Protein analyzer and protein analysis detection method

Abstract

The application relates to the technical field of medical equipment, and discloses a protein analyzer and a protein analysis detection method. The first communication port to the sixth communication port are integrated through a flow path switching device, and are respectively communicated with a sampling needle, a first pump, a quantitative ring, a reagent container, a third pump and a detector; the single flow path switching device is used to realize the switching of multiple flow paths such as sampling, sample feeding and elution analysis, the number of flow path switching elements is effectively reduced, the pipeline connection is simplified, the flow path dead zone is reduced, sample residue and cross contamination are inhibited, and the flow path response speed is improved. Through cooperation of the pumps and the flow path switching device, sampling, mixing, sample feeding and elution analysis and other operations are efficiently connected in time sequence, the overall experiment time is shortened, and the detection efficiency is improved.

Description

Technical Field

[0001] This invention relates to the field of medical device technology, specifically to a protein analyzer and a protein analysis and detection method. Background Technology

[0002] In the field of medical devices, protein analyzers are used to quantitatively detect specific proteins (such as glycated hemoglobin) in blood samples. Generally, protein analyzers use a plunger pump to drive multiple eluents through a quantitative loop, propelling the mixed blood sample through a heating wire for heating. After separation by a chromatography column, the sample is analyzed by turbidimetric or colorimetric methods in a detection cell to obtain the concentration value of the specific protein. This process typically involves auxiliary components such as a mixing cell and injection needle, and requires multiple directional valves and power components to complete operations such as sample injection, mixing, washing, and elution analysis. This results in a complex flow path structure, lengthy tubing connections, a large dead zone (dead volume), a high risk of sample residue, and a slow flow path response. Summary of the Invention

[0003] This invention provides a protein analyzer and a protein analysis and detection method to solve the problems of complex flow paths, large dead zones, and low detection efficiency mentioned above.

[0004] This invention provides a protein analyzer, comprising a sampling needle, a quantitative loop, a first pump, a second pump, a mixing chamber, a flow path switching device, at least one reagent container, a third pump, and a detector. The sampling needle is used to draw blood samples and inject them into the mixing chamber. The second pump, connected to the mixing chamber, is used to inject a hemolytic agent into the mixing chamber. The flow path switching device includes a first connection port, a second connection port, a third connection port, a fourth connection port, a fifth connection port, and a sixth connection port. The sampling needle is connected to the first connection port, and the first pump is connected to the second connection port. When the flow path switching device switches the connection between the first and second connection ports, the sampling needle, the first connection port, the second connection port, and the first pump constitute a first detection flow path. One end of the quantitative loop is connected to the third connection port, and the other end of the quantitative loop is connected to the fourth connection port. When the flow path switching device switches the connection between the first and third connection ports, and the fourth connection port is connected to the second connection port, the sampling needle, the first connection port, the third connection port, the quantitative loop, the fourth connection port, the second connection port, and the first pump constitute a second detection flow path. The reagent container is connected to the fifth connection port, the detector inlet is connected to the sixth connection port, and the third pump is installed on the pipeline between the reagent container and the fifth connection port. When the flow path switching device switches the fifth connection port to the third connection port and the fourth connection port to the sixth connection port, the reagent container, the third pump, the fifth connection port, the third connection port, the quantitative loop, the fourth connection port, the sixth connection port and the detector constitute the third detection flow path.

[0005] This configuration integrates the first to sixth connecting ports via a flow path switching device, connecting them to the sampling needle, first pump, quantitative loop, reagent container, third pump, and detector, respectively. A single flow path switching device enables switching between multiple flow paths for sampling, injection, and elution analysis, effectively reducing the number of flow path switching components, simplifying tubing connections, minimizing dead zones, suppressing sample residue and cross-contamination, and improving flow path response speed. Simultaneously, the first pump drives the sampling needle to aspirate blood samples via the first detection flow path and injects the sample into the quantitative loop via the second detection flow path. The second pump injects hemolytic agent into the mixing chamber, and the third pump independently drives the reagent through the third detection flow path, delivering it along with the sample in the quantitative loop to the detector for elution analysis. Through the coordination of each pump and the flow path switching device, sampling, mixing, injection, and elution analysis operations are efficiently sequentially connected, shortening the overall experimental time and improving detection efficiency. Furthermore, the coordination of the quantitative loop with the flow path switching device and detector ensures accurate quantification of the sample in the quantitative loop before elution and delivery to the detector, improving detection accuracy and repeatability.

