Aggregation of nanoparticles in endosomes and lysosomes produces surface-enhanced Raman spectroscopy

Abstract. The purpose of this study was to explore the use of surface-enhanced Raman spectroscopy (SERS) to image the distribution of epidermal growth factor receptor (EGFR) in cells. To accomplish this task, 30-nm gold nanoparticles (AuNPs) tagged with antibodies to EGFR (1012  per mL) were incubated with cells (106  per mL) of the A431 human epidermoid carcinoma and normal human bronchial epithelial cell lines. Using the 632.8-nm excitation line of a He-Ne laser, Raman spectroscopy measurements were performed using a point mapping scheme. Normal cells show little to no enhancement. SERS signals were observed inside the cytoplasm of A431 cells with an overall enhancement of 4 to 7 orders of magnitude. Raman intensity maps of the 1450 and 1583  cm−1 peaks correlate well with the expected distribution of EGFR and AuNPs, aggregated following uptake by endosomes and lysosomes. Spectral features from tyrosine and tryptophan residues dominate the SERS signals.

[1]  D. Neal,et al.  EPIDERMAL GROWTH-FACTOR RECEPTOR - A MARKER OF EARLY RELAPSE IN BREAST-CANCER AND TUMOR STAGE PROGRESSION IN BLADDER-CANCER - INTERACTIONS WITH NEU , 1989 .

[2]  N. Shah,et al.  Surface-enhanced Raman spectroscopy. , 2008, Annual review of analytical chemistry.

[3]  P. Vandenabeele,et al.  Reference database of Raman spectra of biological molecules , 2007 .

[4]  A. Eisbruch,et al.  Analysis of the epidermal growth factor receptor gene in fresh human head and neck tumors. , 1987, Cancer research.

[5]  May D. Wang,et al.  In vivo tumor targeting and spectroscopic detection with surface-enhanced Raman nanoparticle tags , 2008, Nature Biotechnology.

[6]  Gérald Thouand,et al.  Protein Interactions Investigated by the Raman Spectroscopy for Biosensor Applications , 2012 .

[7]  J. Mendelsohn,et al.  Epidermal growth factor receptor expression in human lung carcinomas defined by a monoclonal antibody. , 1987, Journal of the National Cancer Institute.

[8]  B. Ozanne,et al.  The use of egf receptors and an alveolar differentiation marker to predict a poor survival in lung cancer , 1989 .

[9]  B. Gusterson,et al.  Over‐expression of the EGF receptor is a hallmark of squamous cell carcinomas , 1986, The Journal of pathology.

[10]  D. Appleton,et al.  Presence of epidermal growth factor receptor as an indicator of poor prognosis in patients with breast cancer. , 1985, Journal of clinical pathology.

[11]  R. Buck,et al.  Surface-enhanced Raman spectroscopic investigation of human immunoglobulin G adsorbed on a silver electrode , 1989 .

[12]  Konstantin V. Sokolov,et al.  A Far-red Fluorescent Contrast Agent to Image Epidermal Growth Factor Receptor Expression , 2004, Photochemistry and photobiology.

[13]  Wei Qian,et al.  Cancer cells assemble and align gold nanorods conjugated to antibodies to produce highly enhanced, sharp, and polarized surface Raman spectra: a potential cancer diagnostic marker. , 2007, Nano letters.

[14]  A. Smith,et al.  Imaging EGFR distribution using surface-enhanced Raman spectroscopy , 2009, BiOS.

[15]  J. Hillier,et al.  A study of the nucleation and growth processes in the synthesis of colloidal gold , 1951 .

[16]  K. Soo,et al.  Early diagnosis of oral cancer based on the surface plasmon resonance of gold nanoparticles , 2007, International journal of nanomedicine.

[17]  T. Libermann,et al.  Expression of epidermal growth factor receptors in human brain tumors. , 1984, Cancer research.

[18]  S. Gambhir,et al.  Noninvasive molecular imaging of small living subjects using Raman spectroscopy , 2008, Proceedings of the National Academy of Sciences.

[19]  G. Frens.  Controlled Nucleation for the Regulation of the Particle Size in Monodisperse Gold Suspensions , 1973 .

[20]  Kort Travis,et al.  Plasmon resonance coupling of metal nanoparticles for molecular imaging of carcinogenesis in vivo. , 2007, Journal of biomedical optics.

[21]  Bong-Hyun Jun,et al.  Nanoparticle probes with surface enhanced Raman spectroscopic tags for cellular cancer targeting. , 2006, Analytical chemistry.

[22]  Remy Cromer,et al.  SERS nanoparticles: a new optical detection modality for cancer diagnosis. , 2007, Nanomedicine.