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Zakharov A.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Smirnov I.V.,Moscow State University | Serebryakova M.V.,Research Institute of Physicochemical Medicine | Dronina M.A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | And 7 more authors.
Molecular Biology | Year: 2011

Expression of recombinant antibodies in mammalian cells is one of the key problems in immuno-biotechnology. Alternatively, expression of a broad panel of antibodies and of their fragments may be effectively performed in yeast cells. We obtained expression strains of the methylotrophic yeast Pichia pastoris producing single-chain human catalytic antibody A17 (A.17scFv), Fab-fragment (A.17Fab), and full-size light chain (A.17Lch). These antibodies were characterized in terms of functional activity. The capacity to specifically bind and transform organophosphorus compounds has been demonstrated for A.17scFv and A.17Fab. The loss of activity of the antibody light chain when expressed alone indicates that the active site is formed by both heavy and light chains of the antibody. We determined the reversible constant Kd and the first order constant (k2) of the reaction of the covalent modification of A.17scFv and A.17Fab by irreversible inhibitor of the serine proteases p-nitrophenyl 8-methyl-8-azobicyclo[3.2.1]phosphonate (phosphonate X). Calculated values indicate that activity of the antibodies expressed in yeast is similar to the full-size antibody A17 and to the single-chain antibody A. 17 expressed in CHO and E. coli cells, respectively. © 2011 Pleiades Publishing, Ltd.

Blagodatskikh I.V.,RAS Nesmeyanov Institute of Organoelement Compounds | Tikhonov V.E.,RAS Nesmeyanov Institute of Organoelement Compounds | Postnikov V.A.,Research Institute of Physicochemical Medicine | Kobitskaya (Ivanova) E.M.,RAS Nesmeyanov Institute of Organoelement Compounds | And 2 more authors.
Nanotechnologies in Russia | Year: 2014

Acrylamide-based hydrogel nanoparticles, which had hydrodynamic radii in aqueous dispersions of about 100-180 nm, were prepared by miniemulsion polymerization. The redox initiation at the interfacial boundary made it possible to conduct synthesis at an ambient temperature with a high rate and resulted in latexes of enhanced aggregative stability. The original method for nanoparticle functionalization by aminophenylboronic acid was elaborated for their application as affinity sorbents for 1,2-diols. The ability to reversibly bind ribonucleoside (inosine) and glycated protein (hemoglobin) has been demonstrated. The binding capacity relative to inosine and glycated hemoglobin was determined as a function of the phenylboronate group content. © 2014 Pleiades Publishing, Ltd.

Savinova T.A.,National Research Center < > | Il'Ina E.N.,Research Institute of Physicochemical Medicine | Sidorenko S.V.,Research Institute of Childhood Infections
Molecular Genetics, Microbiology and Virology | Year: 2010

Despite the growing level of resistance to Streptococcus pneumoniae infections, β-lactam antibiotics remain the drugs of choice treating these infections. The resistance of S. pneumoniae to these preparations is mediated by modifications of penicillin-binding proteins (PBPs), which are the targets of antibiotics action. The new approach to detecting mutations in the PBP 1A, 2B, and 2X genes based on the minise-quencing reaction followed by matrix-assisted laser desorption/ionization time of flight (MALDI-ToF) mass spectrometry has been developed in the present study. The evaluation of the prevalence of these mutations in clinical S. pneumoniae isolates (n = 194) with different levels of susceptibility to beta-lactam antibiotics has been performed. In summary, 24 different combinations of mutations (genotypes) have been detected in PBPs. All penicillin-susceptible isolates (n = 49, MIC ≤ 0.06 μg/ml) were characterized by the absence of mutations in all analyzed loci. In PBPs, mutations were detected in 133 (91.7%) out of 145 S. pneumoniae isolates with reduced susceptibility to penicillin (MIC > 0.06 μg/ml), which indicates the high diagnostic sensitivity of this approach. Isolates with MIC → 4 μg/ml (n = 20) possessed multiple mutations in all analyzed genes, which confirms the cumulative effects of the formation of penicillin resistance. At the same time, no association between the presence of mutations in PBP genes and decreased susceptibility to cefotaxime was shown, which makes it possible to suggest significant differences in molecular mechanisms of penicillins and cephalosporins resistance. The suggested method of S. pneumoniae genotyping is appropriate for the individual screening of the susceptibility of isolates to penicillin and the molecular monitoring of the resistance determinants in population. © 2010 Allerton Press, Inc.

Gritskova I.A.,Lomonosov Moscow University of Fine Chemical Technology | Kopylov V.M.,RAS Institute of Chemistry | Simakova G.A.,Lomonosov Moscow University of Fine Chemical Technology | Gusev S.A.,Research Institute of Physicochemical Medicine | And 2 more authors.
Polymer Science - Series B | Year: 2010

The properties of polymer suspensions obtained with the use of organosilicon surfactants of various structures are compared. The polymer suspensions are characterized by a narrow particle size distribution and contain functional groups on their surface. © 2010 Pleiades Publishing, Ltd.

Borovskaya A.D.,Research Institute of Physicochemical Medicine | Il'ina E.N.,Research Institute of Physicochemical Medicine | Savinova T.A.,National Agency for Clinical Pharmacology and Pharmacy | Sidorenko S.V.,National Agency for Clinical Pharmacology and Pharmacy | Govorun V.M.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry
Russian Journal of Bioorganic Chemistry | Year: 2011

Modern phenotypic and genetic methods (with the exception of multilocus sequence typing) do not provide a reliable differentiation of α-hemolytic streptococci of the Mitis group. In this study, we obtained MALDI mass spectra for 28 clinical isolates that were initially identified by standard methods as Streptococcus pheumoniae. The isolate distribution that was obtained in the course of clustering correlated well with the data on multilocus typing. These studied isolates were attributed by this typing to two closely related species: S. pneumoniae and S. mitis. A classification model that differentiated α-hemolytic streptococci with 100% sensitivity and 94.6% specificity was developed on the basis of a genetic algorithm. Two peaks that were different for the S. mitis and S. pneumoniae isolates were revealed by the analysis of statistical diagrams of peak areas. © 2011 Pleiades Publishing, Ltd.

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