Laboratory of molecular immunology

Department of immunology

Head: Sergey Deev, corresponding member of the academy of sciences
deyev@mail.ibch.ru+7(495)429-88-10

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Molecular Immunology Laboratory at the Moscow River.

NamePositionContacts
Sergey Deev, corresponding member of the academy of sciencesdepart. dir.deyev@mail.ibch.ru+7(495)429-88-10
Galina Proshkina, Ph.D.s. r. f.+7(499)724-71-88
Ekaterina Lebedenko, Ph.D.s. r. f.elebedenko@mail.ru+7(495)3306392, +7(926)2417030, +7(499)1510178
Aleksej Shul'ga, Ph.D.s. r. f.schulga@gmail.com
Maxim Nikitin, Ph.D.s. r. f.max.nikitin@gmail.com+7(495)330-63-92
Ekaterina Souslova, Ph.D.r. f.souslova@gmail.com
Yuri Khodarovichr. f.khodarovich@mail.ru+7(495)330-64-65
German Kagarlitskiyj. r. f.webdiver@inbox.ru
Elena Shramova, Ph.D.j. r. f.fei@psha.org.ru+7(916)950-35-49
Oleg Stremovskijj. r. f.ostr@mail.ru
Kristina Mironovaj. r. f.kgobova@gmail.com+7(495)3353788
Tatiana Zdobnova, Ph.D.j. r. f.t.zdobnova@mail.ru+7(495)3306392
Victoria Shipunova, Ph.D.j. r. f.viktoriya.shipunova@phystech.edu+7(985)2519909
Elena KonovalovaPhD stud.elena.ko.mail@gmail.com+7()
Ivan ZelepukinPhD stud.ivan.zelepukin@gmail.com+7()
Olga ShilovaPhD stud.olchernykh@yandex.ru+7()
Yaroslav MoiseevPhD stud.biotech.moiseev@gmail.com
Dariya Kiselevastud.darkiseleva@mail.ru
Polina Kotelnikovastud.kotelnikova@phystech.edu+7()

Former members:

Taras Balandinj. r. f.
Sergej Lukashj. r. f.
Boris Veryugineng.boris.veryugin@gmail.com
Dmitrij Karpenkores. eng.
Ol'ga Korol'chukres. eng.olg.kor@gmail.com

Selected publications

  1. Shipunova V.O., Nikitin M.P., Nikitin P.I., Deyev S.M. (2016). MPQ-cytometry: a magnetism-based method for quantification of nanoparticle-cell interactions. Nanoscale , [+]

    Precise quantification of interactions between nanoparticles and living cells is among the imperative tasks for research in nanobiotechnology, nanotoxicology and biomedicine. To meet the challenge, a rapid method called MPQ-cytometry is developed, which measures the integral non-linear response produced by magnetically labeled nanoparticles in a cell sample with an original magnetic particle quantification (MPQ) technique. MPQ-cytometry provides a sensitivity limit 0.33 ng of nanoparticles and is devoid of a background signal present in many label-based assays. Each measurement takes only a few seconds, and no complicated sample preparation or data processing is required. The capabilities of the method have been demonstrated by quantification of interactions of iron oxide nanoparticles with eukaryotic cells. The total amount of targeted nanoparticles that specifically recognized the HER2/neu oncomarker on the human cancer cell surface was successfully measured, the specificity of interaction permitting the detection of HER2/neu positive cells in a cell mixture. Moreover, it has been shown that MPQ-cytometry analysis of a HER2/neu-specific iron oxide nanoparticle interaction with six cell lines of different tissue origins quantitatively reflects the HER2/neu status of the cells. High correlation of MPQ-cytometry data with those obtained by three other commonly used in molecular and cell biology methods supports consideration of this method as a prospective alternative for both quantifying cell-bound nanoparticles and estimating the expression level of cell surface antigens. The proposed method does not require expensive sophisticated equipment or highly skilled personnel and it can be easily applied for rapid diagnostics, especially under field conditions.

    ID:1530
  2. Shipunova V.O., Nikitin M.P., Zelepukin I.V., Nikitin P.I., Deyev S.M., Petrov R.V. (2015). A comprehensive study of interactions between lectins and glycoproteins for the development of effective theranostic nanoagents. Dokl. Biochem. Biophys. 464 (1), 315–8 [+]

    A comprehensive study of the interactions between lectins and glycoproteins possessing different glycosylation profiles in the composition of nanoparticles was carried out in order to find specifically interacting protein pairs for the creation of novel classes of multifunctional nanoagets that based on protein-assisted selfassembly. We obtained information about specific interactions of certain lectins with selected glycoproteins as well as about the ability of certain monosaccharides to competitively inhibit binding of glycoproteins with lectins. These protein-mediated interactions may be involved in the formulation of self-assembled nanoparticles for therapy and diagnostics of various diseases.

