Glycoprofiling is used by the biopharmaceutical industry to analyze the glycans found on glycoproteins being developed as therapeutics. It is also a key assay for the lot release characterization of manufactured glycoproteins both before and after approval.
The process can be broken down as follows:
QAbio Glycan Release
Typically, PNGase F is used to release N-linked glycans. The enzyme works quickly and effectively, cleaving the glycans in native conditions in most cases. The Endo F family of enzymes are also commonly used for glycan release. These three enzymes vary in specificity to the multi-antennary structures found on N-linked glycans. They also differ from PNGase F in that they leave the penultimate glycan, a charged N-acetyl-glucosamine, attached to the protein. Both PNGase F and the Endo F family of enzymes leave the glycan with a free reducing terminii suitable for fluorescent labelling.
For O-linked glycan release, the Ludger Liberate Orela Kit allows the release of glycans that have free reducing terminii suitable for fluorescent labelling. The kit is straightforward and contains a simple method with no special handling techniques as required by the alternative method, hydrazinolysis.