A device for detecting proteins based on magnetophoresis and a detection method thereof
The detection device for protein analysis by magnetophoresis uses magnetic and electric fields to separate proteins. Combined with SDS solution and ultrasonic stirrer, it solves the problem of the difficulty in separating high-abundance low-molecular-weight proteins in traditional electrophoresis methods, and achieves efficient and accurate protein detection.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HUNAN NORMAL UNIVERSITY
- Filing Date
- 2025-12-16
- Publication Date
- 2026-05-29
AI Technical Summary
Traditional electrophoresis detection methods are difficult to efficiently separate and detect high-abundance low-molecular-weight proteins, and suffer from problems such as low sensitivity, poor resolution, and long operation time, which cannot meet the needs of efficient batch analysis.
The detection device uses a magnetophoretic protein analysis method. It separates proteins by magnetic field and accelerates them by electric field. The proteins are charged by combining SDS solution. The device uses a uniform magnetic field and electric field generator to separate and detect the molecular weight of proteins. It is equipped with an ultrasonic stirrer and a temperature control device to ensure uniform mixing and stable temperature.
It enables efficient separation and detection of the abundance and molecular size of proteins with different molecular weights, improves the accuracy and precision of detection, simplifies the operation process, and is suitable for efficient batch analysis.
Smart Images
Figure CN122109257A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the technical field of protein abundance and molecular weight detection, and in particular to a detection device and method for protein based on magnetophoretic analysis. Background Technology
[0002] Protein abundance and molecular weight are core detection indicators in biomedical research, clinical diagnosis, and biopharmaceutical fields. Traditional electrophoresis detection methods rely on gel pore size sieving and staining. Although these methods have been widely promoted and improved, they still have technical bottlenecks for the detection of high-abundance, low-molecular-weight proteins. Specifically, it is difficult to separate high-abundance, low-molecular-weight proteins and extract clearly distinguishable features. This results in problems such as low sensitivity, poor resolution, and long operation time, which cannot meet the needs of efficient batch analysis. This has become a key constraint on the application of protein research and pharmaceuticals. Summary of the Invention
[0003] To address the shortcomings of electrophoretic detection methods in the aforementioned background art, this invention provides a detection device and method for protein analysis based on magnetophoresis.
[0004] The present invention provides a detection device and method for proteins based on magnetophoretic analysis, which adopts the following technical solution: A detection device based on magnetophoresis to analyze proteins includes a pre-magnetophoresis preparation section and a protein magnetophoresis section connected in sequence.
[0005] Furthermore, the pre-magnetophoresis preparation section includes a mixing container containing an SDS solution. The SDS solution is mixed with the protein to make it charged, laying the foundation for subsequent magnetic field separation. The outlet end of the mixing container is connected to a pipe. The pipe passes sequentially through a stirring area equipped with an ultrasonic stirrer and a uniform electric field area equipped with a uniform electric field generator. The ultrasonic vibration achieves efficient mixing of the solution and the electrophoresis buffer, and the electric field accelerates the protein molecules to obtain a stable initial velocity to enter the magnetic field.
[0006] Furthermore, the mixing container is equipped with a temperature control device, which includes a heating wire wound around the outer wall of the mixing container. The heating wire is connected to the temperature control unit and the power supply through a circuit. A temperature sensor electrically connected to the temperature control unit is installed inside the mixing container to maintain the reaction temperature inside the mixing container and improve the stability of the detection results. A paddle stirrer is also installed inside the mixing container.
[0007] Furthermore, the ultrasonic stirrer is arranged around the pipe section pre-filled with electrophoresis buffer, and an injector is installed at the front end of the pipe. The injector can pre-fill the electrophoresis buffer in the pipe and provide an interface for the subsequent cleaning process.
[0008] Furthermore, the protein magnetophoresis section includes a separation chamber, which is set in a uniform magnetic field region formed by a uniform magnetic field generator. The Lorentz force is used to deflect and separate protein complexes of different molecular weights. A detector is installed on the inner wall of the separation chamber and connected to an external analysis device. The detector collects data and the analysis device is used to analyze and obtain information on the relative size and abundance of protein molecular weights.
