Method of determining lipoprotein concentration in solution using light scattering

Patent metadata

Field	Value
Publication number	US-2020378891-A1
Application number	US-201816769918-A
Country	US
Kind code	A1
Filing date	Dec 4, 2018
Priority date	Dec 4, 2017
Publication date	Dec 3, 2020
Grant date	—

Abstract

The invention relates the use of single particle light scattering, preferably interferometric scattering microscopy (also referred to herein as iSCAT), to measure the concentration of a particle in a solution. The invention furthermore relates to the use of light scattering to detect lipoprotein particles in a sample, and to related diagnostic and treatment methods.

First claim

1 . A method of measuring the concentration of a particle in solution, comprising contacting the solution with a surface and detecting the binding of said particle to the surface using light scattering.

2 . A method as claimed in claim 1 , wherein said surface is passivated, activated, coated, treated or derivatised.

3 . A method as claimed in claim 1 , wherein the detection binding of the particle to the surface is repeated after one or more time intervals.

4 . A method as claimed in claim 3 , wherein the repeated measurements permit calculation of the initial rate of binding of the particle to the surface, and optionally the initial binding rate of the particle is compared to the initial binding rate for a known concentration of the particle.

5 . (canceled)

6 . A method as claimed in claim 1 wherein the measurements permit calculation of the constant binding rate of the particle, and optionally this is compared to the constant binding rate for a known concentration of the particle.

7 . A method as claimed in claim 1 , for measuring the concentration of particle a solution, said method comprising:
i) contacting the solution with a surface; ii) detecting the particle binding to the surface as visualised by light scattering; iii) repeating the detection step and calculating the changes in binding rate and/or initial binding rate over time of the particle to the surface; iv) providing a calibration curve of initial binding rate against concentration based on data from solutions of known particle concentration; v) using the calibration curve in step (iv) to convert the initial binding rate recorded for the sample in step (iii) to the concentration of the particle in solution.

8 . The method as claimed in claim 1 , wherein the concentration is absolute concentration.

9 . The method as claimed in claim 1 , wherein said particle is also contacted with a measuring solution.

10 . The method as claimed in claim 1 , wherein said solution is contacted with said surface in the presence of a calibrant and the binding of said calibrant to said surface is detected using light scattering.

11 . A method for detection of lipoprotein particles in a sample, comprising detecting said particles by light scattering, optionally interferometric scattering microscopy, optionally wherein said detecting comprises determining the size and/or number of particles.

12 . (canceled)

13 . The method of claim 11 , comprising detecting the size and/or number of more than one different type of lipoprotein particle in said sample, and optionally detecting the ratio of different types of lipoprotein particle in said sample.

14 . The method of claim 11 , comprising detecting high density lipoprotein (HDL) particles and/or low density lipoprotein (LDL) particles and/or determining the ratio of HDL to LDL particles in said sample.

15 . (canceled)

16 . A method for diagnosing a disease or condition associated with size and/or number of lipoprotein particles in an individual, or for determining the risk that the individual will develop said disease or condition, comprising detecting the size and/or number of lipoprotein particles in a sample from said individual by light scattering, optionally by interferometric scattering microscopy, wherein the disease is optionally cardiovascular disease.

17 . The method of claim 16 , comprising detecting the size and/or number of more than one different type of lipoprotein particle, or the ratio of different types of lipoprotein particle, in said sample.

18 . The method of claim 16 , comprising detecting high density lipoprotein (HDL) particles and/or low density lipoprotein (LDL) particles, optionally comprising determining the ratio of HDL to LDL particles in said sample.

19 . The method of claim 17 , wherein a decreased number of HDL particles as compared with a control sample or reference sample/level, is indicative that the individual has said disease or condition, or has an increased risk of developing said disease or condition.

20 . The method of claim 17 , wherein an increased number and/or a decreased size of LDL particles as compared with a control sample or reference sample/level, is indicative that the individual has said disease or condition, or has an increased risk of developing said disease or condition.

21 . (canceled)

22 . The method of claim 1 , wherein said solution or sample is a biological sample, optionally obtained from a human individual, wherein optionally said biological sample is blood, plasma, or serum, which is optionally obtained from a human individual.

23 . (canceled)

24 . A method of selecting an individual to whom a substance or composition is to be administered, or to whom a regimen is to be prescribed, wherein said substance or composition or regimen is suitable for treating or preventing a disease or condition associated with size and/or number of lipoprotein particles, comprising detecting the size and/or number of lipoprotein particles in a sample from said individual by a method according to claim 11 , and selecting said patient for said administration or said regimen if the size and/or number of lipoprotein particles detected is indicative of the presence of said disease or condition, or of a risk thereof.

25 . A method of treating or preventing a disease or condition associated with size and/or number of lipoprotein particles in an individual, the method comprising diagnosing or determining risk of said disease or condition in said individual or selecting said individual by a method according to claim 16 , and administering a substance or composition to the individual, or carrying out a regimen thereon, which is effective to treat or prevent said disease or condition in the individual.

26 - 27 . (canceled)

Assignees

• Univ Oxford Innovation Ltd

Inventors

• Andrecka Joanna
• Kukura Philipp
• Young Gavin
• Benesch Justin

Classifications

• G01N2021/513

  Cuvettes for scattering measurements · CPC title

• G01N2015/1454

  using phase shift or interference, e.g. for improving contrast · CPC title

• G01N2015/1021

  Measuring mass of individual particles · CPC title

• G01N33/92 Primary

  involving lipids, e.g. cholesterol {, lipoproteins, or their receptors (steroid hormones G01N33/743)} · CPC title

• G01N33/6887

  from muscle, cartilage or connective tissue · CPC title