Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium.
The recombinant G glycoprotein from the surface of the rabies virus (RVGP) is a promising candidate as a rabies vaccine component and also for diagnostic purposes. In this study, RVGP production by transfected Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 medium) was carried out. The effects of pH and pO(2) were evaluated in batch culture in parallel spinner flasks. The use of a pH equal to 6.3 and a pO(2) of 40% air saturation resulted in the highest RVGP content. These conditions were also used in fed-batch mode, yielding a RVGP content level of 98g/10(7) cells. The main nutrients consumed were glucose, glutamine, asparagine, serine and proline and the major metabolites produced were alanine and ammonia, according to the metabolism studies performed. Since RVGP is a transmembrane protein, two different methods for protein recovery were assessed and compared. Detergent-based cell disruption showed to be more effective than mechanical disruption with glass beads for glycoprotein recovery.
Batista
Fabiana R X
FR
Department of Biotechnological Processes, School of Chemical Engineering, University of Campinas, Campinas, SP, Brazil. rxbatista@gmail.com
Moraes
Angela M
AM
Büntemeyer
Heino
H
Noll
Thomas
T
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D050260:Carbohydrate Metabolism / Q000187:drug effects; D018929:Cell Culture Techniques; D002460:Cell Line / Q000378:metabolism / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004331:Drosophila melanogaster; D006863:Hydrogen-Ion Concentration; D019344:Lactic Acid / Q000378:metabolism; D010101:Oxygen Consumption / Q000187:drug effects; D011820:Rabies virus / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014760:Viral Fusion Proteins / Q000302:isolation & purification* / Q000378:metabolism*
D000596:Amino Acids; D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; D014760:Viral Fusion Proteins; D019344:Lactic Acid