A transfer module for a film transfer apparatus and a film transfer apparatus

Abstract

The utility model discloses a kind of transfer module and membrane transfer instrument for membrane transfer instrument, including transfer slot, negative plate assembly, first membrane transfer water absorption pad, gel matrix, solid phase carrier, second membrane transfer water absorption pad and positive plate assembly are sequentially arranged from bottom to top in the transfer slot, the transfer slot is used to hold buffer solution, negative plate assembly, positive plate assembly are electrically connected with negative electrode connector, positive electrode connector respectively, the negative plate assembly is immersed in buffer solution, and the positive plate assembly is located above the liquid level of buffer solution. The utility model is used for the transfer module of membrane transfer instrument, negative plate assembly is at bottom, positive plate assembly is above and not completely immersed, reduce the contact area of buffer solution and positive plate assembly, reduce electrolytic reaction rate, slow down pH variation and ion consumption, prolong the service life of buffer solution.

Description

Technical Field

[0001] This utility model relates to a biotechnology device, and more particularly to a transfer module for a transfer apparatus and a transfer apparatus. Background Technology

[0002] Protein transfer technology is an important experimental technique in molecular biology and protein research, widely used in fields such as Western blotting and protein-nucleic acid interaction analysis. Its core principle is to transfer electrophoretically separated proteins from a gel matrix to a solid support (such as a nitrocellulose membrane or PVDF membrane) using an electric field, enabling subsequent antibody-specific detection and analysis.

[0003] Traditional protein transfer techniques are divided into two modes: wet transfer and semi-dry transfer. Wet transfer involves immersing the gel and transfer membrane in a buffer solution and applying a constant voltage at a low temperature (typically 4°C) for a long transfer period (several hours to overnight). It is suitable for transferring large molecular weight proteins, but the operation is cumbersome, time-consuming, and requires a large amount of buffer. Application number 2020231569109 discloses a transfer apparatus, a variation of the wet transfer mode. The apparatus includes a transfer stage with at least one set of interlocking slots. Each interlocking slot houses a reaction module, which includes a positive electrode mounting module, a PVDF membrane, a PAGE gel, and a negative electrode mounting module sandwiched together. A dual-channel peristaltic pump corresponding to the reaction module is installed on one side of the transfer stage. The input of the dual-channel peristaltic pump is connected to a reagent bottle, and its output is connected to the positive and negative electrode replenishment reagent inlets on the reaction module. When the transfer process begins, transfer reagent is added to the positive electrode assembly module through the positive electrode replenishment port. Once a certain amount is added, the reagent flows out from the positive electrode permeation holes on the positive electrode electrophoresis plate and adheres to the first transfer filter paper until it becomes saturated and begins to drip, flowing out as waste liquid. Similarly, the same or other reagents are added to the negative electrode assembly through the negative electrode replenishment port. Once a certain amount is added, the reagent flows through the negative electrode permeation holes on the second transfer filter paper, which also flows out as waste liquid when saturated. Compared to traditional wet transfer methods, this transfer apparatus does not require immersing the positive electrode plate, negative electrode plate, gel, transfer membrane, and filter paper in buffer solution. Instead, it uses a peristaltic pump for circulating wetting, but still suffers from high buffer consumption, increasing the manufacturer's operating costs.

[0004] Semi-dry transfer utilizes multiple layers of filter paper soaked in buffer to create a local electric field environment. Buffer consumption is low, but it is limited by the volume of buffer. During long-term transfer or under high voltage, it may generate heat, leading to protein inactivation. Furthermore, if the filter paper, gel, and membrane are not stacked tightly or air bubbles are present, local transfer failure may occur. High molecular weight proteins (such as >150kDa) may be trapped in the gel and cannot be transferred. If the operation time is too long or the buffer is insufficient, the filter paper may dry out, causing the transfer to be interrupted.

[0005] Therefore, the aforementioned wet transfer printing or its modified structures, as well as the semi-dry transfer printing, all have many problems in use, and there is an urgent need to provide a new transfer instrument to solve these technical problems. Utility Model Content

[0006] To address at least one of the problems existing in the prior art described above, particularly the problem of high buffer consumption, this invention proposes a transfer module for a transfer apparatus. This transfer module immerses the negative electrode assembly in the buffer solution, with the positive electrode assembly positioned above the surface of the buffer solution, or immerses the positive electrode assembly in the buffer solution, with the negative electrode assembly positioned above the surface of the buffer solution. This satisfies the buffer solution requirements during the transfer process and overcomes the problem of insufficient buffer solution leading to transfer failure.

[0007] In a first aspect, this utility model provides a transfer module for a transfer apparatus, comprising a transfer tank, wherein a negative electrode assembly, a first transfer absorbent pad, a gel matrix, a solid support, a second transfer absorbent pad, and a positive electrode assembly are arranged sequentially from bottom to top in the transfer tank. The transfer tank is used to hold a buffer solution. The negative electrode assembly and the positive electrode assembly are electrically connected to a negative electrode connector and a positive electrode connector, respectively. The negative electrode assembly is immersed in the buffer solution, and the positive electrode assembly is located above the surface of the buffer solution. Alternatively, the transfer tank may be provided with a negative electrode assembly, a first transfer membrane absorbent pad, a gel matrix, a solid support, a second transfer membrane absorbent pad, and a positive electrode assembly arranged sequentially from top to bottom. The transfer tank is used to hold the buffer solution. The negative electrode assembly and the positive electrode assembly are electrically connected to the negative electrode connector and the positive electrode connector, respectively. The positive electrode assembly is immersed in the buffer solution, and the negative electrode assembly is located above the surface of the buffer solution.

[0008] Immersing the negative electrode assembly in the buffer solution serves two purposes: firstly, it provides sufficient ions to the negative electrode assembly, maintaining a high electric field during the transfer process and maximizing transfer speed and effectiveness; secondly, the transfer process generates significant Joule heat, and immersion in the buffer solution prevents localized overheating that could negatively impact protein activity, thus maintaining the stability of the transfer environment. Furthermore, with the negative electrode assembly at the bottom and the positive electrode assembly above and not fully submerged, the contact area between the buffer solution and the positive electrode assembly is reduced, lowering the electrolysis rate, mitigating pH changes and ion consumption, and extending the buffer solution's lifespan. Moreover, with only the negative electrode assembly immersed in the buffer solution, only 20-50 mL of buffer solution is required for a single transfer, significantly reducing buffer usage compared to the over 400 mL required for traditional wet transfer methods.

[0009] Similarly, immersing the positive electrode assembly in the buffer solution while placing the negative electrode assembly above it achieves a similar effect. Immersing only one of the positive or negative electrode assemblies, rather than both simultaneously, reduces the amount of buffer solution used. Immersion of either the positive or negative electrode assembly in the buffer solution provides a sufficient ionic environment for the transfer process, ensuring transfer speed and effectiveness. Simultaneously, the significant Joule heat generated during the transfer process is transferred to the buffer solution, preventing localized overheating that could lead to protein inactivation.

[0010] Regardless of whether the positive electrode assembly is located above the negative electrode assembly, or vice versa, to facilitate the transfer process, it is preferable to have both the gel matrix and the solid support immersed in the buffer solution. Of course, since the positive electrode assembly is in the buffer solution, the second transfer pad will also be in the buffer solution, acting as a replenishment pad for the first transfer pad. Even if the gel matrix and the solid support are not immersed in the buffer solution, it will not affect the normal operation of the transfer process. Similarly, the same applies when the negative electrode assembly is in the buffer solution.

[0011] Of course, without considering the amount of buffer solution used and / or its lifespan, immersing both the positive and negative electrode components in the buffer solution by increasing the amount of buffer solution will not affect the transfer effect.

[0012] In a preferred embodiment, a limiting block is provided between the negative electrode assembly and the positive electrode assembly to limit the distance between them.

[0013] By setting a limiting block to limit the distance between the negative electrode assembly and the positive electrode assembly, the thickness of the "sandwich" structure composed of the first transfer membrane absorbent pad, the gel matrix, the solid support, and the second transfer membrane absorbent pad is controlled, ensuring the consistency of the pressing height and avoiding excessive compression that could damage the gel matrix and the solid support.

[0014] In a preferred embodiment, the limiting block is disposed in the circumferential direction of the negative electrode assembly and / or the positive electrode assembly, and the space enclosed by the limiting block is used to place the first transfer membrane absorbent pad, the gel matrix, the solid phase carrier, and the second transfer membrane absorbent pad.

[0015] In another preferred embodiment, the limiting block is a first limiting block disposed in the circumferential direction of the negative electrode assembly, and the positive electrode assembly is pressed onto the first limiting block.

[0016] In another preferred embodiment, the limiting block is a second limiting block disposed in the circumferential direction of the positive electrode assembly, and the second limiting block is pressed against the upper end surface of the negative electrode assembly.

[0017] The limiting block can be disposed on the negative electrode assembly, the positive electrode assembly, or both. The limiting block is usually located in the circumferential direction of the negative electrode assembly and / or the positive electrode assembly. The middle area between the negative electrode assembly and the positive electrode assembly is used to place the first transfer membrane absorbent pad, the gel matrix, the solid support, and the second transfer membrane absorbent pad.

