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）。

公开(公告)号：KR1020020017501A

公开(公告)日：2002-03-07

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bcl-2基因的过表达增加CHO细胞中人源化抗体产生和治疗丁酸钠的方法，从而增加动物细胞产物如人源化抗体和细胞因子的单位生产。 构成：通过将过表达bcl-2基因的载体导入到用编码人源化抗体的基因转化的CHO细胞中，获得具有抗细胞凋亡的细胞系，然后加入丁酸钠，从而增加抗体产生。 增加人源化抗体生产的方法包括以下步骤：将过表达人bcl-1基因的载体导入上述重组CHO细胞; 将用载体转化的重组CHO细胞接种到培养基中; 并向其中加入丁酸钠。

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。