2019, Ramos-Gomes, F., Chames, P., Baty, D., Alves, F., Sukhanova, A., Samokhvalov, P., Nabiev, I., Суханова, Алена Владимировна, Самохвалов, Павел Сергеевич, Набиев, Игорь Руфаилович

© 2019 SPIE.Semiconductor quantum dots (QDs) are characterized by orders of magnitude higher multiphoton linear absorption cross-sections compared with conventional organic dyes. Combined with the QD photoluminescence quantum yield approaching 100%, this fact opens great prospects for the twophoton functional tumor imaging with QDs tagged with highly specific recognition molecules. Single-domain antibodies (sdAbs) or "nanobodies" derived from lamas are the smallest high-affinity recognition molecules, which may be tagged with the QDs thus permitting not only solid tumors multiphoton imaging but also rare disseminated cancer cells and micrometastases in the depth of the tissue to be detected. Additionally, unique photostability of QDs enables signal accumulation and significant enhancement of the sensitivity of routine biochemical and immunohistochemical assays to be obtained when the conjugates of QDs, instead of organic dyes, are used.

2019, Ramos-Gomes, Fernanda, Alves, Frauke, Nifontova, Galina, Baryshnikova, Maria, Nabiev, Igor, Sukhanova, Alyona, Нифонтова, Галина Олеговна, Барышникова, Мария Анатольевна, Набиев, Игорь Руфаилович, Суханова, Алена Владимировна

Imaging agents and drug carriers are commonly targeted toward cancer cell through functionalization with specific recognition molecules. Quantum dots (QDs) are fluorescent semiconductor nanocrystals whose extraordinary brightness and photostability make them attractive for direct fluorescent labeling of biomolecules or optical encoding of the membranes and cells. Here, we analyse the cytotoxicity of QD-encoded microcapsules, validate an approach to the activation of the microcapsule's surface for further functionalization with monoclonal antibody Trastuzumab, a humanized monoclonal antibody targeting the extracellular domain of the human epidermal growth factor receptor 2 (HER2) and already in clinical use for the treatment of HER2 positive breast cancer. In addition, we characterize the cell-specific targeting activity of the resultant bio-conjugate by immunofluorescence assay (IFA) and real-time analysis of interaction of the conjugates with live HER2 overexpressing human breast cancer cells. We demonstrate, that encapsulation of QDs into the polymer shell using the layer-by-layer deposition method yields highly fluorescent polyelectrolyte microcapsules with a homogeneous size distribution and biocompatibility upon in vitro treatment of cancer cells. Carbodiimide surface activation ensures optimal disperse and optical characteristics of the QD-encoded microcapsules before antibody conjugation. The prepared conjugates of the microcapsules with cancer-specific monoclonal antibody targeting HER2 provide sufficiently sensitive and specific antibody-mediated binding of the microcapsules with live cancer cells, which demonstrated their potential as prospective cancer cell-targeting agents.

2019, Konopsky, Valery, Petrova, Irina, Nabiev, Igor, Sukhanova, Alyona, Набиев, Игорь Руфаилович, Суханова, Алена Владимировна

Circulating cancer markers are metabolic products found in body fluids of cancer patients, which are specific for a certain type of malignant tumors. Cancer marker detection plays a key role in cancer diagnosis, treatment, and disease monitoring. The growing need for early cancer diagnosis requires quick and sensitive analytical approaches to detection of cancer markers. The approach based on the photonic crystal surface mode (PCSM) detection has been developed as a label-free high-precision biosensing technique. It allows real-time monitoring of molecular and cellular interactions using independent recording of the total internal reflection angle and the excitation angle of the PC surface wave. We used the PC SM technique for simultaneous detection of the ovarian cancer marker cancer antigen 125 and two breast cancer markers, human epidermal growth factor receptor 2 and cancer antigen 15-3. The new assay is based on the real-time flow detection of specific interaction between the antigens and capture antibodies. Its particular advantage is the possibility of multichannel recording with the same chip, which can be used for multiplexed detection of several cancer markers in a single experiment. The developed approach demonstrates high specificity and sensitivity for detection of all three biomarkers.