[0018] In a preferred embodiment, the thickness of the limiting block is the thickness of the first transfer membrane absorbent pad, gel matrix, solid carrier and second transfer membrane absorbent pad after compression when the negative electrode assembly and the positive electrode assembly are pressed together, and the thickness difference before and after compression is 0.1mm-3mm.

[0019] As a more preferred embodiment, the thickness difference before and after compression is 1.5mm-2.5mm.

[0020] As a more preferred embodiment, the thickness difference before and after compression is 1.7mm-2.2mm.

[0021] In a preferred embodiment, the thickness of the first and second transfer membrane absorbent pads is 3-10 mm, the thickness of the gel matrix is ​​0.75-1.5 mm, the thickness of the solid support is 0.1-0.25 mm, and the thickness of the limiting block is 5-15 mm.

[0022] In a more preferred embodiment, the thickness of the first and second transfer membrane absorbent pads is 3-5 mm, the thickness of the gel matrix is ​​1-1.5 mm, the thickness of the solid support is 0.1-0.2 mm, and the thickness of the limiting block is 9-10 mm.

[0023] In a more preferred embodiment, the thickness of the first and second transfer membrane absorbent pads is 5 mm, the thickness of the gel matrix is ​​1-1.5 mm, the thickness of the solid support is 0.2 mm, and the thickness of the limiting block is 9-10 mm.

[0024] In a more preferred embodiment, the thickness of the limiting block is 9.5 mm.

[0025] In a preferred embodiment, the limiting block is an insulating component.

[0026] As a preferred embodiment, the required buffer volume for transferring each gel matrix is ​​15-100 mL.

[0027] As a preferred embodiment, the required buffer volume for transferring each gel matrix is ​​20-50 mL.

[0028] The first and second membrane absorbent pads are made of sponge, polyester fiber, or filter paper.

[0029] The solid support is a nitrocellulose membrane or a PVDF membrane.

[0030] The gel matrix is ​​a polyacrylamide gel.

[0031] Preferably, the gel matrix is ​​sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) or non-denaturing polyacrylamide gel (Native-PAGE).

[0032] SDS-PAGE separates denatured proteins based on molecular weight. Preferred buffer systems include Laemmli (Tris-Glycine) buffer, Tris-Tricine buffer, or Bis-Tris (MOPS / MES) buffer.

[0033] Native-PAGE isolates native proteins under non-denaturing conditions, preserving protein activity and complex structure. In this case, the buffer system is a standard choice, but those skilled in the art can select the appropriate system based on specific circumstances.

[0034] In a preferred embodiment, a cooling module is provided at the bottom of the transfer module, and a temperature conduction plate is provided between the cooling module and the transfer tank. The cooling module cools the transfer tank through the temperature conduction plate.

[0035] By incorporating a cooling module, the transfer tank is kept at a low temperature, further reducing buffer consumption.

[0036] In a preferred embodiment, the cooling module is a water-cooled tank, an air-cooled tank, or a semiconductor cooling chip.

[0037] In a second aspect, this utility model provides a transfer apparatus, characterized in that it includes at least one of the above-mentioned transfer modules, wherein the negative electrode assembly, the first transfer absorbent pad, the gel matrix, the solid carrier, the second transfer absorbent pad, and the positive electrode assembly are pressed together in the transfer tank in the transfer module.

[0038] The transfer apparatus may contain multiple transfer modules that perform transfer operations simultaneously. During the transfer process, the negative electrode assembly, the first transfer absorbent pad, the gel matrix, the solid support, the second transfer absorbent pad, and the positive electrode assembly are pressed together in the transfer tank. The transfer absorbent pad is 100% in contact with the negative / positive electrode assembly, and the distance between the negative and positive electrode assemblies is minimized, which can provide a larger electric field to improve the transfer efficiency.

[0039] In a preferred embodiment, the negative electrode assembly and the positive electrode assembly are clamped together at both ends by insulating fasteners.

[0040] The insulating fasteners here are not limited; any component that can achieve the clamping function can be used in this application.

[0041] In a preferred embodiment, the positive electrode assembly is located above the negative electrode assembly. The positive electrode assembly includes a first positive electrode, and the negative electrode assembly includes a first negative electrode. The first negative electrode is located at the bottom of the transfer tank, and the first positive electrode and the first negative electrode are electrically connected to the positive and negative terminals of the power supply, respectively.

[0042] In a preferred embodiment, the positive electrode assembly further includes a positive electrode mounting plate fixedly connected to the first positive electrode plate. The first positive electrode plate is located at one end near the negative electrode assembly. The positive electrode mounting plate has a central hole, and a first compression cap is installed in the hole. A first rigid pressure head is provided at the top of the first compression cap, and a first elastic pressure head is provided at the bottom of the first compression cap. One end of the first elastic pressure head is connected to the bottom end of the first rigid pressure head, and the other end of the first elastic pressure head is connected to the first positive electrode plate. By applying external force to the first rigid pressure head, the pressing process between the negative electrode assembly and the positive electrode assembly is completed.

[0043] After the negative electrode assembly, the first transfer membrane absorbent pad, the gel matrix, the solid support, the second transfer membrane absorbent pad, and the positive electrode assembly are placed in sequence, the positive electrode assembly moves downward by applying external force to the first rigid pressure head, and applies force to the first transfer membrane absorbent pad, the gel matrix, the solid support, and the second transfer membrane absorbent pad, thereby achieving the pressing operation.

[0044] The external force here can be the cover set above the transfer tank. After the cover is closed on the transfer tank, the first rigid pressure head is forced to move the positive electrode plate assembly downward to achieve the pressing process.

[0045] In a preferred embodiment, one end of the first negative electrode plate extends toward the surface of the buffer solution and is provided with a negative electrode connection end, through which the first negative electrode plate is connected to the negative terminal of the power supply.

[0046] In a preferred embodiment, the temperature conduction plate at the bottom of the transfer tank is electrically connected to the negative terminal connector via a negative terminal wiring screw.

[0047] In a preferred embodiment, the negative electrode assembly is located above the positive electrode assembly. The negative electrode assembly includes a second negative electrode, and the positive electrode assembly includes a second positive electrode. The second positive electrode is located at the bottom of the transfer tank, and the second positive electrode and the second negative electrode are electrically connected to the positive and negative terminals of the power supply, respectively.

[0048] In a preferred embodiment, the negative electrode assembly further includes a negative electrode mounting plate fixedly connected to the second negative electrode plate. The second negative electrode plate is located at one end near the positive electrode assembly. The negative electrode mounting plate has a central hole, and a second compression cap is installed in the hole. A second rigid pressure head is provided at the top of the second compression cap, and a second elastic pressure head is provided at the bottom of the second compression cap. One end of the second elastic pressure head is connected to the bottom end of the second rigid pressure head, and the other end of the second elastic pressure head is connected to the second negative electrode plate. By applying external force to the second rigid pressure head, the compression process between the negative electrode assembly and the positive electrode assembly is completed.

[0049] In a preferred embodiment, one end of the second positive electrode plate extends toward the surface of the buffer solution and is provided with a positive electrode connection terminal, and the second positive electrode plate is connected to the positive terminal of the power supply through the positive electrode connection terminal.

[0050] In a preferred embodiment, the temperature conduction plate at the bottom of the transfer tank is electrically connected to the positive terminal connector via a positive terminal screw.

[0051] In a preferred embodiment, the transfer apparatus further includes an upper cover and a lower base. One end of the upper cover is hinged to the lower base, and the other end of the upper cover is provided with a locking plate. The lower base is provided with a locking element at the corresponding position of the locking plate. The locking plate and the locking element are fastened together. The lower base is provided with a groove, and the transfer module is located in the groove.

[0052] The transfer module is located on the lower base. When the positive electrode assembly is above the negative electrode assembly, when the locking plate of the upper cover is engaged with the locking member of the lower base, the upper cover acts on the first rigid pressure head of the positive electrode assembly, applying a downward force to the positive electrode assembly, causing the positive electrode assembly and the negative electrode assembly to move closer together and press tightly. When the negative electrode assembly is above the positive electrode assembly, when the locking plate of the upper cover is engaged with the locking member of the lower base, the upper cover acts on the second rigid pressure head of the negative electrode assembly, applying a downward force to the negative electrode assembly, causing the positive electrode assembly and the negative electrode assembly to move closer together and press tightly.

[0053] In a preferred embodiment, the upper cover is provided with a positive electrode base and a negative electrode base, the positive electrode plate assembly corresponds to the positive electrode base, and the negative electrode connection end corresponds to the negative electrode base; or, the negative electrode plate assembly corresponds to the negative electrode base, and the positive electrode connection end corresponds to the positive electrode base.

[0054] Both the positive and negative bases are located on the upper cover. When the positive plate assembly comes into contact with the positive base, the positive plate assembly is connected to the positive terminal of the power supply; when the negative plate assembly comes into contact with the negative base, the negative plate assembly is connected to the negative terminal of the power supply.

