dominik Escher
IPC分类号： G01N33/53 , C40B20/00 , C12P21/02
CPC分类号： C07K16/005 , A61K2039/505 , C07K16/00 , C07K16/14 , C07K2317/622 , C07K2317/80 , C07K2318/10 , C07K2319/00 , C12N15/1055 , C12N15/1093
摘要： A method for the isolation of CDRs in a defined framework that is stable and soluble in reducing environment is described as well as thus obtainable scFv. Starting from such scFv with defined framework a scFv library can be generated wherein the framework is conserved while at least one complementary determining region (CDR) is randomized. Such library, e.g. in yeast cells, is suitable for screening for antibody/CDR-interactions or for screening for antibodies.

4. 发明申请

US20060068388A1 Method for the identification of modulators of a secretase activity 审中-公开
标题翻译：识别分泌酶活性调节剂的方法
公开(公告)号：US20060068388A1
公开(公告)日：2006-03-30
申请号：US10512001
申请日：2002-04-18
申请人： Alcide Barberis , Oliver Middendorp , Urs Luthi
发明人： Alcide Barberis , Oliver Middendorp , Urs Luthi
IPC分类号： C12Q1/68 , G01N33/53
CPC分类号： G01N33/573 , C12Q1/025 , C12Q1/37 , C12Q1/6897 , G01N33/6896 , G01N2333/96472 , G01N2500/00 , G01N2500/10
摘要： The present invention relates to a cellular screening method for the identification of modulators of secretases. Said method comprises contacting suitable host cells with a test substance wherein said suitable host cells comprise: a) a fusion protein secretory protein, a membrane anchor domain and a secrease cleavage sequence, b) a protein comprising a secretase activity recognising said cleavage sequence of said fusion protein and a) at least one reporter gene under control of a transcriptional activation system wherein said transcriptional activation system is regulated by the release of said secretory protein from said fusion protein by said secretase activity and its subsequent secretion then culturing said cells under suitable conditions such that said reporter gene allowing detection and/or survival of cells is only expressed or repressed in a manner that is dependent on an altered secretase activity due to said test substance.
摘要翻译：本发明涉及用于鉴定分泌物调节剂的细胞筛选方法。所述方法包括使合适的宿主细胞与测试物质接触，其中所述合适的宿主细胞包含：a）融合蛋白分泌蛋白，膜锚结构域和分泌酶切割序列，b）包含识别所述融合蛋白和a）在转录激活系统控制下的至少一个报告基因，其中所述转录激活系统通过所述分泌酶活性从所述融合蛋白释放所述分泌蛋白及其随后的分泌来调节，然后在合适的条件下培养所述细胞使得允许细胞的检测和/或存活的所述报告基因仅以取决于由于所述测试物质而改变的分泌酶活性的方式表达或抑制。

5. 发明申请

US20070298466A1 Method For The Construction Of Randomized Gene Sequence Libraries In Cells 有权
标题翻译：随机基因序列库在细胞中的构建方法
公开(公告)号：US20070298466A1
公开(公告)日：2007-12-27
申请号：US10552219
申请日：2003-04-11
申请人： Claudia Scharer-Brodbeck
发明人： Claudia Scharer-Brodbeck
IPC分类号： C12P19/34 , C12N1/00 , C12N1/15 , C12N15/63 , C12N15/65
CPC分类号： C07K14/39 , C07K16/00 , C07K16/005 , C12N15/1082 , C12N15/65
摘要： An in vivo method for the construction of randomized gene libraries and/or domain replacement in gene libraries by homologous recombination using a Kluyveromyces lactis killer toxin, in particular the (γ-subunit of the K. lactis killer toxin, as negative selection marker is described. The use of the (γ-subunit of K. lactis as negative selectable marker increases the percentage of randomized clones.
摘要翻译：描述了通过使用克鲁维酵母乳杆菌杀伤毒素，特别是（乳酸克鲁维酵母杀伤毒素的γ-亚基）作为阴性选择标记的同源重组在基因文库中构建随机基因文库和/或域替换的体内方法使用乳酸克鲁维酵母的γ-亚基作为阴性选择标记增加了随机克隆的百分比。

