Revealing membrane alteration in cellsoverexpressing CA IX and EGFR by Surface-Enhanced Raman Scattering

Year: 2019

Authors: Rusciano G., Sasso E., Capaccio A., Zambrano N., Sasso A.

Autors Affiliation: Univ Naples Federico II, Dept Phys E Pancini, Complesso Univ Monte S Angelo,Via Cintia, I-80126 Naples, Italy; Natl Res Council CNR, Natl Inst Opt INO, Via Campi Flegrei 34, I-80078 Pozzuoli, NA, Italy; Univ Naples Federico II, Dept Mol Med & Med Biotechnol, Via S Pansini 5, I-80131 Naples, Italy; CEINGE Adv Biotechnol SCaRL, Via G Salvatore 486, I-80145 Naples, Italy; Nouscom SRL, Rome, Italy

Abstract: Sensitive detection of altered proteins expression in plasma membranes is of fundamental importance, for both diagnostic and prognostic purposes. Surface-Enhanced Raman Scattering (SERS) has proven to be a quite sensitive approach to detect proteins, even in very diluted samples. However, proteins detection in complex environment, such as the cellular membrane, is still a challenge. Herein, we demonstrate a SERS-based platform to reveal the overexpression of target proteins in cell membranes. As a proof of concept, we implemented ectopic expression of carbonic anhydrase IX (CA IX) and epidermal growth factor receptor (EGFR) in the plasma membrane of the SKOV3 tumor cell line. Our outcomes demonstrate that SERS signals from cells put in contact with a hyperuniform SERS substrate allow highlighting subtle differences in the biochemical composition of cell membranes, normally hidden in spontaneous Raman confocal microscopy. This opens new opportunities for a label-free membrane analysis and bio-sensing in a broader sense.

Journal/Review: SCIENTIFIC REPORTS

Volume: 9      Pages from: 1832-1  to: 1832-10

More Information: This work was supported by FIRB 2012-RBFR12WAPY to G.R., MIUR-PRIN2015 and Associazione Culturale DiSciMuS RFC (Biologia dei tumori ipossici) to NZ. The Authors thank A. La Rocca for help in procedure set-up, F. Monteleone for the generation of the EGFR expression vector, M. Raia, G. Scalia, F. Visconte of the CEINGE general and clinical cytometry facility for FACS analysis, and M.G. De Biasi for critical discussion.
KeyWords: CARBONIC-ANHYDRASE IX; SPECTROSCOPY; PROTEIN
DOI: 10.1038/s41598-018-37997-3

Citations: 11
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