[0055] In a preferred embodiment, both the positive electrode base and the negative electrode base are provided with elastic contact elements at their bottom ends, and the positive electrode base and the negative electrode base are connected to the positive electrode plate assembly and the negative electrode plate assembly respectively through the elastic contact elements.

[0056] In a preferred embodiment, a positive electrode base is provided on the upper cover, the positive electrode plate assembly corresponds to the positive electrode base, and the negative electrode plate assembly is electrically connected to the negative electrode connector through a temperature conduction plate at the bottom of the transfer tank and a negative electrode wiring screw; or, a negative electrode base is provided on the upper cover, the negative electrode plate assembly corresponds to the negative electrode base, and the positive electrode plate assembly is electrically connected to the positive electrode connector through a temperature conduction plate at the bottom of the transfer tank and a positive electrode wiring screw.

[0057] When the positive electrode assembly is above the negative electrode assembly, the negative terminal of the power supply is electrically connected to the temperature conduction plate at the bottom of the transfer tank via a negative terminal wiring screw, and the electric field is conducted to the negative electrode assembly through the bottom of the transfer tank. This generates an electric field between the positive and negative electrode assemblies. Compared to placing the negative electrode base on the upper cover, this avoids the possibility of buffer solution in the transfer tank splashing onto the negative electrode base, making it safer to use. Similarly, when the negative electrode assembly is above the positive electrode assembly, a similar arrangement can achieve the same effect.

[0058] In a preferred embodiment, a control knob screen assembly is provided at the end of the lower base away from the transfer module, for selecting transfer parameters and starting the transfer program.

[0059] In a preferred embodiment, the control knob screen assembly is a circular knob touchscreen with a size of 1.28 inches to 7 inches.

[0060] In a preferred embodiment, the control knob screen assembly is 2.1 inches in size.

[0061] In a preferred embodiment, a main control board is also provided inside the lower base, and the main control board is electrically connected to the power supply, the control knob screen assembly, and the cooling module.

[0062] Beneficial effects: (1) The transfer module of this utility model for a transfer apparatus, by placing the positive electrode assembly above the negative electrode assembly, the negative electrode assembly is immersed in the buffer solution, and the positive electrode assembly is located above the surface of the buffer solution, or by placing the negative electrode assembly above the positive electrode assembly, the positive electrode assembly is immersed in the buffer solution, and the negative electrode assembly is located above the surface of the buffer solution, on the one hand, provides sufficient ions for the transfer process, maintains a high electric field state for a long time during the transfer process, and maximizes the transfer speed and transfer effect; on the other hand, only the negative electrode assembly or the positive electrode assembly is immersed in the buffer solution, and the buffer solution required for a single transfer is 20-50 mL, which reduces the use of buffer solution. Compared with the traditional wet transfer, the amount of buffer solution used is only 1 / 4 of that of the traditional wet transfer.

[0063] (2) The transfer module of this utility model for the transfer instrument has only the negative electrode assembly / positive electrode assembly located in the buffer solution, which reduces the contact area between the buffer solution and the positive electrode assembly / negative electrode assembly, reduces the electrolysis reaction rate, slows down pH changes and ion consumption, and extends the service life of the buffer solution.

[0064] (3) The transfer module of the present invention is used in the transfer instrument. Its negative electrode assembly / positive electrode assembly is immersed in the buffer solution, so that the large amount of Joule heat generated during the transfer process is transferred to the buffer solution, which has a cooling effect and avoids local overheating that leads to protein inactivation.

[0065] (4) In the transfer film apparatus of this utility model, the positive electrode plate assembly and the negative electrode plate assembly are pressed together evenly to ensure that the pressing distance at all positions is completely consistent, thus improving the overall accuracy and making the electric field uniformity more stable.

[0066] (5) The transfer apparatus of this utility model is easy to operate and can be widely promoted.

[0067] The following will further explain the concept, specific structure and technical effects of this utility model in conjunction with the accompanying drawings, so as to fully understand the purpose, features and effects of this utility model. Attached Figure Description

[0068] To more clearly illustrate the technical solutions of the embodiments of this utility model, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0069] Figure 1 This is a graph showing the transfer results of a transfer experiment using a marker. Figure 2 This is an assembly diagram of the film transfer device in Embodiment 1 of this utility model; Figure 3 This is a schematic diagram of the transfer apparatus in Embodiment 1 of this utility model; Figure 4 This is a schematic diagram of the connection structure between the strike plate and the strike fastener in the prior art; Figure 5 This is a schematic diagram of the transfer module in Embodiment 1 of this utility model; Figure 6 This is a diagram showing the positional arrangement of the transfer tank, negative electrode plate assembly, and positive electrode plate assembly in Embodiment 1 of this utility model; Figure 7 yes Figure 6 Exploded view; Figure 8 This is a schematic diagram of the transfer apparatus in Embodiment 2 of this utility model.

[0070] The components are: 1-upper cover, 2-lower base, 3-slot, 4-transfer module, 5-control knob screen assembly, 6-transfer slot, 8-first transfer film absorbent pad, 9-gel matrix, 10-solid carrier, 11-second transfer film absorbent pad, 12-positive electrode plate assembly, 13-limiting block, 14-negative electrode connection end, 15-negative electrode base, 16-first compression cover, 17-first rigid pressure head, 18-elastic pressure head, 19-positive electrode base, 20-elastic contact element, 21-cooling module, 22-temperature conduction plate, 23-first negative electrode plate, 25-first positive electrode plate, 26-positive electrode mounting plate, 27-negative electrode wiring screw, 28-locking plate, 29-locking element. Detailed Implementation

[0071] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this utility model. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of this utility model.

[0072] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, the term "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0073] In this application, unless otherwise expressly specified and limited, the terms "installation," "connection," "joining," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components, unless otherwise expressly limited. Those skilled in the art can understand the specific meaning of the above terms in this application based on the specific circumstances.

[0074] If an element is referred to as being "fixed to" or "set on" another element, it can be directly on the other element or there may be an intervening element. If an element is considered to be "connected to" another element, it can be directly connected to the other element or there may be an intervening element. If so, the terms "vertical," "horizontal," "upper," "lower," "left," "right," and similar expressions used in this application are for illustrative purposes only and do not represent the only possible implementation.

[0075] Existing protein transfer technologies are divided into two modes: wet transfer and semi-dry transfer. Semi-dry transfer utilizes multiple layers of filter paper soaked in buffer solution to create a localized electric field environment. However, due to the limited volume of buffer solution, after prolonged transfer, the buffer may become insufficient, causing the filter paper to dry out and interrupting the transfer. Wet transfer is more widely used. In wet transfer, the positive electrode, transfer membrane, gel, and negative electrode are typically placed vertically, i.e., stacked side-by-side. Wet transfer requires all four components—positive, negative, gel, and transfer membrane—to be immersed in buffer solution, resulting in high buffer consumption and increased production costs.

[0076] The inventors of this application have changed their approach. Unlike the vertical placement in existing wet transfer printing, they have cleverly set up a horizontal placement mode. That is, the positive electrode plate, transfer film, gel and negative electrode plate are not stacked side by side, but stacked vertically. The negative electrode plate / positive electrode plate located at the bottom is placed in the buffer solution. This not only ensures the supply of buffer solution during the transfer process, but also overcomes the problem of excessive buffer solution consumption caused by the negative electrode plate and positive electrode plate being completely immersed in the buffer solution.

[0077] This application first provides a transfer module for a transfer apparatus, including a transfer tank 6. Within the transfer tank 6, from bottom to top, are arranged a negative electrode assembly, a first transfer absorbent pad 8, a gel matrix 9, a solid support 10, a second transfer absorbent pad 11, and a positive electrode assembly 12. The transfer tank 6 holds a buffer solution. The negative electrode assembly and the positive electrode assembly 12 are electrically connected to the negative electrode connector and the positive electrode connector, respectively. The negative electrode assembly is immersed in the buffer solution, and the positive electrode assembly 12 is positioned above the surface of the buffer solution. Because the negative electrode assembly is immersed in the buffer solution, the first transfer absorbent pad 8 is in close contact with the negative electrode assembly and is therefore necessarily located within the buffer solution. The first transfer absorbent pad 8 serves to replenish the second transfer absorbent pad 11, preventing the absorbent pad from drying out and affecting the normal progress of the transfer process. Of course, to ensure the smooth progress of the transfer process, it is preferable to immerse both the gel matrix 9 and the solid support 10 in the buffer solution, so that the transfer process is maintained in a high electric field state.

[0078] The above-described transfer module is for the case where the negative electrode assembly is located below the positive electrode assembly 12. As an alternative, the positive electrode assembly 12 can also be located below the negative electrode assembly. In this case, the positive electrode assembly 12 is located in the buffer solution, and the negative electrode assembly is located above the page of the buffer solution.

