公开(公告)号：US20110117643A1

公开(公告)日：2011-05-19

申请号：US13002814

申请日：2009-07-07

申请人： Keun Soo Kim ,  Hye Kyung Ju ,  Choon Ju Jeon ,  Jae Seob Kim

发明人： Keun Soo Kim ,  Hye Kyung Ju ,  Choon Ju Jeon ,  Jae Seob Kim

IPC分类号： C12N15/85

CPC分类号： C12N15/85 ,  C12N9/003 ,  C12N15/8509 ,  C12N2800/107

摘要： The present invention relates to a recombinant expression vector for an animal cell containing a dihydrofolate reductase (DHFR) coding nucleotide sequence operatively linked to a DHFR promoter, to an animal cell line transformed by the vector, and to a method for preparing a target protein using the same. As compared with existing animal cell expression vectors, the vector of the present invention enables an effective screening of a cell line clone in which foreign genes are amplified together with DHFR genes even at a much lower methotrexate concentration. The present invention exhibits excellent effects in cell line preparation as high-productivity cell lines can be ensured in a short time through the use of a lower concentration of methotrexate in the process of protein production cell line establishment.

摘要翻译： 本发明涉及用于动物细胞的重组表达载体，其含有编码与DHFR启动子有效连接的二氢叶酸还原酶（DHFR）的核苷酸序列，与载体转化的动物细胞系，以及使用 一样。 与现有的动物细胞表达载体相比，本发明的载体能够有效筛选外源基因与DHFR基因一起扩增的细胞系克隆，即使在低得多的甲氨蝶呤浓度下也是如此。 本发明在蛋白质生产细胞系建立过程中通过使用较低浓度的甲氨蝶呤可以在短时间内确保高生产率细胞系，从而在细胞系制备中表现出优异的效果。

公开(公告)号：US08426570B2

公开(公告)日：2013-04-23

申请号：US13002814

申请日：2009-07-07

申请人： Keun Soo Kim ,  Hye Kyung Ju ,  Choon Ju Jeon ,  Jae Seob Kim

发明人： Keun Soo Kim ,  Hye Kyung Ju ,  Choon Ju Jeon ,  Jae Seob Kim

IPC分类号： C07H21/02 ,  C07H21/04 ,  C12N5/00 ,  C12N5/02 ,  C12N15/00

CPC分类号： C12N15/85 ,  C12N9/003 ,  C12N15/8509 ,  C12N2800/107

摘要： The present invention relates to a recombinant expression vector for an animal cell containing a dihydrofolate reductase (DHFR) coding nucleotide sequence operatively linked to a DHFR promoter, to an animal cell line transformed by the vector, and to a method for preparing a target protein using the same. As compared with existing animal cell expression vectors, the vector of the present invention enables an effective screening of a cell line clone in which foreign genes are amplified together with DHFR genes even at a much lower methotrexate concentration. The present invention exhibits excellent effects in cell line preparation as high-productivity cell lines can be ensured in a short time through the use of a lower concentration of methotrexate in the process of protein production cell line establishment.

摘要翻译： 本发明涉及用于动物细胞的重组表达载体，其含有编码与DHFR启动子有效连接的二氢叶酸还原酶（DHFR）的核苷酸序列，与载体转化的动物细胞系，以及使用 一样。 与现有的动物细胞表达载体相比，本发明的载体能够有效筛选外源基因与DHFR基因一起扩增的细胞系克隆，即使在低得多的甲氨蝶呤浓度下也是如此。 本发明在蛋白质生产细胞系建立过程中通过使用较低浓度的甲氨蝶呤可以在短时间内确保高生产率细胞系，从而在细胞系制备中表现出优异的效果。

