公开(公告)号：KR100397791B1

公开(公告)日：2003-09-13

申请号：KR1020000050868

申请日：2000-08-30

Applicant: 한국과학기술원

Inventor： 이균민 ,  김노수 ,  홍효정

IPC: C12P21/08

Abstract: PURPOSE: Provided is a method for increasing humanized antibody production in CHO cells by overexpression of bcl-2 gene and treatment of sodium butyrate, thereby increasing unit production of animal cell products, such as humanized antibodies and cytokines. CONSTITUTION: The antibody production is increased by introducing a vector which overexpresses bcl-2 gene into CHO cells transformed with a gene encoding a humanized antibody to obtain a cell line having apoptosis resistance, then adding sodium butyrate. The method for increasing the production of humanized antibodies comprises the steps of: introducing a vector which overexpressing human bcl-1 gene into the recombinant CHO cells above; inoculating the recombinant CHO cells transformed with the vector to a medium; and adding sodium butyrate thereto.

Abstract translation: 目的：提供通过过量表达bcl-2基因和处理丁酸钠来增加CHO细胞中人源化抗体产生的方法，从而增加动物细胞产物如人源化抗体和细胞因子的单位产量。 构成：通过向用编码人源化抗体的基因转化的CHO细胞中导入过量表达bcl-2基因的载体，以获得具有抗细胞凋亡性的细胞系，然后加入丁酸钠，增加抗体产量。 增加人源化抗体产生的方法包括以下步骤：将过量表达人bcl-1基因的载体导入上述重组CHO细胞中; 将用该载体转化的重组CHO细胞接种到培养基上; 并向其中加入丁酸钠。

公开(公告)号：KR1020030060010A

公开(公告)日：2003-07-12

申请号：KR1020020000603

申请日：2002-01-05

Applicant: 한국과학기술원

Inventor： 이균민 ,  이석규

IPC: C12N5/16

CPC classification number: C12P21/02 ,  C07K14/4747 ,  C12N2510/00 ,  C12N2510/02

Abstract: PURPOSE: Provided is a transformed CHO cell line which inhibits the apoptosis due to the over-expression of the anti-apoptosis protein, thereby heightening the cellular viability. This enhances the productivity of the target proteins, and maintains the integrity of the cell membrane. CONSTITUTION: A CHO cell line comprises genes transformed into the anti-apoptosis, and lacks the DHFR gene. The anti-apoptosis genes are selected from the group consisting of BcI-2, E1B-19K, BcI-W, McI-1, and IAP. A method of preparing the transformed CHO cell line comprises the steps of: adapting the CHO dhfr(-) cell line attached-cultured in a medium containing serum to the float-culture using a serum free medium; and introducing a vector containing anti-apoptosis genes to the adapted cell line, and selecting the CHO host cell line where the anti-apoptosis proteins are over-expressed.

Abstract translation: 目的：提供由于抗凋亡蛋白过度表达而抑制细胞凋亡的转化CHO细胞系，从而提高细胞活力。 这提高了目标蛋白质的生产力，并保持细胞膜的完整性。 构成：CHO细胞系包含转化为抗细胞凋亡的基因，缺乏DHFR基因。 抗凋亡基因选自BcI-2，E1B-19K，BcI-W，McI-1和IAP。 制备转化的CHO细胞系的方法包括以下步骤：使用无血清培养基将在含有血清的培养基中附着培养的CHO dhfr（ - ）细胞系适应于浮游培养物; 并将含有抗凋亡基因的载体引入适应的细胞系，并选择其中过度表达抗凋亡蛋白的CHO宿主细胞系。

