Immunomagnetic compositions for the ph-specific capture of extracellular vesicles

Patent metadata

Field	Value
Publication number	US-2023272120-A1
Application number	US-202117998177-A
Country	US
Kind code	A1
Filing date	May 10, 2021
Priority date	May 8, 2020
Publication date	Aug 31, 2023
Grant date	—

Abstract

The present disclosure provides immunomagnetic compositions and their methods of use, in particular magnetic particles conjugated to peptides that bind and capture extracellular vesicles.

First claim

1 . An immunomagnetic composition comprising a population of magnetic particles, wherein each magnetic particle is conjugated to at least one pH responsive extracellular vesicle-binding peptide.

2 .- 4 . (canceled)

5 . The immunomagnetic composition of claim 1 , wherein the population of magnetic particles has an average particle size ranging from about 1 nm to about 100 microns.

6 . (canceled)

7 . The immunomagnetic composition of claim 1 , wherein the magnetic particle comprises a magnetic element selected from iron, nickel, and cobalt, or oxide compounds thereof.

8 . The immunomagnetic composition of claim 1 , wherein a graphene oxide nanomaterial is covalently bound to a surface of the magnetic particle.

9 . (canceled)

10 . The immunomagnetic composition of claim 1 , wherein the peptide is conjugated or covalently bound to a surface of the magnetic particle.

11 .- 13 . (canceled)

14 . The immunomagnetic composition of claim 1 , wherein the magnetic particle comprises one or more streptavidin groups.

15 . The immunomagnetic composition of claim 14 , wherein the peptide comprises a biotinylated residue, and wherein the biotinylated residue is bound to one streptavidin group.

16 . The immunomagnetic composition of claim 1 , wherein the peptide comprises an amino acid sequence having at least 65%80% identity with an amino acid sequence selected from SEQ ID NO: 1 to 21.

17 .- 19 . (canceled)

20 . The immunomagnetic composition of claim 1 , wherein the peptide comprises an amino acid sequence selected from SEQ ID NO: 1 to 21.

21 . The immunomagnetic composition of claim 1 , wherein the peptide consists of an amino acid sequence having at least 80% identity with an amino acid sequence selected from SEQ ID NO: 1 to 21.

22 .- 24 . (canceled)

25 . The immunomagnetic composition of claim 1 , wherein the peptide consists of an amino acid sequence selected from SEQ ID NO: 1 to 21.

26 . A method for isolating a population of extracellular vesicles from a medium the method comprising:
contacting the medium with an immunomagnetic composition of claim 1 and optionally an aqueous solution to form a mixture; adjusting the pH of the mixture to within a range of about 3 to about 5 to bind the population of magnetic particles to the population of extracellular vesicles; and collecting the population of extracellular vesicles bound to the magnetic particles by applying a magnetic field.

27 . The method of claim 26 , wherein the extracellular vesicles comprise ectosomes, microvesicles (MV), microparticles (MP), exosomes, apoptotic bodies, large oncosomes, exophers, enveloped viruses, and exomeres.

28 .- 29 . (canceled)

30 . The method of claim 26 , wherein the biological fluid comprises cell culture medium, plasma, serum, urine, saliva, tears, perilymph fluid, milk, cerebrospinal fluid, blood, or a plant derived fluid.

31 . The method of claim 26 , further comprising contacting the population of extracellular vesicles bound to the population of magnetic particles with an agent such that the agent is encapsulated within the extracellular vesicles.

32 . The method of claim 31 , wherein the agent comprises a therapeutic agent, an anti-cancer agent, a vaccine, an immunotherapy agent, or a regenerative therapy agent.

33 .- 36 . (canceled)

37 . The method of claim 26 , further comprising contacting the population of extracellular vesicles bound to the population of magnetic particles with a surface modifying agent.

38 .- 40 . (canceled)

41 . The method of claim 39 , wherein the component of the lipid bilayer comprises a membrane protein.

42 . The method of claim 26 , further comprising releasing the population of extracellular vesicles from the magnetic particles by adjusting the pH within a pH range of about 7 to about 9.

43 . A population of extracellular vesicles isolated by the method of claim 26 .

44 . A pharmaceutical composition comprising the population of extracellular vesicles of claim 43 and a pharmaceutically acceptable excipient.

45 . A kit comprising:
an immunomagnetic composition of claim 1 ; and an aqueous solution for dispersing the immunomagnetic composition.

Assignees

• Univ Kansas

Inventors

• He Mei

Classifications

• C07K17/06 Primary

  attached to the carrier via a bridging agent · CPC title

• A61K9/127

  Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant · CPC title

• A61K35/28 Primary

  Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells · CPC title

• B03C2201/18

  Magnetic separation whereby the particles are suspended in a liquid · CPC title

• B03C2201/26

  for use in medical or biological applications · CPC title