CRP capture/detection of gram positive bacteria

Patent metadata

Field	Value
Publication number	US-11718651-B2
Application number	US-202117318343-A
Country	US
Kind code	B2
Filing date	May 12, 2021
Priority date	Dec 18, 2013
Publication date	Aug 8, 2023
Grant date	Aug 8, 2023

Abstract

Described herein are engineered microbe-targeting molecules, microbe-targeting articles, kits comprising the same, and uses thereof. Such microbe-targeting molecules, microbe-targeting articles, or the kits comprising the same can bind or capture of a microbe or microbial matter thereof, and can thus be used in various applications, such as diagnosis or treatment of an infection caused by microbes in a subject or any environmental surface.

First claim

What is claimed is:

1. A method for determining the presence or absence of a microbe in a sample, comprising:
(i) contacting a test sample with a microbe-targeting molecule; and
(ii) detecting binding of a microbe to the microbe-targeting molecule,
wherein the microbe is present in the test sample if binding is detected and wherein the microbe-targeting molecule comprises:
a. at least one first domain comprising at least a portion of a c-reactive protein (CRP);
b. at least one second domain comprising at least a portion of a domain selected from the group consisting of:
(i) Fc region of an immunoglobulin;
(ii) microbe-binding domain of a microbe-binding protein, wherein the microbe-binding protein is not CRP;
(iii) neck region of a lectin;
(iv) a detectable label;
(v) domain for conjugation to surface of a carrier scaffold;
(vi) pattern recognition receptor domain of CRP; and
(vii) any combinations of (i)-(vi); and

c. a linker conjugating the first and second domains, and
wherein the microbe-targeting molecule is a multimeric molecule.

2. The method of claim 1 , wherein said detecting step comprises an enzyme-linked immunosorbent assay (ELISA), a fluorescent linked immunosorbent assay (FLISA), immunofluorescent microscopy, hybridization, fluorescence in situ hybridization (FISH), antibody-based imaging, a radiological detection assay, a chemical detection assay, an enzymatic detection assay, an optical detection assay, an electrochemical assay, cell culture, or any combinations thereof.

3. The method of claim 1 , wherein the microbe-targeting molecule is conjugated with a detectable label.

4. The method of claim 1 , wherein the microbe-bound microbe-targeting molecule is detectable without prior cell culture.

5. The method of claim 1 , wherein the contacting step comprises flowing the test sample through a channel, wherein the channel is coated with the microbe-targeting molecules.

6. The method of claim 1 , wherein the contacting step comprises flowing the test sample and microbe-targeting molecule through a channel.

7. The method of claim 1 , wherein said detecting in step (ii) comprises contacting the sample with a labeling molecule conjugated with a detectable label.

8. The method of claim 1 , wherein the multimeric molecule is formed by: interactions between the linkers of different molecules forming the multimeric molecule; or the linker and the second domain of different molecules forming the multimeric molecule; or the second domains of different molecules forming the multimeric molecule.

9. The method of claim 1 , wherein the Fc region comprises at least one mutation.

10. The method of claim 1 , wherein the microbe-binding protein is a carbohydrate binding protein.

11. The method of claim 10 , wherein the microbe-binding domain is a carbohydrate recognition domain (CRD) of the carbohydrate binding protein.

12. The method of claim 11 , wherein the CRD is from a lectin or ficolin.

13. The method of claim 1 , wherein the second domain comprises at least a portion of Fc region of an immunoglobulin and at least a portion of the neck region of a lectin.

14. The method of claim 1 , wherein the detectable molecule is selected from the group consisting of biotin, fluorophore, luminescent or bioluminescent marker, a radiolabel, an enzyme, an enzyme substrate, a quantum dot, an imaging agent, a gold particle, and any combinations thereof.

15. The method of claim 1 , wherein the domain for conjugation to surface of a carrier scaffold comprises an amino group, a N-substituted amino group, a carboxyl group, a carbonyl group, an acid anhydride group, an aldehyde group, a hydroxyl group, an epoxy group, a thiol, a disulfide group, an alkenyl group, a hydrazine group, a hydrazide group, a semicarbazide group, a thiosemicarbazide group, one partner of a binding pair, an amide group, an aryl group, an ester group, an ether group, a glycidyl group, a halo group, a hydride group, an isocyanate group, an urea group, or an urethane group.

16. The method of claim 1 , wherein the microbe-targeting molecule is conjugated to a surface of a carrier scaffold.

Assignees

• Harvard College

Inventors

• Watters Alexander L
• Ingber Donald E
• Cartwright Mark J
• Super Michael
• Rottman Martin
• Murray Evangelia
• Dusel Brendon

Classifications

• C07K14/4737 Primary

  C-reactive protein · CPC title

• A61K47/42

  Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein (oligopeptides having up to five amino acids {A61K47/183}; polyamino acids A61K47/34) · CPC title

• C07K14/4726

  Lectins · CPC title

• G01N33/569 Primary

  for microorganisms, e.g. protozoa, bacteria, viruses · CPC title

• G01N33/56911

  Bacteria · CPC title