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Figure 2 | Microbial Cell Factories

Figure 2

From: Fed-batch fermentation of GM-CSF-producing glycoengineered Pichia pastoris under controlled specific growth rate

Figure 2

SDS-PAGE analysis of GS115, GS115mGM-CSF and Man5mGM-CSF culture supernatant untreated (-) or treated with PNGase F (+). Mouse GM-CSF appears as three molecular species ranging in molecular mass from approximately 14 to 18 kDa. Treatment with PNGase F showed that the upper two bands correspond to mGM-CSF carrying two and one N-glycans, while the minor lower band (indicated with an arrow) is non-N-glycosylated mGM-CSF. The extra band indicated with an asterisk in both "+" lanes is PNGase F. The two bands marked with a bracket in the "+" lanes are highly N-glycosylated endogenous P. pastoris glycoproteins that, due to their extensive and heterogeneous modification with carbohydrates, do not present as one single band on SDS-PAGE. Upon de-N-glycosylation, however, this heterogeneity is eliminated, resulting in two clearly defined bands.

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Microbial Cell Factories

ISSN: 1475-2859

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