Magnetic capture of a target from a fluid

Patent metadata

Field	Value
Publication number	US-10357780-B2
Application number	US-201515522686-A
Country	US
Kind code	B2
Filing date	Oct 27, 2015
Priority date	Oct 27, 2014
Publication date	Jul 23, 2019
Grant date	Jul 23, 2019

Abstract

Disclosed herein is an improved method for magnetic capture of target molecules (e.g., microbes) in a fluid. Kits and solid substrates for carrying the method described herein are also provided. In some embodiments, the methods, kits, and solid substrates described herein are optimized for separation and/or detection of microbes and microbe-associated molecular pattern (MAMP) (including, e.g., but not limited to, a cell component of microbes, lipopolysaccharides (LPS), and/or endotoxin).

First claim

What is claimed is:

1. A method of capturing at least one target from a fluid comprising:
introducing a fluid and target-binding magnetic particles to a magnetic separation chamber in the presence of a magnetic field gradient (a gradient of a magnetic field),
wherein at least a portion of a fluid-contact surface of the magnetic separation chamber comprises magnetic field gradient concentrating particles distributed thereon and substantially aligned along magnetic flux lines of the magnetic field,
wherein the magnetic field gradient concentrating particles act as local magnetic field gradient concentrators and attracts at least a portion of the target-binding magnetic particles to the magnetic field gradient concentrating particles in the presence of the magnetic field gradient, thereby capturing a target bound on the target-binding magnetic particles from the fluid,
wherein the target-binding magnetic particles are microbe-binding magnetic particles, and
wherein the microbe-binding magnetic particles comprise on their surface microbe-binding molecules, and
wherein the microbe-binding molecule is selected from the group consisting of opsonins and lectins.

2. The method of claim 1 , wherein the magnetic field gradient concentrating particles form magnetic micro- or nano-structures on said at least a portion of the fluid-contact surface of the magnetic separation chamber.

3. The method of claim 1 , wherein the diameter of the target-binding magnetic particles is no more than 250 nm.

4. The method of claim 1 , wherein at least 50% area or higher of said at least a portion of the fluid-contact surface comprises the magnetic field gradient concentrating particles distributed thereon.

5. The method of claim 1 , wherein the fluid is flowed through the magnetic separation chamber at a flow rate of about 1 ml/hr to about 10 L/hr.

6. The method of claim 1 , wherein the magnetic separation chamber comprises a channel, a microfluidic channel, a sample well, a microtiter plate, a slide, a flask, a tube, a nanotube, a fiber, a filter, a membrane, a scaffold, an extracorporeal device, a mixer, a hollow fiber, or any combinations thereof.

7. The method of claim 1 , wherein the fluid is a biological fluid obtained or derived from a subject, a fluid or specimen obtained from an environmental source, a fluid from a cell culture, a microbe colony, or any combinations thereof.

8. The method of claim 1 , wherein the target-binding magnetic particles are paramagnetic or superparamagnetic particles.

9. The method of claim 1 , wherein the microbe-binding molecule comprises at least a microbial-binding portion of C-type lectins, collectins, ficolins, receptor-based lectins, lectins from the shrimp Marsupenaeus japonicas, non-C-type lectins, lipopolysaccharide (LPS)-binding proteins, endotoxin-binding proteins, peptidoglycan-binding proteins, or any combinations thereof; or wherein the microbe-binding molecule is selected from the group consisting of mannose-binding lectin (MBL), surfactant protein A, surfactant protein D, collectin 11, L-ficolin, ficolin A, DC-SIGN, DC-SIGNR, SIGNR1, macrophage mannose receptor 1, dectin-1, dectin-2, lectin A, lectin B, lectin C, wheat germ agglutinin, CD14, MD2, lipopolysaccharide-binding protein (LBP), limulus anti-LPS factor (LAL-F), mammalian peptidoglycan recognition protein-1 (PGRP-1), PGRP-2, PGRP-3, PGRP-4, C-reactive protein (CRP), and any combinations thereof.

10. The method of claim 1 , wherein the microbe-binding molecule is selected from the group consisting of MBL (mannose binding lectin), FcMBL (IgG Fc fused to mannose binding lectin), AKT-FcMBL (IgG Fc-fused to mannose binding lectin with the N-terminal amino acid tripeptide of sequence AKT (alanine, lysine, threonine)), and any combination thereof.

11. The method of claim 1 , wherein the microbe-binding molecule comprises an amino acid sequence selected from the group consisting of SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, and any combination thereof.

12. A method of capturing at least one target from a fluid comprising:
introducing a fluid comprising target-binding magnetic particles to a magnetic separation chamber in the presence of a magnetic field gradient (a gradient of a magnetic field), wherein at least a portion of a fluid-contact surface of the magnetic separation chamber comprises magnetic field gradient concentrating particles distributed thereon and aligned along with magnetic flux lines of the magnetic field,
wherein the magnetic field gradient concentrating particles act as local magnetic field gradient concentrators, thereby attracting at least a portion of target-bound target-binding magnetic particles to the magnetic field gradient concentrating particles in the presence of the magnetic field gradient,
wherein the target-binding magnetic particles are microbe-binding magnetic particles,
wherein the microbe-binding magnetic particles comprise on their surface microbe-binding molecules, and
wherein the microbe-binding molecule is selected from the group consisting of opsonins and lectins.

Assignees

• Harvard College

Inventors

• Kang Joo Hun
• Ingber Donald E
• Super Michael

Classifications

• B03C2201/22

  characterised by the magnetic field, e.g. its shape or generation · CPC title

• G01N33/54366

  Apparatus specially adapted for solid-phase testing · CPC title

• B01L2400/043

  magnetic forces · CPC title

• B03C2201/18

  Magnetic separation whereby the particles are suspended in a liquid · CPC title

• B03C1/288

  disposed at the outer circumference of a recipient · CPC title