Protein-free transfection of CHO host cells with an IgG-fusion protein: Selection and characterization of stable high producers and comparison to conventionally transfected clones

Biotechnology and Bioengineering

Volumn 96, Issue 6, 2007, Pages 1118-1126

  Lattenmayer, Christine ^a   Loeschel, Martina ^b   Schriebl, Kornelia ^a   Steinfellner, Willibald ^c   Sterovsky, Thomas ^b   Trummer, Evelyn ^a   Vorauer Uhl, Karola ^c   Müller, Dethardt ^c   Katinger, Hermann ^b,c   Kunert, Renate ^c  

^a Austrian Center of Biopharmaceutical Technology   (Austria)

^b POLYMUN SCIENTIFIC IMMUNBIOLOGISCHE FORSCHUNG GMBH   (Austria)

^c UNIVERSITY OF AGRICULTURAL SCIENCES   (Austria)

Author keywords

Characterization; CHO; EpoFc fusion protein; Nucleofection; Protein free transfection; Serum free

Indexed keywords

NUCLEOFECTION; TRANSFECTANTS; TRANSFECTION;

ANTIBODIES; BODY FLUIDS; CLONING; PROTEIN FOLDING; TRANSCRIPTION;

CELL CULTURE;

HYBRID PROTEIN; IMMUNOGLOBULIN G; LIPOFECTIN; PROTEIN EPOFC; UNCLASSIFIED DRUG;

ANIMAL CELL; ARTICLE; CONTROLLED STUDY; GENETIC PARAMETERS; GENETIC TRANSFECTION; MOLECULAR CLONING; NONHUMAN; NORTHERN BLOTTING; PROTEIN EXPRESSION; SOUTHERN BLOTTING;

ANIMALS; CELL CULTURE TECHNIQUES; CHO CELLS; CRICETINAE; CRICETULUS; CULTURE MEDIA, SERUM-FREE; DNA, COMPLEMENTARY; ERYTHROPOIETIN; GENE DOSAGE; IMMUNOGLOBULIN G; PLASMIDS; RECOMBINANT FUSION PROTEINS; TRANSFECTION;

EID: 34247526431     PISSN: 00063592     EISSN: 10970290     Source Type: Journal    
DOI: 10.1002/bit.21183     Document Type: Article

References (13)

1. Castle P, Robertson JS. 1999. Summary and conclusion. Animal sera, animal sera derivatives and substitutes used in the manufacture of pharmaceuticals. Dev Biol Stand 99:191-196.
2. Derouazi M, Girard P, Van Tilborgh F, Iglesias K, Muller N, Bertschinger M, Wurm FM. 2004. Serum-free large-scale transient transfection of CHO cells. Biotech Bioeng 87(4):537-545.
3. Kuchenbecker M, Rehberger B, Schriek M, Gackowski J, Fehrenbach R. 2005. Establishment of recombinant CHO cell lines under serum-free conditions. Proceedings of the ESACT Meeting , Harrogate, GB.
4. Maniatis T, Fritsch EF, Sambrook J. 1982. Molecular cloning: a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
5. Morris CB, Warburton S. 1994. Serum-screening and selection. In: Doyle G, Newell DG, editors. Cell & tissue culture: Laboratory procedures. New York: Wiley. p 2B: 1.1-1.5.
6. Ozturk SS, Palsson BO. 1990. Loss of antibody productivity during long-term cultivation of a hybridoma cell line in low serum and serum-free media. Hybridoma 9(2):167-175.
7. Puck TT, Cieciura SJ, Robinson A. 1958. Genetics of somatic mammalian cells. III. Long-term cultivation of euploid cells from human and animal subjects. J Exp Med 108(6):945-956.
8. Rosser MP, Xia W, Hartsell S, McCaman M, Zhu Y, Wang S, Harvey S, Bringmann P, Cobb RR. 2005. Transient transfection of CHO-K1-S using serum-free medium in suspension: A rapid mammalian protein expression system. Protein Expr Purif 40(2):237-243.
9. Schifferli KP, Jessee J, Ciccarone VC. 1999. Transfection of Suspension Cultures of CHO Cells. Focus 21(1):16-17.
10. Schriebl K, Trummer E, Lattenmayer C, Weik R, Kunert R, Muller D, Katinger H, Vorauer K. 2006. Bio-chemical characterization of rhEpo-Fc fusion protein expressed in CHO cells. Protein Expr Purif 15:15.
11. Schroder M, Matischak K, Friedl P. 2004. Serum- and protein-free media formulations for the Chinese hamster ovary cell line DUKXB11. J Biotechnol 108(3):279-292.
12. Urlaub G, Chasin LA. 1980. Isolation of Chinese Hamster cell mutants deficient in dihydrofolate reductase activity. Proc Natl Acad Sci USA 77(7):4216-4220.
13. Zanghi J, Fussenegger M, Bailey J. 1999. Serum protects protein-free competent Chinese hamster ovary cells against apoptosis induced by nutrient deprivation in batch culture. Biotech Bioeng 64(1):108-119.