[0006] Secondly, the present invention also provides a protein analysis and detection method, comprising: Sampling steps: The flow path switching device switches the first connection port and the second connection port to connect, the first valve port and the second valve port of the three-way valve are connected, the first pump causes the sampling needle to draw blood samples, and the drawn blood samples are injected into the mixing pool; Mixing step: The first and fifth ports of the five-way valve are connected, and the hemolytic agent is injected into the mixing tank through the second pump. The hemolytic agent and blood sample in the mixing tank are repeatedly aspirated to ensure that they are fully mixed. Sampling procedure: The first pump draws the mixed hemolysin and blood sample from the sampling needle; the flow path switching device switches the first connection port to the third connection port and the fourth connection port to the second connection port, and the first pump injects the mixed hemolysin and blood sample drawn by the sampling needle into the quantitative loop; Detection steps: The flow path switching device switches the fifth connection port to the third connection port and the fourth connection port to the sixth connection port. The reagent in the reagent container and the hemolysin mixed in the quantitative loop and the blood sample are delivered to the detector by the third pump for elution analysis. Attached Figure Description

[0007] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0008] Figure 1 This is a schematic diagram of the flow path of the protein analysis and detection instrument according to an embodiment of the present invention; Figure 2 This is a schematic diagram of the first cleaning flow path for solution filling before sampling, according to an embodiment of the present invention. Figure 3 This is a schematic diagram of the flow path for the second pump to draw and fill the hemolytic agent according to an embodiment of the present invention; Figure 4 This is a schematic diagram of the flow path of the second pump driving the hemolytic agent to clean the sampling needle according to an embodiment of the present invention; Figure 5 This is a schematic diagram of the flow path for filling the hemolytic agent in the first pump according to an embodiment of the present invention; Figure 6 This is a schematic diagram of the flow path of the first pump driving the hemolytic agent to clean the sampling needle and the mixing tank according to an embodiment of the present invention; Figure 7 This is a schematic diagram of the flow path of the second pump driving the hemolytic agent to clean the mixing tank according to an embodiment of the present invention; Figure 8 This is a schematic diagram of the flow path for the second pump to remove the hemolytic agent from the mixing tank according to an embodiment of the present invention; Figure 9 This is a schematic diagram of the sampling flow path of the sampling needle in an embodiment of the present invention; Figure 10 This is a schematic diagram of the flow path for injecting the blood sample into the mixing tank using a sampling needle according to an embodiment of the present invention; Figure 11 This is a schematic diagram of the flow path for mixing blood samples and hemolysing agents in the mixing tank using the second pump according to an embodiment of the present invention; Figure 12 This is a schematic diagram of the flow path for the quantitative loop injection in an embodiment of the present invention; Figure 13 This is a schematic diagram of the detection flow path according to an embodiment of the present invention; Figure 14 This is a flow diagram of the second pump pushing the hemolysin to clean the injection needle after detection, according to an embodiment of the present invention. Figure 15 This is a flow diagram of the first pump pushing the hemolysin to clean the injection needle after detection, according to an embodiment of the present invention. Figure 16 This is a flow diagram of the second pump driving the hemolysin to clean the mixing tank after detection, according to an embodiment of the present invention. Figure 17This is a flow diagram of the second pump removing the hemolytic agent from the mixing tank after detection, according to an embodiment of the present invention.