[0009] Furthermore, the uniform magnetic field generator includes two sets of Helmholtz coils arranged coaxially on both sides of the separation chamber to generate a highly uniform magnetic field. Both sets of Helmholtz coils are externally connected to an adjustment circuit, which is equipped with a digitally controlled power supply and an on / off switch.
[0010] A detection method for a protein detection device based on magnetophoresis analysis, applied to such a device, includes the following steps: Step 1: Injecting the protein solution to be detected into a mixing container containing SDS solution at a preset flow rate; Step 2: Using a temperature control device, adjusting the mixture of protein solution and SDS solution in the container to 30-50℃ and maintaining this temperature, allowing protein molecules to fully bind with SDS and become uniformly charged; Step 3: The mixture enters a pipe section equipped with an ultrasonic stirrer, where it mixes uniformly with the electrophoresis buffer in the pipe under the vibration of the ultrasonic stirrer to form a complex; Step 4: The complex enters a pipe section equipped with a uniform electric field generator, where it is accelerated along the pipe axis under the action of the uniform electric field; Step 5: The accelerated complex enters a separation chamber, where it is deflected perpendicular to the direction of motion under the action of the Lorentz force in the uniform magnetic field region, with complexes of different molecular weights forming different deflection trajectories; Step 6: Using a detector, the arrival position and signal intensity data of each complex are detected in real time and transmitted to an analysis device, which calculates the relative size and abundance of protein molecular weights and outputs the detection results. The above scheme forms a standardized process from protein sample injection, pretreatment, mixing and acceleration to magnetic field separation and data acquisition and analysis. Each step works synergistically, is easy to operate, and improves detection accuracy.
[0011] Preferably, in step six: the analytical device is equipped with a collaborative control platform electrically connected to the uniform electric field generator and the uniform magnetic field generator. The collaborative control platform adjusts the electric field strength and magnetic induction intensity through a matching database to increase the spacing between the deflection trajectories of the complex and improve the problem of overlapping and indistinguishable protein trajectories with similar molecular weights in traditional detection.
[0012] Preferably, after a single test is completed, cleaning fluid is injected through an injector set on the pipeline to rinse the pipeline and then the electrophoresis buffer is precharged again for the next test.
[0013] In summary, the present invention has the following beneficial technical effects: 1. This invention provides a device capable of effectively separating and detecting the abundance and relative molecular weight of proteins with different molecular weights using a variable magnetic field. It integrates mixing, acceleration, separation, detection, and cleaning functions. The device uses SDS solution to eliminate charge differences between protein molecules. After acceleration by an electric field, the proteins enter a uniform magnetic field, are separated according to their different molecular weights, and then analyzed and displayed by a detector, reflecting the abundance and relative molecular weight of each protein. This detection device has a simple structure and is easy to operate. Magnetophoretic analysis technology provides a new detection method for the abundance and relative molecular weight of proteins.
[0014] 2. The present invention provides an ultrasonic stirrer on the pipeline, which acts on the inside of the pipeline through ultrasonic vibration, so as to uniformly mix the electrophoresis buffer and the protein-SDS mixture in the pipeline, maintain charge balance and promote stable separation of the protein-SDS mixture.
[0015] 3. This invention dynamically adjusts electric and magnetic field parameters by matching the database through a collaborative control platform, thereby expanding the deflection trajectory spacing of complexes with different molecular weights. This effectively compensates for the lack of accuracy in detecting small molecular weight, high-abundance proteins, avoids trajectory overlap, and increases the accuracy of molecular weight detection. Attached Figure Description
[0016] Figure 1 This is a schematic diagram of the overall structure of a detection device for protein analysis based on magnetophoresis, according to Embodiment 1 of the invention. Figure 2 This is a schematic diagram of the pre-magnetic electrophoresis preparation part of Embodiment 1 of the invention; Figure 3 This is a schematic diagram of the structure of the protein magnetophoresis portion in Embodiment 1 of the invention; Figure 4 This is a physical model diagram of a detection device based on magnetophoretic protein analysis according to Embodiment 1 of the invention.
[0017] Figure 5 This is a schematic diagram of the pre-magnetic electrophoresis preparation part of Embodiment 3 of the invention; Explanation of reference numerals in the attached drawings: 1. Mixing container; 11. Paddle stirrer; 2. Temperature control device; 21. Heating wire; 22. Temperature control unit; 23. Temperature sensor; 3. Pipe; 31. Injector; 4. Ultrasonic stirrer; 5. Uniform electric field generator; 6. Separation chamber; 61. Detector; 62. Analytical device; 7. Uniform magnetic field generator; 71. Helmholtz coil; 72. Sliding rheostat. Detailed Implementation
[0018] The following combination Figures 1-5 The present invention will be described in further detail below.
