Scientific Poster presentation: Antifoam addition to shake flask cultures of recombinant Pichia pastoris increases yield - Essay Example

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We want to find out if antifoams (Antifoam A (Sigma), Antifoam C (Sigma), J673A (Struktol), P2000 (Fluka) and SB2121 (Struktol)) affect recombinant yield (recombinant green fluorescent protein (GFP)) in shake-flask of P. pastoris and investigate their essential mechanisms.
To specifically find potential result regarding the above concern, we ensure uniform employment of P. pastoris strain X33 throughout the entire experimental procedures. Cells were cultured in shake flasks and tested with antifoams to find their respective and remarkable connection with recombinant protein yield.
A remarkable trend of GFP yield increase was observed when antifoams were incorporated in shake flasks. The concentration was somewhere in the range of 0.4%-1% v/v. Highest yield achieved at the addition of 1% P2000, 1% SB2121, 1% J673A, 0.6% Antifoam A, and 0.8% Antifoam C.
As determined by Bartsch test, those antifoams employed in Figure 1 were tested for their destruction capacity. All antifoams in general were effective at foam destruction. J673A was the most effective at foam destruction and best at increasing GFP yield, while Antifoam C was less destructive.
In general, there was no correlation between kLa and total yield for any of the conditions tested in the study, and density of the cultures, which only imply that kLa may not directly contribute to the increased protein yield of antifoams.
After 12 hours, the DO in the flasks significantly showed no difference in the flasks with or without antifoams. This in general implies that the level of DO content of cultures containing antifoam is a proof that antifoams do not have any direct contributing factor to affect the DO level.
Antifoams do not influence the growth of the cells, but there was enhancement on the level of amount secreted into the medium. Using one-way ...Download file to see next pagesRead More