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公开(公告)号：KR1019960041359A
公开(公告)日：1996-12-19
申请号：KR1019950013818
申请日：1995-05-30
Applicant: 한국과학기술연구원 , 주식회사 녹십자홀딩스
Abstract: 본 발명은 B형 간염 바이러스(hepatitis B virus, HBV)의 표면항원 S에 대한 키메라 항체(chimeric antibody), 이의 발현 벡터, 상기 발현 벡터로 형질전환된 트랜스펙토마(transfectoma) 세포주 및 상기 키메라 항체의 제조방법에 관한 것으로, 본 발명의 트랜스펙토마 세포주 KR12H 및 CS 13으로부터 생산된 항체는 HBV의 표면항원 S에 대한 우수한 항원결합능, 경쟁적 결합능, 친화도 및 HBV 중화효과를 나타낸다.
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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
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）。
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公开(公告)号：KR1020000055980A
公开(公告)日：2000-09-15
申请号：KR1019990004939
申请日：1999-02-12
Applicant: 한국과학기술연구원 , 주식회사 녹십자홀딩스
IPC: A61K39/00
Abstract: PURPOSE: A humanized antibody to surface antigen free-S 2 of hepatitis B virus(HBV) is provided, which maintains similar antigen-binding abilities with existing humanized antibody, but diminishes immune-causing abilities than existing humanized antibody obviously, so which can be used to prevent HBV infection and to treat the chronic hepatitis B. CONSTITUTION: A process for the preparation of humanized antibody to HBV surface antigen free-S 2 comprises of: preparing humanized heavy-chained genes; preparing humanized light-chained genes; preparing representing vector(pCMV-HS2(II)HC) producing humanized heavy-chain; preparing representing vector(pKC-dhfr-HS2(II)) producing humanized light-chain; culturing COS7 cells, and washing with OPTI MEM I; mixing pCMV-HS2(II)HC, pKC-dhfr-HS2(II) and Lipofectamine, adding OPTI MEM I, and pouring the mixture over the COS7 cells, culturing, concentrating, and measuring the concentration of the humanized antibody in COS7 cell; investigating binding ability of humanized antibody to HBV surface antigen free-S2; and investigating binding affinity to antigen of humanized antibody.
Abstract translation: 目的：提供乙型肝炎病毒（HBV）表面抗原自由-S2的人源化抗体，其与现有的人源化抗体保持相似的抗原结合能力，但明显降低免疫原性，比现有的人源化抗体明显降低，因此可以 用于预防HBV感染并治疗慢性乙型肝炎。构成：用于制备HBV表面抗原游离-S2的人源化抗体的方法包括：制备人源化重链基因; 制备人源化轻链基因; 制备表达载体（pCMV-HS2（II）HC）产生人源化重链; 制备产生人源化轻链的载体（pKC-dhfr-HS2（II））; 培养COS7细胞，用OPTI MEM I洗涤; 混合pCMV-HS2（II）HC，pKC-dhfr-HS2（II）和Lipofectamine，加入OPTI MEM I，并将混合物倒在COS7细胞上，培养，浓缩和测量COS7细胞中人源化抗体的浓度; 研究人源化抗体对HBV表面抗原游离-S2的结合能力; 并研究人源化抗体对抗原的结合亲和力。
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Patent Agency Ranking
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
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。
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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
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。
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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
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。
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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
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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公开(公告)号：KR1019990008649A
公开(公告)日：1999-02-05
申请号：KR1019970030696
申请日：1997-07-02
Applicant: 한국과학기술연구원
IPC: C07K16/08
Abstract: 본 발명은 B형 간염 바이러스 (HBV)의 표면항원 프리-S1 (pre-S1)에 결합하는 단일클론항체, 이를 생산하는 하이브리도마 세포주 및 그의 제조방법에 관한 것이다. 구체적으로, 본 발명은 HBV 표면항원 프리-S1 의 43-49번 아미노산을 포함하는 에피토프를 특이적으로 인식하는 생쥐 단일클론항체에 관한 것으로서, 이는 여러 아형 및 변이 HBV 를 중화시킬 수 있어 HBV 감염을 예방하고 치료하는데 널리 사용될 수 있다.
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