[0019] Example 1 Embodiment 1 of the invention discloses a detection device for proteins based on magnetophoresis.
[0020] A detection device based on magnetophoresis to analyze proteins includes a pre-magnetophoresis preparation section and a protein magnetophoresis section connected in sequence.
[0021] Reference Figure 1 , Figure 4 The preparatory part for magnetophoresis includes a mixing container 1 containing an SDS solution, which is sodium dodecyl sulfate solution. After the protein solution is injected into the container, the SDS solution denatures the protein by breaking the peptide chains of the protein molecules, destroying their spatial structure and preventing protein aggregation. At the same time, the SDS molecules bind to the protein molecules, making them carry the same negative charge.
[0022] Combination Figure 2 The mixing container 1 is equipped with a temperature control device 2, which includes a heating wire 21 wound around the outer wall of the mixing container 1. The heating wire 21 is connected to the temperature control unit 22 and the power supply through a circuit. A temperature sensor 23 electrically connected to the temperature control unit 22 is installed inside the mixing container 1. The temperature control device 2 maintains a constant temperature environment of 30-50°C inside the mixing container 1. Under constant temperature conditions, the reaction time between the SDS solution and protein molecules is shortened, avoiding irreversible denaturation of proteins due to excessively high temperature or insufficient binding due to excessively low temperature, thus providing a uniform detection target for subsequent processes.
[0023] As a preferred structure, a paddle agitator 11 is also provided inside the mixing container 1. The rotating paddle agitator makes the mixing uniform and reduces the mixing time.
[0024] The outlet end of the mixing container 1 is connected to a pipe 3, which passes through a mixing area equipped with an ultrasonic stirrer 4 and a uniform electric field area equipped with a uniform electric field generator 5.
[0025] The ultrasonic stirrer 4 is arranged around the section of the pipe 3 pre-filled with electrophoresis buffer. The ultrasonic stirrer 4 acts on the pipe 3 and the internal mixed liquid through ultrasonic vibration, and completes the rapid mixing of the medium without contact and with low damage, which is suitable for the mixing requirements of the liquid in the pipe.
[0026] The main components of the electrophoresis buffer include Tris, glycine, and SDS, which are mainly used to maintain the charge balance of the mixture solution and are suitable for the stable separation of protein molecules.
[0027] Reference Figure 1 , Figure 4The protein magnetophoresis section includes a separation chamber 6, which is set in a uniform magnetic field region formed by a uniform magnetic field generator 7. Detectors 61 are installed on the inner wall of the separation chamber 6 and connected to an external analysis device 62. The detectors 61 are arranged in a matrix and are closely attached to the inner wall of the separation chamber 6. When the accelerated complex is deflected by the magnetic field, it is received by the detectors 61. The analysis device 62 connected to the detectors 61 judges and calculates the abundance and relative molecular mass of various protein molecules based on the position signals of molecules in the solution received by the detectors 61 and the order of receipt.
[0028] The uniform magnetic field generator 7 includes two sets of Helmholtz coils 71 arranged coaxially on both sides of the separation chamber 6. Both sets of Helmholtz coils 71 are externally connected to an adjustment circuit. The adjustment circuit is equipped with a digitally controlled power supply and an on / off switch. The adjustment circuit can be connected in series or in parallel.
[0029] The analysis device 62 is equipped with a collaborative control platform that is electrically connected to the uniform electric field generator 5 and the uniform magnetic field generator 7. After trial operation, the collaborative control platform matches the database with the initial data and adjusts the electric field strength and magnetic induction intensity according to the molecular mass of the target protein through an algorithm to increase the spacing of the complex deflection trajectory, thereby improving the accuracy of the measurement results.
[0030] The adjustment of magnetic induction intensity is mainly achieved by controlling the digital power supply of the regulating circuit. The integrated digital power supply output module replaces the traditional sliding rheostat, realizing precise digital control of the uniform magnetic field.
