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1. 发明公开

EP3194585A1 SORTASE MOLECULES AND USES THEREOF 审中-公开
标题翻译： SORTASE分子及其用途
公开(公告)号：EP3194585A1
公开(公告)日：2017-07-26
申请号：EP15745335.8
申请日：2015-07-21
申请人： Novartis AG
发明人： GUIMARAES, Carla
IPC分类号： C12N9/64
CPC分类号： C12N9/52 , A61K47/65 , C12Y304/2207
摘要： This application provides mutant sortase molecules and methods of making and using them. In a first aspect, disclosed herein, are sortase molecules having one or a combination of mutations. In an embodiment, a sortase molecule is optimized for a parameter of enzyme performance, e.g., Ca++ dependency (or independency) or reaction rate.
摘要翻译：本申请提供了突变分选酶分子以及制备和使用它们的方法。在本文公开的第一方面，是具有一种或多种突变组合的分选酶分子。在一个实施方案中，分选酶分子针对酶性能参数例如Ca ++依赖性（或独立性）或反应速率进行优化。

2. 发明授权

EP2867253B1 METHOD FOR SELECTION AND PRODUCTION OF TAILOR-MADE HIGHLY SELECTIVE AND MULTI-SPECIFIC TARGETING ENTITIES CONTAINING AT LEAST TWO DIFFERENT BINDING ENTITIES AND USES THEREOF 有权转让
标题翻译：与至少两种不同的结合单位以及选择和制作出定制的，高选择性，多特异性靶向单元的方法及其用途
公开(公告)号：EP2867253B1
公开(公告)日：2016-09-14
申请号：EP13730906.8
申请日：2013-06-25
申请人： F. Hoffmann-La Roche AG
发明人： FENN, Sebastian , KOPETZKI, Erhard , TIEFENTHALER, Georg
IPC分类号： C07K16/00 , C12N9/16 , C12N9/52 , C12N9/80 , C12P21/06 , C07K16/46 , C07K16/32
CPC分类号： C07K16/32 , C07K16/468 , C07K2317/24 , C07K2317/52 , C07K2317/55 , C07K2317/64 , C12N9/52 , C12Y304/2207

3. 发明公开

EP2895496A1 METHOD FOR THE PRODUCTION AND SELECTION OF MOLECULES COMPRISING AT LEAST TWO DIFFERENT ENTITIES AND USES THEREOF 有权
标题翻译：对于具有至少两个不同的单位及其生产并选择分子方法及其用途
公开(公告)号：EP2895496A1
公开(公告)日：2015-07-22
申请号：EP13759782.9
申请日：2013-09-12
申请人： F.Hoffmann-La Roche AG
发明人： BECK, Mariel , TIEFENTHALER, Georg
IPC分类号： C07K1/107
CPC分类号： C07K16/468 , C07K1/1075 , C07K16/065 , C07K16/2881 , C07K16/32 , C07K2317/31 , C07K2317/52 , C07K2317/55 , C07K2319/00 , C07K2319/01 , C07K2319/21 , C12N9/52 , C12Y304/2207
摘要： Herein is reported a method for producing a polypeptide comprising at least two polypeptide domains comprising the step of cultivating a cell comprising (a) a nucleic acid encoding a soluble S. aureus sortase A with a C-terminal endoplasmic reticulum retention signal, (b) a nucleic acid encoding a first polypeptide domain comprising at its C-terminus a sortase motif followed by an endoplasmic reticulum retention signal, and (c) a nucleic acid encoding a second polypeptide domain comprising at its N-terminus at least a diglycine, whereby the cell secretes the sortase A conjugate of the first polypeptide domain and the second polypeptide domain, thereby producing a polypeptide comprising at least two polypeptide domains.