公开(公告)号：US20090029387A1

公开(公告)日：2009-01-29

申请号：US11915738

申请日：2006-05-25

申请人： Sang Won Jung ,  Yong Soo Bae ,  Choon Ju Jeon

发明人： Sang Won Jung ,  Yong Soo Bae ,  Choon Ju Jeon

IPC分类号： G01N33/566

CPC分类号： G01N33/5091 ,  C07K7/06

摘要： A method for measuring the activity of cytotoxic T lymphocytes (CTLs) includes preparing peripheral blood mononuclear cells (PBMCs) from blood of an animal; preparing mature dendritic cells by isolating monocytes from the PBMCs, differentiating the monocytes into dendritic cells for presenting an antigen molecule and pulsing dendritic cells with the antigen molecule to obtain the mature dendritic cells; preparing the CTLs as an effector cell by stimulating the PBMCs with the mature dendritic cells to activate and amplify the CTLs; preparing target cells by pulsing the PBMCs, monocytes or B cells with a cytoplasmic transduction peptide (CTP)-antigen complex generated by linking the antigen molecule of step (b) to the CTP; treating the target cells with the effector cells; and analyzing the lysis of the target cells. In addition, a kit for measuring the activity of cytotoxic T lymphocytes is provided.

摘要翻译： 用于测量细胞毒性T淋巴细胞（CTL）的活性的方法包括从动物的血液制备外周血单核细胞（PBMC）; 通过从PBMC分离单核细胞来制备成熟树突状细胞，将单核细胞分化成树突状细胞以呈递抗原分子并用抗原分子脉冲树突状细胞以获得成熟的树突状细胞; 通过用成熟树突状细胞刺激PBMC来激活和扩增CTL来制备CTL作为效应细胞; 通过将步骤（b）的抗原分子与CTP连接而产生的细胞质转导肽（CTP） - 抗原复合物脉冲化PBMC，单核细胞或B细胞来制备靶细胞; 用效应细胞处理靶细胞; 并分析靶细胞的裂解。 此外，还提供了用于测量细胞毒性T淋巴细胞活性的试剂盒。

公开(公告)号：US20090293138A1

公开(公告)日：2009-11-26

申请号：US11915737

申请日：2006-05-27

申请人： Dong Sung Lee ,  Woo Bok Lee ,  Choon Ju Jeon ,  Yong Soo Bae

发明人： Dong Sung Lee ,  Woo Bok Lee ,  Choon Ju Jeon ,  Yong Soo Bae

IPC分类号： A01K67/033 ,  A61K49/00 ,  C12N5/06 ,  A61P35/00

CPC分类号： A01K67/0271 ,  A01K2227/105 ,  A01K2267/0331

摘要： A method for analyzing the prevention and treatment efficacy of a dendritic cell-derived immunotherapeutic for prostate cancer using an animal model carrying tumors expressing a human prostate cancer-specific antigen includes either administering to a normal non-human animal dendritic cells to be analyzed, or administering to a normal non-human animal a cancer cell line expressing the human prostate cancer-specific antigen to induce cancer in the normal non-human animal; administering to the animal the cancer cell line expressing the human prostate cancer-specific antigen to induce cancer in the animal when the dendritic cell administering step was performed, or administering to the animal with cancer dendritic cells to be analyzed when the human prostate cancer-specific antigen expressing cell line administering step was performed; and determining the prevention and treatment efficacy of the dendritic cells as immunotherapeutics for prostate cancer by measuring the formation or growth of cancer cells originated from the cancer cell line in the animal.

摘要翻译： 使用携带表达人前列腺癌特异性抗原的肿瘤的动物模型分析树突状细胞衍生的前列腺癌免疫治疗剂的预防和治疗功效的方法包括向正在分析的正常非人动物树突状细胞施用或 向正常非人动物施用表达人前列腺癌特异性抗原的癌细胞系以在正常非人动物中诱导癌症; 向动物施用表达人前列腺癌特异性抗原的癌细胞系，以在进行树突状细胞施用步骤时在动物中诱导癌症，或者当患有人类前列腺癌特异性的癌症树突状细胞时，对该动物进行分析 进行抗原表达细胞系施用步骤; 并且通过测量源自动物的癌细胞系的癌细胞的形成或生长来确定树突细胞作为前列腺癌的免疫治疗剂的预防和治疗功效。