公开(公告)号：US08961971B2

公开(公告)日：2015-02-24

申请号：US13720573

申请日：2012-12-19

Applicant: Development Center for Biotechnology ,  DCB-USA LLC

Inventor： Yu-Shen Hsu ,  Show-Shan Sheu ,  Ming-I Chang ,  Ming-Chuan Chang ,  Ta-Tung Yuan

IPC: A61K39/00 ,  A61K39/395 ,  A61K38/00 ,  A61K38/04 ,  C07K5/00 ,  C07K7/00 ,  C07K16/00 ,  C07K17/00 ,  C12P21/08 ,  A61K38/10 ,  A61K38/16 ,  A61K38/08 ,  C07K16/46 ,  C07K7/06 ,  C07K7/08 ,  C07K16/28

CPC classification number: C07K16/468 ,  C07K7/06 ,  C07K7/08 ,  C07K16/2809 ,  C07K16/2887 ,  C07K2317/53 ,  C07K2317/622 ,  C07K2317/64 ,  C07K2317/732 ,  C07K2317/74 ,  C07K2317/94

Abstract: This invention relates to bispecific antibodies having combinations of linker and hinge sequences to create linker-hinge interface domains with biological significance. Such linker-hinge interface domains covalently join two molecules, maintain the biological activities of linked molecules (target binding), stabilize the biological characteristics of new molecule (solubility and 4° C. stability), maintain the chemical, biochemical and physical properties (cytotoxicity) of the linked molecules, and modulate the biological characteristics of the linked molecules (activating T-lymphocytes without significant sign of proliferations). Both linker (GGGGS) and hinge (CPPCP) sequences are required to establish functional linker-hinge interface domains as deletion of any of the component resulted in significant lost of T-lymphocyte mediated activity.

Abstract translation: 本发明涉及具有接头和铰链序列组合的双特异性抗体以产生具有生物学意义的接头 - 铰链界面结构域。 这种接头 - 铰链界面域共价连接两个分子，保持连接分子的生物活性（靶结合），稳定新分子的生物学特性（溶解度和4℃稳定性），保持化学，生化和物理性质（细胞毒性 ），并调节连接分子的生物学特性（活化T淋巴细胞而没有显着增殖迹象）。 需要两种接头（GGGGS）和铰链（CPPCP）序列来建立功能性接头 - 铰链界面区域，因为任何组分的缺失导致T淋巴细胞介导的活性的显着丧失。

公开(公告)号：US20150184215A1

公开(公告)日：2015-07-02

申请号：US14584836

申请日：2014-12-29

Applicant: Development Center for Biotechnology

Inventor： Yu-Shen Hsu ,  Show-Shan Sheu ,  Bo-Chin Lei ,  Tsan-Hui Wu

IPC: C12P21/00 ,  C07K1/20 ,  C07K14/34 ,  C07K1/18 ,  C07K1/22

CPC classification number: C07K1/20 ,  C07K14/34 ,  C12P21/02

Abstract: A method for recombinant production of a CRM197 protein includes culturing a recombinant Escherichia coli cell to produce said CRM197 protein, and isolating said CRM197 protein. The recombinant Escherichia coli cell includes an expression vector, which contains a nucleic acid molecule that encodes a fusion protein that includes an E. coli periplasmic signal peptide at N terminal and the CRM197 at C terminal. The CRM197 is encoded by a polynucleotide having the sequence of SEQ ID NO: 1. The E. coli periplasmic signal peptide comprise a pelB leader sequence. The nucleic acid molecule comprises the sequence of SEQ ID NO:2. The Escherichia coli cell is BL21(DE3)pLysS.

Abstract translation: 重组生产CRM197蛋白的方法包括培养重组大肠杆菌细胞以产生所述CRM197蛋白，并分离所述CRM197蛋白。 重组大肠杆菌细胞包含表达载体，其含有编码融合蛋白的核酸分子，所述融合蛋白包括N末端的大肠杆菌周质信号肽和C末端的CRM197。 CRM197由具有SEQ ID NO：1的序列的多核苷酸编码。大肠杆菌周质信号肽包含pelB前导序列。 核酸分子包含SEQ ID NO：2的序列。 大肠杆菌细胞是BL21（DE3）pLysS。

公开(公告)号：US20190330376A1

公开(公告)日：2019-10-31

申请号：US16475121

申请日：2017-12-29

Applicant: Development Center for Biotechnology

Inventor： Chen-Li Chien ,  Gregory Jiann Chen ,  Chuan-Lung Hsu ,  Jei-Hwa Yu ,  Hsien-Yu Tsai ,  Show-Shan Sheu ,  Wei-Jung Chang ,  Chia-Cheng Wu

IPC: C07K16/46 ,  C07K14/745

Abstract: A recombinant protein drug includes a parent protein drug coupled with a modified kininogen-1 peptide. The modified kininogen-1 peptide has the sequence of SEQ ID NO:2 or a homolog having a sequence identity of 80% or higher. The parent protein drug is a bispecific antibody having a first targeting domain linked by a bridging domain with a second targeting domain. The modified kininogen-1 peptide is fused between the first targeting domain and the bridging domain, or between the bridging domain and the second targeting domain. A method for increasing the serum half-life of a protein drug includes constructing a fusion protein comprising the protein drug coupled with a modified kininogen-1 peptide.

公开(公告)号：US20130165629A1

公开(公告)日：2013-06-27

申请号：US13720573

申请日：2012-12-19

Applicant: Development Center for Biotechnology ,  DCB-USA LLC

Inventor： Yu-Shen Hsu ,  Show-Shan Sheu ,  Mingi Chang ,  Ming-Chuan Chang ,  Ta-Tung Yuan

IPC: C07K16/46 ,  C07K7/06 ,  C07K7/08

CPC classification number: C07K16/468 ,  C07K7/06 ,  C07K7/08 ,  C07K16/2809 ,  C07K16/2887 ,  C07K2317/53 ,  C07K2317/622 ,  C07K2317/64 ,  C07K2317/732 ,  C07K2317/74 ,  C07K2317/94

Abstract: This invention relates to bispecific antibodies having combinations of linker and hinge sequences to create linker-hinge interface domains with biological significance. Such linker-hinge interface domains covalently join two molecules, maintain the biological activities of linked molecules (target binding), stabilize the biological characteristics of new molecule (solubility and 4° C. stability), maintain the chemical, biochemical and physical properties (cytotoxicity) of the linked molecules, and modulate the biological characteristics of the linked molecules (activating T-lymphocytes without significant sign of proliferations). Both linker (GGGGS) and hinge (CPPCP) sequences are required to establish functional linker-hinge interface domains as deletion of any of the component resulted in significant lost of T-lymphocyte mediated activity.

Abstract translation: 本发明涉及具有接头和铰链序列组合的双特异性抗体以产生具有生物学意义的接头 - 铰链界面结构域。 这种接头 - 铰链界面域共价连接两个分子，保持连接分子的生物活性（靶结合），稳定新分子的生物学特性（溶解度和4℃稳定性），保持化学，生化和物理性质（细胞毒性 ），并调节连接分子的生物学特性（活化T淋巴细胞而没有显着增殖迹象）。 需要两种接头（GGGGS）和铰链（CPPCP）序列来建立功能性接头 - 铰链界面区域，因为任何组分的缺失导致T淋巴细胞介导的活性的显着丧失。