RU2540312C1 - Method of obtaining magnetic affinity sorbent for separation of recombinant proteins - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: claimed is method of obtaining magnetic affinity sorbent for separation of recombinant proteins. Method includes application of porous silica layer on the surface of magnetic microspheres of volatile ashes from coal combustion and impregnation with solution, containing ions of transition metals. As microspheres fraction of magnetic microspheres, containing 40-41 wt % of glass phase, is used. Application of porous silica layer is realised in hydrothermal conditions. Magnetic microspheres contact with liquid reaction mixture, containing tetraethoxysilane and hexadecyltriammonium bromide as structure-forming agent. After application of silica layer solid phase is separated, washed, thermally processed and activated by boiling in water medium.

EFFECT: invention makes it possible to increase strength of silica envelope on the surface of microspheres without reduction of sorption capacity by medium-size proteins.

7 cl, 2 dwg, 3 tbl, 9 ex

Description

The claimed technical solution relates to affinity sorbents obtained on the basis of technogenic raw materials, in particular magnetic microspheres of volatile energy ashes, and can be used in biochemistry, medical diagnostics and biotechnology for the isolation and detection of recombinant proteins containing polyhistidine sequences.

The expression of recombinant proteins labeled with polyhistidine fragments (from 6 histidine residues or more) is widely used to simplify their isolation from various prokaryotic and eukaryotic systems. For this, metal-ionic affinity sorbents containing chelate-bound transition metal ions (Cu ²⁺ , Ni ²⁺ , Zn ²⁺ , Co ²⁺ ) are used on their surface [Porath J., Carlsson J., Olsson I., Belfrage G. Metal chelate affinity chromatography, a new approach to protein fractionation, Nature 258 (1975) 598-599; Schmitt J., Hess H., Stunnenberg HG Affinity purification of histidine-tagged proteins, Mol. Biol. Rep. 18 (1993) 223-230; Gabert-Porecar V., Menart V. Perspectives of immobilized-metal affinity chromatography, J. Biochem. Biophys. Methods 49 (2001) 335-360]. The adsorption of proteins on metal-chelate sorbents occurs due to the formation of a coordination bond between the immobilized metal ion and electron-donating nitrogen atoms that make up the imidazole histidine ring in the protein molecule.

The improvement of affinity sorbents with coordinatively bound metal ions is carried out in the direction of imparting magnetic properties to them, which makes it possible to manipulate the sorbents using a magnetic field and thus simplifies the procedure for the extraction of recombinant proteins from heterogeneous mixtures. Thus, commercially available magnetically controlled affinity sorbents such as Ni ²⁺ -NTA MABs (Qiagen, Germany) [Schafer F., Romer U., Emmerlich M., Blumer J., Steinert K. Automated high-throughput purification of 6xHis- tagged proteins, J. Biomol. Tech. 13 (2002) 131-142], which consist of a magnetic core based on iron oxides and a polysaccharide shell based on cross-linked agarose with covalently grafted residues of nitrilotriacetic acid (NTA) chelating Ni ^{2+ ions} . Particles of the sorbent Ni ²⁺ -NTA MABs are microspheres with a size of 45-165 μm with a capacity of 5-10 mg / ml for 6His-labeled proteins (~ 20 kDa). Also known are nanoscale (~ 10 nm) metal chelating sorbents with a magnetic core based on FePt [Xu C., Xu K., Gu H., Zhong X., Guo Z., Zheng R., Zhang X., Xu B. Nitrilotriacetic Acid-Modified Magnetic Nanoparticles as a General Agent to Bind Histidine-Tagged Proteins, J. Am. Chem. Soc., 126 (2004) 3392-3393], in which Ni ^{2+ ions are} attached to the surface of FePt nanoparticles also by bonding with nitrilotriacetic acid. The disadvantages of such sorbents are the multi-stage and complexity of their synthesis, which requires the use of a wide range of organic reagents, as well as low chemical stability under conditions of high concentrations of denaturing and chelating reagents used at the sorption stage (EDTA, urea) and sorbent regeneration (imidazole). In particular, when combined with Ni ²⁺ -NTA MABs, an EDTA concentration of not higher than 10 ^-3 M is acceptable. Higher concentrations of EDTA, which are favorable for the isolation of proteins sensitive to the presence of divalent metal cations in the solution, cause the leaching of Ni ^{2+ ions} from the surface media and inactivation of the sorbent. In addition, inactivation of the sorbent also occurs at the stage of desorption during the elution of His-labeled proteins under conditions of high imidazole concentration, which makes it necessary to introduce Me ^{2+ ions} after each adsorption-desorption cycle.