公开(公告)号：KR102140565B1

公开(公告)日：2020-08-03

申请号：KR1020190018104

申请日：2019-02-15

Applicant: 한국과학기술원

Inventor： 이균민 ,  김채린

IPC: C12N5/071 ,  C07K14/51 ,  C07K14/71 ,  C12N9/06

公开(公告)号：KR101724405B1

公开(公告)日：2017-04-11

申请号：KR1020150090348

申请日：2015-06-25

Applicant: 한국과학기술원

Inventor： 이균민 ,  박진형

IPC: C12N5/00 ,  C07K14/475 ,  C07K14/505 ,  C07K14/52 ,  C07K14/745 ,  C07K16/00 ,  C07K19/00

Abstract: 본발명은고도로갈락토실화된단일클론항체생산증가를위한발레르산을포함하는동물세포배양용배지조성물및 이의용도에관한것으로, 재조합단백질생산을위한 CHO-K1 세포배양배지에발레르산(valeric acid) 1.5 mM를세포접종시첨가하여배양하였을경우, 세포성장이억제되고, 단위세포당생산성및 단일클론항체의생산성이크게향상되며, 이와같은생산성증가가대규모배양에서도적용되고, CHO-K1 세포주뿐만아니라 CHO-DG44 세포주에서동일한효과를갖는것을확인하였다. 또한, 생산된단일클론항체의갈락토실화비율이증가되어품질이개선된것을확인함으로써, 본발명의발레르산이첨가된동물세포배양용배지조성물이생산성및 품질개선을위해유용하게이용될수 있다.

Abstract translation: 本发明是高度半乳糖基失火用于动物细胞培养的生产单克隆抗体的培养基包括酸用于增加组合物并且涉及它们的用途，CHO-K1细胞，在用于重组蛋白生产培养基戊酸（戊酸 ）在培养物中1.5毫米的的情况下，当细胞接种，细胞生长被抑制的溶液中加入，在单元电池的生产和在单克隆抗体的生产力显著一方得到改善，另一方面，和在大规模培养相同的增加生产力应用程序，CHO-K1细胞系作为 但是CHO-DG44细胞系具有相同的效果。 另外，在产生的单克隆抗体的失火率去乳杆菌的增加可以通过检查被使用的质量得到了改善，所述戊酸是用于本发明中有用的除了生产率和质量提高的动物细胞培养的组合物的培养基。

公开(公告)号：KR101605885B1

公开(公告)日：2016-04-04

申请号：KR1020140060984

申请日：2014-05-21

Applicant: 한국과학기술원

Inventor： 이균민 ,  하태광

IPC: C12N5/02 ,  C12P21/00

Abstract: 본발명은재조합단백질생산증가및 시알산화강화를알파케토글루타레이트을포함하는세포배양용배지조성물및 이의용도에관한것으로, 구체적으로본 발명의알파케토글루타레이트(alpha-ketoglutarate)를포함하고, 글루타민(glutamine)은비함유된배지조성물은세포의비생산속도및 세포의생존율을높여목적단백질의최대농도를증가시키고, 목적단백질의시알산화를배양후반부까지유지시킬수 효과를나타냄으로써, 종래의기본배지를사용할경우보다재조합단백질이현저히증가하므로, 상기본 발명의배지는동물세포를이용하여치료용재조합단백질을생산하는관련산업분야의산업적이용에유용하게이용될수 있다.

公开(公告)号：KR100999040B1

公开(公告)日：2010-12-09

申请号：KR1020080049783

申请日：2008-05-28

Applicant: 한국과학기술원

Inventor： 이균민 ,  한영규

IPC: C07K14/565 ,  C12N15/00

Abstract: 본 발명은 인간 인터페론 베타(interferon-β)를 발현하는 벡터를 도입한 형질전환 세포주를 이용하여 목적 단백질인 인간 인터페론 베타 단량체의 생산을 증강시키는 방법에 관한 것으로써, 보다 상세하게는 인간 인터페론 베타를 발현하는 유전자를 포함하는 벡터로 형질 전환된 CHO(Chinese hamster ovary) 세포주를 고삼투압 환경, 또는 고삼투압과 저온이 함께 적용된 환경에서 배양하여 인간 인터페론 베타의 집합화 현상(aggregation)을 줄임으로써 인간 인터페론 베타 중합체보다 활성화도가 상대적으로 높은 인간 인터페론 베타 단량체의 생산을 증강시킬 수 있는 방법에 관한 것이다.
인간 인터페론 베타, 재조합 CHO 세포, 고삼투압

