By Mohammed Al-Rubeai
Univ. of Birmingham, united kingdom. textual content offers experiences of the advances in glycans and recombinant proteins. includes an in-depth evaluation of technique and diverse techniques for the advance of glycoprotein construction. Written for learn scholars, scientists, commercial biotechnologists, and biochemical engineers. Expanded-outline structure.
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Extra info for Cell Engineering: Glycosylation
An alternative strategy is to use derivatisation of the reducing terminus by reductive amination with compounds containing hydrophobic groups (Poulter and Burlingame, 1990). , 1991) N-linked glycans. However, it is now used by only a few laboratories on account of its relatively poor sensitivity and signal-to-noise ratio. , 1991). , 1993). However, their presence often prevents the determination of a true glycan profile. Fragment ions are usually produced by glycosidic cleavage, with Y and B ions (see below) being particularly abundant, especially adjacent to HexNAc residues where ions of the type are produced.
1996). 75, a complete separation of two primary corresponding to trisialylated triantennary and tetraantennary trisialylated structure. Further, Guttman (Guttman, 1997) has described the oligosaccharide sequencing by specific enzymatic digestion of N-linked glycan pool from fetuin using carefully designed exoglycosidase matrix containing different mixtures of exoglycosidases, in conjonction with separation of the combined digests by CGE. Comparison of the migration times of the exoglycosidase digest fragments to the maltoligosaccharides of known size, enabled calculation of migration shifts, due to cleavage based on the actual exoglycosidases used.
3. CGE Capillary gel electrophoresis is performed through the addition of an hydrophilic polymer to the running buffer. Oligosaccharides are generally derivatized with a ionic fluorophore which imparts the same charge to all the oligo-saccharides. Coated capillary with no electrosomotic flow are employed. Due to the presence of a sieving medium (e;g; agarose or linear polyacrylamide), the electrophporetic mobility of the oligosaccharides is proportional to the log of their molecular mass. Guttman et al.
Cell Engineering: Glycosylation by Mohammed Al-Rubeai