2019, Orlov, A. V., Pushkarev, A. V., Bragina, V. A., Burenin, A. G., Novichikhin, D. O., Nikitin, P. I., Никитин, Петр Иванович

2022, Orlov, A. V., Znoyko, S. L., Malkerov, J. A., Novichikhin, D. O., Nikitin, P. I., Никитин, Петр Иванович

© 2022 Elsevier LtdWe present an easy-to-use lateral flow immunoassay for rapid, precise and sensitive quantification of one of the most hazardous mycotoxins – ochratoxin A (OTA), which is widely present in food and agricultural commodities. The achieved limit of detection during the 20-min OTA registration is 11 pg/mL. The assay provides accurate results in both low- and high-concentration ranges. That is due to the extraordinary steepness of the linear calibration plot: 5-order dynamic range of concentrations causes almost a 1000-fold change in the signal obtained by electronic detection of magnetic biolabels using their non-linear magnetization. High specificity, repeatability, and reproducibility of the assay have been verified, including measuring OTA in real samples of contaminated corn flour. The developed assay is a promising analytical tool for food and feed safety control; it may become an express, convenient and high-precision alternative to the traditional sophisticated laboratory techniques based on liquid chromatography.

2020, Orlov, A. V., Pushkarev, A. V., Znoyko, S. L., Bragina, V. A., Novichikhin, D. O., Nikitin, P. I., Никитин, Петр Иванович

© 2020 Elsevier B.V.A multiplex label-free biosensor is developed for diagnostics of autoimmune diseases by highly sensitive measuring in human serum both critical characteristics of autoantibody: concentration and native kinetic parameters that reflect autoantibody aggressiveness to the organism's tissues. The biosensor is based on the spectral-correlation interferometry and image processing of a microarray glass biochip, affordable to be single-used in medical applications. Simultaneous 25-min detection and activity characterization of several autoantibodies in the same serum sample have been demonstrated for anti-thyroglobulin (anti-TG) and anti-thyroid peroxidase (anti-TPO) as models. The biosensor offers extremely high sensitivity: limits of detection in serum are 1.7 IU/mL and 6 IU/mL for anti-TPO and anti-TG, respectively. The dynamic range covers the whole range of clinically relevant concentrations of the autoantibodies up to 1000 IU/mL. The developed method of characterization of autoantibody activity by recording the kinetics of their binding with free native antigens is based on autoantibody polyvalency. The measurements in clinical serum samples have shown that the native kinetic parameters are independent of concentration. The proposed biosensor and method of native kinetic registration can be used to develop new criteria for comprehensive diagnostics of autoimmune diseases, based not only on traditional measurements of concentration but also on quantitative evaluation of autoantibody aggressiveness. The developed method can be adapted to other label-free sensors such as those based on the surface plasmon resonance, optical waveguides, etc.

2022, Yaremenko, A. V., Zelepukin, I. V., Ivanov, I. N., Deyev, S. M., Nikitin, P. I., Деев, Сергей Михайлович, Никитин, Петр Иванович

Magnetic nanoparticles are widely used in biomedicine for MRI imaging and anemia treatment. The aging of these nanomaterials in vivo may lead to gradual diminishing of their contrast properties and inducing toxicity. Here, we describe observation of the full lifecycle of 40-nm magnetic particles from their injection to the complete degradation in vivo and associated impact on the organism. We found that in 2 h the nanoparticles were eliminated from the bloodstream, but their initial biodistribution changed over time. In 1 week, a major part of the nanoparticles was transferred to the liver and spleen, where they degraded with a half-life of 21 days. MRI and a magnetic spectral approach revealed preservation of contrast in these organs for more than 1 month. The particle degradation led to the increased number of red blood cells and blood hemoglobin level due to released iron without causing any toxicity in tissues. We also observed an increase in gene expression level of Fe-associated proteins such as transferrin, DMT1, and ferroportin in the liver in response to the iron particle degradation. A deeper understanding of the organism response to the particle degradation can bring new directions to the field of MRI contrast agent design. © 2022, The Author(s).

