What is G0F glycan?
The G0F glycan is an asialo-agalacto-fucosylated biantennary oligosaccharide. Specsheet CofA SDS. Also available labeled: 2AA 2AB APTS. 1463 daltons.
What is a glycan group?
Glycans are chain-like structures that are composed of single sugar molecules (monosaccharides) linked together by chemical bonds.
How does glycosylation affect molecular weight?
Post-translational modifications Extensive glycosylation increases molecular weight, slowing protein migration on a Western blot, but is not accounted for in a molecular weight calculation based on protein sequence (Figure 1 & 2).
How is glycan analysis done?
Glycan structure assessment is challenging due to the isomeric and branched nature of oligosaccharides. The traditional approach involves the use of exoglycosidases to selectively and sequentially release terminal monosaccharides, which produces trimmed glycans to be analyzed by HILIC, CE or MALDI-MS.
What is glycan analysis?
In the development of therapeutic glycoproteins, especially biosimilar products,23 glycan analysis usually involves the use of complementary methods for assessing specific glycosylation attributes, including glycosylation site, glycan structure and abundance.
How much mass does glycosylation add?
N-linked glycosylation sites are estimated to be 2.5 kDa extra per site to the total weight = 20 kDa more.
Does glycosylation affect SDS-PAGE?
The determination of molecular weights (MW) of heavily glycosylated proteins is seriously hampered by the physicochemical characteristics and heterogeneity of the attached carbohydrates. Glycosylation impacts protein migration during SDS-PAGE and SEC analysis.
What are n linked oligosaccharides?
N-linked glycosylation, is the attachment of an oligosaccharide, a carbohydrate consisting of several sugar molecules, sometimes also referred to as glycan, to a nitrogen atom (the amide nitrogen of an asparagine (Asn) residue of a protein), in a process called N-glycosylation, studied in biochemistry.
What is the purpose of glycosylation?
Glycosylation is an important and highly regulated mechanism of secondary protein processing within cells. It plays a critical role in determining protein structure, function and stability. Structurally, glycosylation is known to affect the three dimensional configuration of proteins.
Why is Sialylation important?
Sialylation is essential for early embryonic development and the deletion of UDP-GlcNAc 2-epimerase, a rate-limiting enzyme in sialic acid biosynthesis, is embryonically lethal.
How do you calculate glycosylation?
For detection of glycosylated proteins, two types of methods are generally used: staining and affinity-based methods.
- 2.1. Staining Procedures. A basic, simple method to determine whether a protein is glycosylated is to resolve it on SDS-PAGE and to stain the gel for glycoproteins.
- 2.1. Affinity-Based Procedures.
What percentage of proteins are glycosylated?
Glycosylation is one of the most common post-translational modifications (PTMs) of proteins. At least 50% of human proteins are glycosylated with some estimates being as high as 70%. Glycoprotein analysis requires determining both the sites of glycosylation as well as the glycan structures associated with each site.
How much weight does glycosylation add to a protein?
Why glycosylation is important for recombinant proteins?
The glycosylation of recombinant proteins, especially those destined for potential administration to human subjects, is of critical importance. Glycosylation profoundly affects biological activity, function, clearance from circulation, and crucially, antigenicity.
What is the NIST glyco mass calculator?
The NIST Glyco Mass Calculator. The Glyco MS Calculator is a tool to aid in the analysis of glycoforms, e.g. glycans and glycopeptides. The user inputs the number of residues in the glycan and the program calculates for the masses and elemental composition of the glycan.
What is a g0f glycan?
The G0F glycan is on of the four most common glycans found on monoclonal antibodies, along with the two G1F isomers, and G2F. A mixture of these glycans have been combined in our MAb4 Antibody Reference Panel.
What is the glyco MS calculator?
The Glyco MS Calculator is a tool to aid in the analysis of glycoforms, e.g. glycans and glycopeptides. The user inputs the number of residues in the glycan and the program calculates for the masses and elemental composition of the glycan.
What is the molecular weight of IgG glycan?
Molecular Weight: 1463 Purity: >90% pure as assessed by a combination of 1 H-NMR and HPLC Sources: The G0F glycan is found on many mammalian glycoproteins including human IgG and is a substructure of bi-antennary N-linked oligosaccharides such as A2F, A1F, and NA2F which are widely found on glycoproteins.