4. 发明公开

EP3212770A4 METHODS FOR EFFICIENT DELIVERY OF THERAPEUTIC MOLECULES IN VITRO AND IN VIVO 有权转让
公开(公告)号：EP3212770A4
公开(公告)日：2018-06-13
申请号：EP15855354
申请日：2015-10-29
申请人： MASSACHUSETTS EYE & EAR INFIRMARY
发明人： CHEN ZHENG-YI
IPC分类号： A61K38/46 , C07H21/04 , C07K14/00 , C12N5/00 , C12N9/16 , C12P21/04
CPC分类号： A61K48/0008 , A61K9/1272 , A61K38/00 , A61K38/10 , A61K47/64 , A61K47/6911 , A61K48/0075 , A61K48/0083 , C07J7/009 , C07K7/06 , C07K7/08 , C07K14/463 , C07K14/47 , C07K19/00 , C07K2319/06 , C07K2319/21 , C07K2319/33 , C07K2319/60 , C12N5/062 , C12N9/00 , C12N9/22 , C12N9/52 , C12N15/11 , C12N15/113 , C12N15/87 , C12N15/88 , C12N15/907 , C12N2310/20 , C12N2310/3513 , C12N2510/00 , C12Y304/2207
摘要： Compositions are described for direct protein delivery into multiple cell types in the mammalian inner ear. The compositions are used to deliver protein(s) (such as gene editing factors) editing of genetic mutations associated with deafness or associated disorders thereof. The delivery of genome editing proteins for gene editing and correction of genetic mutations protect or restore hearing from genetic deafness. Methods of treatment include the intracellular delivery of these molecules to a specific therapeutic target.

5. 发明公开

EP2990423A4 METHOD FOR REFINING PROTEIN INCLUDING SELF-CUTTING CASSETTE AND USE THEREOF 有权转让
标题翻译：用自切割盒精制蛋白质的方法及其用途
公开(公告)号：EP2990423A4
公开(公告)日：2016-10-26
申请号：EP14787873
申请日：2014-04-25
申请人： SCRIPPS KOREA ANTIBODY INST
发明人： SONG BYEONG DOO , YUN JEE SUN , CHOI HYO JUNG , KIM HYE IN , LEE EUNG-SUK
IPC分类号： C12N9/52 , C07K1/16 , C07K1/22 , C07K16/00 , C07K19/00 , C12N9/50 , C12N15/10 , C12N15/62 , C12N15/70
CPC分类号： C07K16/00 , C07K1/22 , C07K2319/00 , C07K2319/21 , C07K2319/30 , C07K2319/41 , C07K2319/42 , C07K2319/50 , C12N9/50 , C12N9/52 , C12N15/1093 , C12Y304/2207
摘要： The present invention relates to a self-cutting fusion protein including sequentially a target protein, a peptide consisting of an amino acid sequence of LPXTG, a sortase A cutting function domain, and a tag from the amino terminal; a coding nucleic acid thereof; an expression vector of the present invention including a nucleic acid; and a cell transformed by the expression vector of the present invention. The present invention also relates to a target protein refinement method including the steps of culturing, dissolving, and refining the transformed cell. The present invention relates to a self-cutting fusion protein including a self-cutting cassette which consists of a sortase A cutting function domain and a peptide consisting of an LPXTG amino acid sequence that is a recognition sequence thereof, and is very useful in that the steps of refining a target protein and eliminating the tag thereof can be accomplished by a single refinement step instead of separate steps. Particularly, the invention may be employed in various fields requiring a huge amount of a protein of a high purity in that a target protein existing in an amino terminal enhances the column union ability and the self-cutting ability of a fusion protein, a target protein rid of the tag is obtained with a high purity, a cutting-buffer makes the steps of refining and eliminating the tag be accomplished through a single step, significantly reducing the time and efforts needed for refining, and the accomplishment through a single step reduces the loss of the protein yield. Particularly, the invention is useful for manufacturing a therapeutic antibody-drug complex.