[0009] Explanation of reference numerals in the attached figures: 1-Sampling needle; 2-Quantitative loop; 3-First pump; 4-Second pump; 5-Mixing cell; 6-Flow path switching device; 61-First connecting port; 62-Second connecting port; 63-Third connecting port; 64-Fourth connecting port; 65-Fifth connecting port; 66-Sixth connecting port; 67-Seventh connecting port; 7-Reagent container; 8-Third pump; 9-Detector; 91-Heating wire; 92-Synthesis column; 93-Detection cell; 10-Three-way valve; T1-First valve port; T2-Second valve port; T3-Third valve port; 11-Fourth pump; 12-Five-way valve; K1-First interface; K2-Second interface; K3-Third interface; K4-Fourth interface; K5-Fifth interface; 13-Mixer; 14-Four-way valve; 15-Degassing pump; 16-Filter; 17-Wastewater outlet; 18-Hemolysin container. Detailed Implementation

[0010] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0011] The following is combined Figures 1 to 17 The embodiments of the present invention will be described in detail below.

[0012] According to an embodiment of the present invention, a protein analyzer is provided. The protein analyzer includes a sampling needle 1, a quantitative loop 2, a first pump 3, a second pump 4, a mixing chamber 5, a flow path switching device 6, at least one reagent container 7, a third pump 8, and a detector 9. The sampling needle 1 is used to draw blood samples and inject them into the mixing chamber 5. The second pump 4 is connected to the mixing chamber 5 and is used to inject a hemolytic agent into the mixing chamber 5. The flow path switching device 6 includes a first connection port 61, a second connection port 62, a third connection port 63, a fourth connection port 64, a fifth connection port 65, and a sixth connection port 66. The sampling needle 1 is connected to the first connection port 61, and the first pump 3 is connected to the second connection port 62. When the flow path switching device 6 switches the connection between the first connection port 61 and the second connection port 62, the sampling needle 1, the first connection port 61, the second connection port 62, and the first pump 3 constitute a first detection flow path. One end of the quantitative loop 2 is connected to the third connection port 63, and the other end of the quantitative loop 2 is connected to the fourth connection port 64. When the flow path switching device 6 switches the connection between the first connection port 61 and the third connection port 63, and the fourth connection port 64 and the second connection port 62, the sampling needle 1, the first connection port 61, the third connection port 63, the quantitative loop 2, the fourth connection port 64, the second connection port 62, and the first pump 3 constitute the second detection flow path. The reagent container 7 is connected to the fifth connection port 65, the inlet of the detector 9 is connected to the sixth connection port 66, and the third pump 8 is installed on the pipeline between the reagent container 7 and the fifth connection port 65. When the flow path switching device 6 switches the connection between the fifth connection port 65 and the third connection port 63, and the fourth connection port 64 and the sixth connection port 66, the reagent container 7, the third pump 8, the fifth connection port 65, the third connection port 63, the quantitative loop 2, the fourth connection port 64, the sixth connection port 66, and the detector 9 constitute the third detection flow path.

[0013] The sampling needle 1 can be a needle-shaped structure with a hollow cavity, one end forming a tip for piercing the blood sample container to draw blood samples, and the other end connected to the first communication port 61 through a tube. The sampling needle 1 can also inject the drawn blood sample into the mixing pool 5.

[0014] The mixing tank 5 can be a container with a cavity for receiving blood samples and hemolysing agents and providing a mixing reaction space.

[0015] The two ends of the quantitative loop 2 are connected to the third connecting port 63 and the fourth connecting port 64, respectively, for accommodating a predetermined volume of sample. The quantitative loop 2 can be a ring-shaped tube with a fixed volume, for example, the volume can be, but is not limited to, 5 μL.

[0016] The first pump 3 can be a syringe pump, for example, a 0.5mL syringe pump, used to draw and push liquid.

[0017] The second pump 4 can be an injection pump, for example, a 5mL injection pump, used to inject a hemolytic agent into the mixing tank 5.

[0018] Understandably, the capacity of the syringe pump is not limited to the above-mentioned 0.5mL or 5mL, and can be selected according to needs.

