Continuous processing using plasma

Abstract

To provide a continuous method for producing a product for the prevention and remediation of bacterial contamination, and an apparatus for producing such a product.SOLUTION: An electrode assembly according to the present invention is for exposing an object supported on a moving surface to an electric discharge and includes a) a discharge device capable of generating an electric discharge, and including an electrode having a discharge generation surface capable of generating an electric discharge emanating from an active portion of the surface, the surface being movable to allow generation of an electric discharge from different portions of the surface, b) a barrier closely spaced relative to an inactive region of the electrode, the barrier separating the active portion of the electrode from the inactive portion of the electrode, and c) means for cooling the inactive portion of the electrode.SELECTED DRAWING: Figure 1

Description

TECHNICAL FIELD

[0001] The present invention relates to a method and apparatus for producing a product to which molecules or macromolecules are attached. Such macromolecules include bacteriophages. Accordingly, the product of the method of the present invention is for preventing and ameliorating bacterial contamination of the product or method of the present invention, or materials that come into contact with the product.

BACKGROUND ART

[0002] Bacteriophages ("phages") are viruses that infect and replicate within bacteria. "Lytic" bacteriophages cause bacterial cells to rupture and open (lyse) and be destroyed after replication of virions within the bacteria. When the cells are destroyed, the phage progeny can find new hosts to infect. Bacteriophages are the most abundant living organisms on Earth. They can be found in all environments where bacteria grow. For example, bacteriophages are detected in groundwater and surface water, soil, foods (e.g., sauerkraut, wine), sewage and sludge. They have also been isolated from humans and animals, for example from feces, urine, saliva, spit, rumen and serum. It is known to apply free bacteriophages to products, for example by spraying. However, such treatments are not known to be reliably effective for the prevention or treatment of bacterial contamination.

[0003] Bacteriophages can penetrate different organs and tissues including the central nervous system, and are part of the gut microbiota along with their bacterial hosts. They account for 10 to 80% of total bacterial mortality in aquatic ecosystems and are an important factor limiting bacterial populations.

[0004] Bacteriophages can be dispersed in both solid and liquid culture media. International patent application WO03 / 093462 discloses that immobilized bacteriophages can be covalently bonded to a substrate by chemical or electrical discharge methods, which are not free to diffuse but retain the ability to infect cells, thereby subsequently causing bacterial lysis and releasing free bacteriophages.

[0005] In WO2007 / 072049, a method was developed for using pulsed-field corona discharge, in which particles activated by the discharge were immersed in a bacteriophage solution. Similarly, in WO2012 / 175749, a bacteriophage solution was combined with activated seeds.

[0006] The therapeutic uses of bacteriophages are well known. WO03 / 093462 discloses a method for immobilizing viruses (particularly bacteriophages) while retaining their biological activity for use as antimicrobial agents. Given that the natural environment of bacteriophages is aqueous, it is widely believed that the dehydration stability disclosed in WO03 / 093462 tends toward natural stability in aqueous media.

[0007] WO2012 / 175749 describes small particles to which bacteriophages are attached, and these particles are suitable for combination with food to improve their bacterial contamination as disclosed herein.

[0008] WO2014 / 049008 describes bacteriophage-laden small particles particularly suitable for the treatment or prevention of bacterial infections.

[0009] Wang et al. (2015) [Reference [1] at the end of the specification] showed that low-pressure plasma treatment (a form of discharge treatment) in an oxygen atmosphere for 1 minute enhanced the adhesion of bacteriophages to surfaces.

[0010] Such surface treatments enable the stable bonding of molecules or polymers (including phages) and are called "activation." Surface activation by corona discharge or other plasma-generating electrical methods is well known (e.g., Barton et al. [2]; Canavan et al. [3]; Recek et al. [4]; Kerkeni et al. [5] and others). In industrial applications (e.g., printing), it is well known that such electrical treatments increase the hydrophilicity (wetting properties) of the surface and thus improve adhesion. This increase in adhesion due to increased hydrogen bonding differs from the use of rapid contact between the electrically activated surface and bacteriophages described in the above patents to achieve covalent bonding.