2019, de, Menezes, F. D., dos, Reis, S. R. R., Pinto, S. R., Portilho, F. L., Sukhanova, A., Nabiev, I. R., Суханова, Алена Владимировна, Набиев, Игорь Руфаилович

© 2019 Graphene is one of the crystalline forms of carbon, along with diamond, graphite, carbon nanotubes, and fullerenes, and is considered as a revolutionary and innovating product. The use of a graphene-based nanolabels is one of the latest and most prominent application of graphene, especially in the field of diagnosis and, recently, in loco radiotherapy when coupled with radioisotopes. However, its biological behavior and mutagenicity in different cell or animal models, as well as the in vivo functional activities, are still unrevealed. In this study we have developed by a green route of synthesizing graphene quantum dots (GQDs)and characterized them. We have also developed a methodology for direct radiolabeling of GQDs with radioisotopes.Finally; we have evaluated in vivo biological behavior of GQDs using two different mice models and tested in vitro mutagenicity of GQDs. The results have shown that GQDs were formed with a size range of 160-280 nm, which was confirmed by DRX and Raman spectroscopy analysis, corroborating that the green synthesis is an alternative, environmentally friendly way to produce graphene. The radiolabeling test has shown that stable radiolabeled GQDs can be produced with a high yield (>90%). The in vivo test has demonstrated a ubiquitous behavior when administered to healthy animals, with a high uptake by liver (>26%)and small intestine (>25%). Otherwise, in an inflammation/VEGF hyperexpression animal model (endometriosis), a very peculiar behavior of GQDs was observed, with a high uptake by kidneys (over 85%). The mutagenicity test has demonstrated A:T to G:C substitutions suggesting that GQDs exhibits mutagenic activity.

2019, Samokhvalov, P. S., Volodin, D. O., Bozrova, S. V., Dovzhenko, D. S., Zvaigzne, M. A., Lin'kov, P. A., Nifontova, G. O., Petrova, I. O., Sukhanova, A. V., Nabiev, I. R., Самохвалов, Павел Сергеевич, Нифонтова, Галина Олеговна, Суханова, Алена Владимировна, Набиев, Игорь Руфаилович

© 2019, Pleiades Publishing, Ltd.Abstract: Plasmonic nanoparticles have become a popularly accepted research tool in optoelectronics, photonics, and biomedical applications. The relatively recently appearing semiconductor plasmonic nanoparticles, as opposed to metal ones, are characterized by infrared plasmonic optical transitions and their application has a great future. In this work, the possibility of conversion of semiconductor (excitonic) fluorescence nanocrystals, i.e., quantum dots of the CuInS2 composition, to plasmonic nanoparticles by postsynthetic treatment without changes in the chemical composition of inorganic part of the nanocrystals was demonstrated for the first time ever.

2019, Efimov, A., Agapova, O., Agapov, I., Nifontova, G., Nabiev, I. R., Sukhanova, A., Нифонтова, Галина Олеговна, Набиев, Игорь Руфаилович, Суханова, Алена Владимировна

© 2019, The Author(s). Fluorescent imaging is a widely used technique for detecting and monitoring the distribution, interaction, and transformation processes at molecular, cellular, and tissue level in modern diagnostic and other biomedical applications. Unique photophysical properties of fluorescent semiconductor nanocrystals “quantum dots” (QDs) make them advanced fluorophores for fluorescent labeling of biomolecules or optical encoding of microparticles to be used as bioimaging and theranostic agents in targeted delivery, visualization, diagnostics, and imaging. This paper reports on the results of development of an improved approach to the optical encoding of polyelectrolyte microcapsules with stable, covered with the multifunctional polyethyleneglycol derivatives water-soluble QDs, as well as characterization of the optical properties, morphological and structural properties of the encoded microcapsules. The embedding of QDs into the polymer microcapsule membrane through layer-by-layer deposition on a preliminarily formed polymeric polyelectrolyte shell makes it possible to obtain bright fluorescent particles with an adapted charge and size distribution that are distinctly discernible by flow cytometry as individual homogeneous populations. The fluorescent microcapsules developed can be used in further designing bioimaging and theranostic agents sensitive to various external stimuli along with photoexcitation.