[0031] Reference Figure 3 The adjustment circuit is not limited to the above structure. It can also be configured to include a sliding rheostat 72 connected in series, a power supply and an on / off switch. By changing the resistance of the sliding rheostat 72, the current in the two sets of Helmholtz coils 71 can be adjusted by the precise adjustment function of the sliding rheostat 72, thereby achieving fine control of the magnetic induction intensity of the uniform magnetic field.
[0032] As a preferred structure, valves or on / off switches are installed at the connection points of each connection structure, mainly at the connection point between the outlet end of the mixing container 1 and the pipeline 3, to ensure that each structure achieves its corresponding function.
[0033] The implementation principle of a detection device for protein analysis based on magnetophoresis in Embodiment 1 of the invention is as follows: The protein solution to be detected is poured into a mixing container 1 containing SDS solution. After being heated by a temperature control device 2, it enters a pipe section 3 containing electrophoresis buffer and surrounded by an ultrasonic stirrer 4 for thorough mixing. The mixture enters the pipe section 3 placed in a uniform electric field 5 to accelerate the molecules in the mixture. The accelerated complex enters a separation chamber 6 placed in a uniform magnetic field formed by two sets of Helmholtz coils 71 for deflection. The deflected molecules are detected by a detector 61, and the position signal of the molecules is displayed by an analysis device 62 connected to the detector 61, and the abundance and relative molecular mass of the protein molecules are calculated.
[0034] Example 2 Based on Embodiment 1, Embodiment 2 discloses a detection method for a detection device based on magnetophoretic protein analysis.
[0035] Reference Figure 1 , Figure 4 A detection method for a protein detection device based on magnetophoretic analysis, applied to such a device, includes the following steps: Step 1: Inject the protein solution to be tested into the mixing container 1 containing the SDS solution at a preset flow rate; Step 2: Use temperature control device 2 to adjust the mixture of protein solution and SDS solution in the container to 30-50℃ and maintain the temperature, so that the protein molecules and SDS can fully combine and be uniformly charged. Step 3: The mixture enters the section of pipe 3 equipped with ultrasonic stirrer 4. Under the vibration of ultrasonic stirrer 4, it mixes evenly with the electrophoresis buffer in pipe 3 to form a complex. Step 4: The composite material enters the section of pipe 3 equipped with a uniform electric field generator 5 and is accelerated along the axial direction of pipe 3 under the action of the uniform electric field. Step 5: The accelerated complex enters the separation chamber 6 and deflects along the direction perpendicular to the motion under the action of the Lorentz force in the uniform magnetic field region. Complexes with different molecular masses form different deflection trajectories. The larger the molecular mass, the smaller the deflection radius in the magnetic field; and vice versa.
[0036] Step 6: Use detector 61 to detect the arrival location and signal intensity data of each complex in real time, and transmit it to analysis device 62. Analysis device 62 calculates the relative size and abundance of protein molecular mass and outputs the detection results.
[0037] As a preferred method, in step six, the analysis device 62 is equipped with a collaborative control platform electrically connected to the uniform electric field generator 5 and the uniform magnetic field generator 7. The collaborative control platform matches the database and adjusts the electric field strength and magnetic induction intensity according to the molecular weight of the target protein to increase the spacing between the deflection trajectories of the complexes, so that the spacing between the deflection trajectories of complexes with different molecular weights is significantly expanded to facilitate observation and improve the separation resolution.
[0038] Example 3 Embodiment 3 of the invention discloses a detection device based on magnetophoretic analysis of proteins.
[0039] A detection device for protein analysis based on magnetophoresis includes a pre-magnetophoresis preparation section and a protein magnetophoresis section connected in sequence. The pre-magnetophoresis preparation section includes a mixing container 1 containing SDS solution, and a pipe 3 is connected to the outlet end of the mixing container 1. The pipe 3 passes sequentially through a stirring area equipped with an ultrasonic stirrer 4 and a uniform electric field area equipped with a uniform electric field generator 5. The protein magnetophoresis section includes a separation chamber 6, which is set in a uniform magnetic field area formed by a uniform magnetic field generator 7. A detector 61 is installed on the inner wall of the separation chamber 6 and an external analysis device 62 is connected to it.
[0040] Reference Figure 5 An ultrasonic stirrer 4 is arranged around the section of the pipe 3 pre-filled with electrophoresis buffer, and an injector 31 is installed at the front end of the pipe 3.