[0011] Furthermore, US7,250,195 discloses the generation of corona discharge and its introduction into a vacuum chamber for depositing biomaterials (including amino acids and proteins but not bacteriophages) onto a substrate.

[0012] WO2014 / 134297 discloses a method for covalently bonding bacteriophages to a surface by forming carboxylic acid residues on the surface of a substrate to which bacteriophages can covalently bond.

[0013] In this field, only batch manufacturing methods are practical. For example, the material is placed in a shallow activation vessel, subjected to corona activation from above, and immediately afterward brought into contact with a solution containing the bacteriophage to be immobilized. This method is more or less acceptable for sheet (planar) materials where immobilization occurs on a single surface, but its use is inefficient for filamentous and particulate materials.

[0014] Continuous processing methods for articles using corona discharge are known. For example, US2015 / 0373923 discloses a plasma processing apparatus for treating crops (such as sprouts) to reduce bacterial viability, and US2004 / 0086433 discloses an apparatus with a conveyor for exposing objects to corona discharge. Furthermore, US3,017,339 discloses a method and apparatus for processing thermoplastic tubes with corona discharge. However, no method has been developed for continuous processing of articles using corona discharge to adhere polymers (such as bacteriophages) to articles.

[0015] Therefore, a continuous manufacturing method is needed to produce products with covalently bonded bacteriophages. This is especially true for products containing small particles to which bacteriophages are attached. Methods commonly used in industry (e.g., standard corona discharge treatment) are not suitable for activating small particles. Unlike films, small particles are affected by the force caused by the accumulation of static electricity, and furthermore, typically, blown air is used to cool the electrodes in these industrial processes, which does not affect films but can move the small particles and hinder their activation. [Overview of the project] [Problems that the invention aims to solve]

[0016] The object of the present invention is to provide a continuous method for producing products for the prevention and improvement of bacterial contamination, and an apparatus for producing such products. [Means for solving the problem]

[0017] Therefore, the present invention provides a method for attaching molecules or polymers to an object, the method comprising the following steps: The process of placing an object on or near a surface; A process of controlling the surface potential relative to its surroundings; A process of activating an object by exposing it to an electrical discharge; and A process of bringing an object into contact with molecules or polymers to which it is to be attached. [Brief explanation of the drawing]

[0018] [Figure 1] A schematic diagram of the apparatus 100 described herein is shown, which is used in a process of treating microparticles to which infectious bacteriophages are attached, as also described herein. [Figure 2] As described herein, an isometric view of the apparatus 200 described herein for processing microparticles to allow infectious bacteriophages to adhere to them is shown. [Figure 3] As described herein, a cross-sectional view of the apparatus 200 described herein for processing microparticles to which infectious bacteriophages are attached is shown. [Figure 4] As described herein, a side view of the apparatus 200 described herein for processing microparticles to allow infectious bacteriophages to adhere to them is shown. [Modes for carrying out the invention]

[0019] The object does not need to be in direct contact with the surface, but it needs to be close enough to the surface that the surface's potential directly influences and / or defines the object's potential.

[0020] The potential of a surface, and thus objects in contact with the surface or in proximity to the surface, can be controlled by electrically grounding the surface. Accordingly, the surface can be earthed (grounded). Alternatively, the potential of a surface, and thus objects in contact with the surface or in proximity to the surface, can be controlled by limiting the charge accumulated by the surface and / or objects that are in contact with or associated with the surface. This is done by continuously dissipating the charge, or dissipating the charge when the net charge reaches a predetermined threshold. That is, all or part of the excess negative or positive charge accumulated by the surface can be dissipated by grounding or other means. The dissipated charge can have a potential of up to 10,000 volts per centimeter and can generate an energy of 500 millijoules during discharge.

