Photo-generated carbohydrate arrays and the rapid identification of pathogen-specific antigens and antibodies

• US 8,658,573 B2
• Filed: 09/11/2007
• Issued: 02/25/2014
• Est. Priority Date: 09/11/2006
• Status: Expired due to Fees

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Inventors: Wang, Denong, Carroll, Gregory T., Turro, Nicholas J., Koberstein, Jeffrey T.
• Primary Examiner(s): Babic, Christopher M
• Assistant Examiner(s): Flinders, Jeremy C
• Application Number:
  US11/900,221
• Publication Number:
  US 20100331198A1
• Time in Patent Office: 2,359 Days
• Field of Search: None
• US Class Current: 506/19
• CPC Class Codes: C07H 15/04 attached to an oxygen atom of the saccharide radical
• CPC Class Codes: C40B 40/12 Libraries containing saccharides or polysaccharides, or derivatives thereof
• CPC Class Codes: C40B 50/18 using a particular method of attachment to the solid support
• CPC Class Codes: G01N 2333/32 from Bacillus (G)
• CPC Class Codes: G01N 2400/00 Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
• CPC Class Codes: G01N 2500/02 Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
• CPC Class Codes: G01N 33/56911 Bacteria

Abstract

The invention relates to novel photo-generated carbohydrate arrays and methods of their use to detect the presence of one or more agents in a sample. The invention also relates to a high-throughput strategy to facilitate the identification and immunological characterization of pathogen-specific carbohydrates, including those of Bacillus anthracis. The invention can be used to determine the presence of a pathogen and whether a subject has been exposed to a pathogen, such as by screening for pathogen-specific antibodies.

Claims

1. A microarray comprising:

a surface;

a compound of formula (I);

2. The microarray of claim 1, wherein the agent is a biomarker for a disease.

3. The microarray of claim 1, wherein the agent is an antibody.

4. The microarray of claim 3, wherein the antibody is capable of binding to a specific pathogen.

5. The microarray of claim 4, wherein the pathogen is Bacillus anthracis.

6. The microarray of claim 1, wherein n is an integer from 1 to 20.

7. The microarray of claim 1, wherein the compound of Formula (I) is

8. The microarray of claim 1, wherein at least one carbohydrate is selected from the group consisting of:

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-D-glucose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-α

-D-glucose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-β

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-β

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranoside;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-Dglucopyranosyl-(1→

3)-α

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranosyl-(1→

3)-β

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranosyl-(1→

3)-β

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-Dglucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

or

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-Dglucopyranosyl-(1→

3)-β

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose,

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranose;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-bis[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-tris[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-tetrakis[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

[Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside]₅-BSA;

[Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-bis[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside]₁₀-BSA;

[Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-tris[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside]₁₀-BSA;

glycosides thereof; and

combinations thereof.

9. The microarray of claim 1, wherein the compound of formula (I) forms a self-assembled monolayer.

10. The microarray of claim 1, wherein Y is —

NH₂.

11. The microarray of claim 10, wherein the compound of Formula (III) and the compound of Formula (I) are present in a ratio from about 1:

1 to about 100;

1.

12. The microarray of claim 11, wherein the ratio is from about 5:

1 to about 20;

1.

13. The microarray of claim 1, wherein the compound of Formula (III) is

14. The microarray of claim 1, wherein the array comprises a chip, a plate, a sensor, a slide, a combination thereof, or a portion thereof.

15. The microarray of claim 1, wherein the surface comprises an inorganic material.

16. The microarray of claim 15, wherein the surface comprises silica, glass, quartz, silicon, titanium, titania, gold, or a combination thereof.

17. The microarray of claim 1, wherein the surface comprises an organic material.

18. The microarray of claim 17, wherein the surface comprises polyacrylamide, nylon, polyethylene, polypropylene, PTFE, polycarbonate, polystyrene, poly(tert-butyl acrylate), poly(vinyl alcohol), nitrocellulose, or a combination thereof.

19. The microarray of claim 1, wherein the compound of Formula (III) is

20. The microarray of claim 1, wherein the compound of Formula (I) is

21. The microarray of claim 1, wherein the compound of Formula (I) is

22. The microarray of claim 21, wherein at least one carbohydrate is selected from the group consisting of:

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-D-glucose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-α

-D-glucose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-β

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-β

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranose;

α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranoside;

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-α

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranoside;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-Dglucopyranosyl-(1→

3)-α

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranosyl-(1→

3)-β

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-D-glucopyranosyl-(1→

3)-β

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranose;

L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-Dglucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose;

or

n-Pentenyl 4,6-Dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-β

-Dglucopyranosyl-(1→

3)-β

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-α

-L-rhamnopyranose,

5-(Methoxycarbonyl)-pentyl-4-(3-hydroxy-3-methylbutanamido)-2-O-methyl-4,6-dideoxy-β

-D-glucopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

3)-α

-L-rhamnopyranosyl-(1→

2)-β

-L-rhamnopyranose;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-bis[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-tris[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-tetrakis[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside;

[Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside]₅-BSA;

[Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-bis[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside]₁₀-BSA;

[Methyl 2-O-methyl-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)-tris[4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranosyl-(1→

2)]-4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-D-mannopyranoside]₁₀-BSA;

glycosides thereof; and

combinations thereof.