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RU-2861427-C1 - DEVICE FOR PRODUCING PROTEIN NANOPARTICLES

RU2861427C1RU 2861427 C1RU2861427 C1RU 2861427C1RU-2861427-C1

Abstract

FIELD: biotechnology. SUBSTANCE: provided is a device for producing protein nanoparticles, comprising a heating element, a cooling element, a reaction millitube, and temperature sensors. Wherein the heating element is made in the form of two plates made of a thermally conductive material, which are connected to electrodes configured to be connected to a power source and providing heating of the plates of the heating element. The cooling element is made in the form of two radiators with a fan for cooling the cooling agent of the radiators. The reaction millitube for flow millifluidic production of protein nanoparticles is made of a thermally conductive material and is laid sequentially between the plates of the heating element and between the radiators of the cooling element, wherein a pump is connected to the reaction millitube to supply the reaction mixture to the reaction millitube. The temperature sensors are installed in the heating and cooling elements, controllers are connected to them, which are connected to a control unit. EFFECT: efficient production of protein nanoparticles. 9 cl, 8 dwg

Inventors

  • NIKITIN MAKSIM PETROVICH
  • Drozdov Andrei Sergeevich
  • Korenkov Egor Sergeevich
  • Mochalova Elizaveta Nikitichna

Dates

Publication Date
20260505
Application Date
20241225

Claims (13)

  1. 1. A device for producing protein nanoparticles, comprising:
  2. a heating element made in the form of two plates made of heat-conducting material, which are connected to electrodes designed with the possibility of connection to a power source and providing heating of the plates of the heating element;
  3. a cooling element made in the form of two radiators with a fan for cooling the cooling agent of the radiators;
  4. a reaction millitube for flow-through millifluidic production of protein nanoparticles, made of a heat-conducting material and laid sequentially between the plates of the heating element and between the radiators of the cooling element, wherein a pump is connected to the reaction millitube to ensure the supply of the reaction mixture to the reaction millitube;
  5. temperature sensors installed in the heating and cooling elements, connected to the controllers, which are connected to the control unit.
  6. 2. A device for producing protein nanoparticles according to claim 1, wherein the millitube diameter is from 1-2 mm.
  7. 3. A device for producing protein nanoparticles according to claim 1, wherein the pump is designed to provide a flow rate of the reaction mixture in the reaction millitube of 0.3 to 1 ml/min.
  8. 4. A device for producing protein nanoparticles according to claim 1, wherein the control unit is configured to control the temperature conditions of the heating and cooling elements.
  9. 5. A device for producing protein nanoparticles according to claim 1, wherein the heating of the heating element plates is provided in a temperature range from 45 °C to 100 °C.
  10. 6. A device for producing protein nanoparticles according to claim 1, wherein cooling of the radiators of the cooling element is provided in a temperature range from 1 °C to 35 °C.
  11. 7. A device for producing protein nanoparticles according to claim 1, wherein the heating element plates are made of aluminum.
  12. 8. A device for producing protein nanoparticles according to claim 1, in which water is used as a cooling agent for radiators.
  13. 9. A device for producing protein nanoparticles according to claim 1, wherein the reaction millitube is made of an inert and hydrophobic material.

Description

The invention relates to biomedicine and nanomedicine, and in particular to a device for producing protein nanoparticles for biological or medical applications. The invention can be used in medicine, for example, to produce protein nanoparticles encapsulating agents that are medicinal drugs, such as small molecules (antitumor drugs), inorganic nanoparticles, dyes, or mixtures thereof, used as drug delivery vehicles, contrast agents, or markers for immunochromatographic tests, computed tomography (CT), and magnetic resonance imaging (MRI). Nanoparticles (hereinafter referred to as NPs) are currently widely used in biomedicine, including in therapy, diagnostics, biosensors, bioimaging, and other fields. Among these, the use of nanoparticles as drug carriers is a promising approach for improving the efficacy of therapeutic substances by increasing their solubility, bioavailability, reducing toxicity, etc. Nanoparticles are defined as any objects approximately 100 nanometers in size (1 nm = 1 x 10 -9 m), obtained either chemically/synthetically or naturally. The term "nanoparticles" refers to supramolecular structures consisting of more than one molecule. While nanoparticles are often used to refer to objects up to 100 nm in size, within the scope of this invention, objects larger than 100 nm are also understood—preferably up to 5 μm, but in some cases, sizes up to 100 μm are acceptable. Thus, nanoparticles include both nanoparticles and microparticles, microspheres, and the like. Furthermore, the term refers to particles with the aforementioned dimensions in all dimensions, or in at least one. They are ubiquitous in nature and, in some cases, can be formed by living objects, such as magnetosomes in some bacterial species or exosomes in eukaryotes. Due to their small size, nanoparticles have a number of unique physical, chemical and biological properties that are in demand in various fields of biomedicine. Protein nanoparticles have attracted particular attention due to their inherent biocompatibility, biodegradability and non-toxicity. Nanoparticle production (synthesis) is typically performed in batch reactors, which provide limited control over the parameters of the resulting products and a narrow range of possible batch sizes. In contrast, flow-through systems for NP production, typically based on a microfluidic chip, avoid these drawbacks. However, large-scale devices—milifluidic systems—may offer several advantages over microfluidic systems, such as simpler and less expensive production, increased throughput, and reduced channel clogging. Currently, clinically approved compounds exist, such as paclitaxel (Abraxane), bound to albumin nanoparticles. There are numerous methods for producing protein nanoparticles, typically through individual reactions. Among these, rapid thermal formation (RTF) has been developed for producing protein nanoparticles. It has been shown that intense but rapid heating can promote the formation of protein nanoparticles while minimizing impact on protein function and structure. However, for such batch processes, scaling up the reaction while maintaining control over their physicochemical parameters is a complex task, significantly hindering the translation from laboratory to clinical or industrial scale methods. Therefore, the authors sought to implement the BTF method in a continuous flow system, as it typically offers significantly better scalability and parameter control. Such flow systems typically utilize a microfluidic chip as a core component. Most often, the processes are based on variations of coacervation or desolvation methods, aimed at rapidly and efficiently mixing the protein with an antisolvent or coagulation trigger. Mixing is facilitated by various physical phenomena, such as ultrasound, alternating electromagnetic fields, turbulent flows within oil droplets, and the specific geometry of the chip. Creating a device for producing these nanoparticles without using microfluidic chips is challenging due to the microscopic nature of the channels, making their production relatively complex and expensive. Furthermore, their use may be associated with limited throughput and an increased risk of channel clogging. The same principles used for microfluidic systems can be partially transferred to millifluidic systems, which are flow-through systems with typical dimensions of 1 mm. Millifluidic systems have been successfully used to produce various types of nanoparticles, such as colloidal gold, liposomes, and polymer nanoparticles. However, information on the millifluidic method for producing protein nanoparticles is limited. In the paper “Millifluidic Synthesis of Biocompatible Protein-Loaded Nanocapsules” (Gwenael Bonfante, Fumiyasu Awai, Takaya Kubo, Hiroshi Segawa, Soo Hyeon Kim, Anthony Genot, Sylvain Chambon. Materials Today Sustainability, publication date: 03.09.2024, https://hal.science/hal-04685938/document), bovine serum albumin (BSA) nanoparticles are obtained usi