[0021] Control of the surface potential can be conveniently achieved by providing surface conductivity. Accordingly, to accommodate grounding, the surface, or an object attached to the surface or a portion thereof, may be electrically conductive. Preferably, the surface, or an object to which the surface is attached or a portion of the surface, is partially or entirely metallic, and thus inherently electrically conductive.

[0022] The discharge may be a corona discharge. The generation of the corona discharge requires a source of electrical resistance in the apparatus containing the corona source, which can be provided from the electrode, around the grounding / treatment region, or both.

[0023] The corona discharge generating electrode may be made of ceramic. Ceramic electrodes are advantageous and suitable for generating corona discharges, particularly when the treatment platform is electrically conductive, i.e., contains metallic elements or conductive materials. In addition, ceramic electrodes are resistant to degradation by high voltages, so they advantageously have an extended service life.

[0024] The use of ceramic as the resistance around the electrode advantageously distributes the discharge force uniformly while achieving effective and reliable treatment.

[0025] Alternatively, a metal electrode with a source of electrical resistance attached to the ground area may be used in applications where the processing effect needs to travel over a larger distance.

[0026] Furthermore, a small electrical resistance is provided by using a conveyor belt included in this apparatus or used in this manner. This resistance is advantageous because it helps to maintain the effectiveness of the processing evenly.

[0027] Alternatively, the potential of the surface may be controlled by a second electrode positioned to counteract or regulate the potential caused by the surface and any object placed thereon being exposed to a discharge.

[0028] Preferably, this additional electrode is positioned on the opposite side of the object and / or surface from the power source.

[0029] In this way, the potential of an object on or in contact with a surface is also controlled by means of controlling the electrical (electrostatic) potential of the surface. This can also be true when an object is indirectly in contact with a surface, and its potential can also be controlled in coordination with the surface potential.

[0030] The surface is typically a moving surface, thereby enabling the continuous adhesion of molecules / polymers to an object. The surface can be a conveyor platform. It is a moving surface suitable for transporting objects along a predetermined path. Preferably, the surface is a conveyor belt. Therefore, a second additional electrode for controlling the electrostatic potential of the surface and the objects on it may be placed beneath the conveyor platform.

[0031] Therefore, the present invention is particularly suitable for continuous processing. Continuous processing is in contrast to batch processing. In batch processing, a fixed amount of material or input substrate is processed by a method having predetermined start and end points. In contrast, in continuous processing, the amount of material or material to be processed is determined by the amount of available material and the time the process is performed. Once the processing process and the flow of input and output are stable, the product of the method may be produced for an indeterminate period until the continuous process is stopped.

[0032] Consequently, electrostatic effects are not as important or harmful to batch processes because the opportunities for static charge accumulation are significantly less. Furthermore, while continuous belt-driven processes require flexible surfaces, batch processes can be built around rigid (i.e., inflexible) surfaces such as metal plates. Additionally, because electrical activation is only used intermittently, the opportunities for accumulated charge to dissipate naturally are significantly greater.

[0033] To date, the processing of articles using electrical discharge has been carried out on a wide variety of substrate materials and objects without adverse effects on object positioning due to excess charge. Generally, the size and / or mass of the processed object was sufficient to negate any forces caused by the presence or accumulation of charge.

[0034] Therefore, it was unexpected that the processing of particles using electrical discharge would be substantially prevented by the particles becoming charged during the processing process. The movement of all or part of the group of particles being processed reduces the reliability of the processing, and the processing is less than expected and / or required.

[0035] Therefore, the inventors have developed a method and apparatus of the present invention that enables control of potential, resulting in the advantage of eliminating, reducing, or otherwise controlling electrostatic effects that may act on the object being processed. Small objects are disproportionately affected by static effects, in that static charge can cause small particles to repel or attract each other, leading to uncontrolled dispersion or uncontrolled aggregation. In either case, efficient processing of the particles becomes difficult or impossible. Particles with excessively high charge are repelled by discharge to the extent that they do not pass through the corona and are not activated. Furthermore, the accumulation of static electricity can disperse small particles in a random and unpredictable manner, thereby preventing efficient activation by discharge. Such effects are particularly pronounced and therefore unfavorable when the particles are formed in a single layer.

