Epidermal growth factor receptor-targeted lipid nanoparticles retain self-assembled nanostructures and provide high specificity

Abstract

Next generation drug delivery utilising nanoparticles incorporates active targeting to specific sites. In this work, we combined targeting with the inherent advantages of self-assembled lipid nanoparticles containing internal nano-structures. Epidermal growth factor receptor (EGFR)-targeting, PEGylated lipid nanoparticles using phytantriol and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-PEG-maleimide amphiphiles were created. The self-assembled lipid nanoparticles presented here have internal lyotropic liquid crystalline nano-structures, verified by synchrotron small angle X-ray scattering and cryo-transmission electron microscopy, that offer the potential of high drug loading and enhanced cell penetration. Anti-EGFR Fab' fragments were conjugated to the surface of nanoparticles via a maleimide-thiol reaction at a high conjugation efficiency and retained specificity following conjugation to the nanoparticles. The conjugated nanoparticles were demonstrated to have high affinity for an EGFR target in a ligand binding assay.

Cubosomes: The Next Generation of Smart Lipid Nanoparticles?

“…As the majority of stabilizers are believed to predominantly cover the outer cubosome surface, they are crucial for targeting cells of interest using cubosomes. Examples to date include chemical modifications of F108 to add biotin [67] or folate, [32] antibody fragment coupling to PEGylated lipids, [68] protein labelling, [69] copper free click functional phospholipids [70] and coating cubosomes with Poly-ε-lysine [35] . These can be broadly summarized as either biological or chemical approaches to targeting and are all at an early development stage.…”

“…Their targeting ability was tested via a surface plasmon resonance binding affinity assay to a fusion protein (sEGFR501.Fc) with high affinity for anti-EGFR ligands, however, there is no biological data. [68] Protein modification of cubosomes is a more costly approach, although one advantage is that cubosomes may act as a stabilizer for some proteins. Annexin V binds to apoptotic cell membranes and in fluorescent form is already used for cellular staining assays.…”

Cryo‐electron microscopy and cryo‐electron tomography of nanoparticles

“…A cryo‐TEM study of intermediates in the assembly pathway of cubosomes led to a model for their growth pathway starting with unilamellar vesicles . Cryo‐TEM has also been used to demonstrate the preservation of cubosomes after addition of targeting ligands …”

Nanostructure and cytotoxicity of self-assembled monoolein–capric acid lyotropic liquid crystalline nanoparticles

“…[1][2][3] The polymorphism of MO self-assembly has been studied extensively. Self-assembled MO nds application as carriers in drug delivery, [6][7][8][9][10][11][12][13] and in gene therapy, [14][15][16] cancer therapy, [17][18][19][20] protein crystallisation, [21][22][23][24][25][26] and MRI imaging. 1).…”