Enhanced targeting platform
A platform technology provides particle and nucleic acid conjugates, and compositions thereof, with enhanced targeting to cells, tissues, organs. The particles and nucleic acids and other deliverables contain a synthetic binding protein such as a polypeptide monobody covalently conjugated to the surface of the particle or the nucleic acid, for linking a targeting agent to the particle's surface or the nucleic acid. The particles and nucleic acids and other deliverables optionally contain an antibody non-covalently conjugated to the binding protein, via an Fc domain of the antibody. The particles can include therapeutic agents, diagnostic agents, prophylactic agents, or a combination thereof, to be delivered to desired cells, tissues, and/or organs. The particles and nucleic acids and other deliverables can be used in a wide array of applications including, but not limited to, ex vivo perfusion of mammalian organs and in vivo disease treatment.
1. A composition comprising a particle functionalized with one or more monobodies, wherein the one or more monobodies comprises:
(i) a first surface comprising a targeting agent binding site capable of binding to an Fc domain of an antibody, and
(ii) a second surface comprising a chemical moiety through which the one or more monobodies is conjugated to the particle's surface,
wherein the one or more monobodies each comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, or a variant thereof comprising at least 90% sequence identity to SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4,
with or without the N-terminal tag, the C-terminal tag, or both.
2. The composition of claim 1 , further comprising an antibody bound to the targeting agent binding site of the one or more monobodies via the Fc domain of the antibody.
3. The composition of claim 1 , wherein the one or more monobodies each comprises an immunoglobulin fold and no disulfide bonds.
4. The composition of claim 1 , wherein the conjugation of the one or more monobodies to the particle's surface comprises a covalent linkage, wherein the covalent linkage comprises the structure:
—X—Ra—Y— Formula I
wherein,
X and Y, independently, contain between 3 and 90 atoms, inclusive, between 3 and 85 atoms, inclusive, between 3 and 80 atoms, inclusive, between 3 and 70 atoms, inclusive, between 3 and 60 atoms, inclusive, between 3 and 50 atoms, inclusive, between 3 and 40 atoms, inclusive, between 3 and 30 atoms, inclusive, or between 3 and 20 atoms, inclusive, and
Ra comprises a 3-thiopyrrolidine-2,5-dione moiety, 3-aminopyrrolidine-2,5-dione moiety, 3-thiomaleimide moiety, 3-aminomaleimide moiety, a triazole moiety, a carbamate, oxime ether, hydrazone, a carbonyl, imine, sulfonamide, azo, dialkyl dialkoxysilane, diaryl dialkoxysilane, orthoester, acetal, aconityl, β-thiopropionate, phosphoramidate, trityl, vinyl ether, polyketal, or a combination thereof.
5. The composition of claim 1 , wherein the chemical moiety is an amino acid selected from the group consisting of cysteine, lysine, ornithine, arginine, serine, threonine, tyrosine, and a combination thereof.
6. The composition of claim 1 , wherein the chemical moiety is a cysteine.
7. The composition of claim 1 , wherein the particle is an expression vector.
8. The composition of claim 1 , further comprises a pharmaceutically acceptable carrier.
9. A method of delivering a particle to a subject in need thereof comprising
(i) administering to the subject, or
(ii) administering to an organ, tissue, or cell to be transplanted to the subject,
an effective amount of the composition of claim 1 and an antibody.
10. The method of claim 9 , wherein the antibody is administered prior to or co-administered with administering the composition.
11. The method of claim 9 , wherein administration to the organ occurs ex vivo.
12. The composition claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, with or without the N-terminal tag, the C-terminal tag, or both.
13. The composition of claim 1 , wherein the particle is a polymeric particle, a liposome, or an inorganic particle.
14. The composition of claim 1 , wherein the particle comprises one or more therapeutic agents, diagnostic agents, prophylactic agents, or a combination thereof.
15. The composition of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, without the N-terminal tag.
16. The composition of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4.