Pointing in the Right Direction: Controlling the Orientation of Proteins on Nanoparticles Improves Targeting Efficiency
| Autor: | Ken W Yong, Christopher J.H. Porter, Angus P. R. Johnston, Moore Z. Chen, Daniel Yuen |
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| Rok vydání: | 2019 |
| Předmět: |
Azides
Immunoconjugates Phenylalanine Receptors Cell Surface Bioengineering 02 engineering and technology Conjugated system Crystallography X-Ray Ligands Cyclooctanes Cell surface receptor Humans General Materials Science Receptor chemistry.chemical_classification Mechanical Engineering General Chemistry Single-Domain Antibodies 021001 nanoscience & nanotechnology Condensed Matter Physics Amino acid ErbB Receptors Single-domain antibody chemistry A549 Cells Cancer cell Biophysics Click chemistry Nanoparticles Click Chemistry Bioorthogonal chemistry 0210 nano-technology Protein Binding |
| Zdroj: | Nano Letters. 19:1827-1831 |
| ISSN: | 1530-6992 1530-6984 |
| DOI: | 10.1021/acs.nanolett.8b04916 |
| Popis: | Protein-conjugated nanoparticles have the potential to precisely deliver therapeutics to target sites in the body by specifically binding to cell surface receptors. To maximize targeting efficiency, the three-dimensional presentation of ligands toward these receptors is crucial. Herein, we demonstrate significantly enhanced targeting of nanoparticles to cancer cells by controlling the protein orientation on the nanoparticle surface. To engineer the point of attachment, we used amber codon reassignment to incorporate a synthetic amino acid, p-azidophenylalanine (azPhe), at specific locations within a single domain antibody (sdAb or nanobody) that recognizes the human epidermal growth factor receptor (EGFR). The azPhe modified sdAb can be tethered to the nanoparticle in a specific orientation using a bioorthogonal click reaction with a strained cyclooctyne. The crystal structure of the sdAb bound to EGFR was used to rationally select sites likely to optimally display the sdAb upon conjugation to a fluorescent nanocrystal (Qdot). Qdots with sdAb attached at the azPhe13 position showed 6 times greater binding affinity to EGFR expressing A549 cells, compared to Qdots with conventionally (succinimidyl ester) conjugated sdAb. As ligand-targeted delivery systems move toward clinical application, this work shows that nanoparticle targeting can be optimized by engineering the site of protein conjugation. |
| Databáze: | OpenAIRE |
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