Engineered microbe-targeting molecules and uses thereof

Patent metadata

Field	Value
Publication number	US-10865235-B2
Application number	US-201916683630-A
Country	US
Kind code	B2
Filing date	Nov 14, 2019
Priority date	Jul 18, 2011
Publication date	Dec 15, 2020
Grant date	Dec 15, 2020

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 plurality of magnetic beads, wherein the magnetic beads are conjugated to a plurality of molecules each comprising a first molecule comprising a carbohydrate recognition domain of a mannose-binding lectin (MBL) linked to an Fc region of an antibody (FcMBL); and
a second molecule comprising a detectable label conjugated to a targeting agent specific for a microbe.

2. The composition of claim 1 , wherein the FcMBL molecules are dimerized.

3. The composition of claim 2 , wherein the FcMBL molecules are dimerized via interactions of their respective Fc regions.

4. The composition of claim 1 , wherein the MBL comprises an amino acid sequence of SEQ ID NO: 2.

5. The composition of claim 1 , wherein the targeting agent comprises a C-type lectin.

6. The composition of claim 1 , wherein the targeting agent comprises a collectin.

7. The composition of claim 1 , wherein the targeting agent comprises mannose-binding lectin.

8. The composition of claim 1 , wherein the targeting agent comprises a carbohydrate recognition domain of mannose-binding lectin, or a fragment thereof, linked to a portion of a Fc region.

9. The composition of claim 1 , wherein the first molecule further comprises the carbohydrate recognition domain and a neck region of the MBL and wherein the Fc region comprises an Fc fragment of human IgG1.

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

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

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

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

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

15. The composition of claim 13 , which further comprises a substrate for the enzyme.

16. The composition of claim 15 , wherein the substrate is 3,3′,5,5′-tetramethylbenzidine.

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

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

Assignees

• Harvard College

Inventors

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

Classifications

• A61K47/68 Primary

  the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment · CPC title

• G01N2333/4724

  Lectins · CPC title

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title

• G01N33/54353

  with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393) · CPC title

• G01N33/56961

  Plant cells or fungi · CPC title