Methods and products related to the improved analysis of carbohydrates
First Claim
1. A method of analyzing a sample containing one or more glycoconjugates, which comprise one or more glycans conjugated to a non-saccharide component, comprising:
- (a) analyzing the glycoconjugates to characterize the glycoconjugates, (b) analyzing the non-saccharide components of the glycoconjugates to characterize the non-saccharide components, (c) separating the glycans from the sample containing one or more glycoconjugates, (d) analyzing the glycans to characterize the glycans, and (e) determining the identity and quantity of all of the glycoforms of the glycoconjugates in the sample with the results obtained from steps (a), (b) and (d) and a computational method.
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Accused Products
Abstract
The invention relates, in part, to the improved analysis of carbohydrates. In particular, the invention relates to the analysis of carbohydrates, such as N-glycans and O-glycans found on proteins and saccharides attached to lipids. Improved methods, therefore, for the study of glycosylation patterns on cells, tissue and body fluids are also provided. Information from the analysis of glycans, such as the glycosylation patterns on cells, tissues and in body fluids, can be used in diagnostic and treatment methods as well as for facilitating the study of the effects of glycosylation/altered glycosylation. Such methods are also provided. Methods are further provided to assess production processes, to assess the purity of samples containing glycoconjugates, and to select glycoconjugates with the desired glycosylation.
132 Citations
312 Claims
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1. A method of analyzing a sample containing one or more glycoconjugates, which comprise one or more glycans conjugated to a non-saccharide component, comprising:
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(a) analyzing the glycoconjugates to characterize the glycoconjugates, (b) analyzing the non-saccharide components of the glycoconjugates to characterize the non-saccharide components, (c) separating the glycans from the sample containing one or more glycoconjugates, (d) analyzing the glycans to characterize the glycans, and (e) determining the identity and quantity of all of the glycoforms of the glycoconjugates in the sample with the results obtained from steps (a), (b) and (d) and a computational method. - View Dependent Claims (180)
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2-70. -70. (canceled)
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71. A method of analyzing a sample containing one or more glycoconjugates, which comprise one or more glycans conjugated to a non-saccharide component, comprising:
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(a) analyzing the glycoconjugates to determine the glycosylation sites and glycosylation site occupancy, (b) separating the glycans from the sample containing one or more glycoconjugates, (c) analyzing the glycans to characterize the glycans, and (d) determining the identity and quantity of all of the glycoforms of the glycoconjugates in the sample, wherein determining the glycosylation sites and glycosylation site occupancy comprises cleaving the glycans from the non-saccharide components and labeling the non-saccharide components at their glycosylation sites of a first portion of the sample, cleaving the glycans from the non-saccharide components of a second portion of the sample, analyzing the first and second portions of the sample of glycoconjugates and comparing the results.
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72-90. -90. (canceled)
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91. A method of analyzing a sample containing one or more carbohydrates, comprising:
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(a) analyzing the carbohydrates with MALDI-MS to determine the monomer composition and relative abundance of the carbohydrates, (b) analyzing the carbohydrates with NMR to determine the monomer composition and linkage abundance of the carbohydrates, (c) and generating constraints from the results of (a) and (b) and solving the constraints with a computational method. - View Dependent Claims (92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107)
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108-146. -146. (canceled)
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147. A method of analyzing a sample containing glycans, comprising:
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analyzing a first portion of the sample, wherein the glycans have been removed, analyzing a second portion of the sample, wherein the second portion of the sample contains intact glycoconjugates, which comprise one or more glycans conjugated to a non-saccharide component, and analyzing a third portion of the sample, wherein the third portion of the sample contains glycans.
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148-163. -163. (canceled)
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164. A method of generating a glycoconjugate library, wherein the glycoconjugate comprises one or more glycans conjugated to a non-saccharide component, comprising:
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cleaving the non-saccharide component of the glycoconjugate in a sample and labeling the non-saccharide component fragments generated with a labeling agent in order to generate a glycoconjugate library, and cleaving the glycans from the non-saccharide components and labeling the non-saccharide components in the sample at the glycosylation sites. - View Dependent Claims (172, 173)
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165-171. -171. (canceled)
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174-179. -179. (canceled)
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181-222. -222. (canceled)
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223. A method of analyzing the total glycome of a sample, comprising:
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(a) analyzing all of the glycans of the sample, and (b) determining a profile of the glycans of the sample, wherein the glycans of the sample are analyzed with a method, comprising;
analyzing the glycans with MALDI-MS to determine the monosaccharide composition and relative abundance of the glycans, and analyzing the glycans with NMR to determine the monosaccharide composition and linkage abundance of the glycans. - View Dependent Claims (296, 297, 298, 299, 311)
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224-295. -295. (canceled)
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300-310. -310. (canceled)
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312-328. -328. (canceled)
Specification