Sequencing of oligosaccharides

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase

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435 22, 435 16, 435 15, 435 13, 435 74, 435 71, 435 14, 435 25, C12Q 134, C12Q 140, C12Q 154, C12Q 126

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active

056679840

DESCRIPTION:

BRIEF SUMMARY
The present invention relates to the analysis of oligosaccharides and more particularly to the form of analysis known as sequencing of oligosaccharides.
According to one aspect of the present invention there is provided apparatus suitable for use in the sequencing of an oligosaccharide which apparatus includes means for selecting a sequencing agent to be applied to an oligosaccharide entity.
According to another aspect of the present invention there is provided a process suitable for use in the sequencing of an oligosaccharide which process includes the use of a means for selecting a sequencing agent to be applied to an oligosaccharide entity.
The oligosaccharide entity may be, for example, an oligosaccharide, or a product of an oligosaccharide, or a species having an oligosaccharide portion. A product of an oligosaccharide may be, for example, a product produced by previously applying a sequencing agent to an oligosaccharide; by way of example, the product may itself be an oligosaccharide.
Oligosaccharides form a class of chemical compounds which are each made up of a number of monosaccharide units linked together by glycosidic bonds. Important sources of naturally occurring oligosaccharides are glycoproteins in which saccharides are found linked to a peptide chain either by an N-glycosidic bond or by an O-glycosidic bond; these oligosaccharides may vary from a few monosaccharide units to highly branched structures containing many (e.g. over 30) monosaccharide units.
The "sequencing" of an oligosaccharide involves deducing certain information concerning the structure of the oligosaccharide such as (i) the type of each monosaccharide unit in the oligosaccharide, (ii) the order in which the monosaccharide units are arranged in the oligosaccharide, (iii) the position of linkages between each of the monosaccharide units (e.g. 1-3, 1-4, etc.), and hence any branching pattern and/or (iv) the orientation of linkage between each of the monosaccharide units (i.e. whether a linkage is an .alpha. linkage or a .beta. linkage).
Where it is desired to obtain as much information as possible regarding the structure of an oligosaccharide then "sequencing" of the oligosaccharide may be carried out to obtain as much information as possible in relation to features (i) to (iv) inclusively immediately hereinbefore disclosed.
An agent which assists in obtaining information in relation to some or all of features (i) to (iv) inclusively on being applied to an oligosaccharide entity may be regarded as a "sequencing agent". By way of example, a sequencing agent may be a physical agent or a chemical agent. Examples of physical sequencing agents are proton n.m.r., carbon-13 n.m.r. and mass spectrometry for molecular weight determinations.
Also, by way of example, a sequencing agent may be capable of causing cleavage of a chemical bond or capable of causing formation of a chemical bond.
Where, for example, a sequencing agent is a chemical reagent (which may be, for example, a chemical reagent or a biochemical reagent) the sequencing agent may be regarded as a sequencing reagent. Examples of sequencing reagents are enzymes (such as exoglycosidases and endoglycosidases) and chemical reagents (e.g. a periodate) capable of effecting chemical cleavage of an oligosaccharide and/or a chemical modification of an oligosaccharide which assists in obtaining information regarding the structure of the oligosaccharide as hereinbefore disclosed.
As hereinbefore disclosed the oligosaccharide entity may be, for example, an oligosaccharide, or a product of an oligosaccharide, or a species having an oligosaccharide portion.
Thus, it is to be understood that, by way of example, an oligosaccharide as such may be subjected to sequencing in accordance with the present invention; by way of further example, as an alternative, a product of an oligosaccharide may be subjected to sequencing in accordance with the present invention. (It will be appreciated that the product may itself be an oligosaccharide.)
Alternatively, for example, an oligosaccharide provided

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Joseph K. Welply; "Sequencing methods for carbohydrates and their biological applications"; TIBTECH; 7 (1989) January; pp. 5-10.
Martin F. Chaplin; "A Rapid and Sensitive Method for the Analysis of Carbohydrate Components in Glycoproteins Using Gas-Liquid Chromatography"; Analytical Biochemistry; 1982; pp. 336-341.
Hardy, Mark R., "Monosaccharide Analysis of Glycoconjugates by High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection," Methods in Enzymology, vol. 179, pp. 76-83 (1989).
Pipkorn et al, Int. J. Peptide Protein Res., 27, 1986, pp. 583-588.

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