Methods and products related to the analysis of polysaccarides
The invention relates to methods and products for characterizing and using polysaccharides. Low molecular weight heparin products and methods of use are described. Methods for characterizing purity and activity of polysaccharide preparations including glycosaminoglycans such as heparin are also described.
1. A method of analyzing a sample comprising:
(A) applying an experimental constraint to a polysaccharide in a sample, to produce a modified polysaccharide, having a signature component,
(B) detecting the presence of the signature component, in the sample as an indication that the polysaccharide is present in the sample, and
(C) determining the presence or absence of the signature component to analyze the sample,
wherein the signature component is ΔUH NAC,6S GH NS,3S,6S; ΔUH NS,6S GH NS,3S,6S ; ΔUH NAC,6S GH NS,3S ; or ΔUH NS,6S GH NS,3S .
2. The method of claim 1 , wherein the experimental constraint applied to the sample is capillary electrophoresis.
3. The method of claim 1 , wherein the experimental constraint applied to the sample is high pressure liquid chromatography.
4. The method of claim 1 , wherein the experimental constraint applied to the sample is gel permeation chromatography.
5. The method of claim 1 , wherein the experimental constraint applied to the sample is nuclear magnetic resonance.
6. The method of claim 1 , wherein the sample is a batch of polysaccharide and the signature component is used to monitor the purity of the batch by determining the amount of signature component in the batch.
7. The method of claim 1 , wherein the method of analysis is a method for monitoring the presence of active components in the sample, and wherein the presence of the signature component in the sample is indicative of an active component in the sample.
8. The method of claim 1 , wherein the method of analysis is a method for determining the amount of active components in the sample by determining the amount of signature component in the sample.
9. The method of claim 1 , wherein the method is performed on at least two samples, and wherein the relative amounts of signature component in each of the at least two samples is determined, wherein the highest relative level of signature component is indicative of the most active sample.
10. The method of claim 1 , further comprising the use of the signature component to identify biologically active molecules.
11. The method of claim 10 , wherein the signature component is used to screen a library.
12. The method of claim 1 , wherein the experimental constraint applied to the sample is digestion with an exoenzyme.
13. The method of claim 1 , wherein the experimental constraint applied to the sample is digestion with an endoenzyme.
14. The method of claim 1 , wherein the experimental constraint applied to the sample is chemical digestion.
15. The method of claim 1 , wherein the experimental constraint applied to the sample is chemical modification.
16. The method of claim 1 , wherein the experimental constraint applied to the sample is modification with an enzyme.
17. The method of claim 1 , wherein the polysaccharide in the sample is compared to a reference database of polysaccharides of identical size as the polysaccharide, wherein the polysaccharides of the reference database have also been subjected to the same experimental constraints as the polysaccharide in the sample, wherein the comparison provides a compositional analysis of the sample polysaccharide.
18. The method of claim 1 , wherein the polysaccharide is a glycosaminoglycan.
19. The method of claim 1 , wherein the polysaccharide is low molecular weight heparin.
20. The method of claim 1 , wherein the polysaccharide is heparin.
21. The method of claim 1 , wherein the polysaccharide is a biotechnologically prepared heparin.
22. The method of claim 1 , wherein the polysaccharide is a chemically modified heparin.
23. The method of claim 1 , wherein the polysaccharide is a synthetic heparin.
24. The method of claim 1 , wherein the polysaccharide is heparan sulfate.
25. The method of claim 1 , wherein the sample is a pharmaceutical product.
26. The method of claim 1 , wherein the sample is a biological sample.
27. The method of claim 1 , wherein the sample is a blood sample.
28. The method of claim 1 , wherein the signature component is ΔUH NAC,6S GH NS,3S,6S .
29. The method of claim 1 , wherein the signature component is ΔUH NS,6S GH NS,3S,6S .
30. The method of claim 1 , wherein the signature component is ΔUH NAC,6S GH NS,3S .
31. The method of claim 1 , wherein the signature component is ΔUH NS,6S GH NS,3S .
32. The method of claim 1 , wherein the experimental constraint applied to the sample is mass spectrometry.
33. The method of claim 32 , wherein the mass spectrometry is MALDI-MS.
34. A method of analyzing a sample comprising:
(A) applying an experimental constraint to a polysaccharide in a sample, to produce a modified polysaccharide, having a signature component,
(B) detecting the presence of the signature component, in the sample as an indication that the polysaccharide is present in the sample, and
(C) determining the presence or absence of the signature component to analyze the sample,
wherein the experimental constraint applied to the sample is gel permeation chromatography or nuclear magnetic resonance, and
wherein the signature component is ΔUH NAC,6S GH NS,3S,6S ; ΔUH NS,6S GH NS,3S,6S ; ΔUH NAC,6S GH NS,3S ; or ΔUH NS,6S GH NS,3S .
35. A method of analyzing a sample comprising:
(A) applying an experimental constraint to a polysaccharide in a sample, to produce a modified polysaccharide, having a signature component,
(B) detecting the presence of the signature component, in the sample as an indication that the polysaccharide is present in the sample, and
(C) determining the presence or absence of the signature component to analyze the sample,
wherein the sample is a batch of polysaccharide and the signature component is used to monitor the purity of the batch by determining the amount of signature component in the batch, and
wherein the signature component is ΔUH NAC,6S GH NS,3S,6S ; ΔUH NS,6S GH NS,3S,6S ; ΔUH NAC,6S GH NS,3S ; or ΔUH NS,6S GH NS,3S .
36. A method of analyzing a sample comprising:
(A) applying an experimental constraint to a polysaccharide in a sample, to produce a modified polysaccharide, having a signature component,
(B) detecting the presence of the signature component, in the sample as an indication that the polysaccharide is present in the sample, and
(C) determining the presence or absence of the signature component to analyze the sample,
wherein the signature component is used to screen a library, and
wherein the signature component is ΔUH NAC,6S GH NS,3S,6S ; ΔUH NS,6S GH NS,3S,6S ; ΔUH NAC,6S GH NS,3S ; or ΔUH NS,6S GH NS,3S .