[0041] After a single test is completed, cleaning fluid is injected through injector 31 on pipe 3 to rinse pipe 3 and then precharge the electrophoresis buffer for the next test.
[0042] The above are all preferred embodiments of the present invention and are not intended to limit the scope of protection of the present invention. Therefore, all equivalent changes made in accordance with the structure, shape and principle of the present invention should be covered within the scope of protection of the present invention.
Claims
1. A detection device for proteins based on magnetophoretic analysis, characterized in that: It includes a pre-magnetophoresis preparation section and a protein magnetophoresis section that are connected in sequence; The pre-magnetization preparation section includes a mixing container (1) containing SDS solution. The outlet end of the mixing container (1) is connected to a pipe (3). The pipe (3) passes through a stirring area equipped with an ultrasonic stirrer (4) and a uniform electric field area equipped with a uniform electric field generator (5) in sequence. The protein magnetophoresis section includes a separation chamber (6), which is set in a uniform magnetic field region formed by a uniform magnetic field generator (7). A detector (61) is installed on the inner wall of the separation chamber (6) and an external analysis device (62) is connected.
2. The detection device for protein analysis based on magnetophoresis according to claim 1, characterized in that: The uniform magnetic field generator (7) includes two sets of Helmholtz coils (71) arranged coaxially on both sides of the separation chamber (6). Both sets of Helmholtz coils (71) are externally connected to an adjustment circuit, which is equipped with a digital control power supply and an on / off switch.
3. The detection device for protein analysis based on magnetophoresis according to claim 1, characterized in that: The ultrasonic stirrer (4) is arranged around the section of the pipe (3) pre-filled with electrophoresis buffer, and an injector (31) is installed at the front end of the pipe (3).
4. The detection device for proteins based on magnetophoretic analysis according to claim 1, characterized in that: The mixing container (1) is provided with a temperature control device (2), which includes an electric heating wire (21) wound around the outer wall of the mixing container (1). The electric heating wire (21) is connected to the temperature control unit (22) and the power supply through a line. The mixing container (1) is provided with a temperature sensor (23) that is electrically connected to the temperature control unit (22).
5. The detection device for proteins based on magnetophoretic analysis according to claim 4, characterized in that: The mixing container (1) is also equipped with a paddle agitator (11).
6. A detection method for a protein detection device based on magnetophoretic analysis, applied to the protein detection device based on magnetophoretic analysis as described in any one of claims 1-5, characterized in that, Includes the following steps: Step 1: Inject the protein solution to be tested into a mixing container (1) containing SDS solution at a preset flow rate; Step 2: Use a temperature control device (2) to adjust the mixture of protein solution and SDS solution in the container to 30-50℃ and maintain the temperature, so that the protein molecules and SDS are fully combined and uniformly charged. Step 3: The mixture enters the section of pipe (3) equipped with an ultrasonic stirrer (4). Under the vibration of the ultrasonic stirrer (4), it mixes evenly with the electrophoresis buffer in the pipe (3) to form a complex. Step 4: The composite material enters the section of pipe (3) equipped with a uniform electric field generator (5) and is accelerated along the axial direction of pipe (3) under the action of the uniform electric field. Step 5: The accelerated complex enters the separation chamber (6) and deflects along the direction perpendicular to the motion under the action of the Lorentz force in the uniform magnetic field region. Complexes with different molecular weights form different deflection trajectories. Step 6: Use detector (61) to detect the arrival location and signal intensity data of each complex in real time, and transmit them to the analysis device (62). The analysis device (62) calculates the relative size and abundance of protein molecular mass and outputs the detection results.
7. The detection method of the detection device based on magnetophoretic protein analysis according to claim 6, characterized in that, In step six: The analysis device (62) is equipped with a collaborative control platform that is electrically connected to the uniform electric field generator (5) and the uniform magnetic field generator (7). The collaborative control platform adjusts the electric field strength and magnetic induction intensity through a matching database.
8. The detection method of the detection device based on magnetophoretic protein analysis according to claim 6, characterized in that: After a single test is completed, cleaning fluid is injected through the injector (31) set on the pipe (3) to rinse the pipe (3) and then the electrophoresis buffer is precharged again for the next test.