To increase the stability of magnetic solid-phase systems with immobilized Me ^{2+ ion} to the effects of higher concentrations of complexing agents (> 10 ^-3 M), a number of works have proposed magnetically controlled sorbents consisting of a magnetic core based on synthetic magnetite or maghemite and a silica shell, in which chelating polydentate ligand, Ni ^{2+ ions} (Ni ²⁺ SiMAG, Chemicell, Berlin, Germany) are incorporated [Frenzel A., Bergemann C., Kohl G., Reinard T. Novel purification system for 6 His-tagged proteins by magnetic affinity separation / / J. Chromatogr. B 793 (2003) 325-329] or Zn ²⁺ [Bele M., Hribar G., Campelj S., Macovec D., Gabert-Porecar V., Zorko M., Gaberscek M., Jamnik J., Venturini P Zn-decorated silicacoated magnetic nanoparticles for protein binding and controlled release // J. Chromatogr. In 867 (2008) 160-164]. The resulting sorbent is characterized by strong binding of Ni ^{2+ ions} in the silica matrix, which allows multiple use of the sorbent without intermediate reactivation. However, the complexity of the process of obtaining a magnetic component increases the cost of the sorbent.

In the work of [L.A. Frank, V.V. Borisova, T.A. Vereshchagina, E.V. Fomenko, A.G. Anshits, I.I. Gitelzon. Isolation of recombinant proteins using affine magnetic sorbents based on microspheres of energy sols // Applied Biochemistry and Microbiology 45 (2009) 237-242] obtained magnetic sorbent Ni ²⁺ -SiO ₂ / MM based on magnetic microspheres of energy scars from burning coal with an immobilized ion Ni ²⁺ on the surface of the silica layer synthesized by the sol-gel method. The absence of the stage of synthesis of the magnetic component greatly simplifies the procedure for producing a composite sorbent, and the relatively low cost of isolating a narrow fraction of microspheres from magnetic concentrates, which are actually waste products of electricity production, and low cost are the advantages of the sorbent.

According to this method, at the first stage, a silica component is obtained in the form of a hydrogel by the interaction of liquid silicate glass with a solution of ammonium nitrate at a selected ratio of NH ₄ ⁺ / Na ⁺ = 1.2 in the presence of nitric acid in order to maintain a predetermined constant value of pH = 1 in the suspension. Then the precipitate is filtered off, washed thoroughly with distilled water, subjected to cation exchange with ammonium salts to release from chemisorbed sodium ions, then it is again filtered off and washed. In the next stage, the silica component is applied to the surface of the magnetic microspheres by mixing the obtained hydrogel with magnetic microspheres in a ratio of 1: 3, mix thoroughly, then dry in an oven at 120 ° C for 6 hours. A magnetic fraction is taken from a dry sample by magnetic separation, which is repeatedly subjected to the procedure of purification of a non-magnetic silica component by washing with distilled water and subsequent wet magnetic separation. The isolated magnetic product is filtered off and dried at 120 ° C. To modify the surface of composite microspheres, the introduction of nickel ions impregnates the microspheres with a 10% solution of nickel acetate in the ratio of 0.6 g of microspheres to 2 ml of a solution of Ni (CH ₃ COO) ₂ . The samples are dried at room temperature, then calcined at 500 ° C for 6 hours, and then subjected to wet magnetic separation and drying at 120 ° C.

As the magnetic core of the composite sorbent, a narrow fraction of magnetic microspheres isolated from flying ashes from burning Irsha-Borodino coal is used, which was characterized by the following composition (wt.%): SiO ₂ - 4.30; Fe ₂ O ₃ - 83.17. The content of the ferrospinel phase in the microspheres was 76.4 wt.%, And silicate glass phase - 16.5 wt.%.