[0036] Furthermore, these electrostatic effects on the object being processed can be caused or aggravated by exposure to external discharge.

[0037] This object provides a substrate for the attachment of molecules or polymers to it. Therefore, the terms "object" and "substrate" are used interchangeably herein.

[0038] The term “substrate” is understood to mean any solid-phase material on which molecules or polymers can be immobilized after exposure to a suitable discharge. The substrate can take many forms, but is a variety of plastics or microbeads, including nylon and any other polymer having amino or carboxyl surface groups, cellulose or other hydroxyl-containing polymers, polystyrene or other similar polymers, magnetic particles, and biological materials. The substrate may also be made from materials commonly used in therapeutic / medical applications. For example, nylon sutures for use in surgery; plastics, lint or gauze materials for treating open wounds; microbeads (which are ingestible); adhesives (e.g., cyanoacrylate); and / or biological materials (e.g., collagen or hyaluronic acid). More preferably, the material may be food or feed particles containing proteins and bulk materials (e.g., cellulose or other plant-derived polymers).

[0039] The term “activated / activated / activated” in this invention is understood to mean the activation of the substrate by reacting it with various chemical groups (leaving surface chemistry capable of binding to bacteriophage heads or capsid groups). The activation of the substrate is achieved by electrical discharge (preferably corona discharge). However, other forms of discharge may be used. In particular, the activation of the substrate can be achieved by applying pulsed corona discharge and / or an electron beam. Contact between molecules or polymers and the activated substrate leads to the formation of covalent bonds between the molecules or polymers and the substrate. Therefore, the use of the term “attached” in the context of this invention means covalent bonding.

[0040] The substrate may be particles. The particles may be 2 mm or less in diameter, preferably 1 mm or less, and most preferably in the range of 1 to 1000 μm. The particles may be substantially spherical.

[0041] Appropriately, the particles may be food pellets. Food pellets are suitable for consumption by humans and / or animals. Preferably, food pellets are suitable for use as animal feed. The pellets may preferably be in the range of 1 mm to 20 mm in height, measured from the surface on which they are placed or supported. The pellets may be substantially spherical, cylindrical, or spherical-cylindrical.

[0042] Furthermore, the object may be a seed (e.g., a plant seed). Seeds can be advantageously handled by the present invention due to the effects of static electricity on the handling of small objects such as plant seeds. Seeds suitable for treatment according to the present invention may be 10 mm or less in diameter, preferably in the range of 0.2 to 5 mm.

[0043] The molecule or polymer may be a biomolecule or an organism within a living body. The biomolecule may be an antibody, cytokine, or other signaling molecule. Alternatively, the molecule or polymer may be a macromeric protein or antibody, or a biological entity such as a virus. Preferably, the polymer is a virus, and most preferably, the polymer is a bacteriophage.

[0044] The term "bacteriophage" in this invention refers to a bacteriophage that infects a specific strain of bacteria. It is understood by those skilled in the art that bacteriophages can recognize and infect specific bacterial strains. Therefore, the substrate may be a material that can be advantageously activated to enable the head-specific binding of complex bacteriophages. Bacteriophages are immobilized via covalent bonds formed between the bacteriophage coat protein and the substrate. More preferably, bacteriophages are immobilized to the substrate via their heads or nucleocapsids by activating the substrate before the addition and coupling of the bacteriophages.

[0045] Therefore, bacteriophages immobilized on a substrate according to the present invention can be used as a “bactericide” to combat strain-specific bacterial infections by inducing selective killing of bacteria through cell lysis by inhibiting bacterial growth. Bacteriophages immobilized on a substrate can also be used as an antimicrobial / disinfectant to “kill” bacterially contaminated materials. Therefore, preferably, the bacteriophages retain their infectivity when immobilized.

