Fate of Antibody-Targeted Ultrasmall Gold Nanoparticles in Cancer Cells after Receptor-Mediated Uptake.
Sangheon HanTomasz ZalKonstantin V SokolovPublished in: ACS nano (2021)
Nanoparticles with ultrasmall sizes (less than 10 nm) offer many advantages in biomedical applications compared to their bigger counterparts, including better intratumoral distribution, improved pharmacokinetics (PK), and efficient body clearance. When functionalized with a biocompatible coating and a target-specific antibody, ultrasmall nanoparticles represent an attractive clinical translation platform. Although there is a tremendous body of work dedicated to PK and the biological effects of various nanoparticles, little is known about the fate of different components of functionalized nanoparticles in a biological environment such as in live cells. Here, we used luminescence properties of 5 nm gold nanoparticles (AuNPs) to study the intracellular trafficking and fate of the AuNPs functionalized with an organic layer consisting of a polyethylene glycol (PEG) coating and epidermal growth factor receptor (EGFR)-targeting antibody. We showed that intracellular uptake of the targeted 5 nm AuNPs results in a strong two-photon luminescence (TPL) that is characterized by broad emission and very short lifetimes compared to the fluorescence of the nanoparticle-conjugated fluorophore-tagged antibody, thereby allowing selective imaging of these components using TPL and two-photon excited fluorescence lifetime microscopy (2P-FLIM). Our results indicate that the nanoparticle's coating is detached from the particle's surface inside cells, leading to formation of nanoparticle clusters with a strong TPL. Furthermore, we observed an optically resolved spatial separation of the gold core and the antibody coating of the particles inside cells. We used data from two-photon microscopy, 2P-FLIM, electron microscopy, and in vitro assays to propose a model of interactions of functionalized 5 nm AuNPs with live cells.
Keyphrases
- induced apoptosis
- gold nanoparticles
- epidermal growth factor receptor
- quantum dots
- cell cycle arrest
- photodynamic therapy
- high resolution
- single molecule
- small cell lung cancer
- high throughput
- cancer therapy
- iron oxide
- drug delivery
- energy transfer
- machine learning
- cell proliferation
- oxidative stress
- optical coherence tomography
- advanced non small cell lung cancer
- high speed
- reactive oxygen species
- ionic liquid
- fluorescent probe
- label free
- data analysis
- iron oxide nanoparticles