In the case of a test isolation of a recombinant green fluorescent protein containing a fragment of 6 repeating histidine residues at the NH ₂ end (6His-CLGFP), the adsorption specific capacity of the indicated affinity sorbent was 1-3.5 mg per gram of sorbent.

The disadvantages of this method of obtaining include the multi-stage creation of a silica layer on the surface of magnetic microspheres and the low adhesive strength of the silicate coating, which causes a decrease in the adsorption capacity of the sorbent in the multi-cycle process of protein sorption-desorption. The method adopted for the prototype.

The task is to simplify the synthesis of the sorbent and increase the adhesive strength of the silica shell on the surface of magnetic microspheres without deterioration of the sorption capacity in relation to medium-sized proteins (20-30 kDa).

This problem is solved as follows. To achieve the technical result when obtaining a sorbent, magnetic microspheres of fly ash from burning coal containing an amorphous silicate phase are used as a magnetic component. A porous silica layer is applied to the surface of the microspheres, into which Ni ²⁺ ions are introduced by impregnation with a nickel salt, followed by its thermal decomposition. In contrast to the prototype, the content of silicate glass phase in the magnetic microspheres is about 40 wt.%, And the creation of a porous silica coating is carried out by hydrothermal synthesis with the combined presence of microspheres, hexadecyltrimethylammonium bromide and tetraethoxysilane in the reaction medium followed by high-temperature processing of the product. Then the synthesis product is subjected to activation by boiling in aqueous media with different pH. Hydrothermal synthesis is carried out in dynamic or static conditions, in an acidic or ammonia-alcohol medium at temperatures of 60-120 ° C for 2-120 hours.

The essence of the proposed technical solution is as follows. Separation of concentrates of magnetic microspheres of energy ashes using a combined scheme, including the stages of particle size classification, gravitational and magnetic separation, allows to obtain magnetic microspherical products of stabilized composition and morphology [Sharonova OM, Anshishch NN, Oruzheinikov AI, Akimochkina G.V., Salanov A.N., Nizovsky A.I., Semenova O.N., Anshits A.G. Composition and morphology of magnetic microspheres of coal power ash of the Ekibastuz and Kuznetsk basins // Chemistry for Sustainable Development 11 (2003) 673-682]. The texture of the magnetic microspheres is formed by ferrospinel crystallites with an admixture of minor phases encapsulated in silicate glass of different compositions. Differences in the content of iron, glass phase, and other components lead to the formation of magnetic globules of various morphologies. Glass phase serves as a kind of protective shell that prevents the oxidation of the magnetic component and its potential dissolution in case of contact with aqueous media. Along with this, silicate glass contains on its surface reactive silanol groups ≡ Si-OH, which can serve as anchor groups for fixing d-metal cations according to the type of coordination interaction or, in case of their low content, for bonding an additional functional SiO ₂ layer _{of a} high surface . Considering that nickel ions have 6 ligand binding centers in their coordination sphere, which can be completely saturated by interaction with oxygen atoms of OH groups at their high density, it is important to ensure the unsaturation of at least 2 Me ²⁺ coordination bonds to preserve the binding proteins at the level of sorbent analogues (up to 3.5 mg / g). As shown in the prototype sorbent preparation method, coordination unsaturation of bonds of incorporated Ni ^{2+ ions} can be achieved by introducing an excess of Ni ^{2+ ions} by impregnating the surface with an aqueous solution of nickel salt followed by its thermal decomposition.

Achieving the claimed benefits is ensured by the choice of magnetic microspheres with a high content of silicate component, a method for producing a silicate coating with optimal porous structure and concentration of surface hydroxyl groups, and a method for depositing Ni ^{2+ ions} .

Figure 1 shows an electron microscopic image of the magnetic microspheres of the fraction -0.063 + 0.05 mm.

Figure 2 shows the X-ray diffraction patterns of the reference MCM-41 (1) and modified microspheres (2).

The invention is demonstrated by the following examples.

EXAMPLE 1. To obtain an affinity sorbent, a narrow fraction of magnetic microspheres is used (Fig. 1), isolated from flying ashes from burning Ekibastuz coal, the characteristics of which are shown in Table 1. The conditions for producing the sorbent are shown in Table 2.