    ID:1322
  3. Generalova A.N., Kochneva I.K., Khaydukov E.V., Semchishen V.A., Guller A.E., Nechaev A.V., Shekhter A.B., Zubov V.P., Zvyagin A.V., Deyev S.M. (2015). Submicron polyacrolein particles in situ embedded with upconversion nanoparticles for bioassay. Nanoscale 7 (5), 1709–17 [+]

    We report a new surface modification approach of upconversion nanoparticles (UCNPs) structured as inorganic hosts NaYF4 codoped with Yb(3+) and Er(3+) based on their encapsulation in a two-stage process of precipitation polymerization of acrolein under alkaline conditions in the presence of UCNPs. The use of tetramethylammonium hydroxide both as an initiator of acrolein polymerization and as an agent for UCNP hydrophilization made it possible to increase the polyacrolein yield up to 90%. This approach enabled the facile, lossless embedment of UCNPs into the polymer particles suitable for bioassay. These particles are readily dispersible in aqueous and physiological buffers, exhibiting excellent photoluminescence properties, chemical stability, and also allow the control of particle diameters. The feasibility of the as-produced photoluminescent polymer particles mean-sized 260 nm for in vivo optical whole-animal imaging was also demonstrated using a home-built epi-luminescence imaging system.

    ID:1431
  4. Guller A.E., Generalova A.N., Petersen E.V., Nechaev A.V., Trusova I.A., Landyshev N.N., Nadort A., Grebenik E.A., Deyev S.M., Shekhter A.B., Zvyagin A.V. (2015). Cytotoxicity and non-specific cellular uptake of bare and surface-modified upconversion nanoparticles in human skin cells. Nano Research 8 (5), 1546–1562 [+]

    The cytotoxicity and non-specific cellular uptake of the most popular composition of upconversion nanoparticle (UCNP), NaYF4:Yb3+:Er3+, is reported using normal human skin cells, including dermal fibroblasts and immortalized human epidermal linear keratinocytes (HaCaT). A new hydrophilization reaction of as-synthesized UCNPs based on tetramethylammonium hydroxide (TMAH) enabled evaluation of the intrinsic cytotoxicity of bare UCNPs. The cytotoxicity effects of the UCNP surface-coating and polystyrene host were investigated over the concentration range 62.5–125 μg/mL with 24-h incubation, using a MTT test and optical microscopy. The fibroblast viability was not compromised by UCNPs, whereas the viability of keratinocytes varied from 52% ± 4% to 100% ± 10% than the control group, depending on the surface modification. Bare UCNPs reduced the keratinocyte viability to 76% ± 3%, while exhibiting profound non-specific cellular uptake. Hydrophilic poly(D,L-lactide)- and poly(maleic anhydride-alt-1-octadecene)-coated UCNPs were found to be least cytotoxic among the polymer-coated UCNPs, and were readily internalized by human skin cells. Polystyrene microbeads impregnated with UCNPs remained nontoxic. Surprisingly, no correlation was found between UCNP cytotoxicity and the internalization level in cells, although the latter ranged broadly from 0.03% to 59%, benchmarked against 100% uptake level of TMAH-UCNPs.

    https://static-content.springer.com/image/art%3A10.1007%2Fs12274-014-0641-6/MediaObjects/12274_2014_641_Fig1_HTML.gif
    ID:1430
  5. Поляновский О.Л., Лебеденко Е.Н., Деев С.М. (2013). ErbB-онкогены - мишени моноклональных антител. Биохимия 77 (3), 289–311 ID:872
  6. Shipunova V.O., Nikitin M.P., Lizunova A.A., Ermakova M.A., Deyev S.M., Petrov R.V. (2013). Polyethyleneimine-coated magnetic nanoparticles for cell labeling and modification. Dokl. Biochem. Biophys. 452 (1), 245–7 ID:1266
  7. Aghayeva U.F., Nikitin M.P., Lukash S.V., Deyev S.M. (2013). Denaturation-resistant bifunctional colloidal superstructures assembled via the proteinaceous barnase-barstar interface. ACS Nano 7 (2), 950–61 [+]

    To date, a number of biomolecule-mediated nanoparticle self-assembly systems have been developed that are amenable to controllable disassembly under relatively gentle conditions. However, for some applications such as design of self-assembled multifunctional theragnostic agents, high stability of the assembled structures can be of primary importance. Here, we report extraordinarily high durability of protein-assisted nanoparticle self-assembly systems yielding bifunctional colloidal superstructures resistant to extreme denaturing conditions intolerable for most proteins (e.g., high concentrations of chaotropic agents, high temperature). Among the tested systems (barnase-barstar (BBS), streptavidin-biotin, antibody-antigen, and protein A-immunoglobulin), the BBS is notable due to the combination of its high resistance to severe chemical perturbation and unique advantages offered by genetic engineering of this entirely protein-based system. Comparison of the self-assembly systems shows that whereas in all cases the preassembled structures proved essentially resistant to extreme conditions, the ability of the complementary biomolecular pairs to mediate assembly of the initial biomolecule-particle conjugates differs substantially in these conditions.