6. 发明公开

EP2994530A2 PROTEIN MODIFICATION OF LIVING CELLS USING SORTASE 审中-公开
标题翻译：利用SORTASE蛋白修饰活细胞
公开(公告)号：EP2994530A2
公开(公告)日：2016-03-16
申请号：EP14795120.6
申请日：2014-05-09
申请人： Whitehead Institute for Biomedical Research
发明人： SWEE, Lee, Kim , PLOEGH, Hidde, L.
IPC分类号： C12N9/48
CPC分类号： C12N5/0006 , A61K39/0011 , A61K47/48338 , A61K47/65 , A61K2039/5158 , C12N5/0636 , C12N5/0638 , C12N9/50 , C12Y304/2207
摘要： Non-genetically engineered mammalian cells modified by sortase-mediated conjugation of an agent thereto are provided. Methods of conjugating agents to non- genetically engineered mammalian cells using sortase are provided. Methods of using cells, e.g., for diagnostic and/or therapeutic purposes, are provided.
摘要翻译：提供了通过分离酶介导的试剂缀合而修饰的非基因工程哺乳动物细胞。提供了使用分选酶将试剂与非基因工程哺乳动物细胞缀合的方法。提供了使用细胞的方法，例如用于诊断和/或治疗目的。

7. 发明公开

EP2990423A1 METHOD FOR REFINING PROTEIN INCLUDING SELF-CUTTING CASSETTE AND USE THEREOF 有权转让
标题翻译：方法精制蛋白自切割带及其用途
公开(公告)号：EP2990423A1
公开(公告)日：2016-03-02
申请号：EP14787873.0
申请日：2014-04-25
申请人： Scripps Korea Antibody Institute
发明人： SONG, Byeong Doo , YUN, Jee Sun , CHOI, Hyo Jung , KIM, Hye In , LEE, Eung-Suk
IPC分类号： C07K19/00 , C12N15/62 , C12N15/70 , C07K1/16
CPC分类号： C07K16/00 , C07K1/22 , C07K2319/00 , C07K2319/21 , C07K2319/30 , C07K2319/41 , C07K2319/42 , C07K2319/50 , C12N9/50 , C12N9/52 , C12N15/1093 , C12Y304/2207
摘要： The present invention relates to a self-cutting fusion protein including sequentially a target protein, a peptide consisting of an amino acid sequence of LPXTG, a sortase A cutting function domain, and a tag from the amino terminal; a coding nucleic acid thereof; an expression vector of the present invention including a nucleic acid; and a cell transformed by the expression vector of the present invention. The present invention also relates to a target protein refinement method including the steps of culturing, dissolving, and refining the transformed cell. The present invention relates to a self-cutting fusion protein including a self-cutting cassette which consists of a sortase A cutting function domain and a peptide consisting of an LPXTG amino acid sequence that is a recognition sequence thereof, and is very useful in that the steps of refining a target protein and eliminating the tag thereof can be accomplished by a single refinement step instead of separate steps. Particularly, the invention may be employed in various fields requiring a huge amount of a protein of a high purity in that a target protein existing in an amino terminal enhances the column union ability and the self-cutting ability of a fusion protein, a target protein rid of the tag is obtained with a high purity, a cutting-buffer makes the steps of refining and eliminating the tag be accomplished through a single step, significantly reducing the time and efforts needed for refining, and the accomplishment through a single step reduces the loss of the protein yield. Particularly, the invention is useful for manufacturing a therapeutic antibody-drug complex.
摘要翻译：本发明涉及一种自攻融合蛋白，依次包括靶蛋白，在LPXTG，一个分选酶A切割功能域组成的肽的氨基酸序列，并且从氨基末端标签; 一个编码它们的核酸; 本发明的包括一个核酸的表达载体的; 和细胞通过本发明的表达载体转化。因此，本发明涉及一种靶蛋白的改进方法，其包括培养的步骤，溶解，并改进转化的细胞。本发明涉及一种自攻融合蛋白包括自切割盒，其besteht分选酶的切割功能结构域和LPXTG氨基酸序列组成并的肽是其识别序列，并且是在DASS模具非常有用炼目标蛋白和消除标签它们的步骤可以由一个单一的细化步骤，而不是单独的步骤中完成。特别是，本发明可以在各种领域中，需要巨大的高纯度的蛋白质的量，DASS在氨基末端存在于靶蛋白可以采用增强列联合能力和融合蛋白的自切割能力，靶蛋白去掉标签获得具有高纯度的，切割缓冲器使得精炼和消除标签的步骤，通过一个单一的步骤中完成，显着地减少所需的用于精炼的时间和努力，并通过一个单一的步骤中完成降低了蛋白质产量的损失。具体地说，本发明是用于制造治疗性抗体 - 药物复合物是有用的。