Table 1 Characteristics of the initial fraction of magnetic microspheres Fraction, mm / bulk density The content of elements (calculated on oxides), wt.% The content of the ferrospin phase, wt.% The content of glass phase, wt.% Specific surface, m ² / g SiO ₂ Al ₂ O ₃ Fe ₂ O ₃ -0.063 + 0.05 / 1.5 g / cm ³ 26.48 9.44 59.82 48.8 41.2 1,0

The synthesis of a porous silica coating on the surface of magnetic microspheres is carried out by the "template" method in an aqueous acidic medium in a stationary mode at 80 ° C for 120 hours. The reagents were used in the following molar ratio: 100 H ₂ O: 0.9 HCl: 0.11 hexadecyltrimethylammonium bromide (HTABr): 0.13 tetraethoxysilane (TEOS) with a volume ratio of solid phase (microspheres): liquid phase = 1:20. At the end of the synthesis, the microspheres are separated and washed with distilled water until a negative reaction to chloride ions with a solution of 0.1 n AgNO ₃ , dried at 80 ° C, calcined for 4 hours at 540 ° C. According to x-ray phase analysis (Fig. 2) and low-temperature nitrogen adsorption, the modified microspheres contain a phase of mesoporous silicate with a pore diameter of 36 Å, similar to MCM-41 [Meynen, V., Cool, P., Vansant, EF Verified syntheses of mesoporous materials // Micropor. Mesopor. Mater. 125 (2009) 170-223]. A weakly expressed reflex in the region of the main peak of the MCM-41 in the diffractogram of the obtained material (Fig. 2, X-ray diffraction pattern 2) indicates a low content of the synthesized silicate component and / or low ordering of its structure in comparison with the reference sample MCM-41. Based on a comparison of the specific surface area of the obtained material (~ 10 m ² / g) and MCM-41 (~ 1000 m ² / g), the estimated content of mesoporous SiO ₂ in the composition of the microspherical composite material was about 1 wt.%.

The surface activation of the mesoporous coating of the microspheres is carried out by boiling in water for 4 hours.

The magnetic sorbent is tested using the example of isolating a recombinant protein of 27 kDa in size, which contains a fragment of six repeating histidine residues at the NH ₂ end - the green fluorescent protein of the jellyfish Clytia gregaria (6His-CLGFP). Recombinant E. coli BL21-gold cells expressing this protein are suspended in Tris-HCl saline buffer (0.02 M Tris HCl, pH 7, 0.15 M NaCl - TBS) and destroyed using a UD-20 ultrasonic disintegrator (Techpan, Poland ) (6 pulses of 20 sec, 0 ° C). The mixture was centrifuged and the resulting supernatant (“lysate”) containing 6His-CLGFP was used to test samples of the sorbent.

Into a test tube with a weighed portion of sorbent particles add 1 ml of “lysate” and mix for 2 minutes. Sorbent particles are fixed to the wall of the tube using a magnet, and the protein solution is removed with a pipette and the absorption spectrum is recorded on a UVIKON 943 spectrophotometer (Kontron Instruments, Italy). The sorption capacity of the sorbent is estimated by the difference in the optical densities of the solutions at a wavelength of 485 nm before and after absorption using the molar extinction value of 6His-CLGFP in the green region (E ₄₈₅ = 63 · 10 ³ M ^-1 cm ^-1 ) [SV Markova, LP Burakova, LA Frank, S. Golz, KA Korostileva, ES Vysotski. Green-fluorescent protein from the bioluminescent jellyfish Clytia gregaria: cDNA cloning, expression, and characterization of novel recombinant protein // Photochem. Photobiol Sci. 9 (2010) 757-765].

The sorbent without the introduction of Ni ²⁺ cations ^is characterized by protein sorption activity not exceeding 0.6 mg / g, which is most likely associated with the participation of iron ions, which entered the structure of mesoporous silicate during synthesis under acidic conditions, which contribute to the leaching of iron from magnetic microspheres in the liquid phase. For comparison, table 2 shows the results of the sorption capacity of the sorbent taken as a prototype.

