Bioprocess parameters of cell growth and human mu opioid receptor expression in recombinant Drosophila S2 cell cultures in a bioreactor.
The paper describes a recombinant Schneider 2 (rS2) cell culture and protein expression in a bioreactor. S2 cells were transfected with a plasmid containing a fusion protein (human mu opioid receptor, hMOR, and green fluorescent protein, EGFP) under the control of inducible metallothionein promoter. A bioprocess in a bioreactor with 5% dissolved oxygen, 27 degrees C and 120 rpm enabled the cell culture to attain 5.3x10(7 )viable cells/mL at 96 h. The induction decreased the cell multiplication (2.5x10(7) viable cells/mL at 72 h). Glutamine and glucose and low levels of lactate were consumed. A fast recombinant protein synthesis took place and, at 6 h of induction, 2x10(4) receptors/cell could be detected by a functional binding assay. Fluorescence measurements showed a progressive increase of recombinant protein expression with a maximal value of 1.26x10(5) fluo counts/s at 24 h of induction. The data shown in this paper indicate a practical and scaleable cell culture bioprocess procedure for the preparation of recombinant proteins expressed in S2 cells.
Brillet
Karl
K
Biotechnologie, Integrité du Genome, Transcription et Signalisation, UMR 7100, Ecole Supérieure de Biotechnologie de Strasbourg, Bld Sébastien Brant, BP 10413, 67400, Illkirch, France.
da Conceição
Mateus Menegues
MM
Pattus
Franc
F
Pereira
Carlos Augusto
CA
D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D004330:Drosophila / Q000502:physiology*; D029721:Drosophila Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D017450:Receptors, Opioid, mu / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis
D029721:Drosophila Proteins; D017450:Receptors, Opioid, mu; D011994:Recombinant Proteins