Methods for isolating a target analyte from a heterogeneous sample

• US 8,841,104 B2
• Filed: 04/21/2011
• Issued: 09/23/2014
• Est. Priority Date: 04/21/2010
• Status: Active Grant

• Current Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Original Assignee: nanoMR, Inc. (Genting Bhd.)
• Inventors: Dryga, Sergey A., Esch, Victor C., Clarizia, Lisa-Jo Ann, Adams, Eddie W., Ung, Thearith H., Sitdikov, Ravil A.
• Primary Examiner(s): Hines, Ja'Na
• Application Number:
  US13/091,510
• Publication Number:
  US 20110262925A1
• Time in Patent Office: 1,251 Days
• US Class Current: 435/173.9
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: C07K 16/1267 from Gram-positive bacteria
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/143 Magnetism, e.g. magnetic label
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 2446/90 characterised by small molecule linker used to couple immunoreagents to magnetic particles
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/54333 Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction
• CPC Class Codes: G01N 33/5434 using magnetic particle immunoreagent carriers which constitute new materials per se
• CPC Class Codes: Y10T 436/25 including sample preparation
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

The invention generally relates to methods of using compositions that include sets of magnetic particles, members of each set being conjugated to an antibody specific for a pathogen, and magnets to isolate a pathogen from a body fluid sample.

Claims

1. A method for isolating pathogen from a heterogeneous sample, the method comprising:

providing a vessel that contains a heterogeneous sample comprising pathogen, the vessel being coupled to a microfluidic device that comprises a channel and a magnet;

introducing to the sample a cocktail comprising a plurality of sets of magnetic particles, members of different sets being conjugated to different antibodies that are specific for different pathogen, wherein at least two of the sets are provided at different concentrations, thereby forming a mixture;

flowing the mixture from the vessel through a channel in the microfluidic device and toward a magnet in the microfluidic device; and

separating particle bound pathogen from other components in the sample by using the magnet to retain pathogen bound to magnetic particles while unbound sample components are not retained by the magnet.

2. The method of claim 1, further comprising characterizing the pathogen.

3. The method of claim 2, wherein characterizing comprises identifying the pathogen.

4. The method of claim 3, wherein identifying is selected from sequencing nucleic acid derived from the pathogen or amplifying the nucleic acid.

5. The method of claim 1, wherein the sample is a blood sample.

6. The method of claim 1, wherein each set is present at a concentration designed for detection of a specific pathogen in said sample.

7. The method of claim 1, wherein the particles are differently labeled.

8. The method of claim 7, wherein the label is an optical label.

9. The method of claim 8, wherein the optical label is a fluorescent label.

10. The method of claim 1, wherein the antibodies are monoclonal antibodies.

11. The method of claim 1, wherein the antibodies are polyclonal antibodies.

12. The method of claim 1, wherein the pathogen are gram positive bacteria.

13. The method of claim 1, wherein the pathogen are gram negative bacteria.

14. The method of claim 1, wherein the pathogen are gram positive and gram negative bacteria.