2019, Ivanov, A. E., Pushkarev, A. V., Orlov, A. V., Nikitin, M. P., Nikitin, P. I., Никитин, Петр Иванович

© 2018 Elsevier B.V. Three types of polymer-coated glass slides were prepared by acylation of their γ–aminopropyl derivatives with succinic anhydride or poly(p-nitrophenyl acylate) (PNPA) at various reaction times. The variations in duration of chemical adsorption of PNPA allowed control over nano-corrugation of the reactive surfaces employed for biosensing. The visually smooth and optically clear polymer-coated slides were used for immobilization of a conjugate of bovine serum albumin with chloramphenicol (CAP) and label-free measurement of concentration of this antibiotic. The sensor chips prepared in this way provided strong biosensing response due to fast biomolecular interactions and efficient competition of the target analyte during indirect immunoassay. The kinetics of formation and dissociation of immune complexes “conjugate-antibody” on the sensor chip surface was registered by the label-free methods of spectral-correlation and spectral-phase interferometry. The average thickness of immune complex adsorption layers was in the range of 5–12 nm. The highest value of kinetic association constant (kon = 5 × 104 M−1s−1) and the lowest equilibrium dissociation constant (Kdiss = 2 × 10-8 M−1) were achieved with the chips originated in the PNPA-coated slides, which were produced under short (2 min) polymer chemisorption and exhibited nano-corrugation almost unchanged with respect to the parent glass, according to the atomic-force microscopy. The detection limit for CAP detection was 80 pg/mL, and the dynamic range was 5 orders. The proposed methods for fabrication of sensor chips with nano-corrugated surface, the results of their characterization, as well as the developed assay can be employed for high-throughput recording of multiple molecular interactions by spectral interferometry and in a wide range of biosensing platforms for registration of biologically active compounds in medicine, veterinary, food control and ecological monitoring.

2019, Yaremenko, A. V., Babenyshev, A. V., Balalaeva, I. V., Nikitin, M. P., Zelepukin, I. V., Shipunova, V. O., Nikitin, P. I., Deyev, S. M., Никитин, Петр Иванович, Деев, Сергей Михайлович

© The Royal Society of Chemistry. Delivery of particle-based theranostic agents via their transportation on the surfaces of red blood cells, commonly referred to as RBC-hitchhiking, has historically been developed as a promising strategy for increasing the extremely poor blood circulation lifetime, primarily, of the large-sized sub-micron agents. Here, we show for the first time that RBC-hitchhiking can be extremely efficient for nanoparticle delivery and tumor treatment even in those cases when no circulation prolongation is observed. Specifically, we demonstrate that RBC-hitchhiking of certain small 100 nm particles, unlike that of the conventional sub-micron ones, can boost the delivery of non-targeted particles to lungs up to a record high value of 120-fold (and up to 40% of the injected dose). To achieve this remarkable result, we screened sub-200 nm nanoparticles of different sizes, polymer coatings and ζ-potentials and identified particles with the optimal RBC adsorption/desorption behavior. Furthermore, we demonstrated that such RBC-mediated rerouting of particles to lungs can be used to fight pulmonary metastases of aggressive melanoma B16-F1. Our findings could change the general paradigm of drug delivery for cancer treatment with RBC-hitchhiking. It is not the blood circulation lifetime that is the key factor for nanoparticle efficiency, but rather the complexation of nanoparticles with the RBC. The demonstrated technology could become a valuable tool for development of new strategies based on small nanoparticles for the treatment of aggressive and small-cell types of cancer as well as other lung diseases.

2020, Yaremenko, A. V., Yuryev, M. V., Mirkasymov, A. B., Sokolov, I. L., Zelepukin, I. V., Deyev, S. M., Nikitin, P. I., Деев, Сергей Михайлович, Никитин, Петр Иванович