[0046] The surface may also include cavities or indentations for accommodating particles. The openings of the cavities or indentations preferably face the source of the discharge when the particles within them are activated by the discharge. Preferably, the indentations or cavities are sized to accommodate a single particle.

[0047] Preferably, the particles contained by the cavity or indentation have a diameter of 1 cm or more. The force required to process larger particles is significantly higher than that for smaller particles. This is because the force requirement follows an inverse square method for linear increase in size.

[0048] Therefore, the surface may be made of indentations that accommodate the particles. Some of the particles protrude from the cavity or indentation onto the surface of the surface. Thus, the linear range that the corona discharge must cover to effectively treat the protruding portions of the particles can be significantly lower. In this way, a significant and effective portion of the particles can be activated without having to treat the entire particle. Therefore, a lower power discharge source can be used. Such lower power discharge sources are more inexpensive, which is advantageous to obtain, run, and maintain.

[0049] The present invention also provides an apparatus for processing an object by attaching molecules or polymers, comprising: A surface for supporting the particles to be processed; Means for controlling the potential of the surface relative to its surroundings; A discharge device capable of generating an electrical discharge.

[0050] The present invention also provides the use of the apparatus of the present invention for attaching molecules or polymers to particles.

[0051] The present invention also provides an electrode assembly comprising: The device comprises a discharge device capable of generating an electrical discharge, and further comprises an electrode having a discharge generating surface capable of generating an electrical discharge from an active portion of its surface, wherein the surface is movable to allow the generation of electrical discharges from different portions of the surface.

[0052] Discharges can occur continuously while the discharge-generating surface is moving.

[0053] The advantage of having a movable electrode surface is that the thermal energy that must be dissipated during the electrode's operation is generated in a specific part of the electrode, and this part can be changed during use, allowing for a more even distribution of the thermal energy generated during operation. In this way, hot spots on the electrode are avoided, and the operating life of the electrode is increased along with improved reliability.

[0054] The movable electrode is rotatable, and preferably the electrode is cylindrical. Such a configuration is convenient in that only unidirectional movement is required to produce the advantage of moving the electrode during continuous use.

[0055] The electrode assembly may further include means for cooling the inert portion of the electrode. Preferably, the cooling means is air cooling, and most preferably, the cooling means is forced air cooling.

[0056] The electrode assembly includes a barrier positioned closely spaced relative to the inert region of the electrode, which separates the active portion from the inert portion. This configuration has the advantage of separating the "active" side of the electrode, where the object is activated, from the "inert" side of the electrode, which can be cooled. In this way, the two temperature environments are separated, preventing the cooling mechanism from affecting the processing mechanism on the other side of the barrier.

[0057] The barrier can be made from rubber or plastic, but preferably, the barrier is made from silicone material.

[0058] The barrier may be in contact with the electrode surface or closely separated from the electrode surface, preferably located 10 to 1000 μm from the electrode surface.

[0059] Such materials and configurations have the advantage of allowing the barrier to act as a scraper to remove particles or other substances adhering to the electrode surface. In this way, the electrode can operate continuously for a longer period of time.

[0060] The present invention also provides the use of the electrode assembly of the present invention for attaching molecules or polymers to a substrate.

[0061] The present invention is illustrated in the following specific embodiments with reference to the accompanying drawings. [Examples]

[0062] (Example 1) Figure 1 shows a device 100 for processing microparticles so that infectious bacteriophages can adhere to them.

[0063] The following components of the device are shown in Figure 1: 101 - Hopper containing fine particles that accumulate on a belt; 102- This is a moving belt, which is manufactured from a conductive material or manufactured to be conductive by the addition of a second electrode. This grounds the particles, thus greatly reducing or eliminating the problem of electrostatic charge buildup; 103- The barrier that crosses the belt ensures that one particle (a single layer) enters the corona region; 104- Rotating ceramic electrode, which generates corona to activate the material. The rotating electrode allows cooling of the top surface through airflow; 105 - Air flows over the top of the electrodes to allow cooling; The 106-silicone polymer blade separates the feed from the airflow and scoops up any adhering particles from the electrode surface; 107 - A barrier to prevent liquid from coming into contact with the corona; 108. A device for spraying a phage suspension onto an activated feed, the device having a reservoir for the phage suspension; 109 - Hopper for collecting phage immobilized feed.