[0079] Whether the negative electrode assembly or the positive electrode assembly 12 is immersed in the buffer solution, on the one hand, sufficient ions are provided for the transfer process, ensuring the transfer speed and transfer effect; on the other hand, since the negative electrode assembly, the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, the second transfer membrane absorbent pad 11 and the positive electrode assembly 12 are placed horizontally in the transfer tank, and only the negative electrode assembly or the positive electrode assembly 12 at the bottom is required to be in the buffer solution, while the non-negative electrode assembly and the positive electrode assembly 12 are both in the buffer solution, the use of buffer solution is significantly reduced.

[0080] In operation, the negative electrode assembly and positive electrode assembly 12 in the transfer module are pressed together. This involves the issue of whether the thickness of the first transfer film absorbent pad 8 and the second transfer film absorbent pad 11 matches the pressing distance. In traditional solutions, there are no limiting devices, and the pressing distance is determined by the operator based on experience, which has the following drawbacks: 1) Poor adaptability: Different absorbent pads need to be replaced for gels of different thicknesses (such as 1.0mm, 0.75mm, 1.5mm), or the pressing distance needs to be adjusted, which is cumbersome and costly. 2) Over-compression risk: When the thickness of the absorbent pad does not match the pressing distance, excessive compression may damage the gel or membrane structure. 3) Pressure fluctuations: The pressure is maintained solely by the elasticity of the absorbent pad, and the material deformation after long-term use will lead to pressure decay.

[0081] To address the aforementioned issues, this application provides a limiting block 13 between the negative electrode assembly and the positive electrode assembly 12 to limit the distance between them. The limiting block 13 is positioned along the periphery of the negative electrode assembly and / or the positive electrode assembly 12, and the space enclosed by the limiting block 13 is used to place the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11. One or more limiting blocks 13 may be provided. The limiting block 13 may be provided individually on the negative electrode assembly or the positive electrode assembly 12, or it may be provided on both the negative electrode assembly and the positive electrode assembly 12. The number of limiting blocks 13 may be odd or even. To maintain the stability of the bonding between the negative electrode assembly and the positive electrode assembly 12, the number of limiting blocks 13 is preferably even, and they are symmetrically arranged.

[0082] The limiting block 13 can be a first limiting block disposed in the circumferential direction of the negative electrode assembly, and the positive electrode assembly 12 is pressed onto the first limiting block. Alternatively, the limiting block 13 can be a second limiting block disposed in the circumferential direction of the positive electrode assembly 12, and the second limiting block is pressed onto the upper end surface of the negative electrode assembly.

[0083] Here, "peripheral direction" refers to the direction close to the outer edge (outer edge) of the positive or negative electrode assembly 12. Of course, it is not required that the limiting block 13 be tightly attached to the outer edge of the positive or negative electrode assembly 12; it is only required that the setting of the limiting block 13 does not affect the normal placement of the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11. The first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11 are located within the space enclosed by the limiting block 13. Since the limiting block 13 is located in the periphery of the negative or positive electrode assembly 12, "enclosed space" here refers to the center of the negative or positive electrode assembly 12 or the area far from the outer edge of the negative or positive electrode assembly 12.

[0084] The limiting block 13 is used to limit the distance between the negative electrode assembly and the positive electrode assembly 12. The thickness of the limiting block 13 is the thickness of the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11 after compression when the negative electrode assembly and the positive electrode assembly 12 are pressed together. To avoid the impact of over-compression or improper compression on the absorbent pads and the transfer effect, the thickness difference of the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11 before and after compression is required to be 0.1mm-3mm. Preferably, the thickness difference before and after compression is 1.5mm-2.5mm. More preferably, the thickness difference before and after compression is 1.7mm-2.2mm.

[0085] The thickness difference before and after compression, i.e., the compression amount or compression height, affects the electric field strength. A larger compression amount results in a smaller distance between the positive and negative plates, and a stronger electric field; conversely, a smaller compression amount results in a larger distance between the positive and negative plates, and a weaker electric field. The compression amount also affects the amount of buffer solution used; a larger compression amount requires less buffer solution, but this reduces efficiency. Furthermore, since the gel matrix 9 and the solid support 10 are incompressible, the compression amount should not be too large. In short, the compression amount needs to be controlled within a suitable range. This application indirectly controls the compression amount by controlling the thickness of the limiting block 13.

[0086] When the thickness of the first transfer membrane absorbent pad 8 and the second transfer membrane absorbent pad 11 is 3-10 mm, the thickness of the gel matrix 9 is 0.75-1.5 mm, and the thickness of the solid phase carrier 10 is 0.1-0.25 mm, the thickness of the limiting block 13 is preferably 5-15 mm.

[0087] When the thickness of the first transfer membrane absorbent pad 8 and the second transfer membrane absorbent pad 11 is 3-5 mm, the thickness of the gel matrix 9 is 1-1.5 mm, and the thickness of the solid phase carrier 10 is 0.1-0.2 mm, the thickness of the limiting block 13 is preferably 9-10 mm.

[0088] When the thickness of the first transfer membrane absorbent pad 8 and the second transfer membrane absorbent pad 11 is 5 mm, the thickness of the gel matrix 9 is 1-1.5 mm, and the thickness of the solid phase carrier 10 is 0.2 mm, the thickness of the limiting block 13 is preferably 9-10 mm, and more preferably 9.5 mm.

[0089] To avoid the limiting block 13 affecting the normal electrophoresis process, the limiting block 13 is preferably an insulating component. Of course, when the positive electrode assembly 12 and the negative electrode assembly have insulating shells at their contact points, it is not required whether the limiting block 13 is an insulating component.

[0090] The transfer module provided in this application can greatly reduce the amount of buffer solution used. Considering that the size of the transfer tank 6 affects the amount of buffer solution used, the volume of buffer solution required for transferring each piece of gel matrix is ​​15-100mL. Preferably, it is 20-50mL, at which point the volume of the transfer tank is 283mL (the length, width, and height of the outer contour of the transfer tank are 129mm, 103mm, and 26mm, respectively).

[0091] It should be noted that the volume of buffer solution mentioned here refers to the volume of buffer solution contained in the transfer tank. Before the transfer, the first transfer pad 8 and the second transfer pad 11 need to absorb the buffer solution in advance. The amount of buffer solution absorbed by each pad is approximately 35 mL (the total amount of the first transfer pad 8 and the second transfer pad 11 is approximately 70 mL), which is not considered in the buffer solution volume mentioned here.

[0092] The first membrane absorption pad 8 and the second membrane absorption pad 11 are made of sponge, polyester fiber, or filter paper. The solid carrier 10 is a nitrocellulose membrane or a PVDF membrane.

[0093] During the transfer process, a large amount of Joule heat is generated. The presence of the buffer solution facilitates heat release and avoids protein inactivation caused by excessively high local temperatures. To further enhance the cooling effect and ensure that the transfer tank 6 remains at a low temperature, a cooling module 21 is provided at the bottom of the transfer module 4. A temperature conduction plate 22 is provided between the cooling module 21 and the transfer tank 6, and the cooling module 21 cools the transfer tank 6 through the temperature conduction plate 22. The cooling module 21 here can be a water-cooled tank, an air-cooled tank, or a semiconductor cooling chip. Of course, the choice of the cooling module 21 is not limited, and any component with a similar cooling effect can be used in this application.

[0094] This application also provides a transfer apparatus including the above-mentioned transfer modules. Multiple transfer modules can be configured, and multiple transfer modules can transfer simultaneously to improve transfer efficiency. During normal transfer, the negative electrode assembly, the first transfer absorbent pad 8, the gel matrix 9, the solid support 10, the second transfer absorbent pad 11, and the positive electrode assembly 12 in the transfer module 4 are pressed tightly together within the transfer tank 6. Here, "pressed tightly" means that the negative electrode assembly, the first transfer absorbent pad 8, the gel matrix 9, the solid support 10, the second transfer absorbent pad 11, and the positive electrode assembly 12 are tightly adhered to each other under external force to avoid gaps that could affect the transfer effect.

[0095] The two ends of the negative electrode assembly and the positive electrode assembly 12 are pressed together by insulating fasteners, such as insulating clamps, so that the negative electrode assembly and the positive electrode assembly 12 are close to each other and pressed together, thereby making the first transfer membrane absorbent pad 8, the gel matrix 9, the solid phase carrier 10 and the second transfer membrane absorbent pad 11 located between the negative electrode assembly and the positive electrode assembly 12 stick together tightly.