After surface modification of the sample by applying Ni ²⁺ cations by impregnation with a 10% solution of Ni (CH ₃ COO) ₂ at a ratio of 0.3 g of microspheres: 1 ml of a solution of Ni (CH ₃ COO) ₂ , followed by calcination at 500 ° C for 4 hours (as in the prototype), the adsorption capacity of the obtained sorbent EMT1-H ₂ O-Ni ²⁺ was 1.3 mg / g

EXAMPLE 2. The conditions for obtaining the sorbent (EMT1-H ₂ OS, Ni ²⁺ ) and testing are the same as in example 1, except that the modification of the silicate coating with cations of Ni ^{2+ is} carried out by sorption from a 10% aqueous solution of Ni (CH ₃ COO) ₂ with stirring at 50 ° C for 8 hours, followed by drying and calcination at 500 ° C for 4 hours. As follows from the data in table 2, the sorbent is characterized by a low adsorption capacity for protein (0.7 mg / g), close to the adsorption capacity of the composite carrier without the deposition of nickel ions. This is probably due to the low sorption of Ni ²⁺ cations on the surface of microspheres and / or to the inactivation of the sorbed forms of Ni ²⁺ due to the complete saturation of the coordination bonds of Ni ²⁺ upon interaction with the silanol coating.

EXAMPLE 3. Obtaining a sorbent (EMT1-HCl-Ni ²⁺ ) and its testing is carried out as in example 1, except that the activation of the surface of the mesoporous coating is carried out by boiling in a 1M HCl solution for 4 hours.

EXAMPLE 4. Obtaining the sorbent EMT1-NH ₄ ⁺ -Ni ²⁺ , surface modification and testing was performed as in example 1, except that the surface activation of the mesoporous coating of the microspheres is carried out by boiling in a 1M solution of NH ₄ OH for 4 hours.

EXAMPLE 5. The synthesis of the mesoporous silicate coating on the surface of the microspheres is carried out in an ammonia-alcohol solution in a dynamic mode in a Teflon autoclave for 2 hours at a temperature of 120 ° C using reagents in the following molar ratio: 1 TEOS: 0.2 HTABr: 22 NH ₃ : 52 EtOH: 475 H ₂ O, with a volume ratio of solid phase: liquid phase = 1:20. The rotation of the autoclave, fixed perpendicular to the axis of the shaft, was carried out at a speed of 30 rpm After the synthesis, the separated solid phase is washed, dried at 80 ° C, then calcined at 540 ° C for 4 hours. Activation, modification of the SiO ₂ shell with Ni cations and testing are carried out according to the method described in example 1.

EXAMPLE 6. Obtaining a sorbent (EMT4-H ₂ O-Ni ²⁺ ) includes the same stages as in example 5, except that the synthesis of the mesoporous coating is carried out in a static mode at a temperature of 80 ° C for 120 hours, and activation, modification of the silica shell and testing are carried out, as in example 1.

EXAMPLE 7. Obtaining a sorbent (EMT-7-NH ⁴⁺ -Ni ²⁺ ) is carried out, as in example 5, but at a temperature of 60 ° C for 72 hours. The activation and modification of the silica shell is carried out, as in example 4, and the testing of the sorbent, as in example 1.

EXAMPLE 8. Obtaining a sorbent (EMT5-NH ₄ ⁺ -Ni ²⁺ ) is carried out, as in example 5, only at a temperature of 80 ° C for 15 hours. The activation and modification of the silica shell is carried out according to the method described in example 4. Testing of the sorbent is carried out as in example 1.

EXAMPLE 9. Obtaining a sorbent (EMT8-NH ₄ ⁺ -Ni ²⁺ ) is carried out, as in example 5, only at a temperature of 60 ° C for 15 hours. Activation and modification of the silicate coating is carried out as in example 4, and testing as in example 1. To elute the adsorbed protein, the particles are washed with phosphate buffer containing 0.25 M imidazole. As a result, the target protein was eluted almost completely. After processing the sorbent with an imidazole solution, the amount of protein transferred to the solution was 4.3 mg / g of sorbent. The observed protein loss (0.1 mg / g) is within the limits of the determination error.