[0064] Thus, during operation, the moving (conveyor) belt 102 has fine particles deposited on it from the hopper 101. The particles are 2 mm or less in size, and the electrostatic effect can cause them to move during diffusion or disperse in an uncontrolled manner. The particles deposited on the belt move under the barrier 103 and travel laterally across the belt 102, forming clumps of particles on the belt 102, flattening into a single layer in preparation for bulk processing of the particles.

[0065] The conveyor surface is manufactured from a conductive material, or its potential is manipulated using a second electrode. This serves to ground or earth the particles, eliminating static electricity and thereby preventing the movement of fine particles on the conveyor due to static electricity. This effect is particularly pronounced when the particles are formed in a single layer.

[0066] A single layer of particles moving along the conveyor then passes through the corona discharge 110, causing activation of the particle surface. This activation generates reactive free radicals on the particle surface.

[0067] The electrode that generates the corona discharge 104 is preferably ceramic. This is because the conveyor belt is conductive, and therefore ceramic electrodes are preferable due to their compatibility. The ceramic electrode 104 is cylindrical and rotates while generating the corona discharge 110. The lowest part of the curved surface of the cylindrical electrode 104 is positioned laterally in the direction of movement of the conveyor belt and faces the particles traveling on the conveyor belt 102. The electrode generates the corona discharge from this lowest part of the surface facing the conveyor belt.

[0068] The resonant rotation of the cylindrical electrode 104, while generating corona discharge, causes the electrode surface to rotate, thereby constantly changing the active portion of the electrode as previously inactive parts of the electrode become used. The generation of corona discharge generates heat, causing the temperature of the active portion of the electrode to rise. The rotation of the electrode allows the previously active portion of electrode 104 to move into the cooling region 105, where air flows over the inactive upper portion of electrode 104, cooling it before that portion of the electrode becomes active again.

[0069] Furthermore, the silicone polymer blades 106 are positioned closely and parallel to each other along the length of the curved cylindrical surface of the ceramic electrode. The presence of these silicone polymer blades 106 allows for the separation of the atmosphere on the "active" side of the electrode from the atmosphere on the "inactive" side. In this way, the effectiveness of the cooling system through which air passes on the "inactive" side of the electrode is improved, and the cooling air passing through the "inactive" side is prevented from blowing onto or affecting the fragile single layer of particulate matter generated on the conveyor belt facing the "active" side of the electrode. In addition, the arrangement of the blades 106, closely and parallel to these curved surfaces of the electrode 104, ensures that any particles that may have adhered to the electrode surface for static charge or other reasons are scooped up from the rotating electrode during the processing.

[0070] Next, the activated particles on the conveyor pass through the opening of screen 107 and enter the spray area where a suspension of bacteriophages in the carrier liquid is sprayed onto the activated particles from reservoir 108. By spraying the bacteriophages onto the activated particles, the surface of the bacteriophages reacts with free radicals present on the surface of the activated particles, causing the bacteriophages to covalently bond to the particles. Bacteriophages attached in this way can retain their infectivity and become significantly less susceptible to dehydration, chemical activation, or predation. The suspension is 1 × 10⁶ per 1 ml. 4 ~1 × 10 12 The suspension contains [number] bacteriophages. The suspension is applied by passing it through a spray nozzle, forming a spray with droplets of average diameter Y μm. A pump is used to apply the suspension at a flow rate of Y ml per second. The spray nozzle is positioned Y mm away from activated phages while the bacteriophage-containing spray is being applied.

