Quantitative investigation of compartmentalized dynamics of ErbB2 targeting gold nanorods in live cells by single molecule spectroscopy.
Understanding the diffusion dynamics and receptor uptake mechanism of nanoparticles in cancer cells is crucial to the rational design of multifunctional nanoprobes for targeting and delivery. In this report, for the first time, we quantify the localization and evaluate the diffusion times of Herceptin-conjugated gold nanorods (H-GNRs) in different cell organelles by fluorescence correlation spectroscopy (FCS) and examine the endocytic diffusion of H-GNRs in live ErbB2 overexpressing SK-BR-3 cells. First, by colocalizing H-GNRs in different cellular organelles depicted by the respective markers, we demonstrate that H-GNRs colocalize with the endosome and lysosome but not with the Golgi apparatus. Our study shows that Herceptin-conjugated GNRs have similar intracellular localization characteristics as Herceptin-ErbB2 complex, with a higher concentration found in the endosome (72 +/- 20.6 nM) than lysosome (9.4 +/- 4.2 nM) after internalization. The demonstrated approach and findings not only lay the foundations for a quantitative understanding of the fate of nanoparticle-based targeting but also provide new insights into the rational design of nanoparticle delivery systems for effective treatment.
Chen
Jiji
J
Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47907, USA.
Irudayaraj
Joseph
J
D001943:Breast Neoplasms / Q000378:metabolism*; D003460:Crystallization / Q000379:methods; D016503:Drug Delivery Systems / Q000379:methods*; D005956:Glucose-6-Phosphate Isomerase; D006046:Gold / Q000737:chemistry*; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry; D008422:Materials Testing; D008968:Molecular Conformation; D015336:Molecular Probe Techniques; D050997:Nanomedicine / Q000379:methods; D049329:Nanostructures / Q000737:chemistry; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis*; D013499:Surface Properties
D005956:Glucose-6-Phosphate Isomerase; D006046:Gold; D046911:Macromolecular Substances; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human