Cell Engineering: Glycosylation by Mohammed Al-Rubeai

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By Mohammed Al-Rubeai

Univ. of Birmingham, united kingdom. textual content presents stories of the advances in glycans and recombinant proteins. comprises an in-depth review of technique and diverse ways for the advance of glycoprotein construction. Written for examine scholars, scientists, commercial biotechnologists, and biochemical engineers. Expanded-outline structure.

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Cell Engineering: Glycosylation

Univ. of Birmingham, united kingdom. textual content presents stories of the advances in glycans and recombinant proteins. includes an in-depth evaluation of technique and numerous techniques for the development of glycoprotein construction. Written for study scholars, scientists, business biotechnologists, and biochemical engineers.

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1963) is required to produce the necessary volatility. The technique is ideally coupled with on-line GC/MS and, when combined with specific derivatization techniques, as described below, provides the main methods for the determination of constituent monosaccharide composition and linkage of larger carbohydrates. Compounds as involatile as monosaccharide phosphates can be examined if suitably derivatised (Harvey and Horning, 1973). , 1989). However, column temperatures in the region of 400oC are needed for the larger compounds and, above this temperature, most compounds decompose.

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. These ions tend to eliminate methanol if there is a free methoxy group in the 3-position of the GlcNAc residue but, otherwise, there is little in the fragmentation spectrum that yields much more information of linkage.

The analysis of glycans by this technique falls into a number of stages including release of glycans form the glycoprotein, the 2-AB labelling of the glycans, the initial HPLC profiling, enzymatic sequencing of glycans and conformation of structures by mass spectrometry which are summarised in the diagram shown in Figure 1. The techniques may be adapted according to the source and purity of the (glyco)proteins under study and of the techniques available for their analysis as will be discussed below HPLC techniques for glycan analysis are now very sensitive and can be readily combined with further analysis by a number of techniques.

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