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Biomedical subjects

Tatiana B Tennikova

Publications and source records attributed to Tatiana B Tennikova.

8 recordsLinked to original sources

Development of multifunctional polymer-mineral composite materials for bone tissue engineering.

The main goal of this article is the development of a novel approach to construct multifunctional composite scaffolds for bone tissue engineering. For this purpose, different kinds of mineral macroporous supports, water-soluble aldehyde-containing copolymers of N-vinylpyrrolidone, as well as different nonspecific and biospecific ligands governing cell adhesion and growth have been used. The composite materials were tested initially for cytotoxicity in cell culture experiments using a model cell line.

Adsorption↗

Chromatographic investigation of macromolecular affinity interactions.

High-performance monolithic disk affinity chromatography was applied to the investigation of formation of complexes between (1) complementary polyriboadenylic and polyribouridylic acids, e.g. poly(A) and poly(U), respectively, (2) poly(A) and synthetic polycation poly(allylamine), pAA. Polyriboadenylic acid and poly(allylamine) were immobilized on macroporous disks (CIM disks). Quantitative parameters of affinity interactions between macromolecules were established using frontal analysis at different flow rates.

Adsorption↗

Affinity processes realized on high-flow-through methacrylate-based macroporous monoliths.

The technology for preparation of rigid macroporous polymers suggested in the late 1980s has become a powerful instrument for the development of a novel scientific and practical field. At present, monolithic stationary phases are widely used in the processes of bioseparation (chromatography), bioconversion (enzyme reactors) as well as in other processes based on interphase mass distribution (for example, solid phase peptide and oligonucleotide synthesis). Bioaffinity modes of suggested dynamic methods are very promising for their use in different analytical processes (immunological, ecological, medical and other types of analytical monitoring), preparative isolation of blood proteins such as myoglobin, hemoglobin, immunoglobulins, etc. and also recombinant products directly from cell supernatants or lysates. For the first time, it has been shown that bioaffinity pairing with participation of immobilized on carefully designed rigid supports is very fast and the whole process of affinity separation can be realized within second time scale. The principle of bioaffinity recognition is generaly at the construction of biological reactors (for example, enzyme reactors). Improved kinetics of biocatalized reactions is explained by a minimal influence on the surface of the used sorbent. Very perspective field is the use of discussed monoliths for solid phase chemical synthesis of fragments of biological macromolecules (peptides and oligonucleotides). Several examples of these applications will be presented and discussed.

Chromatography, Liquid↗

Monolithic peptidyl sorbents for comparison of affinity properties of plasminogen activators.

Plasminogen activators are the proteases which convert plasminogen into plasmin dissolving, in its turn, the major component of blood clots, fibrin. They are extremely useful in heart attack therapy. Modern and most appropriate way of scaled up production of these valuable proteins is gene engineering. In this case, a separation and a purification of target product become the important steps of the whole process. Recently developed affinity chromatography on short monolithic columns seems to be a very attractive method for these purposes. High speed of a process prevents the protein's denaturation due to temperature or/and solvents influence. The better mass transfer mechanism (convection rather than diffusion) allows considering only biospecific complexing as time limiting step. Specificity of several synthetic peptides to plasminogen activators have been studied by affinity chromatography on short monolithic columns. Peptide ligands were synthesized by conventional solid phase peptide synthesis (SPPS). The immobilization procedure was carried out as a one step process at static conditions. The results of quantitative evaluation of such affinity interactions were compared with those established for plasminogen that is the natural affinity counterpart to both proteases. Additionally, some of investigated peptides were synthesized directly on GMA-EDMA disks and their affinity properties were compared with those established for the case of immobilized ligands. The possibility of using of synthetic peptidyl ligands for plasminogen activators isolation from native cell supernatant and model protein mixtures has been demonstrated.

Adsorption↗

Effect of experimental conditions on strong biocomplimentary pairing in high-performance monolithic disk affinity chromatography.

The effect of flow-rate on quantitatively determined binding parameters for several biocomplementary pairs in affinity mode high-performance monolithic disk affinity chromatography (HPMDAC) has been investigated using frontal analysis approach. Affinity interactions were evaluated from linearized adsorption isotherms and dynamic dissociation constants of the complexes K(diss.) and the theoretical adsorption capacities Q(max) were calculated. HPMDAC isolation of a typical protein trypsin from both buffered solution and artificial mixture as well as biospecific extraction of antibodies against bovine serum albumin and recombinant protein G from such complex mixtures as blood serum and cellular lysate were examined. Immobilized counterparts soybean trypsin inhibitor, bovine serum albumin, and human immunoglobulin G were used in chromatographic experiments. The maximum adsorption capacities obtained at different flow-rates were compared with those determined at static conditions. The dependence of quantitative parameters on the surface density of immobilized ligands has also been explored. Finally, a series of experiments was carried out to evaluate the dependence of dynamic affinity binding on temperature for two complementary pairs.

Affinity Labels↗

Multifunctional fractionation of polyclonal antibodies by immunoaffinity high-performance monolithic disk chromatography.

High-performance monolithic disk chromatography (HPMDC), including its affinity mode, is a very efficient method for fast separations of biological molecules of different sizes and shapes. In this paper, protein and peptide ligands, immobilized on the inner surface of thin, monolithic supports (Convective Interaction Media or CIM disks), have been used to develop methods for fast, quantitative affinity fractionation of pools of polyclonal antibodies from blood sera of rabbits, immunized with complex protein-peptide conjugates. The combination of several disks with different affinity functionalities in the same cartridge enables the separation of different antibodies to be achieved within a few minutes. The apparent dissociation constants of affinity complexes were determined by frontal analysis. Variation of elution flow rate over a broad range does not affect the affinity separation characteristics. Indifferent synthetic peptides used as biocompatible spacers do not change the affinity properties of the ligands. The highly reproducible results of immunoaffinity HPMDC are compared with data obtained by widely used enzyme-linked immunosorbent assay.

Amino Acid Sequence↗

Chromatographic reactors based on biological activity.

In the last decade there were many papers published on the study of enzyme catalyzed reactions performed in so-called chromatographic reactors. The attractive feature of such systems is that during the course of the reaction the compounds are already separated, which can drive the reaction beyond the thermodynamic equilibrium as well as remove putative inhibitors. In this chapter, an overview of such chromatographic bioreactor systems is given. Besides, some immobilization techniques to improve enzyme activity are discussed together with modern chromatographic supports with improved hydrodynamic characteristics to be used in this context.

Biocompatible Materials↗