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A method of determining one or more glycosylation markers of disease comprises obtaining a diseased sample and a control sample, wherein the diseased sample is a sample from a patient diagnosed with disease and the control sample is a sample from a healthy control; releasing a diseased glycan pool of total glycoproteins from the diseased sample and a control glycan pool of total glycoproteins from the control sample without purifying the glycoproteins and without exposing the diseased sample and the control sample to hydrazinolysis wherein the total glycoproteins from the disease and the total glycoproteins from the control sample are immobilized in a high throughput format; measuring a diseased glycoprofile of the diseased glycan pool and a control glycoprofile of the control glycan pool using chromatography, mass spectrometry or a combination thereof; comparing the diseased glycoprofile and the control glycoprofiles to determine said one or more glycosylation markers of disease. A method for diagnosing and monitoring disease in a subject comprising obtaining a sample of body fluid or a body tissue of the subject; releasing a glycan pool of total glycoproteins from the sample without purifying the glycoproteins and exposing the sample to hydrazinolysis; measuring a glycoprofile of the glycan pool. Also provided are methods of glycan releasing compatible with a high throughput format and for analysis of glycosylation from 2D-PAGE spots.

Type

Patent

Publication Date

02/11/2006