CHO transfectants produce large amounts of recombinant protein in suspension culture

被引：11

作者：

Schutt, C ^{[1
]}

Furll, B ^{[1
]}

Stelter, F ^{[1
]}

Jack, RS ^{[1
]}

Witt, S ^{[1
]}

机构：

[1] BIOMETEC GMBH GREIFSWALD,GREIFSWALD,GERMANY

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1997年 / 204卷 / 01期

关键词：

minifermenter; CHO transfectant; spheroid; suspension culture;

D O I：

10.1016/S0022-1759(97)00027-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Chinese hamster ovary (CHO) cells transfected with various genes are widely used as adherent cell monolayers to produce recombinant proteins. In this report we present a new culture technique for CHO cells transfected with the vector pPOL-DHFR-CD14 using a minifermenter (miniPERM, Heraeus) for the production of recombinant human endotoxin receptor CD14 (rCD14). The transfectants were cultured for 12-17 days under serum-free conditions and formed spheroids. From this system we harvested supernatants containing up to 3.1 mg/ml recombinant CD14 (rCD14). This represents a 200-fold increase of rCD14 yield compared to conventional adherent CHO cell culture.

引用

收藏

页码：99 / 102

页数：4

相关论文

共 15 条

[1]

AUDRAN R, 1994, CANCER IMMUNOL IMMUN, V39, P299, DOI 10.1007/s002620050129

[2] IN-VITRO PRODUCTION OF MONOCLONAL-ANTIBODIES IN HIGH-CONCENTRATION IN A NEW AND EASY TO HANDLE MODULAR MINIFERMENTER [J].

FALKENBERG, FW ;

WEICHERT, H ;

KRANE, M ;

BARTELS, I ;

PALME, M ;

NAGELS, HO ;

FIEBIG, H .

JOURNAL OF IMMUNOLOGICAL METHODS, 1995, 179 (01) :13-29

[3] AN ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR THE QUANTIFICATION OF SOLUBILIZED CD14 IN BIOLOGICAL-FLUIDS [J].

GRUNWALD, U ;

KRUGER, C ;

WESTERMANN, J ;

LUKOWSKY, A ;

EHLERS, M ;

SCHUTT, C .

JOURNAL OF IMMUNOLOGICAL METHODS, 1992, 155 (02) :225-232

[4] OXYGEN-CONSUMPTION RATE AND MITOCHONDRIAL DENSITY IN HUMAN-MELANOMA MONOLAYER-CULTURES AND MULTICELLULAR SPHEROIDS [J].

HYSTAD, ME ;

ROFSTAD, EK .

INTERNATIONAL JOURNAL OF CANCER, 1994, 57 (04) :532-537

[5] BOTH MEMBRANE-BOUND AND SOLUBLE FORMS OF CD14 BIND TO GRAM-NEGATIVE BACTERIA [J].

JACK, RS ;

GRUNWALD, U ;

STELTER, F ;

WORKALEMAHU, G ;

SCHUTT, C .

EUROPEAN JOURNAL OF IMMUNOLOGY, 1995, 25 (05) :1436-1441

[6] BETA-GALACTOSIDASE ACTIVITY IN TRANSFECTED LTK(-) CELLS IS DIFFERENTIALLY REGULATED IN MONOLAYER AND IN SPHEROID CULTURES [J].

KLUNDER, I ;

HULSER, DF .

EXPERIMENTAL CELL RESEARCH, 1993, 207 (01) :155-162

[7] RELEASE FROM A HUMAN MONOCYTE-LIKE CELL-LINE OF 2 DIFFERENT SOLUBLE FORMS OF THE LIPOPOLYSACCHARIDE RECEPTOR, CD14 [J].

LABETA, MO ;

DURIEUX, JJ ;

FERNANDEZ, N ;

HERRMANN, R ;

FERRARA, P .

EUROPEAN JOURNAL OF IMMUNOLOGY, 1993, 23 (09) :2144-2151

[8] MAINTENANCE OF PHENOBARBITAL-INDUCIBLE CYP2B GENE-EXPRESSION IN C57BL/6 MOUSE HEPATOCYTES IN PRIMARY CULTURE AS SPHEROIDS [J].

NEMOTO, N ;

SAKURAI, J ;

FUNAE, Y .

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1995, 316 (01) :362-369

[9] DRUG AND RADIATION-RESISTANCE IN SPHEROIDS - CELL CONTACT AND KINETICS [J].

OLIVE, PL ;

DURAND, RE .

CANCER AND METASTASIS REVIEWS, 1994, 13 (02) :121-138

[10] COLLAGENS, INTEGRINS AND THE MESENCHYMAL DRIFT IN GLIOBLASTOMAS - A COMPARISON OF BIOPSY SPECIMENS, SPHEROID AND EARLY MONOLAYER-CULTURES [J].

PAULUS, W ;

HUETTNER, C ;

TONN, JC .

INTERNATIONAL JOURNAL OF CANCER, 1994, 58 (06) :841-846