CHO transfectants produce large amounts of recombinant protein in suspension culture

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 p...

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Bibliographic Details
Published inJournal of immunological methods Vol. 204; no. 1; pp. 99 - 102
Main Authors Schütt, Christine, Fürll, Birgitt, Stelter, Felix, Jack, Robert S, Witt, Sabine
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 12.05.1997
Subjects
Animals
Cell Culture Techniques
CHO Cells - metabolism
CHO transfectant
Cloning, Molecular - methods
Cricetinae
Humans
Lipopolysaccharide Receptors - biosynthesis
Lipopolysaccharide Receptors - genetics
Minifermenter
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Spheroid
Suspension culture
Transfection
rCD14, recombinant CD14
Suspension culture
CHO, chinese hamster ovary
MTX, methotrexate
PBS, phospate buffered saline
Minifermenter
CHO transfectant
LPS, lipopolysaccharide
Spheroid
EDTA, ethylenediamine tetraacetic acid
DHFR, dihydrofolate reductase
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Summary: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.
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ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(97)00027-6