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    헤파타이티스 B 바이러스의 pre-S2 표면항원에 대한 키메라 항체를 생산하는 세포주와 그 항체
    23.
    发明授权
    헤파타이티스 B 바이러스의 pre-S2 표면항원에 대한 키메라 항체를 생산하는 세포주와 그 항체 失效
    用于生产HBV PRE-S2的CHIMERA抗体的细胞系

    公开(公告)号：KR1019950009841B1

    公开(公告)日：1995-08-29

    申请号：KR1019920007593

    申请日：1992-05-04

    Applicant: 한국과학기술연구원

    Inventor： 한문희 ,  홍효정 ,  박성섭 ,  류춘제 ,  진병래

    IPC: C12N15/51

    Abstract: The chimeric antibody to pre-S2 of HBV (I) of subtype adr is prepared by; A (for heavy chain) (1) cutting DNA of aLys-30 of 8.9 kb with BanHI and BglII for HuCr1, (2) inserting that fragment into plasmid PSV2-neo to get PVS2neo-hCr1, (3) cutting DNA of pMHG-S2 of 9.4 kb and adding EcoRI linker, and (4) inserting that fragment into PVS2neo-hCr1 to get plasmid pHS2-neo (KCTC 0031BP); (for light chain) (5) extracting hygro gene from pSVhygro, (6) cutting pHuCk with EcoRI to get HuCk gene, (7) cutting pMGK-S2 with XmnI to get MuVk and Ek genes and adding XhoI linker, (8) inserting genes of (5), (6), (7) into plasmid pBluescript SK+ to get plasmid pLS2-hygro, C (KCTC 0032BP), (for transfectoma) (9) transfecting DNA fragments of PvuI treated pHS2-neo and SpeI/NotI treated pLS2-hygro to get A-44 (KCTC 0033BP). From A-44 transfectoma, 55kd size of heavy chain and 25kd of light chain of (1) were purified.

    Abstract translation: 亚型adr的HBV（I）前S2的嵌合抗体通过以下方法制备： A（用于重链）（1）用BamHI和BglII切割用于HuCr1的8.9kb的Lys-30 DNA，（2）将该片段插入质粒PSV2-neo以获得PVS2neo-hCr1，（3）切割pMHG- S2，9.4kb，加入EcoRI接头，（4）将该片段插入到PVS2neo-hCr1中，得到质粒pHS2-neo（KCTC 0031BP）; （轻链）（5）从pSVhygro提取hygro基因，（6）用EcoRI切割pHuCk得到HuCk基因，（7）用XmnI切割pMGK-S2以获得MuVk和Ek基因并加入XhoI接头，（8）插入 （5），（6），（7）的基因转化到质粒pBluescript SK +中以获得质粒pLS2-hygro，C（KCTC 0032BP）（用于转染瘤）（9）转染PvuI处理的pHS2-neo和SpeI / NotI的DNA片段 治疗的pLS2-hygro得到A-44（KCTC 0033BP）。 从A-44转染瘤，纯化了55kd大小的重链和25kd的轻链（1）。

    B형간염바이러스의다른두항원을동시에인식하는이중특이항체및그의제조방법
    26.
    发明授权
    B형간염바이러스의다른두항원을동시에인식하는이중특이항체및그의제조방법 失效
    乙型肝炎病毒BISPECIFIC抗体及其制备方法

    公开(公告)号：KR100267058B1

    公开(公告)日：2000-09-15

    申请号：KR1019970076922

    申请日：1997-12-29

    Applicant: 한국과학기술연구원

    Inventor： 홍효정 ,  류춘제 ,  박성섭

    IPC: C07K16/08

    Abstract: PURPOSE: A bispecific antibody being capable of recognizing antigens of hepatitis B virus and a producing method thereof are provided, which can recognize hepatitis B virus antigens which are different each other, and is thus effectively used in the prevention and treatment of hepatitis. CONSTITUTION: The bispecific antibody TBSIg recognizing antigens of hepatitis B virus contains amino acid sequence represented by sequence ID No. 12 and comprises a S antigen of hepatitis B virus recognizing region, link peptides, a pre-S2 of hepatitis B virus recognizing region, and human antibody Fc region. An expression vector pCMV-dhfr-TBSIg contains the bispecific TBSIg gene represented by sequence ID No. 11. The transformant E. coli(KCTC 8861P) is produced by transforming with the expression vector pCMV-dhfr-TBSIg. The CHO cell line BSG-18H produces the bispecific antibody TBSIg. The method for producing the bispecific antibody TBSIg comprises the steps of: transforming CHO cell line with the expression vector pCMV-dhfr-TBSIg; selecting CHO cell line capable of mass producing the bispecific antibody by incubating the transformed CHO cell line in a medium containing MTX; transferring the CHO cell line into a serum free medium and incubating it; and subjecting the fermented culture to column chromatography to obtain the bispecific antibody. The sequences ID. No. 11 and 12 above are described as in the description.