[0096] In the case where the positive electrode assembly 12 is located above the negative electrode assembly, the positive electrode assembly 12 can be a first positive electrode 25, and the negative electrode assembly can be a first negative electrode 23. The first negative electrode 23 is located at the bottom of the transfer tank 6, and the first positive electrode 25 and the first negative electrode 23 are electrically connected to the positive and negative terminals of the power supply, respectively. After the power is turned on, a stable electric field is formed between the first positive electrode 25 and the first negative electrode 23 for the film transfer operation. In addition to the first positive electrode plate 25, the positive electrode assembly 12 may also include a positive electrode mounting plate 26 fixedly connected to the first positive electrode plate 25. The first positive electrode plate 25 is located at one end near the negative electrode assembly. The positive electrode mounting plate 26 has a central hole, in which a first compression cap 16 is installed. A first rigid pressure head 17 is provided at the top of the first compression cap 16, and a first elastic pressure head 18 is provided at the bottom of the first compression cap 16. One end of the first elastic pressure head 18 is connected to the bottom end of the first rigid pressure head 17, and the other end of the first elastic pressure head 18 is connected to the first positive electrode plate 25. By applying external force to the first rigid pressure head 17, the pressing process between the negative electrode assembly and the positive electrode assembly 12 is completed. When external force is applied to the first rigid pressure head 17, it causes the positive electrode mounting plate 26 to move downward, and the first positive electrode plate 25 moves downward accordingly to contact the negative electrode assembly and complete the pressing operation.

[0097] The first elastic pressure head 18 uses a spring as its component. The function of setting up the first elastic pressure head 18 here is as follows: Provides constant elastic force to control the mechanical movement of the positive electrode assembly 12 toward the negative electrode assembly; When the force applied to the first elastic pressure head 18 disappears, the positive electrode plate assembly 12 returns to its initial position; It can serve as an elastic contact that enables electrical connection between the positive terminal of the power supply and the first positive plate 25, and ensures the stability of the conductive connection through elastic pressure.

[0098] There are two configuration options for the negative electrode assembly. In the first option, one end of the first negative electrode 23 extends towards the surface of the buffer solution and is equipped with a negative electrode connection terminal 14. The first negative electrode 23 is connected to the negative terminal of the power supply through the negative electrode connection terminal 14. In the second option, a temperature conduction plate 22 is provided at the bottom of the transfer tank 6, and a negative electrode connection screw 27 is provided on the temperature conduction plate 22. The transfer tank 6 is electrically connected to the negative electrode connector through the negative electrode connection screw 27. The negative terminal of the power supply is conducted to the temperature conduction plate 22 through the negative electrode connection screw 27, and the temperature conduction plate 22 further conducts the conductivity to the transfer tank 6 and the first negative electrode 23 located at the bottom of the transfer tank 6.

[0099] In the case where the negative electrode assembly is located above the positive electrode assembly 12, the negative electrode assembly can be a second negative electrode assembly. The positive electrode assembly 12 includes a second positive electrode assembly located at the bottom of the transfer tank 6. The second positive electrode assembly and the second negative electrode assembly are electrically connected to the positive and negative terminals of the power supply, respectively. In addition to the second negative electrode assembly, the negative electrode assembly may also include a negative electrode mounting plate fixedly connected to the second negative electrode assembly. The second negative electrode assembly is located at one end near the positive electrode assembly 12. The negative electrode mounting plate has a central hole, and a second compression cap is installed in the hole. A second rigid pressure head is provided at the top of the second compression cap, and a second elastic pressure head is provided at the bottom of the second compression cap. One end of the second elastic pressure head is connected to the bottom end of the second rigid pressure head, and the other end of the second elastic pressure head is connected to the second negative electrode assembly. By applying external force to the second rigid pressure head, the compression process between the negative electrode assembly and the positive electrode assembly 12 is completed.

[0100] There are two configuration options for the positive electrode assembly. The first option involves a positive electrode connection terminal extending from one end of the second positive electrode towards the surface of the buffer solution, connecting the second positive electrode to the positive terminal of the power supply. The second option involves a temperature conduction plate 22 at the bottom of the transfer tank 6, with a positive electrode connection screw mounted on it. The transfer tank 6 is then electrically connected to the positive electrode connector via this screw.

[0101] The above-mentioned transfer device also includes an upper cover 1 and a lower base 2. One end of the upper cover 1 is hinged to the lower base 2, and the other end of the upper cover 1 is provided with a locking plate 28. The lower base 2 is provided with a locking fastener 29 at the corresponding position of the locking plate. The locking plate 28 and the locking fastener 29 are fastened together. The lower base 2 is provided with a groove 3, and the transfer module 4 is located in the groove 3.

[0102] The upper cover 1 may be provided with a positive electrode base 19 and a negative electrode base 15. The positive electrode plate assembly 12 corresponds to the positive electrode base 19, and the negative electrode connection end 14 corresponds to the negative electrode base 15. If the positive electrode plate assembly 12 is located below the negative electrode plate assembly, the negative electrode plate assembly corresponds to the negative electrode base 15, and the positive electrode connection end corresponds to the positive electrode base 19.

[0103] In addition to the above scenarios, when the positive electrode assembly 12 is located above the negative electrode assembly, the upper cover 1 can be provided with a separate positive electrode base 19, without a negative electrode base 15. In this case, the positive electrode assembly 12 corresponds to the positive electrode base 19, and the negative electrode assembly is electrically connected to the negative electrode connector through the temperature conduction plate 22 at the bottom of the transfer groove 6 and the negative electrode wiring screw 27. Similarly, when the negative electrode assembly is located above the positive electrode assembly 12, the upper cover can be provided with a separate negative electrode base 15, without a positive electrode base 19. The negative electrode assembly corresponds to the negative electrode base 15, and the positive electrode assembly 12 is electrically connected to the positive electrode connector through the temperature conduction plate 22 at the bottom of the transfer groove 6 and the positive electrode wiring screw.

[0104] A control knob screen assembly 5 is located at the end of the lower base 2 furthest from the transfer module. This assembly is used to select transfer parameters and initiate the transfer process. The control knob screen assembly 5 is a circular rotary touchscreen. By rotating the circular rotary touchscreen, transfer parameters can be selected, and then clicking "OK" starts the transfer process. Preferably, the size of the control knob screen assembly 5 is 1.28 inches to 7 inches. More preferably, the size is 2.1 inches.

[0105] The lower base 2 also houses the main control board, which is electrically connected to the power supply, the control knob screen assembly 5, and the cooling module 21.

[0106] Experimental Example 1 The effects of selecting different thicknesses for the first transfer membrane absorbent pad 8, gel matrix 9, solid carrier 10, second transfer membrane absorbent pad 11, and limiting block 13 on the transfer membrane effect were investigated.

[0107] The first transfer absorbent pad 8, gel matrix 9, solid support 10, and second transfer absorbent pad 11 form a "sandwich" structure. This section discusses the influence of the thickness of each component on the transfer effect under a specific "sandwich" structure. Specifically, the first transfer absorbent pad 8 and the second transfer absorbent pad 11 are sponges with a thickness of 3-10 mm; the gel matrix 9 is sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) with a thickness of 1 or 1.5 mm; the solid support 10 is a nitrocellulose membrane with a thickness of 0.2 mm; the limiting block 13 has a thickness of 5-15 mm; and the selected buffer system is the Tris-Glycine buffer system (purchased from Yamei, model TF102). Combinations of different thicknesses are shown in Table 1.

[0108] In Table 1, Δ represents the compression amount, where Δ = absorbent pad thickness × 2 + gel matrix thickness + 0.2 - limiting block thickness.

[0109] Table 1 The experimental procedure is as follows: 1. Sample loading and electrophoresis steps. Add the marker (WJ103, purchased from Yaxin, molecular weight 10-250 kDa) to each test group (refer to Table 1) to the sample loading wells of gel matrix 9 (each gel matrix contains 15 sample loading wells), with a volume of 2 μL added to each well. Perform SDS-PAGE electrophoresis at a constant voltage until the bromophenol blue indicator reaches the bottom of the gel. The electrophoresis process is then complete, and 15 bands can be obtained for each gel matrix.

[0110] 2. Assemble the "sandwich" structure. Assemble the electrophoretically extracted gel matrix 9, the first transfer absorbent pad 8, the solid support 10, and the second transfer absorbent pad 11 into a "sandwich" structure in the following order: cathode plate - first transfer absorbent pad 8 - gel matrix 9 - solid support 10 - second transfer absorbent pad 11 - anode plate. The thickness of each component is shown in Table 1. Transfer experiments were conducted, and the results are as follows: Figure 1 As shown. The experiment was repeated 5 times for each test group.

[0111] Since each gel matrix 9 can produce 15 bands, and each band has 11 lanes (10kDa, 15kDa, 20kDa, 25kDa, 35kDa, 40kDa, 50kDa, 70kDa, 100kDa, 150kDa, 250kDa), the total number of markers in each gel matrix 9 is 165.

[0112] 3. Detection and analysis steps.

[0113] After the transfer is complete, disassemble the "sandwich" structure and visually inspect the following items: 1) Check each of the 15 wells of gel matrix 9 for any residue of 250 kDa marker; 2) Check the clarity and integrity of the 165 marker strips on each solid carrier 10; 3) Check the second membrane absorbent pad 11 (the absorbent pad near the positive electrode) for any residual traces of marker penetration.

[0114] 4. Calculation method The pressure distribution uniformity, film transfer efficiency, and structural damage rate were calculated for each test group.

[0115] (1) Pressure Uniformity (PU) PU(%)=[1-N_ng_total / (5*165)]*100% N_ng_total / : The total number of markers on the solid support 10 that were judged as NG in the 5 experiments of this test group. The criteria for judging NG are light color, incomplete, thin, and blurry.

