Recombinant expression of glycoproteins is of great interest, particularly in the production of therapeutic glycoproteins such as immunoglobulins and hormones for the biopharmaceutical market. Each expression system has its advantages and disadvantages (1). Some factors to be aware of when choosing an expression system are:
- Bacterial systems cannot produce a protein with mammalian-type glycosylation.
- Yeast primarily modifies glycoproteins with only high mannose N-glycans.
- Plant and insect cell lines can modify N-glycans with a core α1-3 fucose residue not found in humans.
- Plants can also add a non-human xylose residue to the N-glycan.
- Some mammalian cell lines can add the non-human epitopes Gal α1-3 Gal or N-glycolylneuraminic acid (NGNA) to glycoproteins.
- Betenbaugh M.J., et al (2004) Curr Opin Struct Biol. 14(5):601-6. PMID: 15465322
- Will a mammalian glycoprotein be correctly glycosylated and folded if expressed in bacteria?
- Are heterologous proteins expressed with their canonical glycans, or with the glycans typical of the host?
- What are the differences between mammalian, yeast, insect (baculovirus), and plant cell expression systems?
- Will each different protein expressed in a given system carry the same type of glycans?
- Will glycoproteins produced in yeast or plants be recognized by mammalian cells?
- What is the importance of the α-Gal and NGNA epitopes?
- Is it possible to predict whether a protein is N- or O-glycosylated?
- Which proteins are glycosylated?
- O-Glycosidase (P0733)
- O-Glycosidase Application Note 1 (P0733)
- Endo D Removal Magnetic Chitin Bead Protocol (P0742)
- Endo F2 Reaction Protocol (P0772)
- Endo H/Endo Hf Protocol
- Endo S Removal Magnetic Chitin Bead Protocol (P0741)
- Endo-α-N-Acetylgalactosaminidase Application Note 1
- Glycan SPE C18 and Graphitized Carbon Protocols (P0710)
- Glycoproteomics: Buffer Exchange Protocols (P0710)
- Glycoproteomics: Buffer Exchange Protocols (P0711)
- Intact Protein LS-ESI-TOF Protocol (P0710)
- PNGase F Protocol
- Protocol for α1-3,6 Galactosidase (P0731)
- Rapid PNGase F (non-reducing format) (P0711) Reaction Protocol
- Rapid PNGase F (non-reducing format) (P0711) SDS-PAGE Protocol
- Rapid PNGase F Antibody Standard Protocol (P6043)
- Rapid PNGase F by SDS-PAGE Protocol (P0710)
- Rapid PNGase F Protocols (P0710)
- Reaction Conditions for Endo D (P0742)
- Reaction Conditions for Endo S (P0741)
- Reaction Conditions for PNGase A (P0707)
- Reaction Conditions for Remove-iT® PNGase F (P0706)
- Reaction Protocols for Protein Deglycosylation Mix II (P6044)
- Removal of Endo F2 by Magnetic Beads (P0772)
- Removal of terminal N-acetylglucosamine from the biantennary N-linked sugars of IgG
- Remove-iT® PNGase F Magnetic Chitin Bead Protocol (P0706)
- RNase B Deglycosylation Protocol (P7817)
- Typical Reaction Conditions (P0732)
- Typical Reaction Conditions for a1-2,3,6 Mannosidase (P0768)
- Typical Reaction Conditions for Endo F3 Protocol (P0771)
- Typical Reaction Conditions for α1-2 Fucosidase (P0724)
- Typical Reaction Conditions for α1-2, 3, 4, 6 Fucosidase (P0748)
- Typical Reaction Conditions for α1-2,3 Mannosidase (P0729)
- Typical Reaction Conditions for α1-3, 4 Fucosidase (P0769)
- Typical Reaction Conditions for α1-3, 4, 6 Galactosidase (P0747)
- Typical Reaction Conditions for α1-3,6 Galactosidase (P0731)
- Typical Reaction Conditions for α1-6 Mannosidase (P0727)
- Typical Reaction Conditions for α2-3 Neuraminidase S (P0743)
- Typical Reaction Conditions for α2-3,6,8 Neuraminidase (P0720)
- Typical Reaction Conditions for α2-3,6,8,9 Neuraminidase A (P0722)
- Typical Reaction Conditions for β1-3 Galactosidase (P0726)
- Typical Reaction Conditions for β1-3,4 Galactosidase Reaction Protocol (P0746)
- Typical Reaction Conditions for β1-4 Galactosidase (P0730)
- Typical Reaction Conditions for β1-4 Galactosidase S (P0745)
- Typical Reaction Conditions for β-N-Acetylglucosaminidase S (P0744)
- Typical Reaction Conditions for β-N-Acetylhexosaminidasef (P0721)
- Typical Reaction Conditions α-N-Acetylgalactosaminidase (P0734)
The Structure, Function and Importance of Carbohydrates
Read about the structure, function, and importance of Carbohydrates from biology experts at NEB.
- Glycobiology Unit Conversion Chart
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Learn about glycobiology and its importance in clinical and diagnostic applications in this episode of NEB TV. Also, hear more about how NEB is setting the bar for product quality in this rapidly growing field.
Learn about the core sequences and common modifications of N-linked and O-linked glycans in this video. Analysis of these glycans can be accomplished with the use of deglycosylation enzymes, which can provide complete sugar removal with no protein degradation.
Behind the Paper: An engineered Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
Minyong and Jim summarize their recent Nature Communications publication describing selective capture of N-glycans and N-glycopeptides by an engineered high affinity Fbs1 carbohydrate binding protein.