    Abstract translation: 目的：提供能够识别乙型肝炎病毒抗原的双特异性抗体及其制备方法，其可以识别彼此不同的乙型肝炎病毒抗原，因此有效地用于预防和治疗肝炎。 构成：识别乙型肝炎病毒抗原的双特异性抗体TBSIg含有由序列号12表示的氨基酸序列，包含乙型肝炎病毒识别区域的S抗原，链接肽，乙型肝炎病毒识别区域的前S2位点，以及 人抗体Fc区。 表达载体pCMV-dhfr-TBSIg含有由序列号11表示的双特异性TBSIg基因。通过用表达载体pCMV-dhfr-TBSIg转化产生转化体大肠杆菌（KCTC 8861P）。 CHO细胞系BSG-18H产生双特异性抗体TBSIg。 制备双特异性抗体TBSIg的方法包括以下步骤：用表达载体pCMV-dhfr-TBSIg转化CHO细胞系; 选择能够通过在含有MTX的培养基中培养转化的CHO细胞系来大量产生双特异性抗体的CHO细胞系; 将CHO细胞系转移到无血清培养基中并孵育; 并将发酵的培养物进行柱色谱以获得双特异性抗体。 序列ID。 上述11和12描述如下。

    B형간염바이러스의표면항원프리-S1에대한인간화항체및이의제조방법
    27.
    发明公开
    B형간염바이러스의표면항원프리-S1에대한인간화항체및이의제조방법 失效
    乙型肝炎病毒表面抗原预处理人类抗原的制备

    公开(公告)号：KR1020000033009A

    公开(公告)日：2000-06-15

    申请号：KR1019980049664

    申请日：1998-11-19

    Applicant: 주식회사 녹십자홀딩스 ,  한국과학기술연구원

    Inventor： 홍효정 ,  류춘제 ,  허향숙

    IPC: C12N15/51 ,  C12N15/62

    CPC classification number: C12N15/70 ,  A61K2039/505 ,  C07K14/02 ,  C12N15/62

    Abstract: PURPOSE: Humanized antibody on surface antigen pre-S1 of hepatitis B virus (HBV) is provided which is derived from mouse monoclonal antibody (KR 359) The humanized antibody maintains the affinity on the antigen but decreases the antigenecity of mouse antibody on the human body by changing amino acid residues specific for mouse into human specific one. CONSTITUTION: Humanized variable region of heavy chain is derived from variable region of heavy chain of human antibody DP67 substituted into mouse framework residues and complementarity-determining region (CDR)1, and 3 and part of 2. Complete humanized heavy chain is constructed by joining leader sequence for secretion, constant region of heavy chain from pRC/CMV-HC-HuS, and humanized variable region. Humanized variable region of light chain is derived from variable region of light chain of human antibody DPK9 substituted into mouse framework residues and complementarity-determining region (CDR)1, 2, and 3. Complete humanized light chain is constructed by joining leader sequence for secretion, constant region of heavy chain from pKC-hr-HuS, and humanized variable region. Each complete chain is cloned into expression vector, pRc/CMV, and transfected into COS7 cell for the production of antibody.

    Abstract translation: 目的：提供源自小鼠单克隆抗体（KR359）的乙型肝炎病毒（HBV）表面抗原前S1上的人源化抗体人源化抗体对抗原保持亲和力，但降低小鼠抗体对人体的抗原性 通过将特异性小鼠的氨基酸残基改变为人特异性的。 构成：重链人源化可变区源自人抗体DP67的重链可变区，被取代成小鼠框架残基和互补决定区（CDR）1，3和部分2。完全人源化重链通过连接 pRC / CMV-HC-HuS重链序列，人源化可变区。 轻链的人源化可变区源自人抗体DPK9的轻链可变区，其被取代成小鼠框架残基和互补决定区（CDR）1,2和3.完全人源化轻链是通过连接前导序列分泌构建的 ，来自pKC-hr-HuS的重链恒定区和人源化可变区。 将每条完整链克隆到表达载体pRc / CMV中，并转染到COS7细胞中以产生抗体。

