Nanoallergens and uses thereof
Embodiments of the present disclosure provide a nanoparticle based platform, and nanoallergens for identifying, evaluating and studying allergen mimotopes as multiple copies of a single mimotope or various combinations on the same particle. The nanoparticle is extremely versatile and allows multivalent binding to IgEs specific to a variety of mimotopes, simulating allergen proteins. Nanoparticles can include various molecular ratios of components. For example, the nanoallergens can include about 0.1-40% mimotope-lipid conjugate and about 60-99.9% lipid. The mimotope-lipid conjugate includes a mimotope, a first linker, and lipid molecule. Nanoallergens can be used in in vitro and in vivo applications to identify a specific patient's sensitivity to a set of epitopes and predict a symptomatic clinical response, identify allergen epitopes through blind screening peptide sequences from allergen protein, and in a clinical application similar to a scratch test.
1. A liposomal nanoparticle comprising:
about 0.1 mol % to about 20 mol % of a synmimotope-lipid conjugate;
about 2 mol % to about 10 mol % of a polyethylene glycol-lipid (PEG-lipid) conjugate; and
about 80 mol % to about 97 mol % of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC);
wherein
the nanoparticle comprises a spherical lipid bilayer comprising the phospholipid and the synmimotope-lipid conjugate, the spherical lipid bilayer having an interior surface and an exterior surface;
the exterior surface of the spherical lipid bilayer comprises the PEG-lipid conjugate wherein PEG moieties of the PEG-lipid conjugate form a coating over the exterior surface of the nanoparticle, and one or more synmimotope moieties of synmimotope-lipid conjugates protrude above the coating formed by the PEG moieties;
wherein the synmimotope-lipid conjugate comprises a conjugate of Formula I:
A-B-C-D-E-(F)n (I)
wherein
A is a synmimotope moiety of the synmimotope-lipid conjugate, wherein the synmimotope moiety is a hapten moiety comprising 2,4-dinitrophenol (DNP);
B is a first linker, wherein the first linker comprises two ethylene glycol moieties covalently attached to the synmimotope moiety (A) and second linker (C) by amide bonds; and
C is the second linker, wherein the second linker (C) comprises three lysine amino acids covalently attached by amide bonds to the first linker (B); and
D is a third linker, wherein the third linker (D) comprises three ethylene glycol 6 moieties attached covalently at distal ends by amide bonds to the second linker (C) and tag (E);
E is the tag, wherein the tag comprises a tryptophan residue;
F is a palmitoyl moiety covalently attached to the tag (E) by an amide bond; and
n is 2;
wherein the diameter of the nanoparticle is about 20 nm to about 2 μm.
2. The nanoparticle of claim 1 wherein the PEG-lipid conjugate comprises about 10 to about 200 ethylene glycol residues and the lipid is a (C 8 -C 22 )acyl moiety or a phospholipid, wherein the ethylene glycol residues and the lipid are optionally linked by an amide bond.
3. The nanoparticle of claim 1 wherein the nanoparticle has a diameter of about 20 nm to about 300 nm.
4. The nanoparticle of claim 1 wherein the nanoparticle further comprises a plurality of allergen epitopes protruding above the coating formed by the PEG moieties, wherein the plurality of allergen epitopes is homogeneous or heterogeneous.
5. The nanoparticle of claim 1 wherein the nanoparticle comprises:
about 2 mol % synmimotope-lipid conjugate;
about 5 mol % PEG-lipid conjugate; and
about 93 mol % 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).
6. The nanoparticle of claim 1 further comprising about 1 mol % to about 35 mol % cholesterol, with respect to the molar amount of the 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).
7. A liposomal nanoparticle comprising:
about 0.1 mol % to about 20 mol % of a synmimotope-lipid conjugate;
about 2 mol % to about 10 mol % of a polyethylene glycol-lipid (PEG-lipid) conjugate; and
about 80 mol % to about 97 mol % of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC);
wherein
the nanoparticle comprises a spherical lipid bilayer comprising the phospholipid and the synmimotope-lipid conjugate, the spherical lipid bilayer having an interior surface and an exterior surface;
the exterior surface of the spherical lipid bilayer comprises the PEG-lipid conjugate wherein PEG moieties of the PEG-lipid conjugate form a coating over the exterior surface of the nanoparticle, and one or more synmimotope moieties of synmimotope-lipid conjugates protrude above the coating formed by the PEG moieties, wherein the one or more synmimotope moieties protruding above the coating are homogeneous;
wherein the synmimotope-lipid conjugate comprises a conjugate of Formula I:
A-B-C-D-E-(F)n (I)
wherein
A is a synmimotope moiety of the synmimotope-lipid conjugate, wherein the synmimotope moiety is a hapten moiety comprising 2,4-dinitrophenol (DNP);
B is a first linker, wherein the first linker comprises two ethylene glycol moieties covalently attached to the synmimotope moiety (A) and second linker (C) by amide bonds; and
C is the second linker, wherein the second linker (C) comprises three lysine amino acids covalently attached by amide bonds to the first linker (B); and
D is a third linker, wherein the third linker (D) comprises three ethylene glycol 6 moieties attached covalently at distal ends by amide bonds to the second linker (C) and tag (E);
E is the tag, wherein the tag comprises a tryptophan residue;
F is a palmitoyl moiety covalently attached to the tag (E) by an amide bond; and
n is 2;
wherein the diameter of the nanoparticle is about 20 nm to about 300 nm.