发明公开
EP3049530A2 VERFAHREN UND VORRICHTUNG ZUR ZELLFREIEN PROTEINSYNTHESE IN GEGENWART EINES CASPASE-INHIBITORS
审中-公开
方法和设备,在一个半胱氨酸蛋白酶抑制剂的存在无细胞蛋白质合成
- 专利标题: VERFAHREN UND VORRICHTUNG ZUR ZELLFREIEN PROTEINSYNTHESE IN GEGENWART EINES CASPASE-INHIBITORS
- 专利标题(英): Method and device for cell-free protein synthesis in the presence of a caspase inhibitor
- 专利标题(中): 方法和设备,在一个半胱氨酸蛋白酶抑制剂的存在无细胞蛋白质合成
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申请号: EP14777515.9申请日: 2014-09-17
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公开(公告)号: EP3049530A2公开(公告)日: 2016-08-03
- 发明人: KUBICK, Stefan , STECH, Marlitt , WÜSTENHAGEN, Doreen , QUAST, Robert
- 申请人: Fraunhofer Gesellschaft zur Förderung der angewandten Forschung E.V.
- 申请人地址: Hansastrasse 27C 80686 München DE
- 专利权人: Fraunhofer Gesellschaft zur Förderung der angewandten Forschung E.V.
- 当前专利权人: Fraunhofer Gesellschaft zur Förderung der angewandten Forschung E.V.
- 当前专利权人地址: Hansastrasse 27C 80686 München DE
- 代理机构: v. Bezold & Partner Patentanwälte - PartG mbB
- 优先权: DE102013015977 20130925; DE102013020900 20131211
- 国际公布: WO2015043729 20150402
- 主分类号: C12P21/02
- IPC分类号: C12P21/02 ; C12M1/00 ; C07K5/083
摘要:
The invention relates to an improved cell-free protein synthesis method using a eukaryotic cell lysate in the presence of a caspase inhibitor, to a device for carrying out said method, and to the use of a caspase inhibitor for increasing the protein yield or for increasing the stability of the synthesized proteins in such a cell-free protein synthesis method using a eukaryotic cell lysate. In a preferred embodiment of the invention, the cell-free protein synthesis method is carried out as a continuous method in a device which comprises at least two compartments separated by a dialysis membrane. The translation reaction takes place in at least one first compartment, the reaction compartment, and during the translation reaction i) reactants are diffused from at least one other compartment, the supply and discharge compartment, into the reaction compartment, and ii) reaction byproducts are diffused from the reaction compartment into the supply and discharge compartment by means of the dialysis membrane. According to the invention, the caspase inhibitor is provided at least in the reaction compartment but can also be preferably provided in the supply and discharge compartment in order to supply unused inhibitors to the reaction compartment.
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