© 2020 Elsevier B.V.Blood circulation is the key parameter that determines the in vivo efficiency of nanoagents. Despite clinical success of the stealth liposomal agents with their inert and shielded surfaces, a great number of non-stealth nanomaterials is being developed due to their potential of enhanced functionality. By harnessing surface phenomena, such agents can offer advanced control over drug release through intricately designed nanopores, catalysis-propelled motion, computer-like analysis of several disease markers for precise target identification, etc. However, investigation of pharmacokinetic behavior of these agents becomes a great challenge due to ultra-short circulation (usually around several minutes) and impossibility to use the invasive blood-sampling techniques. Accordingly, the data on circulation of such agents has been scarce and irregular. Here, we demonstrate high-throughput capabilities of the developed magnetic particle quantification technique for nanoparticle circulation measurements and present a comprehensive investigation of factors that affect blood circulation of the non-stealth nanoparticles. Namely, we studied the following 9 factors: particle size, zeta-potential, coating, injection dose, repetitive administration, induction of anesthesia, mice strain, absence/presence of tumors, tumor size. Our fundamental findings demonstrate potential ways to extend the half-life of the agents in blood thereby giving them a better chance of achieving their goal in the organism. The study will be valuable for design of the next generation nanomaterials with advanced biomedical functionality.

2022, Mashkovich, E. A., Deryabin, M. S., Kurin, V. V., Bakunov, M. I., Zelepukin, I. V., Lipey, N. A., Popov, A. A., Shipunova, V. O., Yu. , Griaznova, O., Nikitin, P. I., Kabashin, A. V., Deyev, S. M., Zvyagin, A. V., Попов, Антон Александрович, Грязнова, Ольга Юрьевна, Никитин, Петр Иванович, Кабашин, Андрей Викторович, Деев, Сергей Михайлович

© 2021 Elsevier B.V.Nanomaterials with controllable biodegradation properties respond to the main challenge of cancer nanomedicine to minimise side effects and maximise the delivery efficacy to tumours. These biodegradation properties vary from clear aqueous solutions to protein-abundant biological fluids. A photoacoustic method suitable for in vitro quantification of highly scattering colloids with optical absorption properties is introduced and demonstrated by determination of the degradation rate of laser-synthesized silicon nanoparticles (Si NPs) in turbid serum solutions. In vitro screening of a variety of polymer surface-coatings of Si NPs revealed a stand-alone property of polyallylamine (PAA) to accelerate the Si NP dissolution. PAA-coated Si NP half-life was measured ∼ 100-min in aqueous solutions and slowed down to ∼ 24 h in serum. As-produced PAA-coated Si NPs appeared suitable for blockade of the mononuclear phagocyte system. Pre-treatment with PAA-Si NPs caused 1.4-times reduced uptake of magnetic particles by human THP-1 cells in vitro and a 13-fold increase of the magnetic particle delivery to the B16-F1 xenograft tumours in vivo. The demonstrated photoacoustic method is believed to facilitate design and screening of biodegradable materials suitable for in vivo applications such as controlled drug release.

2022, Bragina, V. A., Khomyakova, E., Orlov, A. V., Znoyko, S. L., Nikitin, P. I., Никитин, Петр Иванович

© 2022 by the authors. Licensee MDPI, Basel, Switzerland.Extracellular vesicles (EVs) are promising agents for liquid biopsy—a non-invasive approach for the diagnosis of cancer and evaluation of therapy response. However, EV potential is limited by the lack of sufficiently sensitive, time-, and cost-efficient methods for their registration. This research aimed at developing a highly sensitive and easy-to-use immunochromatographic tool based on magnetic nanoparticles for EV quantification. The tool is demonstrated by detection of EVs isolated from cell culture supernatants and various body fluids using characteristic biomarkers, CD9 and CD81, and a tumor-associated marker—epithelial cell adhesion molecules. The detection limit of 3.7 × 105 EV/µL is one to two orders better than the most sensitive traditional lateral flow system and commercial ELISA kits. The detection specificity is ensured by an isotype control line on the test strip. The tool’s advantages are due to the spatial quantification of EV-bound magnetic nanolabels within the strip volume by an original electronic technique. The inexpensive tool, promising for liquid biopsy in daily clinical routines, can be extended to other relevant biomarkers.

2022, Orlov, A. V., Novichikhin, D. O., Malkerov, Y. A., Nikitin, P. I., Gorshkov, B. G., Guteneva, N. V., Никитин, Петр Иванович