Methods of measuring structural and functional changes of a biomolecular composition

Patent metadata

Field	Value
Publication number	US-11221287-B2
Application number	US-201715846430-A
Country	US
Kind code	B2
Filing date	Dec 19, 2017
Priority date	Dec 19, 2016
Publication date	Jan 11, 2022
Grant date	Jan 11, 2022

Abstract

This invention relates to a method for measuring viscosity of a fluid using particle diffusometry (PD). The method finds practical applications in detecting structural and functional changes of a biomolecular composition by comparing the viscosity change as compared with the standard of the biomolecular composition. This method may also find uses in clinical diagnosis and quality control of clinical biological medicines, food and feeds during the process of manufacturing, distribution and consumption.

First claim

What is claimed is:

1. A method for measuring viscosity or a change in viscosity of a liquid comprising biomolecules, the method comprising:
preparing a testing solution that contains the liquid and uniformly sized particles having a uniform size within a range of about 50 to about 1,000 nm in diameter;
capturing a plurality of images of the particles in the testing solution over a period of time;
identifying interrogation regions within each of the plurality of images;
correlating the interrogation regions of a sequential pair of first and second images of the plurality of images (cross-correlation) and correlating the interrogation regions of the first image with itself (autocorrelation);
determining a diffusion coefficient of the particles based on the cross-correlation and the autocorrelation; and
calculating a viscosity of the liquid from the diffusion coefficient with Einstein's diffusion equation.

2. The method of claim 1 , wherein the uniformly sized particles are made of structurally stable materials.

3. The method of claim 1 , wherein the biomolecules are selected from the group consisting of nucleic acid, amino acid, lipid, peptide, protein, antibody, enzyme, carbohydrate, DNA, RNA, polysaccharide, oligonucleotide, oligosaccharide, proteoglycans, and glycoprotein.

4. The method of claim 3 , wherein the biomolecules are biological therapeutics.

5. The method of claim 1 , wherein the liquid is a biomolecular composition and the method is used for measuring structural and functional changes of the biomolecular composition from a standard of the biomolecular composition, the method comprising:
measuring viscosity of the biomolecular composition;
measuring viscosity of the standard of the biomolecular composition; and
comparing viscosity of the biomolecular composition with that of the standard of the biomolecular composition, wherein finding of a substantial difference suggests structural and functional changes of the biomolecular composition.

6. A method for measuring structural and functional changes of a biomolecular composition from a standard of the biomolecular composition, the method comprising:
preparing a testing solution using the biomolecular composition with added uniformly sized particles having a uniform size within a range of about 50 to about 1,000 nm in diameter;
capturing a plurality of images of the particles of the testing solution over a period of time;
identifying interrogation regions within each of the plurality of images;
correlating the interrogation regions of a sequential pair of first and second images of the plurality of images (cross-correlation) and correlating the interrogation regions of the first image with itself (autocorrelation);
determining a diffusion coefficient based on the cross-correlation and the autocorrelation;
calculating a viscosity of the biomolecular composition from the diffusion coefficient with Einstein's diffusion equation;
obtaining a viscosity of the standard of the biomolecular composition by repeating the preparing, the capturing, the identifying, the correlating, the determining, and the calculating steps; and
comparing viscosity of the biomolecular composition and that of the standard of the biomolecular composition, wherein finding of a substantial difference suggests structural and functional changes of the biomolecular composition.

7. The method of claim 6 , wherein the biomolecular composition comprises a biomolecule selected from the group consisting of nucleic acid, amino acid, lipid, peptide, protein, antibody, enzyme, carbohydrate, DNA, RNA, polysaccharide, oligonucleotide, oligosaccharide, proteoglycans, and glycoprotein.

8. The method of claim 6 , wherein the biomolecular composition comprises a combination of two or more of biomolecules selected from the group consisting of nucleic acid, amino acid, lipid, peptide, protein, antibody, enzyme, carbohydrate, DNA, RNA, polysaccharide, oligonucleotide, oligosaccharide, proteoglycans, and glycoprotein.

9. The method of claim 8 , wherein the biomolecular composition is prepared by combining individual components.

10. The method of claim 6 , wherein the biomolecular composition is derived from bodily fluids, cell cultures, environmental samples, air samples, water samples, soil samples, or other matrices that contain biomolecules.

11. The method of claim 6 , wherein the biomolecular composition is derived from a living organism including prokaryotic cells, eukaryotic cells, viruses, or prions.

12. The method of claim 6 , wherein the biomolecular composition is a therapeutic for treatment of a disease.

13. The method of claim 12 , wherein the biomolecular composition is an insulin formulation.

14. The method of claim 12 , wherein the biomolecular composition is an enzyme or an antibody formulation.

15. The method of claim 12 , wherein the biomolecular composition is a peptide, protein or glycoprotein formulation.

16. A method for detecting presence of bacterial, viral, protozoa, fungal, or other parasitic contamination of a liquid, the method comprising:
preparing a testing solution using the liquid with added uniformly sized particles having a uniform size within a range of about 50 to about 1,000 nm in diameter;
capturing a plurality of images of the micro particles of the testing solution over a period of time;
identifying interrogation regions within each of the plurality of images;
correlating the interrogation regions of a sequential pair of first and second images of the plurality of images (cross-correlation) and correlating the interrogation regions of the first image with itself (autocorrelation);
determining a diffusion coefficient of the particles based on the cross-correlation and the autocorrelation;
calculating a viscosity of the liquid from the diffusion coefficient with Einstein's diffusion equation;
obtaining a viscosity of a non-contaminated standard of the liquid by repeating the preparing, the capturing, the identifying, the correlating, the determining, and the calculating steps; and
comparing the viscosities of the liquid and the non-contaminated standard of the liquid, wherein finding of a substantial difference suggests presence of bacterial, viral, protozoa, fungal, or other parasitic contamination of the liquid.

17. The method of claim 16 , wherein the liquid is for human or animal consumption.

18. The method of claim 16 , wherein the liquid is selected from the group consisting of biological medicine, water, waste water of any source, fruit juice, vegetable juice, liquid food, and a liquid waste from a food or feed processing.

19. The method of claim 16 , wherein the method is used for quality control of biological medicines, food and feeds during the process of manufacturing, distribution and consumption.

20. The method of claim 19 , wherein the biological medicine is a biotherapeutic formulation.

Assignees

• Purdue Research Foundation

Inventors

• Wereley Steven Truitt
• Kinzer-Ursem Tamara Lea
• Clayton Katherine Noel
• Linnes Jacqueline Callihan
• Lee Donghoon
• Moehling Taylor Jo

Classifications

• G01N13/00 Primary

  Investigating surface or boundary effects, e.g. wetting power; Investigating diffusion effects; Analysing materials by determining surface, boundary, or diffusion effects (scanning-probe techniques or apparatus G01Q) · CPC title

• G01N2013/003

  Diffusion; diffusivity between liquids · CPC title

• G01N2011/006

  Determining flow properties indirectly by measuring other parameters of the system · CPC title

• G01N2015/0053

  in liquids, e.g. trouble · CPC title

• G01N2011/008

  optical properties · CPC title