    B형간염바이러스의표면항원프리-S1에대한인간화항체및이의제조방법
    28.
    发明公开
    B형간염바이러스의표면항원프리-S1에대한인간화항체및이의제조방법 失效
    乙型肝炎病毒表面抗原预处理人类抗原的制备

    公开(公告)号：KR1020000033008A

    公开(公告)日：2000-06-15

    申请号：KR1019980049663

    申请日：1998-11-19

    Applicant: 주식회사 녹십자홀딩스 ,  한국과학기술연구원

    Inventor： 홍효정 ,  류춘제 ,  허향숙

    IPC: C12N15/51 ,  C12N15/62

    CPC classification number: C07K16/082 ,  C07K2317/24 ,  C07K2317/56 ,  C07K2317/92

    Abstract: PURPOSE: Humanized antibody on surface antigen pre-S1 of hepatitis B virus (HBV) is provided which is derived from mouse monoclonal antibody (KR 127), The humanized antibody maintains the affinity on the antigen but decreases the antigenecity of mouse antibody on the human body by changing amino acid residues specific for mouse into human specific one. CONSTITUTION: Humanized variable region of heavy chain is derived from variable region of heavy chain of human antibody DP7 substituted into mouse framework residues and complementarity-determining region (CDR)1 and 3, and part of 2. Complete humanized heavy chain is constructed by joining leader sequence for secretion, constant region of heavy chain from pRC/CMV-HC-HuS, and humanized variable region. Humanized variable region of light chain is derived from variable region of light chain of human antibody DPK12 substituted into mouse framework residues and complementarity-determining region (CDR)1, 2, and 3. Complete humanized light chain is constructed by joining leader sequence for secretion, constant region of heavy chain from pKC-hr-HuS, and humanized variable region. Each complete chain is cloned into expression vector, pRc/CMV, and transfected into COS7 cell for the production of antibody.

    Abstract translation: 目的：提供源自小鼠单克隆抗体（KR 127）的乙型肝炎病毒（HBV）表面抗原前S1上的人源化抗体，人源化抗体对抗原保持亲和力，但降低小鼠抗体对人的抗原性 通过将特异性的小鼠的氨基酸残基改变成人特异性的身体。 构成：重链人源化可变区源自人抗体DP7的重链可变区，取代于小鼠框架残基和互补决定区（CDR）1和3中，部分为2.完全人源化重链通过连接 pRC / CMV-HC-HuS重链序列，人源化可变区。 轻链的人源化可变区源自人抗体DPK12的轻链可变区，被取代成小鼠框架残基和互补决定区（CDR）1,2和3.完全人源化轻链是通过连接前导序列分泌构建的 ，来自pKC-hr-HuS的重链恒定区和人源化可变区。 将每条完整链克隆到表达载体pRc / CMV中，并转染到COS7细胞中以产生抗体。

    B형간염바이러스의다른두항원을동시에인식하는이중특이항체및그의제조방법
    29.
    发明公开
    B형간염바이러스의다른두항원을동시에인식하는이중특이항체및그의제조방법 失效
    双抗体同时识别乙型肝炎病毒的两种抗原及其制备方法

    公开(公告)号：KR1019990056898A

    公开(公告)日：1999-07-15

    申请号：KR1019970076922

    申请日：1997-12-29

    Applicant: 한국과학기술연구원

    Inventor： 홍효정 ,  류춘제 ,  박성섭

    IPC: C07K16/08

    Abstract: 본 발명은 B형 간염 바이러스의 다른 두 항원을 동시에 인식하는 이중특이 항체 및 그의 제조방법에 관한 것이다. 구체적으로, 본 발명은 B형 간염 바이러스 (HBV) 표면항원인 S 항원 및 프리-S2 항원을 동시에 인식하는 이중특이 항체 (bispecific antibody), 그의 유전자, 이를 포함하는 발현 벡터, 상기 항체를 생산하는 세포주 및 이를 이용하여 간염 항체를 얻는 제조방법에 관한 것으로서, 상기 이중특이 항체는 HBV 입자를 효과적으로 인식하여 간염을 예방하고 치료하는데 유용하게 사용될 수 있다.

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