[0116] (2) Transfer efficiency (TE) TE(%)=[1-N_gel_total / (5* 15)]*100% N_gel_total: The total number of pores with 250kDa Marker residue on gel matrix 9 in the 5 experiments of this test group.

[0117] (3) Structural Integrity (SI) SI(%)=(1-N_pad_total / 5)*100% N_pad_total: The number of experiments in this test group where marker residue appeared on the second membrane transfer absorbent pad out of 5 experiments. The sum of the numbers.

[0118] The results of pressure distribution uniformity, film transfer efficiency, and structural integrity rate for test groups 1-16 are shown in Table 2.

[0119] Table 2 By analyzing the results of five repeated experiments in test group 3 and test group 12, it was found that their PU% values ​​showed a good normal distribution, as detailed below.

[0120] Test group 3 (5 / 1.0 / 9, Δ=2.2mm) PU%: 98.0%, 98.3%, 98.2%, 98.5%, 98.1% Mean: 98.22% Standard deviation (Std): ≈0.18 Test group 12 (5 / 1.5 / 10, Δ=1.7mm) PU%: 97.6%, 97.9%, 97.8%, 98.0%, 97.7% Mean: 97.80% Standard deviation (Std): ≈0.15 As shown in Table 2, the compression amount Δ plays a crucial role in the transfer effect. When Δ ∈ [1.5, 3.0] mm, the pressure distribution uniformity, transfer efficiency, and structural integrity are all relatively good. Among them, test group 3 (thickness of the first / second transfer absorbent pad is 5 mm, the thickness of the gel matrix is ​​1 mm, the thickness of the limiting block is 9 mm, and Δ is 2.2) and test group 12 (thickness of the first / second transfer absorbent pad is 5 mm, the thickness of the gel matrix is ​​1.5 mm, the thickness of the limiting block is 10 mm, and Δ is 1.7) have the best transfer effects. When Δ < 1.5 mm, the pressure distribution uniformity and transfer efficiency are slightly reduced due to insufficient pressure. However, when Δ > 3 mm, the pressure is too high, the pressure distribution uniformity decreases sharply, and structural damage occurs, resulting in a deterioration in the overall transfer effect.

[0121] Furthermore, the thickness of the gel matrix also has a fine-tuning effect on the selection of the limiting block. When the thickness of the first / second transfer membrane absorbent pad is 5 mm, the thickness of the gel matrix is ​​1 mm, and the limiting block is 9 mm thick, the former performs better than the latter (test group 4) with a 10 mm thick limiting block. When the thickness of the first / second transfer membrane absorbent pad is 5 mm, the thickness of the gel matrix is ​​1.5 mm, and the limiting block is 10 mm thick, the latter performs better than the latter (test group 11) with a 9 mm thick limiting block.

[0122] Experimental Example 2 The impact of the choice of limiting block thickness on the film transfer effect was examined.

[0123] As described in Experiment 1, the optimal transfer effect was achieved when the thickness of the first / second transfer absorbent pad was 5 mm, the gel matrix thickness was 1 mm, the solid support thickness was 0.2 mm, and the limiting block thickness was 9 mm; and when the thickness of the first / second transfer absorbent pad was 5 mm, the gel matrix thickness was 1.5 mm, the solid support thickness was 0.2 mm, and the limiting block thickness was 10 mm. With the thicknesses of the first / second transfer absorbent pad, gel matrix, and solid support remaining constant, the following comparative experiment was conducted to examine the impact of varying the limiting block thickness between 9 mm and 10 mm on the transfer effect. The specific test method was the same as in Experiment 1, and the test results are shown in Tables 3 and 4. Table 3 corresponds to a first / second transfer membrane absorbent pad thickness of 5 mm, a gel matrix thickness of 1 mm, and a solid support thickness of 0.2 mm; Table 4 corresponds to a first / second transfer membrane absorbent pad thickness of 5 mm, a gel matrix thickness of 1.5 mm, and a solid support thickness of 0.2 mm.

[0124] Table 3 Table 4 As shown in Tables 3 and 4, under the two different conditions of gel matrix thickness of 1.0 mm and 1.5 mm, the pressure distribution uniformity (PU%) and transfer efficiency (TE%) both reach their peak values ​​when the thickness of the limiting block is 9.5 mm. For the case of gel matrix thickness of 1.0 mm, the PU% and TE% values ​​both increase with the increase of the limiting block thickness, reaching their peak values ​​at a thickness of 9.5 mm. At this point, when the limiting block thickness is further increased, the PU% and TE% values ​​both decrease, and the PU% and TE% values ​​at a thickness of 10 mm are both lower than the corresponding values ​​at a thickness of 9 mm.

[0125] When the thickness of the gel matrix is ​​1.5 mm, the changes in its PU% and TE% values ​​are similar to those when it is 1.0 mm. The difference is that when the thickness of the limiting block is greater than 9.5 mm, although the PU% and TE% values ​​decrease slightly, they are still higher than the corresponding values ​​when the thickness is 9.0 mm and 9.2 mm. This indicates that when the thickness of the gel matrix is ​​1.5 mm, it has better compatibility with the thicker limiting block.

[0126] Experimental Example 3 The "sandwich" structure of test group 3 in Experiment Example 1 was subjected to transfer experiments using the transfer method of this application, the traditional wet transfer method, and the semi-dry transfer method. The transfer results are shown in Table 5.

[0127] Table 5 Example 1 like Figure 2 , Figure 3 A film transfer device includes an upper cover 1 and a lower base 2. One end of the upper cover 1 is hinged to the lower base 2, and the other end of the upper cover 1 is provided with a locking plate 28. The lower base 2 is provided with a locking element 29 at a corresponding position on the locking plate 28. When the locking plate 28 and the locking element 29 are engaged, the film transfer device can perform a film transfer operation. Here, a platform is provided at the end of the locking plate 28, and an inverted hook is provided at the top of the locking element 29. The hook is engaged with the platform, completing the connection between the locking plate 28 and the locking element 29.

[0128] The connection between the snap plate 28 and the snap fastener 29 is existing technology. Other alternative structures for the snap plate 28 and snap fastener 29 are as follows: Figure 4 As shown.

[0129] The lower base 2 has a groove 3, in which the transfer module 4 is placed. A control knob screen assembly 5 is provided on the outside of the groove 3 on the lower base 2. The locking plate 28 and the locking fastener 29 are engaged between the control knob screen assembly 5 and the groove 3. The transfer parameters are selected by controlling the knob screen assembly 5 to complete the transfer process under different conditions.

[0130] like Figure 5 , 67. The transfer module 4 includes a transfer tank 6. From bottom to top, the transfer tank 6 contains a negative electrode assembly, a first transfer membrane absorbent pad 8, a gel matrix 9, a solid support 10, a second transfer membrane absorbent pad 11, and a positive electrode assembly 12. The negative electrode assembly and the positive electrode assembly 12 are electrically connected to the negative electrode connector and the positive electrode connector, respectively. Before transfer, the transfer tank 6 needs to be filled with buffer solution, with a volume of 20mL-50mL. At this volume, the negative electrode assembly, the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11 are all immersed in the buffer solution, ensuring sufficient ions near the negative electrode assembly for proper transfer. The positive electrode assembly 12 is located above the surface of the buffer solution and is not immersed in it, reducing the contact area between the positive electrode assembly 12 and the buffer solution, lowering the electrolysis rate, and extending the service life of the buffer solution. The above setup overcomes both the problem of excessive buffer usage in traditional wet transfer and the problem of insufficient buffer after prolonged use in semi-dry transfer, thus balancing transfer efficiency and buffer usage. Alternatively, the amount of buffer can be controlled so that the second transfer pad 11 is not submerged in the buffer, thus not affecting the normal transfer process.

[0131] During the transfer process, the negative electrode assembly, the first transfer absorbent pad 8, the gel matrix 9, the solid support 10, the second transfer absorbent pad 11, and the positive electrode assembly 12 are clamped or pressed together in the transfer tank 6. The negative electrode assembly and the positive electrode assembly 12 can be clamped at both ends by insulating fasteners or by applying external force.

[0132] In the transfer tank 6, the components are placed horizontally in the transfer apparatus. The preparation process of the "sandwich" structure, consisting of the negative electrode assembly, the first transfer absorbent pad 8, the gel matrix 9, the solid support 10, the second transfer absorbent pad 11, and the positive electrode assembly 12, and the subsequent transfer operation are carried out in the same device without the need for transfer, greatly simplifying the operation. In traditional wet transfer, the transfer tank 6 is placed vertically. Before being placed in the transfer tank 6, the negative electrode assembly, the first transfer absorbent pad 8, the gel matrix 9, the solid support 10, the second transfer absorbent pad 11, and the positive electrode assembly 12 first complete the assembly of the "sandwich" structure, and then place them vertically in the transfer tank 6 after assembly.

