Methods for isolating a target analyte from a heterogeneous sample

• US 9,696,302 B2
• Filed: 09/05/2014
• Issued: 07/04/2017
• Est. Priority Date: 04/21/2010
• Status: Active Grant

• Current Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Original Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Inventors: Ung, Thearith H., Sitdikov, Ravil A., Dryga, Sergey A., Esch, Victor C., Clarizia, Lisa-Jo Ann, Adams, Eddie W.
• Primary Examiner(s): Hines, Ja'Na
• Application Number:
  US14/478,692
• Publication Number:
  US 20150212079A1
• Time in Patent Office: 1,033 Days
• 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 fluidic device that comprises a channel and a magnet;

introducing to the sample in the vessel a cocktail comprising a plurality of magnetic particles, conjugated to antibodies that bind said pathogen, thereby forming a mixture comprising antibody-bound pathogen particles;

flowing the mixture from the vessel through a channel in the fluidic device such that said magnetic particles attach to said magnet; and

separating antibody-bound pathogen attached to said magnet from other components in the sample.

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

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

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

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

6. The method of claim 1, wherein the particles are differentially labeled.

7. The method of claim 6, wherein the particles comprise is an optical label.

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

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

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

11. The method of claim 1, wherein the pathogen is gram positive bacteria.

12. The method of claim 1, wherein the pathogen is gram negative bacteria.

13. The method of claim 1, wherein the pathogen is a virus.