Methods and compositions for sustained immunotherapy
This disclosure provides methods of making functionalized PEG iron oxide nanoparticles.
1. A method for making peptide MHC (pMHC)-functionalized PEG iron oxide nanoparticles comprising
thermally decomposing iron acetyl acetonate in the presence of functionalized PEG molecules and benzyl ether, to obtain functionalized PEG iron oxide nanoparticles, wherein the thermal decomposition occurs at a temperature from about 80 to about 300° C.; and
providing a pMHC, and
contacting the functionalized PEG iron oxide nanoparticles with the pMHC.
2. The method of claim 1 , wherein the iron oxide nanoparticle is water-soluble.
3. The method of claim 1 , wherein the thermal decomposition comprises a single-step reaction.
4. The method of claim 1 , wherein the temperature for the thermal decomposition is about 80 to about 200° C., or about 80 to about 150° C., or about 100 to about 250° C., or about 100 to about 200° C., or about 150 to about 250° C.
5. The method of claim 1 , wherein the thermal decomposition is carried out for about 1 to about 2 hours.
6. The method of claim 1 , wherein the nanoparticles are stable at about 4° C. in PBS without any detectable degradation or aggregation.
7. The method of claim 6 , wherein the nanoparticles are stable for at least 6 months.
8. The method of claim 1 , wherein the method further comprises purifying the nanoparticles with a magnetic column.
9. The method of claim 1 , wherein the functionalized PEG molecules are maleimide functionalized and the pMHC encodes a cysteine at its carboxy-terminal end.
10. The method of claim 1 , wherein the functionalized PEG molecules are less than about 5 kilodaltons.
11. The method of claim 9 , wherein the maleimide functionalized PEG molecules are methoxy-PEG molecules.
12. The method of claim 1 , wherein the MHC of the pMHC comprises an MHC class I molecule.
13. The method of claim 1 , wherein the MHC of the pMHC comprises an MHC class II molecule.
14. The method of claim 1 wherein the peptide of the pMHC comprises an epitope of an autoantigen involved in an autoimmune response.
15. The method of claim 14 , wherein the autoantigen involved in an autoimmune response is an autoantigen involved in a human autoimmune response.
16. The method of claim 1 , wherein the functionalized PEG iron oxide nanoparticles is less than about 100 nm in diameter.