InsectSelect™ BSD Kit with Sf9 cells *DISCONTINUED* - FAQs

How do protein glycosylation patterns compare between Pichia pastoris yeast, insect cells (Sf9, Sf21, HighFive) and mammalian cells?

Pichia glycosylation consists mostly of short chain Man(10) GlcNAc residues. Compared to other yeast like S. cerevisiae which tend to hyperglycosylate, Pichia glycosylation is closer to the typical mammalian high-mannose glycosylation pattern. O-linked oligosaccharides are present but are not major components of the total soluble glycoprotein of Pichia. Standard N-linked or O-linked glycoslation amino acid consensus sequences must be present in the expressed protein in order for glycosylation to take place in Pichia.

In insect cells infected with baculovirus, the nature of N-linked glycosylation is dependent on the protein expressed, the host cell line used and the length of time of infection of cells with baculovirus. Baculovirus infection alters normal glycosylation and oligosaccharide processing characteristics of insect cells. N-linked glycosylation is generally of the high-mannose type. O-linked glycosylation is, although not identical, similar to mammalian cells depending on localization and type of protein.

We also offer the Mimic insect cell line. Mimic cells are transgenic insect cells that have been engineered to produce recombinant proteins with terminally sialylated N-glycans like those found in mammalian systems. These are identical to the SfSWT-1 cells developed by Donald L. Jarvis. This cell line is suitable for expressing recombinant glycoproteins with baculovirus and other insect expression systems, including the Bac-to-Bac Expression System and the InsectSelect system.

References for glycosylation in Pichia:
1) Grinna LS, et al. Size distribution and general structural features of N-linked oligosaccharides from the methylotrophic yeast, Pichia pastoris. Yeast. 1989 Mar-Apr;5(2):107-15.
2) Juge N, et al. Overexpression, purification, and characterization of recombinant barley alpha-amylases 1 and 2 secreted by the methylotrophic yeast Pichia pastoris. Protein Expr Purif. 1996 Sep;8(2):204-14.
3) Trimble RB, et al.Structure of oligosaccharides on Saccharomyces SUC2 invertase secreted by the methylotrophic yeast Pichia pastoris. J Biol Chem. 1991 Dec 5;266(34):22807-17.
4) Tschopp JF, et al.Expression of the lacZ gene from two methanol-regulated promoters in Pichia pastoris. Nucleic Acids Res. 1987 May 11;15(9):3859-76.