Engineered microbe-targeting molecules and uses thereof

Patent metadata

Field	Value
Publication number	US-9593160-B2
Application number	US-201214233553-A
Country	US
Kind code	B2
Filing date	Jul 18, 2012
Priority date	Jul 18, 2011
Publication date	Mar 14, 2017
Grant date	Mar 14, 2017

Abstract

Described herein are engineered microbe-targeting or microbe-binding molecules, kits comprising the same and uses thereof. Some particular embodiments of the microbe-targeting or microbe-binding molecules comprise a carbohydrate recognition domain of mannose-binding lectin, or a fragment thereof, linked to a portion of a Fc region. In some embodiments, the microbe-targeting molecules or microbe-binding molecules can be conjugated to a substrate, e.g., a magnetic microbead, forming a microbe-targeting substrate (e.g., a microbe-targeting magnetic microbead). Such microbe-targeting molecules and/or substrates and the kits comprising the same can bind and/or capture of a microbe and/or microbial matter thereof, and can thus be used in various applications, e.g., diagnosis and/or treatment of an infection caused by microbes such as sepsis in a subject or any environmental surface. Microbe-targeting molecules and/or substrates can be regenerated after use by washing with a low pH buffer or buffer in which calcium is insoluble.

First claim

What is claimed is:

1. A composition comprising:
a. a microbe;
b. a first molecule comprising an unlabeled engineered microbe-targeting molecule which comprises a lectin or carbohydrate-binding portion thereof; and
c. a second, different, molecule comprising a detectable label conjugated to a targeting agent specific for the microbe.

2. The composition of claim 1 , wherein the engineered microbe-targeting molecule is present on a substrate.

3. The composition of claim 1 , wherein the engineered microbe-targeting molecule is covalently linked to a substrate.

4. The composition of claim 1 , wherein the lectin is a C-type lectin.

5. The composition of claim 1 , wherein the lectin is a collectin.

6. The composition of claim 1 , wherein the lectin is mannose-binding lectin.

7. The composition of claim 1 , wherein the unlabeled engineered microbe-targeting molecule comprises a carbohydrate recognition domain of mannose-binding lectin, or a fragment thereof, linked to a portion of a Fc region.

8. The composition of claim 1 , wherein the engineered microbe-targeting molecule comprises a carbohydrate recognition domain and neck region of a mannose-binding lectin fused to the C-terminal of a Fc fragment of human IgG1.

9. The composition of claim 8 , wherein the portion of the Fc region is linked N-terminal of the carbohydrate recognition domain.

10. The composition of claim 1 , wherein the lectin comprises a carbohydrate recognition domain having the sequence of SEQ ID NO: 4 or a fragment thereof.

11. The composition of claim 1 , wherein the lectin comprises a carbohydrate recognition domain having a sequence having at least 90% identity to the sequence of SEQ ID NO: 4.

12. The composition of claim 1 , wherein the detectable label comprises an enzyme.

13. The composition of claim 12 , wherein the enzyme is horseradish peroxidase (HRP).

14. The composition of claim 12 , which further comprises a substrate for the enzyme.

15. The composition of claim 14 , wherein the substrate is TMB (3,3′,5,5′-tetramethylbenzidine).

16. The composition of claim 1 , wherein the targeting agent specific for the microbe comprises an engineered microbe-targeting molecule or an antibody.

17. The composition of claim 1 , wherein the targeting agent specific for the microbe comprises MBL.

18. A kit comprising:
a. a first molecule comprising an unlabeled engineered microbe-targeting molecule which comprises a lectin or carbohydrate-binding portion thereof; and
b. one or more containers containing a second, different molecule comprising a detectable label conjugated to a targeting agent specific for a microbe.

Assignees

• Ingber Donald E
• Super Michael
• Way Jeffrey Charles
• Cartwright Mark J
• Berthet Julia B
• Super Dinah R
• Rottman Martin M
• Watters Alexander
• Harvard College

Inventors

• Ingber Donald E
• Super Michael
• Way Jeffrey Charles
• Cartwright Mark J
• Berthet Julia B
• Super Dinah R
• Rottman Martin M
• Watters Alexander

Classifications

• A61P31/04

  Antibacterial agents · CPC title

• C07K2319/33

  fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies · CPC title

• G01N33/56938

  Staphylococcus · CPC title

• G01N33/56911

  Bacteria · CPC title

• G01N2333/42

  Lectins, e.g. concanavalin, phytohaemagglutinin · CPC title