Use of polyamines as resolving agents for capillary electrophoresis of labeled glycans using gels

Patent metadata

Field	Value
Publication number	US-11988633-B2
Application number	US-201816644467-A
Country	US
Kind code	B2
Filing date	Sep 20, 2018
Priority date	Sep 22, 2017
Publication date	May 21, 2024
Grant date	May 21, 2024

Abstract

The present disclosure provides methods of improving the resolution of labeled glycans in capillary electrophoresis techniques using a gel as a sieving matrix, by incorporating polyamines in the gel.

First claim

The invention claimed is:

1. A method of improving resolution in separating labeled glycans in a sample by capillary electrophoresis analysis, wherein said separation takes place in a gel, wherein said gel comprises a polyamine having a structure of any of Structure 1

wherein R 1 , R 2 , and R 3 are independently selected from H and CH 3 ,
Structure 2

wherein each of R 1 and R 2 is independently selected from H and CH 3 , and said polyamine is N,N,N′,N′-Tetramethyl-1,3-propanediamine,
Structure 3

wherein each of R 1 and R 2 is independently selected from H and CH 3 ,
Structure 4

wherein each of R 1 -R 3 is independently selected from H and CH 3 ,
Structure 5

wherein each of R 1 -R 4 can independently be H or CH 3 ,
Structure 6,

wherein each of R 1 -R 6 can independently be H or CH 3 ,
Structure 7

wherein each of R 1 -R 6 is independently H or CH 3 , or
Structure 8

wherein each of R 1 -R 7 is independently H or CH 3 , provided that, when said separation takes place in a capillary containing the gel and the gel is of polyethylene oxide (“PEO”), the capillary is coated.

2. The method of method 1, wherein said polyamine is of Structure 4 and is 1,4,7-Trimethyl-1,4,7-triazacyclononane.

3. The method of claim 1 , wherein said polyamine is of Structure 1 and is 1, 2-Ethanediamine, N1-(2-aminoethyl) or 1, 2-Ethanediamine, N1-[2-(dimethylamino) ethyl]-N1, N2, N2-trimethyl-.

4. The method of claim 3 , wherein said polyamine is 1, 2-Ethanediamine, N1-[2-(dimethylamino) ethyl]-N1, N2, N2-trimethyl-.

5. The method of claim 1 , wherein said glycans have been labeled by being reacted with 8-aminopyrene-1,3,6-trisulfonic acid (“APTS”), or 8-Aminonaphthalene-1,3,6-trisulfonic acid disodium salt (“ANTS”).

6. The method of claim 5 , wherein the glycans have been labeled with APTS and are MAN-5 glycan and A1F glycan.

7. The method of claim 5 , wherein the glycans have been labeled with ANTS and are MAN-5 glycan and A1F glycan.

8. A composition for capillary electrophoretic separation of labeled glycans, said composition comprising a gel suitable for capillary gel electrophoresis of labeled glycans, which gel comprises at least one polyamine of any of the following structures:
Structure 1

wherein each of R 1 -R 3 are independently selected from H and CH 3 ,
Structure 2

wherein R 1 and R 2 are independently selected from H and CH 3 , and said polyamine is N,N,N′,N′-Tetramethyl-1,3-propanediamine,
Structure 3

wherein each R 1 and R 2 are independently selected from H and CH 3 ,
Structure 4

wherein each of R 1 -R 3 is independently selected from H and CH 3 ,
Structure 5

wherein each of R 1 -R 4 can independently be H or CH 3 ,
Structure 6,

wherein each of R 1 -R 6 can independently be H or CH 3 ,
Structure 7

wherein each of R 1 -R 6 is independently H or CH 3 , or
Structure 8

wherein each of R 1 -R 7 is independently H or CH 3 ,
provided that, when the capillary electrophoresis separation takes place in a capillary containing the gel and the gel is of polyethylene oxide (“PEO”), the capillary is coated.

9. The composition of claim 8 , wherein said polyamine is of Structure 4 and is 1,4,7-Trimethyl-1,4,7-triazacyclononane.

10. The composition of claim 8 , wherein said polyamine is of Structure 1 and is 1, 2-Ethanediamine, N1-(2-aminoethyl) or 1, 2-Ethanediamine, N1-[2-(dimethylamino) ethyl]-N1, N2, N2-trimethyl-.

11. The composition of claim 8 , wherein said polyamine is N,N,N′,N″,N″-Pentamethyldiethylenetriamine or N,N,N′,N′-Tetramethyl-1,3-propanediamine.

12. The composition of claim 8 , wherein said gel suitable for capillary gel electrophoresis of labeled glycans comprises polyethylene oxide.

13. A cartridge or capillary loaded with a gel for capillary electrophoretic separation of labeled glycans, which gel comprises at least one polyamine of any of the following structures:
Structure 1

wherein each of R 1 -R 3 are independently selected from H and CH 3 ,
Structure 2

wherein R 1 and R 2 are independently selected from H and CH 3 , and said polyamine is N,N,N′,N′-Tetramethyl-1,3-propanediamine,
Structure 3

wherein each R 1 and R 2 are independently selected from H and CH 3 ,
Structure 4

wherein each of R 1 -R 3 is independently selected from H and CH 3 ,
Structure 5

wherein each of R 1 -R 4 is independently selected from H and CH 3 ,
Structure 6,

wherein each of R 1 -R 6 is independently selected from H and CH 3 ,
Structure 7

wherein each of R 1 -R 6 is independently H or CH 3 , or
Structure 8

wherein each of R 1 -R 7 is independently H or CH 3 , provided that, when the cartridge or capillary is a capillary and the gel is of polyethylene oxide (“PEO”), the capillary is coated.

14. The cartridge or capillary of claim 13 , wherein said at least one polyamine

Assignees

• Agilent Technologies Inc

Inventors

• Limjap Fielito U
• Kimzey Michael J
• Haxo Francis T

Classifications

• G01N27/44747 Primary

  Composition of gel or of carrier mixture · CPC title

• C08B37/0003

  General processes for their isolation or fractionation, e.g. purification or extraction from biomass · CPC title

• C08B37/0087

  Glucomannans or galactomannans; Tara or tara gum, i.e. D-mannose and D-galactose units, e.g. from Cesalpinia spinosa; Tamarind gum, i.e. D-galactose, D-glucose and D-xylose units, e.g. from Tamarindus indica; Gum Arabic, i.e. L-arabinose, L-rhamnose, D-galactose and D-glucuronic acid units, e.g. from Acacia Senegal or Acacia Seyal; Derivatives thereof · CPC title

• C08G73/0206

  Polyalkylene(poly)amines · CPC title

• C08G73/0273

  Polyamines containing heterocyclic moieties in the main chain · CPC title