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Fluorescence techniques used to measure interactions between hydroxyapatite nanoparticles and epidermal growth factor receptors
Author(s) -
Kathawala Mustafa H.,
Khoo Stella P. K.,
Sudhaharan Thankiah,
Zhao Xinxin,
Say Chye Loo Joachim,
Ahmed Sohail,
Woei Ng Kee
Publication year - 2015
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201400404
Subject(s) - förster resonance energy transfer , photobleaching , biophysics , nanoparticle , chinese hamster ovary cell , fluorescence lifetime imaging microscopy , fluorescence , epidermal growth factor , chemistry , receptor , confocal microscopy , nanotechnology , materials science , microbiology and biotechnology , biochemistry , biology , physics , quantum mechanics
The potential applications of nanomaterials in therapeutics are immense and to fully explore this potential, it is important to understand the interaction of nanoparticles with cellular components. To examine the interaction between nanoparticles and cell membrane receptors, this report describes the use of advanced fluorescence techniques to measure interactions between hydroxyapatite (HA) nanoparticles and epidermal growth factor receptors (EGFRs), as a model system. FITC‐labelled HA nanoparticles and monomeric red fluorescent protein (mRFP)‐conjugated EGFRs expressed in Chinese hamster ovary cells (CHO‐K1) were generated and their interaction measured using acceptor photobleaching‐fluorescence resonance energy transfer (AP‐FRET) and fluorescence lifetime imaging microscopy‐fluorescence resonance energy transfer (FLIM‐FRET). Results confirmed that hydroxyapatite nanoparticles not only interacted with EGFR but also attenuated downstream EGFR signalling, possibly by hindering normal dimerization of EGFR. Furthermore, the extent of signal attenuation suggested correlation with specific surface area of the nanoparticles, whereby greater specific surface area resulted in greater downstream signal attenuation. This novel demonstration establishes fluorescence techniques as a viable method to study nanoparticle interactions with proteins such as cell surface receptors. The approach described herein can be extended to study interactions between any fluorescently labelled nanoparticle‐biomolecule pair.