Capture of pathogenic and non-pathogenic biopolymers and bioparticles

• US 9,260,536 B2
• Filed: 07/20/2012
• Issued: 02/16/2016
• Est. Priority Date: 07/22/2011
• Status: Active Grant

• Current Assignee: Stellan Hjerten
• Original Assignee: Stellan Hjerten
• Inventors: Hjerten, Stellan
• Primary Examiner(s): Skowronek, Karlheinz R
• Assistant Examiner(s): TELLER, ROY R
• Application Number:
  US14/233,491
• Publication Number:
  US 20140178439A1
• Time in Patent Office: 1,306 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/729 Agar; Agarose; Agaropectin
• CPC Class Codes: A61K 31/765 Polymers containing oxygen
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: B01D 15/327 with hydrophobic interaction
• CPC Class Codes: B01J 20/287 Non-polar phases; Reversed phases
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3212 consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3255 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
• CPC Class Codes: B01J 20/3274 Proteins, nucleic acids, polysaccharides, antibodies or antigens
• CPC Class Codes: C08B 37/12 Agar-agar; Derivatives thereof not used
• CPC Class Codes: G01N 2030/8813 biological materials
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 33/6803 General methods of protein analysis not limited to specific proteins or families of proteins

Abstract

A method of preparing a capturing agent for a selected biopolymer or bioparticle, includes the steps of:

• a) selecting a desired biopolymer or bioparticle, and
• b) providing a support matrix modified to have, at predetermined conditions, such a net negative surface charge and net surface hydrophobicity in relation to the net negative surface charge and the net surface hydrophobicity of the selected biopolymer or bioparticle that the modified support matrix is capable of selective interaction and capture of the biopolymer or bioparticle through a resulting net hydrophobic interaction being the actual hydrophobic interaction minus the actual electrostatic repulsion. The use of such a capturing agent as a therapeutically active substance, as well as in chromatography and diagnosis are also disclosed.

Claims

1. A method of preparing a capturing agent for a selected biopolymer or bioparticle, comprising the steps of:

a) selecting a desired biopolymer or bioparticle, and

b) providing a support matrix modified to have, at predetermined conditions, such a net negative surface charge and net surface hydrophobicity in relation to the net negative surface charge and the net surface hydrophobicity of the selected biopolymer or bioparticle that the modified support matrix is capable of selective interaction and capture of the biopolymer or bioparticle through a resulting net hydrophobic interaction being the actual hydrophobic interaction minus the actual electrostatic repulsion.

2. The method according to claim 1, wherein at least one of the net surface charge of the modified support matrix and the net surface hydrophobicity of the support matrix is in the range of from about 20% lower to about 20% higher than that of the selected biopolymer or bioparticle.

3. The method according to claim 1, wherein providing the modified support matrix comprises screening support matrices for selective interaction with the selected biopolymer or bioparticle by varying at least one of the parameters net surface charge and net surface hydrophobicity of the support matrices.

4. The method according to claim 3, which comprises keeping the net surface charge constant and varying the net surface hydrophobicity.

5. The method according to claim 3, which comprises keeping the net surface hydrophobicity constant and varying the net surface charge.

6. The method according to claim 3, which comprises varying the net surface hydrophobicity of an at least substantially non-charged support matrix to select a support matrix having a hydrophobicity such that the selected biopolymer or bioparticle weakly binds thereto, adding a low negative net surface charge to the support matrix, and, optionally, adding a small amount of additional hydrophobicity to the support matrix.

7. The method according to claim 1, wherein providing the modified support matrix comprises the step of measuring at least one of the parameters net surface charge and net surface hydrophobicity of the selected biopolymer or bioparticle.

8. The method according to claim 7, wherein the net surface charge is measured by free zone electrophoresis, and the net surface hydrophobicity is measured by hydrophobic-interaction chromatography or by a precipitation technique.

9. The method according to claim 1, further comprising the step of isolating the selected biopolymer or bioparticle from a biological fluid.

10. The method according to claim 9, wherein the step of isolating the selected biopolymer or bioparticle comprises:

a) purification by anion-exchange chromatography or zone electrophoresis; and

b) hydrophobic-interaction chromatography.

11. The method according to claim 1, wherein the selected biopolymer is a protein or a polypeptide.

12. The method according to claim 1, wherein the selected biopolymer or bioparticle is a pathogen.

13. The method according to claim 1, wherein the biopolymer or bioparticle has a biological activity, and said predetermined conditions include pH and ionic strength values substantially corresponding to those of the milieu where the biopolymer or bioparticle exhibits its biological activity.

14. The method according to claim 1, wherein the modified support matrix comprises an inert non-charged hydrophilic matrix supporting (i) hydrophilic groups or ligands which are negatively charged at said predetermined conditions, and (ii) hydrophobic groups or ligands.

15. The method according to claim 14, wherein the hydrophilic matrix is mixed with at least one vegetable extract, preferably selected from fucoindans, alginates, laminarins, and polyphenols, which are known to activate the immune system.

16. The method according to claim 14, wherein the hydrophilic support matrix comprises a polymer or a particle selected from agarose, dextran, cellulose, starch, guar gum, polyacrylamide, polyvinylalcohol, and coated silica beads.

17. The method according to claim 1, wherein the modified support matrix comprises a protein or polypeptide.

18. The method according to claim 1, wherein the modified support matrix comprises a bioparticle or a nanoparticle.

19. The method according to claim 14, which comprises the sequential steps of providing the inert non-charged hydrophilic matrix, coupling hydrophilic, negatively charged ligands to the matrix, and coupling hydrophobic ligands to the matrix.

20. The method according to claim 19, wherein the negatively charged ligands and the hydrophobic ligands comprise molecular chains of substantially the same length.

21. The method according to claim 14, wherein the modified support matrix further comprises at least one group or ligand capable of selectively binding to the biopolymer or bioparticle.

22. The method according to claim 21, wherein the selectively binding group or ligand comprises a carbohydrate binding group or a carbohydrate-interacting group in lectins.

28. A method of treating a subject, which comprises extracorporeal treatment of the subject'"'"'s blood with a capturing agent obtainable by the method according to claim 1.

23. A therapeutically active substance, comprising a support matrix modified to have, at predetermined conditions, such a net negative surface charge and net surface hydrophobicity in relation to the net negative surface charge and net surface hydrophobicity of a selected biopolymer or bioparticle that the modified support matrix is capable of selective interaction with and capture of the biopolymer of bioparticle through the net interaction created by the net hydrophobic interaction minus the net electrostatic repulsion.

24. The therapeutically active substance according to claim 23, which is selected from an antibiotic;

an antiviral agent;

an antifungal agent;

an anti-malarial agent; and

an anticancer agent.

25. A pharmaceutical composition, comprising a therapeutically active substance according to claim 23.

27. A method of treating a subject, which comprises administering to the subject a therapeutically active substance according to claim 23.

26. An antimicrobial composition, comprising a support matrix modified to have, at predetermined conditions, such a net negative surface charge and net surface hydrophobicity in relation to the net negative surface charge and the net surface hydrophobicity of a selected biopolymer or bioparticle, including microorganisms, that the support matrix is capable of selective interaction with and capture of the biopolymer or bioparticle through the net interaction created by the net hydrophobic interaction minus the net electrostatic repulsion.