Method and system for detecting a target within a population of molecules

Patent metadata

Field	Value
Publication number	US-10175246-B2
Application number	US-201715420159-A
Country	US
Kind code	B2
Filing date	Jan 31, 2017
Priority date	Oct 27, 2006
Publication date	Jan 8, 2019
Grant date	Jan 8, 2019

Abstract

A method of detecting a target within a population of molecules comprising: contacting a plurality of labeled probe molecules with the population of molecules potentially containing a target of the probe molecules; acquiring a probe specific signal emitted by said labeled probe molecules that bound to said target together with a background signal; preferentially modulating said probe specific signal by at least one of modulating said acquisition and modulating an emission of said probe specific signal; and detecting said probe specific signal over said background signal using said preferential modulation.

First claim

What is claimed is:

1. A system for analysis of a population of molecules, wherein the population of molecules contains a plurality of probe molecules including at least one labeled probe molecule, wherein the plurality of probe molecules is in contact with a population of molecules potentially containing a target comprising a plurality of target molecules, wherein at least some of the plurality of probe molecules selectively bind to at least some target molecules, so that the at least one labeled probe molecule is bound to at least a portion of the target, said system comprising:
a vessel adapted to contain the population of molecules;
a fluorescent excitation source adapted to direct an excitation beam in a first direction through said vessel, the beam configured to cause at the least one labeled probe molecule to emit a probe specific signal;
a detector adapted to detect said probe specific signal and produce a detection output accordingly;
a signal modulator configured to perform a preferential modulation to at least one of the probe specific signal and said detection output, wherein said signal modulator is configured to create a cyclic magnetic field gradient using two electromagnetic poles driven with a frequency of cycling of current between the two poles, wherein said poles are configured to achieve an increase in said magnetic field gradient outside said poles, wherein said magnetic field gradient is configured to increase a local concentration of probe molecules with bound target molecules to form a cloud of condensed particles, wherein said magnetic field gradient causes the cloud of condensed target molecules with bound magnetic particles to cross a path of the fluorescence excitation energy beam as the cloud of condensed target molecules with bound magnetic particles crosses between said electromagnetic poles in a cyclical manner, wherein said detection output includes data regarding at least one of the following:
at least one of said at least one labeled probe molecule bound to at least a portion of the target; and
the cloud of condensed particles; and

wherein the detection includes detecting by analyzing a signal provided by the excitation beam when there is relative movement between the excitation beam and the probe molecules, wherein said relative movement results in the probe molecules being positioned in and out of the excitation beam.

2. The system of claim 1 , wherein at least one of the at least one labeled probe molecule is each attached to a magnetic particle, and wherein said signal modulator is configured to perform the preferential modulation.

3. The system according to claim 1 , wherein said preferential modulation includes moving the at least one labeled probe molecule bound to at least a portion of the target in a second direction, in and out of the excitation beam.

4. The system according to claim 2 , wherein said magnetic field gradient causes said relative movement between the excitation beam and the bound probe molecules, wherein said modulator is also configured to increase sensitivity to said probe specific signal over a background signal.

5. The system according to claim 1 , wherein said modulator has two magnetic poles configured to increase the local concentration of the probe molecules with attached magnetic particles to form the cloud.

6. The system according to claim 5 , wherein said magnetic field gradient causes relative movement between the excitation beam and the probe molecules with attached magnetic particles.

7. The system according to claim 1 , wherein said modulator is also configured to increase sensitivity to said probe specific signal over a background signal.

8. The system according to claim 1 , wherein at least some of the probe molecules are each attached to a magnetic particle, and wherein said modulator is configured to apply a magnetic force to form a cloud of condensed particles and attached magnetic particles in a narrow path.

9. The system according to claim 8 , wherein said modulator is also configured to cause the cloud to cross a path of the excitation energy beam.

10. The system according to claim 1 , wherein at least one of said at least one labeled probe molecule is each attached to a magnetic particle, and wherein a single one of said two electromagnetic poles is configured to increase a local concentration of probe molecules with bound target molecules to form a cloud of condensed particles and magnetic particles in a narrow path by application of an external magnetic force thereto.

11. The system according to claim 5 , wherein said two magnetic poles include pole tips configured to increase the field gradient outside said poles.

12. The system according to claim 11 , wherein each of said pole tips has a parabolic-shape.

13. The system according to claim 1 , wherein at least some of said probe molecules include at least one of nucleic acids, peptides, proteins, antibodies and ion chelators.

14. The system according to claim 1 , wherein at least some of said plurality of target molecules include at least one molecule type selected from the group consisting of a nucleic acid sequence, an amino acid sequence, a carbohydrate, a carbohydrate sequence, an ion, and a feature of a protein determined by non-primary structure.

15. The system according to claim 1 , wherein at least some of said probe molecules include at least one of nucleic acids, peptides, proteins, antibodies, and ion chelators; and
wherein at least some of said plurality of target molecules include at least one molecule type selected from the group consisting of a nucleic acid sequence, an amino acid sequence, a carbohydrate, a carbohydrate sequence, an ion, and a feature of a protein determined by non-primary structure.

16. The system according to claim 1 , wherein said probe molecules bound to said target molecules include at least one protein molecule bound to one protein target.

17. The system according to claim 1 , wherein said probe molecules bound to said target molecules include at least one nucleic acid molecule bound to a nucleic acid target.

18. The system according to claim 1 , wherein said probe molecules bound to said target molecules include at least one antibody bound to an epitope.

19. The system according to claim 1 , wherein the at least one labeled probe molecule each has a fluorescent label.

20. The system according to claim 19 , wherein at least one of the at least one probe molecule having a fluorescent label is each attached to a magnetic particle.

21. The system according to claim 1 wherein at least a first portion of said probe molecules each have a fluorescent label and are bound to a target and wherein at least a second portion of said probe molecules are each attached to a magnetic particle and are bound to the same target.

22. The system according to claim 21 , wherein the probe molecules bound to the target molecules include at least one protein molecule bound to one protein target.

23. The system according to claim 21 , wherein the probe molecules bound to the target molecules include at least one nucleic acid molecule bound to a nucleic acid target.

24. The method of claim 1 , wherein said detecting includes detecting a probe specific signal originating from at least one label attached to at least one probe.

25. The method of claim 22 , wherein at least one of said at least one probe molecule is attached to a magnetic particle.

Assignees

• Univ Ramot

Inventors

• Arie Ady
• Danielli Amos

Classifications

• G01N33/582 Primary

  with fluorescent label · CPC title

• G01N21/6428 Primary

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• C12Q1/6818

  involving interaction of two or more labels, e.g. resonant energy transfer · CPC title

• G01N33/54333 Primary

  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 title

• G01N21/1717

  with a modulation of one or more physical properties of the sample during the optical investigation, e.g. electro-reflectance · CPC title