[0019] The flow path switching device 6 is a multi-way valve structure used to switch the connection status of various flow paths inside the protein analyzer.

[0020] The flow path switching device 6 may include a valve body and a valve core. The valve body has a first connecting port 61, a second connecting port 62, a third connecting port 63, a fourth connecting port 64, a fifth connecting port 65, and a sixth connecting port 66. Each connecting port is connected to a corresponding component through a pipeline. The valve core can rotate or move relative to the valve body to change the connection relationship between the connecting ports.

[0021] Specifically, the first connection port 61 can be connected to the sampling needle 1 via a tubing, and the second connection port 62 can be connected to the first pump 3 via a tubing. The third connection port 63 and the fourth connection port 64 can be connected to both ends of the quantitative loop 2 via tubing, respectively. The fifth connection port 65 can be connected to the reagent container 7 via a tubing (the third pump 8 is located on this tubing), and the sixth connection port 66 can be connected to the inlet of the detector 9 via a tubing.

[0022] For example, the flow path switching device 6 can be a four-position seven-way rotary valve with four working positions and seven communication ports. In addition to the first communication port 61 to the sixth communication port 66 mentioned above, it can also include a seventh communication port 67 (described below). The seventh communication port 67 can be connected to the hemolytic agent container 18 through a pipeline.

[0023] The valve core of the four-position seven-way rotary valve can switch to different working positions by rotating a certain angle (e.g., 60°). In each working position, the flow channel inside the valve core selectively connects different ports to form different flow paths.

[0024] For example, in the first working position, the first connecting port 61 is connected to the second connecting port 62; in the second working position, the first connecting port 61 is connected to the third connecting port 63 and the fourth connecting port 64 is connected to the second connecting port 62; in the third working position, the fifth connecting port 65 is connected to the third connecting port 63 and the fourth connecting port 64 is connected to the sixth connecting port 66; in the fourth working position, the seventh connecting port 67 is connected to the second connecting port 62.

[0025] In another optional embodiment, the flow path switching device 6 can be a valve group composed of multiple solenoid valves, each solenoid valve controlling the opening and closing of the corresponding pipeline, and the connection state switching between each connection port is realized through the coordinated switching action of multiple solenoid valves.

[0026] The flow path switching device 6 integrates multiple connection ports into a single valve body, enabling the switching of multiple flow paths in the protein analyzer, such as sampling, injection, elution analysis, and cleaning, to be completed through only one device. This effectively reduces the number of external pipeline connections and joints, lowers the flow path dead zone (dead volume) and leakage risk, and improves the flow path response speed and structural compactness.

[0027] The reagent container 7 can be a reagent bottle or reagent jar, used to hold the reagents required for elution analysis, such as the first reagent, the second reagent, or the third reagent.

[0028] The third pump 8 can be a plunger pump, used to provide high-pressure driving force to deliver reagents to the detector 9.

[0029] The detector 9 may include a heating wire 91, a synthesis column 92, and a detection cell 93 arranged sequentially from upstream to downstream. The heating wire 91 is used to heat the liquid in the flow path to a suitable temperature, the synthesis column 92 is used to elute and separate the sample, and the detection cell 93 is used to measure the concentration of the complex solution by turbidimetric method.