2016, Sizova, S. V., Oleinikov, V. A., Bouchonville, N., Molinari, M., Samokhvalov, P. S., Sukhanova, A., Nabiev, I., Суханова, Алена Владимировна, Набиев, Игорь Руфаилович, Олейников, Владимир Александрович

The integration of novel nanomaterials with highly functional biological molecules has numerous advanced applications, including optoelectronics, biosensing, imaging, and energy harvesting. This review summarizes recent progress in understanding the mechanisms of energy transfer between semiconductor nanocrystal (so-called quantum dots [QDs]) and photosensitive proteins in heterostructures, such as hybrids of semiconductor nanocrystals with purple membranes containing bacteriorhodopsin (bR) or with photosynthetic reaction centers (RCs). Understanding of these mechanisms should enable prediction of the possible ways to improve the biological function of biomolecules by means of their assembling with QDs and develop novel functional materials with controlled photonic properties and applications. The possible mechanisms of energy transfer from QDs to photochromic biomolecules are discussed and correlated with experimental data. The principles of hybrid structures engineering, donor/acceptor parameters affecting both energy transfer efficiency and biological function, and functionality of these hybrid structures are described. New nanobiohybrid materials are shown to have advanced implications for optoelectronics, photonics, and photovoltaics due to the ability of nanocomponents of these materials for efficient energy harvesting, conversion, and transfer of additional energy to Biosystems, thus making them working more efficiently.

2019, Poly, S., Lambert, E., Ewald, M., Karaulov, A., Sukhanova, A., Bozrova, S., Nabiev, I., Суханова, Алена Владимировна, Набиев, Игорь Руфаилович

© Copyright © 2019 Sukhanova, Poly, Bozrova, Lambert, Ewald, Karaulov, Molinari and Nabiev.Nanoparticles attract much interest as fluorescent labels for diagnostic and therapeutic tools, although their applications are often hindered by size- and shape-dependent cytotoxicity. This cytotoxicity is related not only to the leak of toxic metals from nanoparticles into a biological solution, but also to molecular cytotoxicity effects determined by the formation of a protein corona, appearance of an altered protein conformation leading to exposure of cryptic epitopes and cooperative effects involved in the interaction of proteins and peptides with nanoparticles. In the last case, nanoparticles may serve, depending on their nature, as centers of self-association or fibrillation of proteins and peptides, provoking amyloid-like proteinopathies, or as inhibitors of self-association of proteins, or they can self-assemble on biopolymers as on templates. In this study, human insulin protein was used to analyze nanoparticle-induced proteinopathy in physiological conditions. It is known that human insulin may form amyloid fibers, but only under extreme experimental conditions (very low pH and high temperatures). Here, we have shown that the quantum dots (QDs) may induce amyloid-like fibrillation of human insulin under physiological conditions through a complex process strongly dependent on the size and surface charge of QDs. The insulin molecular structure and fibril morphology have been shown to be modified at different stages of its fibrillation, which has been proved by comparative analysis of the data obtained using circular dichroism, dynamic light scattering, amyloid-specific thioflavin T (ThT) assay, transmission electron microscopy, and high-speed atomic force microscopy. We have found important roles of the QD size and surface charge in the destabilization of the insulin structure and the subsequent fibrillation. Remodeling of the insulin secondary structure accompanied by remarkable increase in the rate of formation of amyloid-like fibrils under physiologically normal conditions was observed when the protein was incubated with QDs of exact specific diameter coated with slightly negative specific polyethylene glycol (PEG) derivatives. Strongly negatively or slightly positively charged PEG-modified QDs of the same specific diameter or QDs of bigger or smaller diameters had no effect on insulin fibrillation. The observed effects pave the way to the control of amyloidosis proteinopathy by varying the nanoparticle size and surface charge.