[0071] Once the bacteriophage covalently binds to the particle, the conveyor sends the particle to the storage hopper 109.

[0072] (Example 2) The apparatus 200 for processing microparticles to allow infectious bacteriophages to adhere to them is shown in Figures 2 and 3.

[0073] The following components of the device are shown in Figures 2 and 3: 202 is a moving belt, the potential of which and the potential of the items placed on it are controlled via a second / opposite electrode 210. This grounds the particles, thus significantly reducing or eliminating the problem of electrostatic accumulation. 204- Rotating ceramic electrode, which generates corona to activate the material. The rotating electrode allows cooling of the top surface through airflow; 205 - Cooling pipe, which supplies / flows air over the top of the electrodes to enable cooling; The 206-silicone polymer blade separates the feed from the airflow and scoops up any adhering particles from the electrode surface; 207 - A barrier to prevent liquid from coming into contact with the coronavirus; 210-counter electrode; and 212 - High-voltage (HV) cable.

[0074] Thus, during operation, the moving (conveyor) belt 202 has fine particles deposited on it from a hopper or other container (not shown). The particles are of a size such that electrostatic effects can move them during diffusion or disperse them in an uncontrolled manner. The particles deposited on the belt move under a barrier (not shown) and travel laterally across the belt 202, forming clumps of particles on the belt 202, flattening them into a single layer in preparation for bulk processing of the particles.

[0075] The potential on the conveyor surface is manipulated using a second electrode that acts as a counter electrode. This helps control the electrostatic charge generated by the particles, thereby controlling or preventing the electrostatic movement of fine particles on the conveyor. This effect is particularly pronounced when the particles are formed in a single layer.

[0076] A single layer of particles traveling along a conveyor belt then passes through a corona discharge, which causes activation of the particle surface. This activation generates reactive free radicals on the particle surface.

[0077] The electrode that generates the corona discharge 204 is made of ceramic. The ceramic electrode 204 is cylindrical and rotates while generating the corona discharge. The lowest part of the curved surface of the cylindrical electrode 204 is positioned laterally to the direction of movement of the conveyor belt and faces the particles traveling on the conveyor belt 202. The electrode generates the corona discharge from this lowest part of its surface facing the conveyor belt. The electrical energy that generates the corona discharge is supplied via a high-voltage (HV) cable 212. The HV cable is kept at a safe distance of 60 mm from other conductive materials.

[0078] As the cylindrical electrode 204 rotates due to resonant rotation while generating corona discharge, the electrode surface rotates, causing previously inert portions of the electrode to be used, and the active portion of the electrode to constantly change. The generation of corona discharge generates heat, causing the temperature of the active portion of the electrode to rise. The rotation of the electrode allows the previously active portion of electrode 204 to move closer to the cooling pipe 205, where air flows over the inert upper portion of electrode 204, cooling it before that portion of the electrode becomes active again.

[0079] Furthermore, the silicone polymer blades 206 are positioned closely and parallel to each other along the length of the curved cylindrical surface of the ceramic electrode. The presence of these silicone polymer blades 206 allows for the separation of the atmosphere on the "active" side of the electrode from the atmosphere on the "inert" side. In this way, the effectiveness of the cooling system through which air passes on the "inert" side of the electrode is improved, and the cooling air passing through the "inert" side is prevented from blowing onto or affecting the fragile single layer of particulate matter generated on the conveyor belt facing the "active" side of the electrode. In addition, the arrangement of the blades 206, closely and parallel to these curved surfaces of the electrode 204, ensures that any particles that may have adhered to the electrode surface for static charge or other reasons are scooped up from the rotating electrode during the processing.