[0030] During protein analysis and detection, refer to Figure 9 and Figure 10 The flow path switching device 6 switches the connection between the first connection port 61 and the second connection port 62. The first pump 3 drives the sampling needle 1 to draw blood samples through the first detection flow path, and the sampling needle 1 injects the drawn blood samples into the mixing tank 5. (Refer to...) Figure 11 The second pump 4 injects a hemolytic agent into the mixing tank 5. The blood sample and the hemolytic agent react in the mixing tank 5 to form a complex. (Refer to...) Figure 12 The flow path switching device 6 switches the connection between the first connecting port 61 and the third connecting port 63, and the fourth connecting port 64 and the second connecting port 62. The first pump 3 drives the sampling needle 1 through the second detection flow path to draw the mixed sample from the mixing cell 5 and inject the mixed sample into the metering loop 2. The metering loop 2 contains a set volume of mixed sample, for example, 5 μL of mixed sample, and the first pump 3 can extract a sample volume exceeding the volume of the metering loop 2, for example, 50 μL, to keep the mixed sample inside the metering loop 2 clean. (Refer to...) Figure 13 The flow path switching device 6 switches the fifth connection port 65 to the third connection port 63, and the fourth connection port 64 to the sixth connection port 66. The third pump 8 delivers the reagent in the reagent container 7 and the mixed sample in the quantitative loop 2 to the detector 9 through the third detection flow path. The reagent and sample are heated by the heating wire 91 and eluted by the synthesis column 92 in sequence, and then turbidity detection is performed in the detection cell 93 to obtain the concentration value of the specific protein.

[0031] In the above scheme, the flow path switching device 6 integrates the first connecting port 61 to the sixth connecting port 66, and uses a single flow path switching device 6 to switch between multiple flow paths such as sampling, injection, and elution analysis. This effectively reduces the number of flow path switching components, simplifies pipeline connections, reduces flow path dead zones, suppresses sample residue and cross-contamination, and improves flow path response speed. The first pump 3 achieves sampling and injection through the first and second detection flow paths respectively, making sampling and injection operations efficiently connected in sequence and shortening the overall experimental time. The quantitative loop 2 works in conjunction with the flow path switching device 6 and the detector 9 to ensure that the sample is accurately quantified in the quantitative loop 2 and then eluted by reagents before being delivered to the detector 9, which helps to improve detection accuracy and repeatability. The third pump 8 independently drives the reagent through the third detection flow path to be delivered to the detector 9 along with the sample in the quantitative loop 2, ensuring the stability and reliability of the elution analysis process. At the same time, it ensures that reagent delivery does not interfere with the sample sampling, mixing, and injection processes, further improving detection efficiency. Meanwhile, since the relative positions of each connection port on the valve body are fixed, the pipeline connection length is shortened, which helps to reduce sample residue in the pipeline and reduce the risk of cross-contamination.

[0032] In some embodiments, refer to Figure 2 The protein analyzer also includes a three-way valve 10 and a fourth pump 11. The first valve port T1 of the three-way valve 10 is connected to the second connecting port 62 via a pipeline, the second valve port T2 of the three-way valve 10 is connected to the sampling needle 1 via a pipeline, and the third valve port T3 of the three-way valve 10 is connected to the inlet of the fourth pump 11 via a pipeline. The outlet of the fourth pump 11 is used to connect to the waste liquid outlet. When the first valve port T1 of the three-way valve 10 is connected to the second valve port T2, the sampling needle 1 is connected to the second connecting port 62. The flow path switching device 6 switches the fifth connecting port 65 to the second connecting port 62. When the first valve port T1 of the three-way valve 10 is connected to the third valve port T3, the reagent container 7, the third pump 8, the fifth connecting port 65, the second connecting port 62, the three-way valve 10, and the fourth pump 11 constitute the first filling and cleaning flow path.

[0033] The three-way valve 10 can be an electromagnetic directional valve with three valve ports. The valve body has a movable valve core inside, and selective connection or disconnection between different valve ports can be achieved by changing the position of the valve core.

[0034] When the flow path switching device 6 switches the fifth connection port 65 to the second connection port 62, and the first valve port T1 of the three-way valve 10 is connected to the second valve port T2, the third pump 8 draws the reagent from the reagent container 7, and under the action of the fourth pump 11, the reagent fills the entire first filling and cleaning flow path, realizing the filling of the reagent, the cleaning of the flow path and the removal of air bubbles.