6. 发明申请

US20060019238A1 Method for identifying protein-protein interactions 审中-公开
标题翻译：识别蛋白质 - 蛋白质相互作用的方法
公开(公告)号：US20060019238A1
公开(公告)日：2006-01-26
申请号：US10515249
申请日：2003-05-21
申请人： David Urech , Peter Lichtlen , Alcide Barberis
发明人： David Urech , Peter Lichtlen , Alcide Barberis
IPC分类号： C12Q1/70 , C12Q1/68 , G01N33/53 , C12P21/06 , C12N9/12 , G01N33/567
CPC分类号： C12Q1/02 , C07K2319/00 , C12N9/12 , C12N15/1055 , C12Q1/6897
摘要： The properties of yeast help, a type I ER membrane protein which is involved in the unfolded protein response (UPR), have been exploited to develop â. system for the detection and study of interactions between extracellular and/or membrane proteins. In the system, proteins of interest are fused to the lumenal N-terminus of a truncated Ire1p. A specific interaction between two partners may be visualized through dimerization of the Ire1p moiety which, either, directly or indirectly, results in a detection means, for example, the expression of a selectable reporter gene. Depending on the type of reporter gene used, its expression can positively or negatively influence cell growth, thus allowing selection of both stimulation and inhibition of protein-protein interactions. The system presented here can also be used to study intracellular protein interactions.
摘要翻译：酵母帮助的性质，涉及解折叠蛋白反应（UPR）的I型ER膜蛋白已被开发利用。用于检测和研究细胞外和/或膜蛋白之间相互作用的系统。在该系统中，所关注的蛋白质融合到截短的Ire1p的内腔N-末端。可以通过Ire1p部分的二聚化可视化两个配偶体之间的特异性相互作用，其直接或间接地产生检测手段，例如选择性报道基因的表达。根据使用的报告基因的类型，其表达可以积极或消极地影响细胞生长，从而允许选择刺激和抑制蛋白质 - 蛋白质相互作用。这里介绍的系统也可用于研究细胞内蛋白质的相互作用。

7. 发明申请

US20100137150A1 Sequence Based Engineering and Optimization of Single Chain Antibodies 有权
标题翻译：基于序列的工程和单链抗体的优化
公开(公告)号：US20100137150A1
公开(公告)日：2010-06-03
申请号：US12530756
申请日：2008-03-12
申请人： David M. Urech , Leonardo Borras
发明人： David M. Urech , Leonardo Borras
IPC分类号： C40B30/02
CPC分类号： C07K16/00 , C07K16/241 , C07K2317/56 , C07K2317/565 , C07K2317/567 , C07K2317/622 , C07K2317/94 , G06F19/18
摘要： The invention provides methods of using sequence based analysis and rational strategies to modify and improve the structural and biophysical properties of single chain antibodies (scFvs), including stability, solubility, and antigen binding affinity. These methods and strategies can be used individually or in combination. The methods of the present invention also include the use of a database comprising scFv sequences from an experimentally screened scFv library of antibodies that have been selected to have superior solubility and stability. The invention also provides methods of using the properties found for these selected antibodies in a general approach for reshaping scFv antibodies to improve stability and solubility properties of a single chain antibody fragment.
摘要翻译：本发明提供了使用基于序列的分析和合理策略来修饰和改善单链抗体（scFv）的结构和生物物理性质（包括稳定性，溶解度和抗原结合亲和力）的方法。这些方法和策略可以单独使用或组合使用。本发明的方法还包括使用包含已被选择具有优异溶解度和稳定性的抗体的实验筛选的scFv文库的scFv序列的数据库。本发明还提供了在通用方法中使用针对这些所选抗体发现的性质的方法，用于重塑scFv抗体以改善单链抗体片段的稳定性和溶解性质。

8. 发明授权

US07566528B2 Method for the identification and/or validation of receptor tyrosine kinase inhibitors 失效
标题翻译：用于鉴定和/或验证受体酪氨酸激酶抑制剂的方法
公开(公告)号：US07566528B2
公开(公告)日：2009-07-28
申请号：US11112344
申请日：2005-04-22
申请人： Tea Gunde , Catherine Berset , Alcide Barberis
发明人： Tea Gunde , Catherine Berset , Alcide Barberis
IPC分类号： C12Q1/02 , C12N1/19 , C12N1/38
CPC分类号： C12Q1/485 , G01N2500/00
摘要： An in vivo method for the identification and/or validation of receptor tyrosine kinase inhibitors is described. Said method is characterized by the following steps: providing host cells comprising a nucleic acid construct encoding a peptide which comprises a tyrosine kinase domain of a receptor tyrosine kinase wherein said peptide lacks a transmembrane domain or a functional fragment thereof and said tyrosine kinase activity in the cytoplasma leads to proliferation arrest, contacting said host cells with a candidate compound and identification of inhibitors of said tyrosine kinase activity by cultivation of said host cells under suitable conditions such that the modulation of the tyrosine kinase activity by the candidate compound leads to cell growth.
摘要翻译：描述了用于鉴定和/或验证受体酪氨酸激酶抑制剂的体内方法。所述方法的特征在于以下步骤：提供宿主细胞，其包含编码包含受体酪氨酸激酶的酪氨酸激酶结构域的肽的核酸构建体，其中所述肽缺少跨膜结构域或其功能片段，并且所述酪氨酸激酶活性在细胞质导致增殖停滞，通过在合适的条件下培养所述宿主细胞，使所述宿主细胞与候选化合物接触并鉴定所述酪氨酸激酶活性的抑制剂，使得由候选化合物调节酪氨酸激酶活性导致细胞生长。

