Mutant Protein-directed 3D Plasmon Graphene Oxide For Cell SERS Imaging

Seeking a versatile nanoplatform for multimodal bioimaging and therapeutics is a challengeable task.A general complementary bottom-up bio-nanotechnology for controlling 3D supramolecular coassembly has been proposed.Coengineering proof-on-concept of supermolecular architecture can be demonstrated via genetic engineered protein dimer and plasmonic engineered GO.Incorporation of anisotropic plasmonic nanoparticles as interlayer among the GO 3D supramolecular architecture can provide covalent conjugation sites and simultaneously endow tunable optical properties of GO ranging from ultraviolet to near infrared region.Interestingly,precise designing specific two-site mutation of plasmid is favorable for giving organized coassembly instead of random networks of GO,which contributes to giving superior enhanced Raman imaging behaviour for tracking cancer cell.Unexpected penetration into cell nucleus via the sub-micro 3D supramolecular coassembly exhibits excellent nucleus therapeutics potential of cancer cell.Delivery of protein into cell holds longstanding attentions due to its enormous medical applications,e.g.,cancer therapy,vaccination and regenerative medicine.However,promoting therapeutic protein to subcellular compartments including cytosol,nucleus,and mitochondria,is quite challengeable task due to its difficulty in escaping endosomal pathway avoiding being trafficked through endomembrane compartments.Thus,preparing efficient nanocarriers,such as polymeric-based,lipid-mediated,inorganic-based and protein-mediated nanocarriers,for protein intracellular delivery have been attracting more and more interests.Complementary inorganic-based nanocarrier of protein and nanoparticles protein and nanoparticles allows incorporation of biofunctionality into the nanostructure and nanoscaffold for loading protein.Self-assembly of protein and inorganic nanoparticles via non-covalent conjugation based on hydrophobic adsorption,host-guest encapsulation or electrostatic attraction have been mainly relied on natural complementary interaction.Herein,we propose coengineering assembled approach for giving programmed synthesis of protein-nanoparticle supramolecular architecture.We demonstrated the proposal via a representative example of genetic engineered protein and plasmonic engineered inorganic/organic polymeric-graphene oxide.As a flexible 2D material,graphene or graphene oxide(GO)has been extensively applied in biosensing and biomedical applications due to its water-dispersibility,low cytotoxicity,large surface area as compared to carbon nanotubes.Assembling flexible 2D graphene sheets into 3D architectures promoted extensive applications of graphene-based materials in energy,sensing,biological field due to their superior properties.Thus,some advanced technologies including template-guidance,chemical vapor deposition,colloidal-spheres synthesis.solvothermal,sol-gel synthesis,have been developed for fabricating graphene 3D architecture.Investigations on graphene or graphene oxide 3D architecture have been focusing on fabrication and applications of 3D random interconnected graphene-based networks;in contrast,fabricating organized architecture,e.g.,lamellar or liquid crystal,appears more challengeable due to its well-stacked highly-ordered multiple layer.Until now,investigations on GO lamellar scaffolds only have been limited on polymer-guided technology;however,there is no report on engineered protein-graphene oxide supramolecular coassembly so far.In this thesis,in order to control GO 3D supramolecular architecture(GO 3D SA),plasmonic engineering giving metallic interlayer for conjugating GO and genetic engineered protein have been designed.The pathway solves the several critical problems of GO-biomedical applications.(?)inorganic/organic 3D hybrids as a stabilized capsule provides efficient paths for access and diffusion of ions and molecules,and intracellular delivery of therapeutic protein.(?)genetic engineered protein instead of wild-type protein for coassembling with nanoparticles extends limitation of selective specific proteins to give supramolecular architecture.(?)anisotropic plasmonic interlayer can avoid entanglement of GO and give tunable optical ability for tracking cancer cell.(IV)efficient drug loading featured a stimuli-responsive drug release from the supramolecular coassembly can be achieved.