公开(公告)号：KR1020090123622A

公开(公告)日：2009-12-02

申请号：KR1020080049783

申请日：2008-05-28

Applicant: 한국과학기술원

Inventor： 이균민 ,  한영규

IPC: C07K14/565 ,  C12N15/00

Abstract: PURPOSE: A method for enhancing productivity of human interferon beta monomer by reducing aggregation of the human interferon beta is provided. CONSTITUTION: A productivity of human interferon beta monomer is enhanced by transforming the cell line for expressing human interferon beta and culturing in a medium of hyperosmosis. The cell line is CHO(Chinese hamster ovary), VERO, BHK, HeLa, Cv1, MDCK, 293, 3T3, myeloma, PC12 or WI38 cell. The used cell line is CHO cell in which DHFR gene is deleted. The condition of hyperosmosis is 350~550 mOsm/kg. The medium is serum-free medium. A method for enhancing producitivity of human interferone beta monomer comprises a step of transferring transformed cell line to medium of hyperosmosis to express human interferon beta.

Abstract translation: 目的：提供通过减少人类干扰素β的聚集来提高人类干扰素β单体生产率的方法。 构成：人类干扰素β单体的生产力通过转化用于表达人干扰素β的细胞系并在高渗性培养基中培养来增强。 细胞系是CHO（中国仓鼠卵巢），VERO，BHK，HeLa，Cv1，MDCK，293,3T3，骨髓瘤，PC12或WI38细胞。 所用细胞系是其中缺失DHFR基因的CHO细胞。 高渗症状为350〜550mOsm / kg。 培养基为无血清培养基。 提高人类干扰素β单体的生产率的方法包括将转化的细胞系转移到高渗性培养基以表达人干扰素β的步骤。

公开(公告)号：KR1020090119166A

公开(公告)日：2009-11-19

申请号：KR1020080045044

申请日：2008-05-15

Applicant: 한국과학기술원

Inventor： 이균민 ,  전민경

IPC: A61K39/395 ,  A61K39/00

CPC classification number: Y02A50/412 ,  Y02A50/464 ,  Y02A50/472 ,  Y02A50/476 ,  A61K39/0005 ,  A61K39/145

Abstract: PURPOSE: A method for inducing antigen-specific CTL(Cytotoxic T Lymphocyte) using dendritic cell is provided to effectively use for immunotherapy. CONSTITUTION: A vaccine for immunotherapy contains a same dendritic cell(DC) which is derived from monocyte such as HLA-A(Human Leukocyte Antigen-A). An antigen is antigen which is derived from pathogen including pathogenic bacteria, virus and parasite or cancer antigen. The antigen is peptide, lipopolysaccharide, polysaccharide, glycoprotein or polynucleotide. A cell therapeutic agent contains antigen-specific CTL.

Abstract translation: 目的：提供使用树突状细胞诱导抗原特异性CTL（细胞毒性T淋巴细胞）的方法，以有效用于免疫治疗。 构成：用于免疫治疗的疫苗含有从单核细胞如HLA-A（人类白细胞抗原-A）衍生的相同的树突状细胞（DC）。 抗原是源自病原体的抗原，包括病原菌，病毒和寄生虫或癌抗原。 抗原是肽，脂多糖，多糖，糖蛋白或多核苷酸。 细胞治疗剂含有抗原特异性CTL。

公开(公告)号：KR100434118B1

公开(公告)日：2004-06-04

申请号：KR1020010057353

申请日：2001-09-17

Applicant: 한국과학기술원

Inventor： 이균민 ,  김노수

IPC: C12N15/57

CPC classification number: C12N15/1137 ,  A61K38/00 ,  C12N2310/111 ,  C12Y304/22056