The stability of the sorption properties of the sorbent during repeated use was investigated, the results are shown in Table 3. It was found that the adsorption capacity of the magnetic sorbent EMT8-NH ₄ ⁺ -Ni ^{2+ was} maintained during 3-fold use. After 7-fold use, the capacity decreased by 1.5 times. Under similar conditions, the adsorption capacity of the sorbent prototype decreased by 5 times. The stability of the sorption properties in relation to the green fluorescent protein in a multi-cycle sorption process indicates an increased strength of the silicate coating in comparison with the sorbent obtained by the prototype method.

The above examples confirm the possibility of obtaining a magnetic affinity sorbent with immobilized Ni ^{2+ ions} in the synthesis process containing a smaller number of stages compared to the prototype method. The technical result achieved is also an increased strength of the silicate coating and an adsorption capacity with respect to the recombinant protein of 20-30 kDa (1.3-4.4 mg / g) size comparable with the sorbent prototype.

The implementation of the claimed technical solution is not limited to the given examples. Ferrospheres of other sources with a high content of amorphous silicate phase can be used as a magnetic component. A high affinity of the sorbent for proteins having polyhistidine sequences can be achieved by introducing not only nickel cations, but also other transition metal cations, such as Fe ³⁺ , Cu ²⁺ , Co ²⁺ , Zn ²⁺ . Hydrothermal synthesis of a porous silicate coating can be carried out using other organic structure-forming reagents that require the use of similar synthesis conditions and subsequent heat treatment.

table 2 Conditions for obtaining silica coating on magnetic microspheres and characteristics of sorbents Example (Sample No.) Sample marking Synthesis Conditions S _beats , m ² / g D then Å Adsorption capacity, mg / g The composition of the reaction mixture in the liquid phase, mol T, ° С τ, hour Mode prototype Ni ²⁺ -SiO ₂ / MM Sol-gel method 61 but 1,0 prototype Ni ²⁺ -SiO ₂ / MM Sol-gel method 61 but 3,5 one EMT1-H ₂ O-Ni ² 100 H ₂ O: 0.9 HCl: 0.11 HTABr: 0.13 TEOS 80 120 Static 10.6 36 1.3 2 EMT1-H ₂ OS, Ni ²⁺ 100 H ₂ O: 0.9 HCl: 0.11 HTABr: 0.13 TEOS 80 120 Static 10.6 36 0.7 3 EMT1-HCl-Ni ²⁺ 100 H ₂ O: 0.9 HCl: 0.11 HTABr: 0.13 TEOS 80 120 Static 10.6 36 1,6 four EMT1-NH ₄ ⁺ -Ni ²⁺ 100 H ₂ O: 0.9 HCl: 0.11 HTABr: 0.13 TEOS 80 120 Static 10.6 36 2.0 5 EMT2-H ₂ O-Ni ² 1 TEOS: 0.2 HTABr: 22 NH ₃ : 52 EtOH: 475 H ₂ O 120 2 Dynamic 0.8 polymodal 1.3 6 EMT4-H ₂ O-Ni ² 1 TEOS: 0.2 HTABr: 22 NH ₃ : 52 EtOH: 475 H ₂ O 80 120 Static 3.2 25 2,3 7 EMT7-NH ⁴⁺ -Ni ²⁺ 1 TEOS: 0.2 HTABr: 22 NH ₃ : 52 EtOH: 475 H ₂ O 60 72 Static 7.6 but 2.6 8 EMT5-NH ₄ ⁺ -Ni ²⁺ 1 TEOS: 0.2 HTABr: 22 NH ₃ : 52 EtOH: 475 H ₂ O 80 fifteen Dynamic 2,8 33 2,4 9 EMT8-NH ₄ ⁺ -Ni ²⁺ 1 TEOS: 0.2 HTABr: 22 NH ₃ : 52 EtOH: 475 H ₂ O 60 fifteen Dynamic 8.2 29th 4.4

Table 3 Adsorption properties of the sorbent EMT8-NH ₄ ⁺ -Ni ²⁺ in 7 cycles of protein adsorption-desorption Cycle Adsorption capacity, mg / g The amount of desorbed protein, mg / g I 4.4 4.3 II 4.3 4.2 III 4.3 4.2 IV 4.0 4.1 V 4.3 3,5 VI 3.9 3.3 VII 3.0 2,8