[0133] The first transfer membrane absorbent pad 8 and the second transfer membrane absorbent pad 11 are made of sponge. The solid carrier 10 is a nitrocellulose membrane or a PVDF membrane.

[0134] Four limiting blocks 13 are arranged circumferentially on the negative electrode assembly. The limiting blocks 13 are symmetrically arranged at the four corners of the negative electrode assembly. When the positive electrode assembly 12 abuts against the limiting blocks 13, the pressing process ends. The thickness of the limiting blocks 13 is the compressed thickness of the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11. By setting the limiting blocks 13, the first transfer membrane absorbent pad 8, the gel matrix 9, the solid support 10, and the second transfer membrane absorbent pad 11 always maintain the same compressed thickness. In this embodiment, when the thickness of the first transfer membrane absorbent pad 8 and the second transfer membrane absorbent pad 11 is 5 mm, the thickness of the gel matrix 9 is 1 mm, and the thickness of the solid support 10 is 0.2 mm, the thickness of the limiting blocks 13 is 9.5 mm.

[0135] Of course, the limiting block 13 can also be disposed on the lower surface of the positive electrode assembly 12, with its placement similar to that of the negative electrode assembly, and the limiting block 13 is pressed against the upper end surface of the negative electrode assembly. Alternatively, the limiting block 13 can be disposed on both the negative electrode assembly and the positive electrode assembly 12. In this case, the placement positions of the limiting blocks 13 on the negative electrode assembly and the positive electrode assembly 12 correspond to each other, and the upper and lower limiting blocks 13 abut against each other.

[0136] like Figure 5 , Figure 6 , Figure 7 As shown, the negative electrode assembly includes a first negative electrode plate 23, with a negative electrode connection end 14 extending from one end of the first negative electrode plate 23. The first negative electrode plate 23 is located at the bottom of the transfer groove 6, and the negative electrode connection end 14 is electrically connected to the negative electrode base 15 or the negative electrode connector. The positive electrode assembly 12 includes a first positive electrode plate 25 and a positive electrode mounting plate 26 fixedly connected to the first positive electrode plate 25. The first positive electrode plate 25 is located at one end near the negative electrode assembly. The positive electrode mounting plate 26 has a central opening, and a first compression cover 16 is installed in the opening. A first rigid pressure head 17 is provided at the top of the first compression cover 16, and a first elastic pressure head 18 is provided at the bottom of the first compression cover 16. One end of the first elastic pressure head 18 is connected to the bottom end of the first rigid pressure head 17, and the other end of the first elastic pressure head 18 is connected to the first positive electrode plate 25. By applying external force to the first rigid pressure head 17, the clamping process between the negative electrode assembly and the positive electrode assembly 12 is completed.

[0137] like Figure 3The upper cover 1 is also provided with a positive electrode base 19 and a negative electrode base 15. Both the positive electrode base 19 and the negative electrode base 15 have elastic contact members 20 at their bottom ends. The positive electrode base 19 and the negative electrode base 15 are connected to the positive electrode plate assembly 12 and the negative electrode connection end 14, respectively, through the elastic contact members 20. The positive electrode plate assembly 12 (or the first rigid pressure head 17) corresponds to the positive electrode base 19, and the negative electrode connection end 14 of the negative electrode plate assembly corresponds to the negative electrode base 15. The positive electrode of the power supply is connected to the positive electrode plate assembly 12 through the positive electrode base 19, and the negative electrode of the power supply is connected to the negative electrode plate assembly through the negative electrode base 15, thereby forming a stable electric field between the positive electrode plate assembly 12 and the negative electrode plate assembly.

[0138] The bottom of the transfer module 4 is equipped with a cooling module 21, which can be a water-cooled tank, an air-cooled tank, or a thermoelectric cooler. Optional thermoelectric coolers include Peltier coolers.

[0139] The cooling module 21 is located below the transfer tank 6, and a temperature conduction plate 22 (closely attached to the bottom of the transfer tank 6) is provided between the cooling module 21 and the transfer tank 6. The cooling module 21 cools the transfer tank 6 through the temperature conduction plate 22. By cooling the transfer tank 6 through the cooling module 21, the transfer tank 6 is kept at a low temperature, avoiding local overheating that could lead to protein inactivation and loss of buffer solution.

[0140] The lower base 2, away from the transfer module 4, is equipped with a control knob screen assembly 5 for selecting transfer parameters.

[0141] The lower base 2 also houses the main control board, which is electrically connected to the power supply, the control knob screen assembly 5, and the cooling module.

[0142] Work process: Open the upper cover 1, take out the transfer tank 6, and place it on the operating table; Place the negative electrode assembly in the transfer tank 6, and measure 35 mL of buffer solution into the transfer tank 6. Soak two 5mm sponges in the buffer solution until they are fully saturated. Place a 5mm sponge, a gel matrix 9, a solid support 10, and a 5mm sponge in the transfer tank 6 in sequence; Place the transfer tank 6 back into the lower base 2, and place the positive electrode assembly 12 on the 5mm sponge. Press the upper cover 1 downwards until the latch plate is fastened in the latching member. At this time, the upper cover 1 acts on the first rigid pressure head 17, which drives the positive electrode plate assembly 12 to press onto the negative electrode plate assembly. The first rigid pressure head 17 contacts the positive electrode base 19, and the negative electrode connection end 14 contacts the negative electrode base 15. By controlling the rotary screen assembly 5, setting the film transfer conditions (such as constant current 1A, time 15 minutes), turning on the power, and starting the film transfer; Once the transfer is complete, the transfer machine stops running. Open the upper cover 1 and remove the transfer tank 6. Remove the positive electrode assembly 12, upper sponge, solid support 10, and gel matrix 9 in sequence, and check the membrane surface for bubbles or mechanical damage.

[0143] Example 2 like Figure 8 Unlike Embodiment 1, the upper cover 1 is provided with a positive electrode base 19, and the positive electrode plate assembly 12 corresponds to the positive electrode base 19. The negative electrode plate assembly includes a first negative electrode plate 23, which is located at the bottom of the transfer tank 6. The bottom of the transfer tank 6 is closely attached to a temperature conduction plate 22. Since the temperature conduction plate 22 is a metal plate, it is connected to the negative terminal of the power supply via a negative electrode connection screw 27. The negative terminal of the power supply is conducted to the temperature conduction plate 22 through the negative electrode connection screw 27, and the temperature conduction plate 22 is further conducted to the transfer tank 6 and the first negative electrode plate 23 located at the bottom of the transfer tank 6. At this time, an electric field is generated between the positive electrode plate assembly 12 and the negative electrode plate assembly. Compared with the design of mounting the negative electrode base on the upper cover, the electric field is more stable in this embodiment.

[0144] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0145] The embodiments described above are merely examples of several implementations of this utility model, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this utility model, and these modifications and improvements all fall within the protection scope of this utility model.

Claims

1. A transfer module for a transfer apparatus, characterized in that, The system includes a transfer tank (6), in which a negative electrode assembly, a first transfer membrane absorbent pad (8), a gel matrix (9), a solid support (10), a second transfer membrane absorbent pad (11), and a positive electrode assembly (12) are arranged sequentially from bottom to top. The transfer tank (6) is used to hold a buffer solution. The negative electrode assembly and the positive electrode assembly (12) are electrically connected to the negative electrode connector and the positive electrode connector, respectively. The negative electrode assembly is immersed in the buffer solution, and the positive electrode assembly (12) is located above the surface of the buffer solution. Alternatively, the transfer tank (6) may be provided with a negative electrode plate assembly and a first transfer film absorber in sequence from top to bottom. The membrane consists of a pad (8), a gel matrix (9), a solid support (10), a second transfer membrane absorbent pad (11), and a positive electrode assembly (12). The transfer tank (6) is used to hold the buffer solution. The negative electrode assembly and the positive electrode assembly (12) are electrically connected to the negative electrode connector and the positive electrode connector, respectively. The positive electrode assembly (12) is immersed in the buffer solution, and the negative electrode assembly is located above the surface of the buffer solution.

2. The transfer module for a transfer apparatus according to claim 1, characterized in that, A limiting block (13) is provided between the negative electrode assembly and the positive electrode assembly (12) to limit the distance between the negative electrode assembly and the positive electrode assembly (12).

3. The transfer module for a transfer apparatus according to claim 2, characterized in that, The limiting block (13) is disposed in the circumferential direction of the negative electrode assembly and / or the positive electrode assembly (12), and the space enclosed by the limiting block (13) is used to place the first transfer membrane absorbent pad (8), the gel matrix (9), the solid phase carrier (10) and the second transfer membrane absorbent pad (11).

4. The transfer module for a transfer apparatus according to claim 3, characterized in that, The limiting block (13) is a first limiting block set in the circumferential direction of the negative electrode assembly, and the positive electrode assembly (12) is pressed onto the first limiting block.

5. The transfer module for a transfer apparatus according to claim 3, characterized in that, The limiting block (13) is a second limiting block disposed in the circumferential direction of the positive electrode assembly (12), and the second limiting block is pressed against the upper end surface of the negative electrode assembly.

