Quantification and imaging of HER2 protein using nanocrystals conjugated with single-domain antibodies

1st International Symposium 'Physics, Engineering and Technologies for Bio-Medicine'

医药卫生;物理学;工业技术

Glukhov, S.^1 ; Berestovoy, M.^1 ; Chames, P.^2 ; Baty, D.^2 ; Nabiev, I.^1,3 ; Sukhanova, A.^1,3

Laboratory of Nano-Bioengineering, National Research Nuclear University, MEPhI (Moscow Engineering Physics Institute), Moscow, Russia^1

INSERM U1068, CNRS UMR7258, Cancer Research Center, Institut Paoli-Calmettes and Aix-Marseille, Marseille, France^2

Laboratory of Research in Nanoscience, LRN - EA4682, University of Reims Champagne Ardenne, Reims, France^3

关键词: Emission quantum yield;    Emission spectrums;    Human epidermal growth factor receptor 2 (HER2);    Inorganic nanocrystals;    Integrated diagnosis;    Near infrared region;    Prognostic markers;    Single-domain antibodies;   

Others  :  https://iopscience.iop.org/article/10.1088/1742-6596/784/1/012016/pdf
DOI  :  10.1088/1742-6596/784/1/012016

来源: IOP

【 摘 要 】

This study dealt with quantification and imaging of human epidermal growth factor receptor 2 (HER2), an important prognostic marker for cancer diagnosis and treatment, using specific quantum-dot-based conjugates. Fluorescent inorganic nanocrystals or quantum dots (QDs) are extremely highly resistant to photobleaching and have a high emission quantum yield and a continuous range of emission spectra, from the ultraviolet to the infrared regions. Ultrasmall nanoprobes consisting of highly affine anti-HER2 single-domain antibodies (sdAbs or nanobodies) conjugated with QDs in a strictly oriented manner have been designed. QDs with a fluorescence peak maxima at wavelengths of 562 nm, 569 nm, 570 nm or in the near-infrared region were used. Here, we present our results of ISA quantification of HER2 protein, in situ imaging of HER2 protein on the surface of HER2-positive SK-BR-3 cells in immunohistochemical experiments, and counting of stained with anti-HER2 conjugates HER2-positive SK-BR-3 cells in their mixture with unstained cells of the same culture in flow cytometry experiments. The data demonstrate that the anti-HER2 QD-sdAb conjugates obtained are highly specific and sensitive and could be used in numerous applications for advanced integrated diagnosis.