Abstract: The present invention relates to an antisense nucleotide of caspase-3, an expression vector encoding antisense RNA of caspase-3, and an inhibition method of cellular apoptosis in recombinant Chinese hamster ovary (CHO) cells using the same. More precisely, the present invention relates to an antisense nucleotide composed of a base sequence represented by Sequence ID No. 1, which suppresses the expression of caspase-3, an expression vector containing the above antisense nucleotide and expressing antisense RNA of caspase-3, and an inhibition method of apoptosis of recombinant cells by suppressing the expression of caspase-3 through introducing the above expression vector into the recombinant cells and then expressing antisense RNA of caspase-3. According to the present invention, apoptosis can be inhibited by suppressing the activation of caspase-3, which is related to the apoptosis, and the integrity of target protein produced in the recombinant cells can be enhanced.

Abstract translation: 本发明涉及半胱氨酸天冬氨酸蛋白酶-3的反义核苷酸，编码半胱天冬酶-3的反义RNA的表达载体以及使用其的重组中国仓鼠卵巢（CHO）细胞中细胞凋亡的抑制方法。 更确切地说，本发明涉及由抑制半胱氨酸蛋白酶-3表达的序列ID No.1表示的碱基序列，含有上述反义核苷酸的表达载体和表达胱天蛋白酶-3的反义RNA组成的反义核苷酸， 以及通过将上述表达载体导入重组细胞中，然后表达胱天蛋白酶-3的反义RNA来抑制重组细胞凋亡的抑制方法。 根据本发明，通过抑制与细胞凋亡有关的半胱天冬酶-3的活化可以抑制细胞凋亡，并且可以增强在重组细胞中产生的靶蛋白的完整性。 ＆lt;图像＆GT;

公开(公告)号：KR1020030024290A

公开(公告)日：2003-03-26

申请号：KR1020010057353

申请日：2001-09-17

Applicant: 한국과학기술원

Inventor： 이균민 ,  김노수

IPC: C12N15/57

CPC classification number: C12N15/1137 ,  A61K38/00 ,  C12N2310/111 ,  C12Y304/22056

Abstract: PURPOSE: An antisense nucleotide of caspase-3, an expression vector thereof, and an inhibition method of apoptosis of a cell using the same antisense nucleotide of caspase-3 are provided, thereby the apoptosis of the cell can be effectively inhibited and quality of an interest protein produced by the cell can be improved. CONSTITUTION: The antisense nucleotide which capable of inhibiting the expression of caspase-3 has the nucleotide sequence of SEQ ID NO: 4. The expression vector ASCASP3-200(KCTC 10038BP) contains the antisense nucleotide of SEQ ID NO: 4 and expresses antisense RNA of caspase-3. The method for inhibition of apoptosis of the cell comprises transforming a recombinant cell with the expression vector ASCASP3-200(KCTC 10038BP) and expressing the antisense RNA of caspase-3, wherein the recombinant cell is Chinese hamster ovary(CHO).

Abstract translation: 目的：提供半胱天冬酶-3的反义核苷酸，其表达载体，以及使用相同的半胱天冬酶-3反义核苷酸的细胞凋亡的抑制方法，从而可有效抑制细胞凋亡， 可以改善由细胞产生的兴趣蛋白质。 构成：能够抑制半胱天冬酶-3表达的反义核苷酸具有SEQ ID NO：4的核苷酸序列。表达载体ASCASP3-200（KCTC 10038BP）含有SEQ ID NO：4的反义核苷酸，并表达反义RNA 的caspase-3。 用于抑制细胞凋亡的方法包括用表达载体ASCASP3-200（KCTC 10038BP）转化重组细胞并表达胱天蛋白酶-3的反义RNA，其中重组细胞是中国仓鼠卵巢（CHO）。