8. 发明公开

EP2733153A1 Methods for the preparation of immunoconjugates and uses thereof 审中-公开
标题翻译： Verfahren zur Herstellung von Immunkonjugaten und Verwendungen davon
公开(公告)号：EP2733153A1
公开(公告)日：2014-05-21
申请号：EP12306424.8
申请日：2012-11-15
申请人： INSERM (Institut National de la Santé et de la Recherche Médicale)
发明人： The designation of the inventor has not yet been filed
IPC分类号： C07K16/00 , C12N9/48
CPC分类号： C07K16/00 , A01K67/0278 , A01K2217/072 , A01K2227/105 , A01K2267/01 , C07K2317/50 , C12N9/6472 , C12Y304/2207
摘要： The present invention relates to a fusion protein consisting of a constant domain (CL) of a light chain of an immunoglobulin fused at its C-terminal end to a polypeptide (P) characterized by the sequence: LPXT G ⁢ n wherein n is an integer number and X represents an amino acid other than tryptophan and cysteine.
摘要翻译：本发明涉及一种融合蛋白，其由其C-末端融合的免疫球蛋白的轻链的恒定结构域（CL）与多肽（P）组成，所述多肽（P）的特征在于序列：LPXT G ¢n，其中n为整数，X表示不同于色氨酸和半胱氨酸的氨基酸。

9. 发明公开

EP3411492A1 THIOL-BASED DEEP EUTECTIC SOLVENT 审中-公开
公开(公告)号：EP3411492A1
公开(公告)日：2018-12-12
申请号：EP17703394.1
申请日：2017-02-03
申请人： F. Hoffmann-La Roche AG
发明人： BOENITZ-DULAT, Mara , SCHATTE, Martin
IPC分类号： C12P21/02 , C07C215/90 , C07C323/12 , C07C323/66
CPC分类号： C12P21/06 , C07C215/40 , C07C323/12 , C07C323/66 , C07K1/02 , C12N9/6472 , C12P21/02 , C12Y304/2207 , G01N33/532
摘要： A deep eutectic solvent consisting of (2-hydroxyethyl) trimethyl ammonium chloride and dithiothreitol in a molar ratio of from 1:2 to 1:3 and from 0% to 10% co-solvent, and methods of enzymatic production of polypeptides using the deep eutectic solvent.

10. 发明公开

EP3353310A1 PROCESS FOR PRODUCING THIOESTERS EMPLOYING A SORTASE A 有权
公开(公告)号：EP3353310A1
公开(公告)日：2018-08-01
申请号：EP16775567.7
申请日：2016-09-22
申请人： F. Hoffmann-La Roche AG
发明人： SCHATTE, Martin , BOENITZ-DULAT, Mara , BERGMANN, Frank
IPC分类号： C12P11/00 , C12P21/02
CPC分类号： C12N9/52 , A61K47/65 , C12P11/00 , C12P21/02 , C12Y304/2207
摘要： Herein is reported a method for the enzymatic production of a thioester comprising incubating i) a first polypeptide comprising the amino acid sequence LPXTG (SEQ ID NO: 01, wherein X can be any amino acid residue), ii) a second polypeptide that has at its N-terminus a cysteine amino acid residue or is a cysteinyl compound, and iii) a third polypeptide with sortase A activity.

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