[0035] This configuration allows for the pre-filling of reagents into the flow path between the third pump 8 and the detector 9 during the power-on preparation phase or before detection. This process removes air bubbles from the flow path and cleans it, ensuring the accuracy and stability of subsequent elution analysis. Simultaneously, the inclusion of a three-way valve 10 and a fourth pump 11 allows reagent filling and waste liquid discharge to be completed through the same flow path switching device 6 and the fourth pump 11. This further simplifies the number of flow path components, reduces flow path complexity, and ensures that flow path filling and sample detection processes do not interfere with each other, thus improving the flexibility and efficiency of flow path switching.

[0036] In some embodiments, the protein analyzer further includes a hemolysin container 18. The flow path switching device 6 also includes a seventh connection port 67, which is connected to the hemolysin container 18.

[0037] The hemolytic agent container 18 can be a reagent bottle or reagent tank for holding the hemolytic agent, which is used to mix and react with the blood sample to lyse the red blood cells in the blood sample and generate a complex that can be detected by turbidimetric analysis.

[0038] When the flow path switching device 6 switches the seventh connection port 67 to the second connection port 62, and the first valve port T1 of the three-way valve 10 is connected to the second valve port T2, the hemolytic agent container 18, the seventh connection port 67, the second connection port 62, the three-way valve 10 and the first pump 3 constitute the second filling and cleaning flow path.

[0039] Reference Figure 5 The first pump 3 draws the hemolytic agent from the hemolytic agent container 18 into the first pump 3 through the second filling and cleaning flow path. For example, if the first pump 3 is a 0.5 mL syringe pump, the first pump 3 can draw 0.5 mL of hemolytic agent. Subsequently, refer to Figure 6 The flow path switching device 6 switches the first connection port 61 to the second connection port 62, and the first pump 3 pushes out the hemolytic agent inside. The hemolytic agent flows out sequentially through the second connection port 62, the three-way valve 10, and the sampling needle 1 to clean the inner wall of the sampling needle 1. Further, the hemolytic agent flows from the outlet of the mixing tank 5 to the waste liquid outlet after passing through the mixing tank 5 to clean the mixing tank 5.

[0040] By connecting the seventh port 67 to the hemolytic agent container 18, the hemolytic agent can be directly connected to the first pump 3 via the flow path switching device 6, simplifying the pipeline connection from the hemolytic agent to the first pump 3, reducing the number of joints, and lowering the risk of leakage. Simultaneously, the first pump 3 is used for filling the hemolytic agent and cleaning the sampling needle 1 and mixing tank 5, reducing the need for a dedicated cleaning pump, lowering equipment costs, and reducing flow path complexity.

[0041] In some embodiments, the protein analyzer further includes a five-way valve 12. The five-way valve 12 has a first interface K1, a second interface K2, a third interface K3, a fourth interface K4, and a fifth interface K5, and can be an electromagnetic directional valve, which can selectively connect different interfaces by rotating or moving the valve core.

[0042] The second pump 4 is connected to the first port K1 of the five-way valve 12. The hemolytic agent container 18 is connected to the second port K2 of the five-way valve 12 through a pipeline. The seventh port 67 is connected to the third port K3 of the five-way valve 12. The fourth port K4 of the five-way valve 12 is connected to the sampling needle 1 through a pipeline. The fifth port K5 of the five-way valve 12 is connected to the mixing tank 5 through a pipeline.

[0043] When the second port K2 of the five-way valve 12 is connected to the third port K3, the seventh port 67 is connected to the hemolytic agent container 18, so that the hemolytic agent in the hemolytic agent container 18 can enter the flow path switching device 6 through the seventh port 67, thereby forming the above-mentioned second filling and cleaning flow path.

[0044] Reference Figure 3 When the first port K1 of the five-way valve 12 is connected to the second port K2, the hemolytic agent container 18, the second port K2, the first port K1 and the second pump 4 constitute the third filling and cleaning flow path, and the second pump 4 can draw the hemolytic agent into its own pump chamber.

[0045] Reference Figure 4 When the first port K1 of the five-way valve 12 is connected to the fourth port K4, the second pump 4, the first port K1, the fourth port K4 and the sampling needle 1 form the fourth filling and cleaning flow path. The second pump 4 pushes out the hemolysing agent inside, and the hemolysing agent flows out through the outer wall of the sampling needle 1 to clean the outer wall of the sampling needle 1.