9. 发明申请

US20090311251A1 SCFV ANTIBODIES WHICH PASS EPITHELIAL AND/OR ENDOTHELIAL LAYERS 有权
标题翻译：通过上皮和/或内皮层的SCFV抗体
公开(公告)号：US20090311251A1
公开(公告)日：2009-12-17
申请号：US12307875
申请日：2007-07-10
申请人： Adrian Auf Der Maur , Alcide Barberis , David M. Urech , Peter Lichtlen
发明人： Adrian Auf Der Maur , Alcide Barberis , David M. Urech , Peter Lichtlen
IPC分类号： A61K39/395 , G01N33/53 , C07K16/00
CPC分类号： C07K16/18 , A61K39/39591 , A61K2039/505 , A61K2039/54 , C07K16/22 , C07K16/241 , C07K2317/24 , C07K2317/622
摘要： scFv antibodies which specifically bind selected antigens and are obtainable by a method comprising (i) selecting from a pool of soluble and stable antibody frameworks a soluble and stable framework matching best the framework of a non-human antibody against the antigen with a certain binding specificity, (ii) either providing said soluble and stable framework with CDRs that bind specifically to said antigen, or mutating the framework of said non-human antibody towards the sequence of said soluble and stable framework, to generate scFv antibodies, (iii) testing the generated antibody for solubility and stability, and testing the generated antibody for antigen binding, and (iv) selecting an scFV that is soluble, stable and binds to the antigen specifically. Also provided are pharmaceutical compositions comprising said scFv antibody, methods of treatment and diagnosis for diseases related to over expression of antigens that are specifically bound by said antibody.
摘要翻译： scFv抗体，其特异性结合所选择的抗原，并且可通过以下方法获得：（i）从可溶性和稳定的抗体框架库中选择可溶性和稳定的框架，以最佳匹配抗人抗体的框架与一定的结合特异性提供与所述抗原特异性结合的CDR提供所述可溶性和稳定的框架，或者将所述非人抗体的框架朝向所述可溶性和稳定框架的序列突变以产生scFv抗体，（iii）测试产生抗体的溶解度和稳定性，并测试所产生的抗体的抗原结合，和（iv）选择可溶性，稳定并特异性结合抗原的scFV。还提供了包含所述scFv抗体的药物组合物，与由所述抗体特异性结合的抗原过度表达相关疾病的治疗和诊断方法。

10. 发明申请

US20080233110A1 Intrabodies with defined framework that is stable in a reducing environment and applications thereof 审中-公开
标题翻译：具有在还原环境中稳定的具有框架的体内抗体及其应用
公开(公告)号：US20080233110A1
公开(公告)日：2008-09-25
申请号：US11894097
申请日：2007-08-20
申请人： Adrian Auf Der Maur , Alcide Barberis , Dominik Escher
发明人： Adrian Auf Der Maur , Alcide Barberis , Dominik Escher
IPC分类号： A61K39/395 , C07K16/00 , C40B30/06 , C40B50/00 , C40B40/10
CPC分类号： C07K16/005 , A61K2039/505 , C07K16/00 , C07K16/14 , C07K2317/622 , C07K2317/80 , C07K2318/10 , C07K2319/00 , C12N15/1055 , C12N15/1093
摘要： A method for the isolation of CDRs in a defined framework that is stable and soluble in reducing environment is described as well as thus obtainable scFv. Starting from such scFv with defined framework a scFv library can be generated wherein the framework is conserved while at least one complementary determining region (CDR) is randomized. Such library, e.g. in yeast cells, is suitable for screening for antibody/CDR-interactions or for screening for antibodies.
摘要翻译：描述了在限定的框架中分离CDR的方法，其稳定和可溶于还原环境，以及由此获得的scFv。从具有定义框架的这种scFv开始，可以生成scFv文库，其中框架是保守的，而至少一个互补决定区（CDR）是随机的。这样的文库，例如在酵母细胞中，适用于筛选抗体/ CDR相互作用或筛选抗体。

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