Self-assembling graphene oxide-protein matrix
The present invention relates to a stable self-assembling graphene oxide-protein matrix comprising a disordered protein (DP) and graphene oxide (GO), wherein the DP has an opposite charge to the GO, further wherein the graphene oxide-protein matrix is in the form of a 3D structure having a lumen defined by a membrane having an inner and outer surface. The invention further relates to methods and kits for preparing such a graphene oxide-protein matrix and its uses.
1 . A graphene oxide-protein matrix comprising a disordered protein (DP) and graphene oxide (GO), wherein the DP has an opposite charge to the GO, further wherein the graphene oxide-protein matrix is in the form of a 3D structure having a lumen defined by a membrane having an inner and outer surface wherein the membrane comprises the graphene oxide protein matrix and at least partially surrounds the lumen.
2 . The graphene oxide-protein matrix of claim 1 , wherein the 3D structure is
a. multilamellar
b. a tube; or
c. a membrane, a multi-lamella tube, a tubular network, a sphere, a cavity, a sac, or a vesicle optionally wherein the 3D structure:
d. comprises bifurcations;
e. is biocompatible; and/or
f. is permeable.
3 . The graphene oxide-protein matrix of claim 1 , wherein;
a. the internal diameter of the lumen of 3D structure is at least 10 pm; and/or
b. the membrane of the 3D structure is from about 5 pm to about 50 pm thick; and/or
c. wherein the lumen comprises one or more apertures.
4 . The graphene oxide-protein matrix of claim 1 , wherein the DP is positively-charged and the GO is negatively charged.
5 . The graphene oxide-protein matrix of claim 1 , wherein the disordered protein comprises a polypeptide having a repeating amino acid sequence motif.
6 . The graphene oxide-protein matrix of claim 1 , wherein the disordered protein is;
a. Resilin, optionally comprising the sequence MSKGP-(GRGDQPYQ)n, wherein n is greater than
b. ELR-IK 24 , optionally comprising the sequence MESLLP-(VPGIG VPGIG VPGKG VPGIG VPGIG) n, wherein n is greater than 5.
7 . The graphene oxide-protein matrix of claim 1 , wherein the DP is or comprises an elastin-like polymer (ELP), optionally wherein the ELP comprises an amino acid motif (VPGXG)n, wherein X is any amino acid apart from proline, and n is any number from 2 to 50.
8 . The graphene oxide-protein matrix of claim 1 , wherein the DP comprises a polymer having one or more repeating units of between 3 and 10 amino acids, optionally wherein at least 50% of the repeating units are hydrophobic.
9 . The graphene oxide-protein matrix of claim 1 , wherein the DP comprises a polymer having one or more repeating units of between 3 and 10 amino acids, optionally wherein at least 15% of the repeating units are positively charged.
10 . The graphene oxide-protein matrix of claim 1 , wherein the matrix further comprises:
a. cells, optionally wherein the cells are human umbilical vascular endothelial cells (hUVECs);
b. additional structures, optionally wherein the additional structures are selected from the group consisting of: vesicles, nanostructures, nano-capsules, growth factors, polysaccharides, exosomes, liposomes or quantum dots.
11 . The graphene oxide-protein matrix of claim 1 , wherein the GO is reduced GO.
12 . A kit for preparing a graphene oxide-protein matrix, the kit comprising;
a. an aqueous solution of a disordered protein (DP)
b. an aqueous solution of graphene oxide (GO)
wherein when the aqueous solution of DP and the aqueous solution of GO are admixed, a graphene oxide-protein matrix in the form of a 3 D structure having a lumen defined by a membrane having an inner and outer surface wherein the membrane comprises the graphene oxide protein matrix and at least partially surrounds the lumen is formed spontaneously, further wherein the DP has an opposite charge to the GO.
13 . A method of preparing a graphene oxide-protein matrix, the method comprising; admixing an aqueous solution of a disordered protein (DP) with an aqueous solution of graphene oxide (GO), wherein the DP has an opposite charge to the GO, further wherein the graphene oxide-protein matrix is in the form of a three-dimensional (3D) structure having a lumen defined by a membrane having an inner and outer surface, wherein the membrane comprises the graphene oxide protein matrix and at least partially surrounds the lumen.
14 . The method of claim 13 , wherein the graphene oxide-protein matrix is prepared using a device comprising;
(a) a reservoir containing the aqueous solution of DP; and
(b) a nozzle in fluid connection with the reservoir of DP
wherein the DP solution is delivered via the nozzle to a reservoir comprising the GO, optionally wherein the nozzle is controlled by a computer.
15 . The method of claim 13 , wherein
a. the aqueous solution of DP is added to the aqueous solution of GO; and/or
b. prior to matrix assembly the DP is comprised of at least 35% random coil structure and at least 5% α-helical structure; and/or
c. the percentage of β-sheet structure DP increases when complexed with GO.
16 . The method of claim 13 , wherein
a. the aqueous solution of DP has a concentration of from about 0.2% wt/vol to about 7.5% wt/vol; and/or
b. the aqueous solution of GO has a concentration of from about 0.01% wt/vol to about 0.5% wt/vol; and/or
c. the ratio of concentration of DP solution to concentration of GO solution is at least 10:1.
17 . The method of claim 13 , wherein;
a. the pH of the DP solution is from about 5 to about 9; and/or
b. the pH of the GO solution is from about pH2 to about pH6.
18 . The method of claim 13 , wherein when the DP has a transition temperature, the 3D structure is formed at or above the transition temperature of the DP.
19 . The method of claim 13 , wherein;
a. prior to admixing the DP and GO solutions, the method further comprises suspending cells in the DP solution; and/or
b. further comprising a step of seeding cells onto the 3D structure during or after admixing the DP and GO solutions.
20 . The method of claim 13 , further comprising a step of adding additional structures to the 3D structure prior to, during or after admixing the DP and GO solutions.