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    • 2. 发明公开
    • 플라스미드 DNA의 분리방법
    • 用于分离用于基因治疗的PLASMID DNA的方法
    • KR1020030038063A
    • 2003-05-16
    • KR1020010069476
    • 2001-11-08
    • 동아쏘시오홀딩스 주식회사
    • 김병문김채영최재훈정문섭이동섭강수형김원배
    • C12N15/10
    • PURPOSE: A process for separating plasmid DNA for gene therapy is provided, thereby separating high quality of plasmid DNA without using any toxic organic solvents and animal origin enzymes. CONSTITUTION: A process for separating plasmid DNA for gene therapy comprises the steps of: (1) alkaline-dissolving cells containing plasmid DNA to remove impurities, such as proteins and lipids; (2) removing DNA derived from a host cell; (3) removing RNA, dimer plasmid and bacterial endotoxin derived from the host cell using a bacterial endotoxin removing buffer solution and anionic exchange chromatography; and (4) purifying it using gel filtration chromatography, wherein the impurities are removed by using ammonium acetate precipitation or phenol-chloroform precipitation; and the DNA derived from the host cell is removed by using polyethylene glycol precipitation or nitro cellulose membrane adsorption.
    • 目的:提供用于分离用于基因治疗的质粒DNA的方法,从而分离高质量的质粒DNA,而不使用任何有毒有机溶剂和动物来源的酶。 构成:用于分离用于基因治疗的质粒DNA的方法包括以下步骤:(1)将含有质粒DNA的细胞碱溶解以除去蛋白质和脂质等杂质; (2)去除源自宿主细胞的DNA; (3)使用细菌内毒素去除缓冲溶液和阴离子交换色谱法除去源自宿主细胞的RNA,二聚体质粒和细菌内毒素; 和(4)使用凝胶过滤色谱纯化,其中通过使用乙酸铵沉淀或苯酚 - 氯仿沉淀除去杂质; 并且通过使用聚乙二醇沉淀或硝基纤维素膜吸附除去来自宿主细胞的DNA。
    • 3. 发明公开
    • 이동성이 있는 분비가능한 동형삼중체 형성능을 가진 재조합 트레일을 코딩하는 유전자를 포함하는 유전자 전달체
    • 包含基因编码可溶性和可共享共转化子的基因递送载体形成重组人TRAIL
    • KR1020070097206A
    • 2007-10-04
    • KR1020060028087
    • 2006-03-28
    • 동아쏘시오홀딩스 주식회사
    • 정문섭김채영김병문
    • C12N15/861C12N15/09A61K35/76A61K9/00
    • C12N15/861A61K9/0019A61K35/76C12N15/09
    • A gene-delivering vector comprising a gene encoding a recombinant human TRAIL(TNF(tumor necrosis factor)-related apoptosis-inducing ligand) having ability of forming soluble and secretable homotrimer is provided to inhibit growth of cancer cells, induce apoptosis of cancer cells, and improve safety of use by eliminating infectious properties from the vector through removing E1 and E3 genes associated with adenovirus replication. A shuttle vector pAdloxTRAIL for producing replication-incompetent adenovirus comprises an expression cassette containing a secretion signal sequence of SEQ ID NO:3, a trimer-forming domain of SEQ ID NO:4 and TRAIL gene of SEQ ID NO:1, a human gene high expression promoter selected from CMV(cytomegalovirus), RSV(Rous sarcoma virus) and beta-actin and loxP gene, wherein E1 and E3 genes required for adenovirus replication are removed from the shuttle vector. A replication-incompetent adenovirus Ad-stTRAIL(KCCM 10743P) produces soluble and secretable homotrimer. An anticancer agent comprises the replication-incompetent adenovirus.
    • 提供了包含编码具有形成可溶性和可分泌同源三聚体的能力的重组人TRAIL(TNF(肿瘤坏死因子)相关凋亡诱导配体)的基因的基因递送载体,以抑制癌细胞的生长,诱导癌细胞的凋亡, 并通过去除与腺病毒复制相关的E1和E3基因,从载体中消除感染性,从而提高使用的安全性。 用于产生复制无能的腺病毒的穿梭载体pAdloxTRAIL包含含有SEQ ID NO:3的分泌信号序列,SEQ ID NO:4的三聚体形成结构域和SEQ ID NO:1的TRAIL基因,人基因 选自CMV(巨细胞病毒),RSV(Rous肉瘤病毒)和β-肌动蛋白和loxP基因的高表达启动子,其中从穿梭载体中除去腺病毒复制所需的E1和E3基因。 复制无能的腺病毒Ad-stTRAIL(KCCM 10743P)产生可溶性和可分泌的同三聚体。 抗癌剂包括无复制性腺病毒。
    • 10. 发明公开
    • 만성 B형 간염의 치료를 위한 DNA 백신 및 그의 제조방법
    • 用于治疗慢性乙型肝炎的DNA疫苗及其制备方法
    • KR1020080015211A
    • 2008-02-19
    • KR1020060076619
    • 2006-08-14
    • 포항공과대학교 산학협력단주식회사 포스코동아쏘시오홀딩스 주식회사주식회사 제넥신주식회사 대웅제약
    • 성영철양세환임세진이창근박수형송만기손종문윤승규김채영김병문이성희김원배
    • A61K48/00
    • A61K48/0058C12N15/90Y10S514/894
    • A poly-antigenic HBV DNA vaccine is provided to induce excellently an antibody reaction and a cell mediated immunity by comprising genes coding a surface antigen of an HBV as well as a core antigen and a polymerase antigen, thereby capable of preventing and treating chronic B-type hepatitis. An HBV DNA vaccine for treating chronic B-type hepatitis comprises genes coding an HBsAg, an L-protein, a core and a polymerase antigen of an HBV, wherein at least one of the genes codes a codon-optimized antigen, the genes coding the HBsAg and L-protein are introduced simultaneously into one expression vector, and the genes coding the core antigen and polymerase are introduced simultaneously into another expression vector. The vaccine further comprises an immunological adjuvant. A method for preparing the HBV DNA vaccine comprises the steps of: (a) codon-optimizing a gene coding an HBsAg, an L-protein, and a core antigen of an HBV; and (b) introducing the codon-optimized gene coding the HBsAg, L-protein, and core antigen and a gene coding a polymerase of the HBV, which is codon-optimized or not codon-optimized, into at least one expression vector. A method for treating chronic B-type hepatitis comprises a step of administering the vaccine.
    • 提供多抗原性HBV DNA疫苗以通过包含编码HBV表面抗原的基因以及核心抗原和聚合酶抗原来极好地诱导抗体反应和细胞介导的免疫,由此能够预防和治疗慢性B- 型肝炎。 用于治疗慢性B型肝炎的HBV DNA疫苗包括编码HBV的HBsAg,L-蛋白,核心和聚合酶抗原的基因,其中至少一个基因编码密码子优化的抗原,编码 将HBsAg和L-蛋白同时引入一个表达载体中,将编码核心抗原和聚合酶的基因同时引入另一种表达载体。 疫苗还包含免疫佐剂。 制备HBV DNA疫苗的方法包括以下步骤:(a)密码子优化编码HBV的HBsAg,L-蛋白和核心抗原的基因; 和(b)将编码HBsAg,L-蛋白和核心抗原的密码子优化的基因和编码经密码子优化或未经密码子优化的HBV聚合酶的基因导入至少一个表达载体中。 一种治疗慢性B型肝炎的方法包括施用疫苗的步骤。