    ID:862
  8. Nikitin M.P., Zdobnova T.A., Lukash S.V., Stremovskiy O.A., Deyev S.M. (2010). Protein-assisted self-assembly of multifunctional nanoparticles. Proc. Natl. Acad. Sci. U.S.A. 107 (13), 5827–32 [+]

    A bioengineering method for self-assembly of multifunctional superstructures with in-advance programmable properties has been proposed. The method employs two unique proteins, barnase and barstar, to rapidly join the structural components together directly in water solutions. The properties of the superstructures can be designed on demand by linking different agents of various sizes and chemical nature, designated for specific goals. As a proof of concept, colloidally stable trifunctional structures have been assembled by binding together magnetic particles, quantum dots, and antibodies using barnase and barstar. The assembly has demonstrated that the bonds between these proteins are strong enough to hold macroscopic (5 nm-3 microm) particles together. Specific interaction of such superstructures with cancer cells resulted in fluorescent labeling of the cells and their responsiveness to magnetic field. The method can be used to join inorganic moieties, organic particles, and single biomolecules for synergistic use in different applications such as biosensors, photonics, and nanomedicine.

    ID:744
  9. (2009). Современные технологии создания неприродных антител для клинического применения. Acta Naturae 1 (1), 30–52 ID:336
  10. Zdobnova T.A., Dorofeev S.G., Tananaev P.N., Vasiliev R.B., Balandin T.G., Edelweiss E.F., Stremovskiy O.A., Balalaeva I.V., Turchin I.V., Lebedenko E.N., Zlomanov V.P., Deyev S.M. (2009). Fluorescent immunolabeling of cancer cells by quantum dots and antibody scFv fragment. J Biomed Opt 14 (2), 021004 [+]

    Semiconductor quantum dots (QDs) coupled with cancer-specific targeting ligands are new promising agents for fluorescent visualization of cancer cells. Human epidermal growth factor receptor 2/neu (HER2/neu), overexpressed on the surface of many cancer cells, is an important target for cancer diagnostics. Antibody scFv fragments as a targeting agent for direct delivery of fluorophores offer significant advantages over full-size antibodies due to their small size, lower cross-reactivity, and immunogenicity. We have used quantum dots linked to anti-HER2/neu 4D5 scFv antibody to label HER2/neu-overexpressing live cells. Labeling of target cells was shown to have high brightness, photostability, and specificity. The results indicate that construction based on quantum dots and scFv antibody can be successfully used for cancer cell visualization.

    ID:332
  11. Schulga A.A., Mechev P.V., Kirpichnikov M.P., Skryabin K.G., Deyev S.M. (2009). Construction of the plasmid-free strain for human growth hormone production. Biochimie 128-129, 148–53 [+]

    The E. coli strain, overproducing human growth hormone (hGH) was made by integration of the hGH gene under the control of T7 promoter into the chromosomal LacZ gene of BL21(DE3) via lambda Red recombineering. The strain gave higher productivity (50 mg·L(-1)·OD550(-1)) and better growth characteristics than the corresponding strain in which the same hGH expression cassette was placed in a plasmid. The protein produced by the plasmid-free strain was purified and characterized to be hGH. The results demonstrates that a plasmid-free recombinant strain having a single-copy gene expression cassette in the chromosome could provide better gene activity regulation, higher productivity, superior growth characteristics, as well as more stringent control of the gene sequence invariance than a plasmid-based strain.

    ID:1669
  12. Deyev S.M., Lebedenko E.N. (2008). Multivalency: the hallmark of antibodies used for optimization of tumor targeting by design. Bioessays 30 (9), 904–18 [+]

    High-precision tumor targeting with conventional therapeutics is based on the concept of the ideal drug as a "magic bullet"; this became possible after techniques were developed for production of monoclonal antibodies (mAbs). Innovative DNA technologies have revolutionized this area and enhanced clinical efficiency of mAbs. The experience of applying small-size recombinant antibodies (monovalent binding fragments and their derivatives) to cancer targeting showed that even high-affinity monovalent interactions provide fast blood clearance but only modest retention time on the target antigen. Conversion of recombinant antibodies into multivalent format increases their functional affinity, decreases dissociation rates for cell-surface and optimizes biodistribution. In addition, it allows the creation of bispecific antibody molecules that can target two different antigens simultaneously and do not exist in nature. Different multimerization strategies used now in antibody engineering make it possible to optimize biodistribution and tumor targeting of recombinant antibody constructs for cancer diagnostics and therapy.

    ID:333

Sergey Deev

  • Russia, Moscow, Ul. Miklukho-Maklaya 16/10 — On the map
  • IBCh RAS, build. 52, office. 558
  • Phone: +7(495)429-88-10
  • E-mail: deyev@mail.ibch.ru