Biofunctionalized nanoparticles and uses thereof in adoptive cell therapy

Patent metadata

Field	Value
Publication number	US-10588986-B2
Application number	US-201615763566-A
Country	US
Kind code	B2
Filing date	Sep 27, 2016
Priority date	Sep 28, 2015
Publication date	Mar 17, 2020
Grant date	Mar 17, 2020

Abstract

The present invention relates to biofunctionalized nanoparticles and uses thereof in adoptive cell therapy. In particular, the present invention relates to a nanoparticle comprising an amount of at least one antigen and an amount of at least one antibody having specificity for a B cell receptor wherein the antigen and antibody are attached to the surface of the nanoparticle.

First claim

The invention claimed is:

1. A nanoparticle,
comprising a streptavidin coating and an amount of at least one antigen and an amount of at least one antibody having specificity for a human or mouse B cell receptor (BCR),
wherein each of the at least one antigen and at least one antibody are attached to the surface of the nanoparticle via a linker having a biotin moiety and forming a streptavidin-biotin complex with the streptavidin coating, and
wherein one of the at least one antigen is ovalbumin or a trimer HIV-GAG peptide, wherein the trimer HIV-GAG peptide has an amino acid sequence identity as set forth in SEQ ID NO:1.

2. The nanoparticle of claim 1 which is in the form of a sphere, made of an organic polymer and having a size in the range of 100 to 500 nanometers.

3. The nanoparticle of claim 1 wherein another antigen of the at least one antigen is viral antigen.

4. The nanoparticle of claim 1 wherein the at least one antigen is a HLA molecule.

5. The nanoparticle of claim 1 wherein the at least one antibody has specificity for the framework region of a kappa or lambda BCR light chain or for the framework region of a BCR heavy chain and is selected from the group consisting of rat anti-mouse kappa light chain (anti-κ mAb clone 187.1), mouse anti-human k light chain (clone G20-361), lambda light-chain (clone JC5-10) and human lambda light chain (clone JDC-12).

6. The nanoparticle of claim 1 wherein the at least one antibody comprises at least 2 or 3 anti-BCR antibodies, wherein each of said at least 2 or 3 anti-BCR antibodies is monobiotinylated and each monobiotinylated antibody is attached to the surface of the nanoparticle as part of a streptavidin-biotin complex.

7. The nanoparticle of claim 1 , wherein the linker is a sulfo-NHS-LC-LC-biotin linker.

8. The nanoparticle of claim 2 , wherein the sphere has a size in the range of 350-450 nanometers.

9. The nanoparticle of claim 2 , wherein the sphere has a size of 400 nanometers.

10. The nanoparticle of claim 2 , wherein the organic polymer comprises polystyrene.

11. The nanoparticle of claim 10 , wherein the sphere has a size of 400 nanometers.

12. A nanoparticle able to be internalized by B cells, comprising
an organic polymer sphere having a size range of 100 to 500 nanometers;
a streptavidin coating;
a plurality of biotin linkers, wherein each biotin linker forms a streptavidin-biotin complex with the streptavidin coating;
at least one antigen capable of eliciting a T-cell response monobiotinylated to a first biotin linker; and
at least one antibody monobiotinylated to a second biotin linker, wherein said at least one antibody comprises specificity for the framework region of a mouse or human kappa or lambda B cell receptor (BCR) light chain or for the framework region of a mouse or human BCR heavy chain and is selected from the group consisting of rat anti-mouse kappa light chain(anti-κ mAb clone 187.1), mouse anti-human k light chain (clone G20-361), lambda light-chain (clone JC5-10) and human lambda light chain (clone JDC-12).

13. The nanoparticle of claim 12 , wherein each of the plurality of biotin linkers is a sulfo-NHS-LC-LC-Biotin linker.

14. The nanoparticle of claim 12 , wherein the organic polymer comprises polystyrene and the sphere has a size range of 350-450 nanometers.

15. The nanoparticle of claim 12 , wherein the organic polymer comprises polystyrene and the sphere has a size of 400 nanometers.

16. The nanoparticle of claim 12 , wherein said at least one antigen is selected from the group consisting of a protein, peptide nucleic acid, DNA plasmid, a tissue preparation and a cell preparation.

17. The nanoparticle of claim 12 , wherein said at least one antigen comprises an antigen selected from the group consisting of a viral antigen, a bacterial antigen, a fungal antigen, a protozoal antigen, a tumor-associated antigen, an auto-antigen, an allergen, a xenoantigen, an alloantigen and an antigenic molecule that is exogenously administered for therapeutic or other purposes and may trigger an unwanted immune response.

18. The nanoparticle of claim 12 , wherein said at least one antigen is ovalbumin or a triem HIV-GAG peptide, wherein the trimer HIV-GAG peptide has an amino acid sequence identity as set forth in SEQ ID NO: 1.

19. A nanoparticle, comprising
a polystyrene sphere having a size of 400 nanometers;
a streptavidin coating;
a plurality of sulfo-NHS-LC-LC-biotin linkers forming streptavidin-biotin complexes with said streptavidin coating;
an antigen, monobiotinylated to a first sulfo-NHS-LC-LC-biotin linker, wherein the antigen is ovalbumin or a trimer HIV-GAG peptide, wherein the trimer HIV-GAG peptide has an amino acid sequence as set forth in SEQ ID NO:1; and
an antibody, monobiotinylated to a second sulfo-NHS-LC-LC-biotin linker, wherein the antibody is specific for the framework region of a mouse or human kappa or lambda B cell receptor (BCR) light chain or for the framework region of a mouse or human BCR heavy chain and is selected from the group consisting of rat anti-mouse kappa light chain(anti-κ mAb clone 187.1), mouse anti-human k light chain (clone G20-361), lambda light-chain (clone JC5-10) and human lambda light chain (clone JDC-12);
wherein said nanoparticle is able to be internalized by B cells and is suitable for generating antigen-specific T cells.

Assignees

• Inst Nat Sante Rech Med
• Ens—Ecole Normale Superieure De Lyon
• Univ Claude Bernard Lyon
• Centre Nat Rech Scient
• Hospice Civils De Lyon
• Ens Ecole Normale Superieure De Lyon

Inventors

• Sicard Antoine
• Thaunat Olivier

Classifications

• A61K47/6939 Primary

  the polymer being a polysaccharide, e.g. starch, chitosan, chitin, cellulose or pectin · CPC title

• A61K2039/55555

  Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N2740/16034

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• A61K2039/625

  binding through the biotin-streptavidin system or similar · CPC title