Asynchronous magnetic bead rotation (AMBR) microviscometer for analysis of analytes

• US 9,983,110 B2
• Filed: 11/04/2014
• Issued: 05/29/2018
• Est. Priority Date: 11/04/2013
• Status: Active Grant

• Current Assignee: Board of Regents of the University of Michigan (University of Michigan)
• Original Assignee: Board of Regents of the University of Michigan (University of Michigan)
• Inventors: Kopelman, Raoul, Burns, Mark A., Li, Yunzi
• Primary Examiner(s): Thomas, David C
• Application Number:
  US14/532,907
• Publication Number:
  US 20150140560A1
• Time in Patent Office: 1,302 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2527/153 Viscosity
• CPC Class Codes: C12Q 2563/137 Metal/ion, e.g. metal label
• CPC Class Codes: G01N 11/10 by moving a body within the material
• CPC Class Codes: G01N 11/165 Sample held between two members substantially perpendicular to axis of rotation, e.g. parallel plate viscometer
• CPC Class Codes: G01N 2011/0086 magnetic properties

Abstract

The disclosure provides a label-free viscosity-based analyte detection system using paramagnetic beads as an asynchronous magnetic bead rotation (AMBR) microviscometer. It is disclosed herein that the bead rotation period is linearly proportional to the viscosity of a solution comprising analytes surrounding the paramagnetic bead. Optical measurement of asynchronous microbead motion determines solution viscosity precisely in microscale volumes, thus allowing an estimate of analyte concentration. The results demonstrate the feasibility of viscosity-based analyte detection using AMBR in microscale aqueous volumes.

Claims

1. A method of detecting a chemical change in state of an analyte in a sample comprising:

(a) contacting the sample with a magnetic particle;

(b) measuring a first instantaneous non-linear rotation rate of the magnetic particle in the sample at a first time; and

(c) measuring a second instantaneous non-linear rotation rate of the magnetic particle in the sample at a second time;

wherein a difference between the first instantaneous rotation rate relative to the second instantaneous rotation rate indicates a change in viscosity in the sample, which indicates the change in state of the analyte in the sample,

wherein the chemical change in state is selected from the group consisting of nucleic acid synthesis, protein synthesis, nucleic acid hydrolysis, nucleic acid ligation, and protein hydrolysis.

2. The method of claim 1 wherein the analyte is a monomer of a biopolymer.

3. The method of claim 1 wherein the analyte is selected from the group consisting of a cell, a nucleic acid, a protein, a carbohydrate, a lipid, and an amino acid.

4. The method of claim 1 wherein the nucleic acid synthesis is by polymerase chain reaction (PCR).

5. The method of claim 1 wherein the protein synthesis is by in vitro translation.

6. The method of claim 1 wherein the protein synthesis occurs in a cell.

7. The method of claim 1 wherein the nucleic acid hydrolysis is catalyzed by a type I or a type II restriction endonuclease.

8. The method of claim 1 wherein the chemical change is formation or loss of a nucleic acid hybrid, a blood clot, or a ligand-receptor interaction, a nucleic acid-protein interaction, a protein-lipid interaction, a protein-carbohydrate interaction, an antibody-antigen interaction.

9. The method of claim 2 wherein the change in viscosity indicates a change in concentration of analyte in the sample.

10. The method of claim 2 wherein the change in viscosity indicates a change in average length of analyte in the sample.

11. The method of claim 1 wherein the change in viscosity is measured in real time.

12. The method of claim 1 wherein the change in viscosity is from about 0.01 centipoise (cP) to about 10 cP.

13. The method of claim 12 wherein the change in viscosity is from about 0.1 cP to about 2 cP.

14. A method of detecting a chemical change in state of an analyte in a sample comprising:

(a) contacting the sample with a magnetic particle;

(b) measuring a first instantaneous non-linear rotation rate of the magnetic particle in the sample at a first time; and

(c) measuring a second instantaneous non-linear rotation rate of the magnetic particle in the sample at a second time;

wherein a difference between the first instantaneous rotation rate relative to the second instantaneous rotation rate indicates a change in viscosity in the sample, which indicates the change in state of the analyte in the sample,

wherein the chemical change in state is catalyzed by an enzyme.

15. The method of claim 14 wherein the analyte is a monomer of a biopolymer.

16. The method of claim 14 wherein the analyte is selected from the group consisting of a cell, a nucleic acid, a protein, a carbohydrate, a lipid, and an amino acid.

17. The method of claim 14 wherein the enzyme is selected from the group consisting of a polymerase, a nuclease, a hydrolase, a lyase, an oxidase, a peptidase, and a ligase.

18. The method of claim 14 wherein the chemical change is formation or loss of a nucleic acid hybrid, a blood clot, or a ligand-receptor interaction, a nucleic acid-protein interaction, a protein-lipid interaction, a protein-carbohydrate interaction, an antibody-antigen interaction.

19. The method of claim 15 wherein the change in viscosity indicates a change in concentration of analyte in the sample.

20. The method of claim 15 wherein the change in viscosity indicates a change in average length of analyte in the sample.

21. The method of claim 14 wherein the change in viscosity is measured in real time.

22. The method of claim 14 wherein the change in viscosity is from about 0.01 centipoise (cP) to about 10 cP.

23. The method of claim 22 wherein the change in viscosity is from about 0.1 cP to about 2 cP.