Methods of making lyophilized compositions comprising albumin-VEGF paclitaxel nanoparticle complexes
Described herein are compositions of antibodies and carrier proteins and methods of making and using the same, in particular, as a cancer therapeutic. Also described are lyophilized compositions of antibodies and carrier proteins and methods of making and using the same, in particular, as a cancer therapeutic.
1. A method of making a lyophilized nanoparticle composition comprising nanoparticle complexes comprising albumin, antibodies having a VEGF-binding portion, and paclitaxel, the method comprising mixing aqueous albumin-paclitaxel nanoparticles with the antibodies in vitro under conditions to form the nanoparticle complexes, and lyophilizing the nanoparticle complexes to form the lyophilized nanoparticle composition, such that when reconstituted with an aqueous solution the nanoparticle complexes have binding specificity for VEGF, wherein said complexes have an average size of less than 1 μm, and wherein the antibodies are associated with the albumin through non-covalent bonds.
2. The method of claim 1 , wherein said nanoparticle complexes have an average size between about 130 nm and about 800 nm.
3. The method of claim 1 , wherein the ratio of albumin-paclitaxel nanoparticle to antibodies is between 10:1 and 10:30.
4. The method of claim 1 , wherein the ratio of albumin-paclitaxel nanoparticle to antibodies is between 10:2 and 10:6.
5. The method of claim 1 , wherein the antibodies are bevacizumab antibodies.
6. The method of claim 5 , wherein said nanoparticle complexes have an average size between about 130 nm and about 800 nm.
7. The method of claim 5 , wherein the ratio of albumin-paclitaxel nanoparticle to antibodies is between 10:1 and 10:30.
8. The method of claim 5 , wherein the ratio of albumin-paclitaxel nanoparticle to antibodies is between 10:2 and 10:6.
9. The method of claim 1 , wherein the paclitaxel is associated with the albumin through non -covalent bonds.
10. The method of claim 5 , wherein the paclitaxel is associated with the albumin through non-covalent bonds.
11. The method of claim 1 , wherein lyophilized nanoparticle composition is stable at about 20° C. to about 25° C. for at least 3 months.
12. The method of claim 1 , wherein each of the nanoparticle complexes comprises between about 100 and about 1000 antibodies.
13. The method of claim 1 , wherein each of the nanoparticle complexes comprises between about 400 and about 800 antibodies.
14. The method of claim 1 , wherein the nanoparticle complexes have a dissociation constant between about 1×10 −11 M and about 1×10 −9 M.
15. The method of claim 1 , wherein the aqueous albumin-paclitaxel nanoparticles are mixed with the antibodies in a solution having a pH of between about 4 and about 8.
16. The method of claim 1 , wherein the aqueous albumin-paclitaxel nanoparticles are incubated with the antibodies at a temperature of about 5° C. to about 60° C.
17. The method of claim 1 , wherein the nanoparticle complexes retain their, size distribution upon lyophilization.