CRP capture/detection of gram positive bacteria

Patent metadata

Field	Value
Publication number	US-10513546-B2
Application number	US-201415105298-A
Country	US
Kind code	B2
Filing date	Dec 18, 2014
Priority date	Dec 18, 2013
Publication date	Dec 24, 2019
Grant date	Dec 24, 2019

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. An assay for determining the presence or absence of a microbe in a test sample, the assay comprising:
(i) contacting a test sample with a microbe-targeting molecule; and
(ii) detecting binding of a microbe to the microbe-targeting molecule, wherein said detecting comprises contacting the sample from (i) with a labeling molecule comprising a detectable label that is conjugated to a c-reactive protein or a microbe binding portion thereof, and
wherein a 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;
(ii) 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

a linker conjugating the first and second domains, and wherein the molecule is a multimeric molecule.

2. The assay 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.

3. An assay for determining the presence or absence of a microbe in a test sample, the assay comprising:
(i) contacting a test sample with a microbe-targeting molecule; and
(ii) detecting binding of a microbe to the microbe-targeting molecule, wherein said detecting comprises contacting the sample from (i) with a labeling molecule comprising a detectable label that is conjugated to a c-reactive protein or a microbe binding portion thereof, and
wherein a 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;
(ii) 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

a linker conjugating the first and second domains, and 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.

4. An assay for determining the presence or absence of a microbe in a test sample, the assay comprising:
(i) contacting a test sample with a microbe-targeting molecule; and
(ii) detecting binding of a microbe to the microbe-targeting molecule, wherein said detecting comprises contacting the sample from (i) with a labeling molecule comprising a detectable label that is conjugated to a c-reactive protein or a microbe binding portion thereof, and
wherein a 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;
(ii) 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

a linker conjugating the first and second domains, and wherein the microbe-targeting molecule is conjugated to a surface of a carrier scaffold.

5. The assay of claim 4 , wherein the carrier scaffold is selected from the group consisting of a nucleic acid scaffold, a protein scaffold, a lipid scaffold, a polymeric scaffold, a dendrimer, a particle, a bead, a nanotube, a microtiter plate, a medical apparatus or implant, a microchip, a filtration device, a membrane, a diagnostic strip, a dipstick, an extracorporeal device, a spiral mixer, a hollow-fiber tube, a living cell, magnetic material, hollow fiber, and any combinations thereof.

Assignees

• Harvard College

Inventors

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

Classifications

• G01N33/6854

  Immunoglobulins · CPC title

• G01N33/569 Primary

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

• C07K14/4726

  Lectins · CPC title

• C07K14/4737 Primary

  C-reactive protein · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title