Methods and products related to the improved analysis of carbohydrates
View Patent ↗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. Improved methods, therefore, for the study of glycosylation patterns on cells, tissue and body fluids are also provided. Information regarding 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 on protein function. Such methods are also provided. Methods are also provided to assess protein production processes, to assess the purity of proteins produced, and to select proteins with the desired glycosylation.
1. A method comprising steps of:
providing a sample containing at least one acidic glycan;
contacting the sample with a mass spectroscopy matrix comprising a perfluorinated ion exchange compound (NAFION®) and 6-aza-2-thiothymine (ATT); and detecting the at least one acidic glycan.
2. The method of claim 1 , wherein the method comprises analyzing the purity of the sample containing at least one acidic glycan.
3. The method claim 1 , wherein the method comprises analyzing the glycans of a sample of one or more cells, a tissue or body fluid from a subject, wherein the sample contains at least one acidic glycan.
4. A high-throughput method of claim 1 , wherein more than one sample containing at least one acidic glycan is analyzed.
5. The method of claim 4 , wherein the more than one sample containing at least one acidic glycan are contained in a 96-well plate.
6. The method of claim 4 , wherein the more than one sample containing at least one acidic glycan are affixed to a membrane.
7. The method of claim 1 , wherein the method comprises analyzing the total glycome of a sample of body fluid, wherein the sample of body fluid contains at least one acidic glycan, comprising:
(a) analyzing the glycans of a sample of body fluid, and
(b) comparing the results from (a) with a known pattern.
8. The method of claim 7 , wherein the sample of body fluid is a sample of serum, plasma, blood, urine, saliva, sputum, tears, CSF, seminal fluid, feces, tissues or cells.
9. The method of claim 7 , wherein the method is a method of diagnosis and the pattern is associated with a diseased state.
10. The method of claim 9 , wherein the pattern associated with a diseased state is a pattern associated with cancer.
11. The method of claim 10 , wherein the cancer is prostate cancer, melanoma, bladder cancer, breast cancer, lymphoma, ovarian cancer, lung cancer, colorectal cancer or head and neck cancer.
12. The method of claim 9 , wherein pattern associated with a diseased state is a pattern associated with an immunological disorder.
13. The method of claim 9 , wherein pattern associated with a diseased state is a pattern associated with a neurodegenerative disease.
14. The method of claim 13 , wherein the neurodegenerative disease is a transmissible spongiform encephalopathy, Alzheimer's disease or neuropathy.
15. The method of claim 9 , wherein pattern associated with a diseased state is a pattern associated with inflammation.
16. The method of claim 9 , wherein pattern associated with a diseased state is a pattern associated with rheumatoid arthritis.
17. The method of claim 9 , wherein pattern associated with a diseased state is a pattern associated with cystic fibrosis.
18. The method of claim 9 , wherein pattern associated with a diseased state is a pattern associated with an infection.
19. The method of claim 18 , wherein the infection is viral or bacterial.
20. The method of claim 7 , wherein the method comprises monitoring prognosis and the known pattern is associated with the prognosis of a disease.
21. The method of claim 7 , wherein the method comprises monitoring drug treatment and the known pattern is associated with the drug treatment.
22. The method of claim 1 , wherein the method comprises generating the complete glycan profile of a sample of body fluid, wherein the body fluid contains at least one acidic glycan, comprising:
(a) analyzing the glycans in the sample, and
(b) identifying the complete glycan profile of the sample.