Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium

Behavior of Wild-type and Transfected S2 Cells Cultured in Two Different Media

“…A decrease in RVGP concentration was noticed after day 5. This behavior was already observed previously [24], and may be attributed to the fact that protein denaturation or aggregation can occur during recovery of transmembrane protein in samples with high cell concentration.…”

“…Non-secreted and secreted recombinant proteins such as the G glycoprotein from rabies virus (RVGP) and the surface antigen of hepatitis B virus (HBsAg), respectively, can be expressed in Drosophila Expression System (DES), both showing to be appropriately processed and biologically active [13,[16][17][18][19][20][21][22][23]. Furthermore, S2AcRVGP2 cells cultivated in IPL-41 medium supplemented with 10 g L −1 glucose, 0.5 g L −1 fructose, 2 g L −1 lactose, 3.5 g L −1 glutamine, 0.6 g L −1 tyrosine, 1.48 g L −1 methionine, 6 g L −1 yeastolate ultrafiltrate, 1% chemically defined lipid concentrate, and 0.1% Pluronic F-68 are able to produce up to 31 ng RVGP L −1 in batch culture provided that dissolved oxygen and pH are controlled at 40% and 6.3, respectively [24].…”

Rabies virus glycoprotein expression in Drosophila S2 cells. I: Design of expression/selection vectors, subpopulations selection and influence of sodium butyrate and culture medium on protein expression

“…In addition, DES has been reported to have advantageous bioprocess characteristics, such as high cell density attained, low cost culture medium needed and continuous bioprocess possibility (Affleck and Walker, 2008;Batista et al, 2008Batista et al, , 2009Bovo et al, 2008;Brillet et al, 2008;Galesi et al, 2008;Iwaki and Castellino, 2008;Jorge et al, 2008;Park et al, 2008;Santos et al, 2007;Swiech et al, 2008aSwiech et al, , 2008b. In previous publications Yokomizo et al, 2007) we have shown that the DES indeed represents a novel and promising approach for the production of the rabies virus glycoprotein (RVGP).…”

Kinetic studies of recombinant rabies virus glycoprotein (RVGP) cDNA transcription and mRNA translation in Drosophila melanogaster S2 cell populations

“…Several complex glycoproteins were already expressed in the S2 cell system, using these promoters (Mallender et al 2001;Zhang et al 2007;Scotter et al 2006;Brillet et al 2006;Kim et al 2005;Johansson et al 2007;Jennings et al 2006;Li et al 2005;Lim et al 2004;Lee et al 2007). Aiming the expression of high levels of RVGP under the control of these promoters in the S2 cell system, for vaccination as well as structure/function evaluation, many studies were already carried out on cell growth and heterologous recombinant protein expression kinetics (Yokomizo et al 2007;Galesi et al 2008;Swiech et al 2008a;Batista et al 2009;Ventini et al 2010;Lemos et al 2009), as well as on metabolism and synthesis of secondary products (Swiech et al 2008b, c) and culture medium formulation and supplementation (Galesi et al 2007;Batista et al 2008Batista et al , 2011Mendonça et al 2008Mendonça et al , 2009. For constitutive RVGP expression using the actin promoter, instead of a gradual and sustained increase of RVGP, we have observed a sharp RVGP increase, at the beginning of the stationary cell growth phase, which could not be associated with the culture system or the cell culture media, pH, oxygen concentration or substrate change (Galesi et al 2008;Ventini et al 2010;Batista et al 2011).…”