Sequencing of oligosaccharides

Details Sequencing of oligosaccharides Sequencing of oligosaccharides

1993-12-08

1995-06-27

McMahon, Timothy M.

Chemical apparatus and process disinfecting, deodorizing, preser

Control element responsive to a sensed operating condition

Sensed condition is operating time and control is operation...

435101, 435291, G01N 3350

Patent

active

054277442

DESCRIPTION:

BRIEF SUMMARY
This is a continuation of PCT/GB92/00831 filed on Apr. 7, 1992 which is now WO 92/19766.
The present invention relates to the analysis of oligosaccharides and more particularly to the form of analysis known as sequencing of oligosaccharides.
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 oligosaccharides are found linked to a peptide chain by either an N-glycosidic bond or by an O-glycosidic bond; these oligosaccharides may vary up to highly branched structures containing many (e.g. over 30) monosaccharide units.
In a sequential analysis of an unknown oligosaccharide structure, a single reaction is performed with a particular reagent on a pure sample of the unknown oligosaccharide and its products analysed. The reagent is chosen such that the product of the reaction will reveal the presence or absence of a particular structural sub-unit or sub-units which may or may not occur in the sample. Based on the results of this analysis, a further reaction may be carried out using a different reagent, either with more of the original pure sample or with the products of the previous reaction. From the analysis of the second reaction further conclusions may be drawn about the structure of the original oligosaccharide sample and the process repeated as often as necessary, until as much information as possible has been extracted about the original oligosaccharide structure with the reagents available or until supplies of the pure sample and derived products have been exhausted. The end point of any analysis of an oligosaccharide structure, whether the analytical technique is sequential or not, is to deduce information concerning the oligosaccharide structure; this information includes, oligosaccharide, (e.g. 1-3, 1-4), and hence any branching pattern, units (i.e. whether a linkage is an alpha linkage or a beta linkage). regarded as a "sequencing agent". Where a sequencing agent is a chemical or biochemical reagent the sequencing agent may be regarded as a sequencing reagent. Examples of sequencing reagents are enzymes and typical enzymes used in sequencing oligosaccharides are listed in the Enzyme Rules Table set out below.


__________________________________________________________________________ Enzyme Rules Table The following is a list of the enzymes commonly used for cleaving monosaccharides from N-linked oligosaccharides and the rules showing which monosaccharides are cleaved by each of these enzymes and from which part of the oligosaccharide structure cleavage can be expected. Monosaccharide Enzyme Cleaved Rules for Cleavage __________________________________________________________________________ 1) Achatina fulica beta mannosidase Mannose 1-beta-4 to any site 2) A.saitoi alpha mannosidase Mannose 1-alpha-2 to any site 3) Jack bean alpha mannosidase Mannose 1. mannose 1-alpha-2 to any site 2. cleaves the 1-alpha-3,6 mannoses if there is no bisect on the middle mannose 4) Jack bean alpha mannosidase Mannose Different from 3) in that it (under arm-specific conditions) will not cleave the mannose 1-alpha-6 case when restricted by a side arm longer than 1 unit out from the middle mannose 5) Bovine testis beta galactosidase Galactose 1-beta-3,4 to any non-branched site 6) Jack bean beta galactosidase Galactose 1-beta-4 to any non-branched site 7) C.lampas beta xylosidase Xylose 1-beta-any to any site 8) S.pneum beta N-acetyl N-acetyl 1. 1-beta-3 to any non-branched hexosaminidase hexosamine site 2. 1-beta-2 mannose 1-alpha-3 or 6 to middle mannose provided the first mannose does not have a bond at atom 6, and also in the case of 1-alpha-6 that the middle mannose is not bisected 9) Jack bean beta N-acetyl N-acetyl 1-beta-any to any site hexosaminidase hexosamine 10) Bovine epididymis alpha fucosidase Fucose

REFERENCES:
patent: 5100778 (1992-03-01), Rademacher et al.
Joseph K. Welply: "Sequencing methods for carbohydrates and their biological applications"; TIBTECH; 7 (1989) Jan.; pp. 5-10.

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