Claims (7)

1. A method of obtaining a magnetic affinity sorbent for the isolation of recombinant proteins, including applying a porous silica layer on the surface of the magnetic microspheres of fly ash from coal combustion and subsequent impregnation with a solution containing transition metal ions, characterized in that the fraction of magnetic microspheres containing 40 is used as microspheres -41 wt.% Glass phase, and the application of the porous silica layer is carried out under hydrothermal conditions, exposing the microspheres to contact with the liquid reaction mixture, with containing tetraethoxysilane and hexadecyltriammonium bromide as a structure-forming agent, followed by isolation of the obtained solid phase, washing, heat treatment and activation by boiling in an aqueous medium. 2. The production method according to claim 1, characterized in that the hydrothermal synthesis is carried out in an aqueous acidic medium in a static mode at a temperature of 80 ° C for 120 hours at a ratio of reagents (mol.): 100 H ₂ O: 0.9 HCl: 0.11 hexadecyltrimethylammonium bromide: 0.13 tetraethoxysilane and a solid to volume ratio (microspheres): liquid phase = 1:20, and the surface of the microspheres is activated by boiling in water for 4 hours, either in a 1 M hydrochloric acid solution or in 1 M solution of ammonium hydroxide. 3. The production method according to claim 1, characterized in that the hydrothermal synthesis is carried out in an ammonia-alcohol medium in a dynamic mode at a temperature of 120 ° C for 2 hours at a reagent ratio (mol.): 1 tetraethoxysilane: 0.2 hexadecyltrimethylammonium bromide: 22 NH ₃ : 52 ethyl alcohol: 475 H ₂ O and a volume ratio of solid phase (microspheres): liquid phase = 1:20, and the surface activation of the microspheres is carried out by boiling in water for 4 hours. 4. The production method according to claim 1, characterized in that the hydrothermal synthesis is carried out in an ammonia-alcohol medium in a static mode at a temperature of 80 ° C for 120 hours at a ratio of reagents (mol.): 1 tetraethoxysilane: 0.2 hexadecyltrimethylammonium bromide: 22 NH ₃ : 52 ethyl alcohol: 475 N ₂ O and a volume ratio of solid phase (microspheres): liquid phase = 1:20, and the surface activation of the microspheres is carried out by boiling in water for 4 hours. 5. The production method according to claim 1, characterized in that the hydrothermal synthesis is carried out in an ammonia-alcohol medium in a static mode at a temperature of 60 ° C for 72 hours at a ratio of reactants (mol.): 1 tetraethoxysilane: 0.2 hexadecyl trimethylammonium bromide: 22 NH ₃ : 52 ethyl alcohol: 475 H ₂ O and a volume ratio of solid phase (microspheres): liquid phase = 1: 20, and surface activation of the microspheres is carried out by boiling in a 1 M NH ₄ OH solution for 4 hours. 6. The production method according to claim 1, characterized in that the hydrothermal synthesis is carried out in an ammonia-alcohol medium in a dynamic mode at a temperature of 80 ° C for 15 hours at a ratio of reactants (mol.): 1 tetraethoxysilane: 0.2 hexadecyltrimethylammonium bromide: 22 NH ₃ : 52 ethyl alcohol: 475 N ₂ O and a volume ratio of solid phase (microspheres): liquid phase = 1:20, and the surface activation of the microspheres is carried out by boiling in a 1 M solution of NH ₄ OH for 4 hours. 7. The production method according to claim 1, characterized in that the hydrothermal synthesis is carried out in an ammonia-alcohol medium in a dynamic mode at a temperature of 60 ° C for 15 hours at a ratio of reagents (mol.): 1 tetraethoxysilane: 0.2 hexadecyltrimethylammonium bromide: 22 NH ₃ : 52 ethyl alcohol: 475 N ₂ O and a volume ratio of solid phase (microspheres): liquid phase = 1:20, and the surface activation of the microspheres is carried out by boiling in a 1 M solution of NH ₄ OH for 4 hours.

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