6. The transfer module for a transfer apparatus according to claim 2, characterized in that, The thickness of the limiting block is the thickness of the first transfer membrane absorbent pad (8), gel matrix (9), solid carrier (10) and the second transfer membrane absorbent pad (11) after compression when the negative electrode assembly and the positive electrode assembly (12) are pressed together. The thickness difference before and after compression is 0.1mm-3mm.

7. The transfer module for a transfer apparatus according to claim 6, characterized in that, The thickness difference before and after compression is 1.5mm-2.5mm.

8. The transfer module for a transfer apparatus according to claim 7, characterized in that, The thickness difference before and after compression is 1.7mm-2.2mm.

9. The transfer module for a transfer apparatus according to claim 6, characterized in that, The thickness of the first transfer membrane absorbent pad (8) and the second transfer membrane absorbent pad (11) is 3-10 mm, the thickness of the gel matrix (9) is 0.75-1.5 mm, the thickness of the solid phase carrier (10) is 0.1-0.25 mm, and the thickness of the limiting block (13) is 5-15 mm.

10. The transfer module for a transfer apparatus according to claim 9, characterized in that, The thickness of the first transfer membrane absorbent pad (8) and the second transfer membrane absorbent pad (11) is 3-5 mm, the thickness of the gel matrix (9) is 1-1.5 mm, the thickness of the solid phase carrier (10) is 0.1-0.2 mm, and the thickness of the limiting block (13) is 9-10 mm.

11. The transfer module for a transfer apparatus according to claim 10, characterized in that, The thickness of the first transfer membrane absorbent pad (8) and the second transfer membrane absorbent pad (11) is 5 mm, the thickness of the gel matrix (9) is 1-1.5 mm, the thickness of the solid phase carrier (10) is 0.2 mm, and the thickness of the limiting block (13) is 9-10 mm.

12. The transfer module for a transfer apparatus according to claim 11, characterized in that, The thickness of the limiting block (13) is 9.5 mm.

13. The transfer module for a transfer apparatus according to any one of claims 2-12, characterized in that, The limiting block (13) is an insulating component.

14. The transfer module for a transfer apparatus according to claim 1, characterized in that, The required buffer volume for transferring each gel matrix is ​​15-100 mL.

15. The transfer module for a transfer apparatus according to claim 14, characterized in that, The required buffer volume for transferring each gel matrix is ​​20-50 mL.

16. The transfer module for a transfer apparatus according to claim 1, characterized in that, The first membrane absorbent pad (8) and the second membrane absorbent pad (11) are made of sponge, polyester fiber or filter paper.

17. The transfer module for a transfer apparatus according to claim 1, characterized in that, The solid support (10) is a nitrocellulose membrane or a PVDF membrane.

18. The transfer module for a transfer apparatus according to claim 1, characterized in that, A cooling module (21) is provided at the bottom of the transfer module (4). A temperature conduction plate (22) is provided between the cooling module (21) and the transfer tank (6). The cooling module (21) cools the transfer tank (6) through the temperature conduction plate (22).

19. The transfer module for a transfer apparatus according to claim 18, characterized in that, The cooling module (21) is a water-cooled tank, an air-cooled tank, or a semiconductor cooling chip.

20. A transfer apparatus, characterized in that, The transfer module includes at least one of the transfer modules according to any one of claims 1-19, wherein the negative electrode assembly, the first transfer membrane absorbent pad (8), the gel matrix (9), the solid support (10), the second transfer membrane absorbent pad (11), and the positive electrode assembly (12) are pressed together in the transfer groove (6).

21. The transfer apparatus according to claim 20, characterized in that, The negative electrode assembly and the positive electrode assembly (12) are clamped together at both ends by insulating fasteners.

22. The transfer apparatus according to claim 20, characterized in that, The positive electrode assembly (12) is located above the negative electrode assembly. The positive electrode assembly (12) includes a first positive electrode (25), and the negative electrode assembly includes a first negative electrode (23). The first negative electrode (23) is located at the bottom of the transfer groove (6). The first positive electrode (25) and the first negative electrode (23) are electrically connected to the positive and negative terminals of the power supply, respectively.

23. The transfer apparatus according to claim 22, characterized in that, The positive electrode assembly (12) also includes a positive electrode mounting plate (26) fixedly connected to the first positive electrode plate (25). The first positive electrode plate (25) is located at one end close to the negative electrode assembly. The positive electrode mounting plate (26) has a central hole, and a first compression cap (16) is installed in the hole. A first rigid pressure head (17) is provided at the top of the first compression cap (16), and a first elastic pressure head (18) is provided at the bottom of the first compression cap (16). One end of the first elastic pressure head (18) is connected to the bottom end of the first rigid pressure head (17), and the other end of the first elastic pressure head (18) is connected to the first positive electrode plate (25). By applying external force to the first rigid pressure head (17), the pressing process between the negative electrode assembly and the positive electrode assembly (12) is completed.

24. The transfer apparatus according to claim 22, characterized in that, One end of the first negative electrode plate (23) extends toward the surface of the buffer solution and is provided with a negative electrode connection end (14). The first negative electrode plate (23) is connected to the negative terminal of the power supply through the negative electrode connection end (14).

25. The transfer apparatus according to claim 22, characterized in that, The temperature conduction plate (22) at the bottom of the transfer tank (6) is electrically connected to the negative terminal connector via the negative terminal wiring screw (27).

26. The transfer apparatus according to claim 20, characterized in that, The negative electrode assembly is located above the positive electrode assembly (12). The negative electrode assembly includes a second negative electrode, and the positive electrode assembly (12) includes a second positive electrode. The second positive electrode is located at the bottom of the transfer groove (6). The second positive electrode and the second negative electrode are electrically connected to the positive and negative terminals of the power supply, respectively.

27. The transfer apparatus according to claim 26, characterized in that, The negative electrode plate assembly also includes a negative electrode mounting plate fixedly connected to the second negative electrode plate. The second negative electrode plate is located at one end close to the positive electrode plate assembly (12). The negative electrode mounting plate has a central hole, and a second compression cap is installed in the hole. A second rigid pressure head is provided at the top of the second compression cap, and a second elastic pressure head is provided at the bottom of the second compression cap. One end of the second elastic pressure head is connected to the bottom end of the second rigid pressure head, and the other end of the second elastic pressure head is connected to the second negative electrode plate. The compression process between the negative electrode plate assembly and the positive electrode plate assembly (12) is completed by applying external force to the second rigid pressure head.

28. The transfer apparatus according to claim 26, characterized in that, One end of the second positive electrode plate extends toward the surface of the buffer solution and is provided with a positive electrode connection terminal. The second positive electrode plate is connected to the positive terminal of the power supply through the positive electrode connection terminal.

29. The transfer apparatus according to claim 26, characterized in that, The temperature conduction plate (22) at the bottom of the transfer tank (6) is electrically connected to the positive terminal connector via a positive terminal screw.

30. The transfer apparatus according to any one of claims 20-29, characterized in that, The transfer printing machine also includes an upper cover (1) and a lower base (2). One end of the upper cover (1) is hinged to the lower base (2). The other end of the upper cover (1) is provided with a locking plate. The lower base (2) is provided with a locking element at the corresponding position of the locking plate. The locking plate and the locking element are fastened together. The lower base (2) is provided with a groove (3). The transfer module (4) is located in the groove (3).

31. The transfer apparatus according to claim 30, characterized in that, The upper cover is provided with a positive electrode base (19) and a negative electrode base (15). The positive electrode plate assembly (12) corresponds to the positive electrode base (19), and the negative electrode connection end (14) corresponds to the negative electrode base (15); or, the negative electrode plate assembly corresponds to the negative electrode base (15), and the positive electrode connection end corresponds to the positive electrode base (19).

32. The transfer apparatus according to claim 30, characterized in that, A positive electrode base (19) is provided on the upper cover, and the positive electrode plate assembly (12) corresponds to the positive electrode base (19). The negative electrode plate assembly is electrically connected to the negative electrode connector through the temperature conduction plate (22) at the bottom of the transfer groove (6) and the negative electrode wiring screw (27); or, a negative electrode base (15) is provided on the upper cover, and the negative electrode plate assembly corresponds to the negative electrode base (15). The positive electrode plate assembly (12) is electrically connected to the positive electrode connector through the temperature conduction plate (22) at the bottom of the transfer groove (6) and the positive electrode wiring screw.

33. The transfer apparatus according to claim 30, characterized in that, The lower base is provided with a control knob screen assembly (5) at the end away from the transfer module, which is used to select the transfer parameters and start the transfer program.

34. The transfer apparatus according to claim 33, characterized in that, The control knob screen assembly (5) is a circular knob touch screen with a size of 1.28 inches to 7 inches.

35. The transfer apparatus according to claim 34, characterized in that, The control knob screen assembly (5) is 2.1 inches in size.

36. The transfer apparatus according to claim 33, characterized in that, The lower base (2) is also equipped with a main control board, which is electrically connected to the power supply, the control knob screen assembly (5), and the cooling module (21).

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