[0080] Next, the activated particles on the conveyor pass through an opening in a screen (not shown) into a spray area where a suspension of bacteriophages in a carrier liquid is sprayed onto the activated particles from a reservoir (not shown). By spraying the bacteriophages onto the activated particles, the surface of the bacteriophages reacts with free radicals present on the surface of the activated particles, causing the bacteriophages to covalently bond to the particles. Bacteriophages attached in this way can retain their infectivity and become significantly less susceptible to dehydration, chemical activation, or predation. The suspension is 1 × 10⁶ per 1 ml. 4 ~1 × 10 12 It contains [number] bacteriophages. The suspension is applied by passing the suspension through a spray nozzle to form a spray that consists of droplets.

[0081] Once the bacteriophage covalently binds to the particle, the conveyor sends the particle to a storage hopper (not shown).

[0082] The present invention thus provides a method and apparatus for treating a substrate to which a substance can adhere, and the use of an apparatus and product provided by applying the method of the present invention using the apparatus.

[0083] (References) [1]C. Wang,D. Sauvageau,A. Elias Immobilization of Active Bacteriophages on Polyhydroxyalkanoate Surfaces. ACS Appl. Mater. Interfaces 2016,8,1128-1138 [2] Barton, D.; Short, RD; Fraser, S.; Bradley, JW The Effect of Ion Energy upon Plasma Polymerization Deposition Rate for Acrylic Acid. Chem. Commun. (Cambridge, U. K) 2003, 7, 348-349. [3] Canavan, HE; ​​Cheng, [4]Recek,N.;Mozetic,M.;Jaganjac,M.;Milkovic,L.;Zarkovic,N.;Vesel,A. Adsorption of Proteins and Cell Adhesion to Plasma Treated Polymer Substrates. Int. J. Polym. Mater. 2014,63,685-691. [5]Kerkeni,A.;Behary,N.;Dhulster,P.;Chihib,N.;Perwuelz,A. Study on the Effect of Plasma Treatment of Woven Polyester Fabrics with Respect to Nisin Adsorption and Antibacterial Activity. J. Appl. Polym. Sci. 2013,129,866-873.

Claims

1. An electrode assembly for exposing an object supported on a moving surface to an electrical discharge, comprising: a) A discharge device capable of generating an electrical discharge, comprising an electrode having a discharge generating surface including an active portion and an inactive portion, wherein the active portion is capable of generating an electrical discharge originating from the active portion of the surface, and the discharge generating surface is movable to allow the generation of such discharge from different portions of the discharge generating surface; b) A barrier positioned in close proximity to the inert portion of the electrode, the barrier separating the active portion of the electrode from the inert portion; c) Means for cooling the inert portion of the electrode.

2. The electrode assembly according to claim 1, wherein the discharge occurs continuously while the discharge generating surface moves.

3. The electrode assembly according to claim 1 or 2, wherein the discharge is a corona discharge.

4. The electrode assembly according to any one of claims 1 to 3, wherein the means for cooling the inert portion of the electrode is air cooling.

5. The electrode assembly according to claim 1, wherein the means for cooling the inert portion of the electrode is forced air cooling.

6. The electrode assembly according to any one of claims 1 to 5, wherein the barrier is manufactured from rubber or plastic.

7. The electrode assembly according to claim 6, wherein the barrier is manufactured from a silicone material.

8. The electrode assembly according to claim 3, wherein the discharge generating surface is ceramic.

9. The electrode assembly according to claim 3, wherein the discharge generating surface is made of metal.

10. A method for exposing an object to an electrical discharge, comprising the step of exposing the object to an electrical discharge generated by an electrode assembly according to any one of claims 1 to 9.

11. A method according to claim 10 for attaching molecules or polymers to an object, a) A step of activating the object by exposing it to the discharge generated by the electrode assembly described in any one of claims 1 to 9, and b) A step of bringing the object into contact with the molecule or polymer to which it is attached.

12. A method according to claim 11 for attaching a molecule or polymer to an object, wherein the molecule or polymer is a bacteriophage.

13. An apparatus for exposing an object to an electrical discharge, comprising an electrode assembly according to any one of claims 1 to 9.

14. Use of the electrode assembly according to any one of claims 1 to 9 for exposing an object to an electrical discharge.

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