[0046] Reference Figure 7 When the first port K1 of the five-way valve 12 is connected to the fifth port K5, the second pump 4, the first port K1, the fifth port K5 and the mixing tank 5 constitute the fifth filling cleaning flow path. The second pump 4 pushes the hemolysing agent in it into the mixing tank 5 to clean the mixing tank 5.

[0047] In one working state, the second pump 4 (e.g., a 5mL syringe pump) first draws the hemolysing agent into the pump chamber through the third filling and cleaning flow path; then, by switching the five-way valve 12 to connect the first port K1 and the fourth port K4, the second pump 4 pushes the hemolysing agent (e.g., 900μL) through the outer wall of the sampling needle 1, and the waste liquid flows to the waste liquid port, thus cleaning the outer wall of the sampling needle 1; then, by switching the five-way valve 12 to connect the first port K1 and the fifth port K5, the second pump 4 pushes the hemolysing agent (e.g., 2mL) through the mixing tank 5, and it flows out through the waste liquid port of the mixing tank 5, thus cleaning the mixing tank 5; after cleaning, the second pump 4 can also remove the hemolysing agent in the mixing tank 5.

[0048] By incorporating a five-way valve 12, the second pump 4 can be reused to perform multiple functions, including hemolytic agent aspiration, cleaning of the outer wall of sampling needle 1, and cleaning of mixing tank 5. This further reduces the number of pumps and valves, simplifying the flow path structure. Simultaneously, the five-way valve 12, in conjunction with the flow path switching device 6 and the three-way valve 10, ensures that the cleaning of the inner and outer walls of sampling needle 1, the cleaning of mixing tank 5, and the switching of the main detection flow path do not interfere with each other. Each cleaning step can be performed independently or prepared in parallel with the main detection process, improving overall experimental efficiency.

[0049] In some embodiments, a mixer 13 may be provided on the pipeline between the fifth interface K5 and the mixing pool 5.

[0050] Mixer 13 is a device for enhancing the mixing effect of fluids, such as a magnetic stirrer, ultrasonic mixer 13, or static mixer 13.

[0051] When the second pump 4 injects the hemolytic agent into the mixing tank 5 through the fifth filling and cleaning flow path, or when the blood sample and the hemolytic agent are mixed in the mixing tank 5, the mixer 13 can enhance the mixing effect of the hemolytic agent and the blood sample, promote the lysis of red blood cells in the blood sample and their full reaction with the hemolytic agent to form a complex. By setting up the mixer 13, the uniformity of mixing between the blood sample and the hemolytic agent is improved, which helps to shorten the reaction time and improve the detection reaction efficiency and repeatability.

[0052] In some embodiments, reagent containers 7 may be provided in three parts, each for holding different reagents, such as a first reagent, a second reagent, and a third reagent. The protein analyzer may also include a four-way valve 14, which has four ports and may be an electromagnetic directional valve.

[0053] The three reagent containers 7 are connected to the three ports of the four-way valve 14 via tubing, and the other port of the four-way valve 14 is connected to the inlet of the third pump 8. By switching the different ports of the four-way valve 14, the third pump 8 can selectively connect to any one of the three reagent containers 7, thereby extracting different reagents for elution analysis.

[0054] During the detection process, the third pump 8 can draw the first reagent through the four-way valve 14 and elute the sample in the quantitative loop 2 through the third detection flow path. When it is necessary to change the reagent, the four-way valve 14 is switched to connect the third pump 8 with the second or third reagent to achieve sequential or gradient elution of different reagents.

[0055] By setting a four-way valve 14, the third pump 8 can be compatible with the supply of multiple reagents, reducing the number of pumps and facilitating the switching of multiple reagents and gradient elution, thereby improving the detection flexibility and applicability of the protein analyzer.

[0056] In some embodiments, a degassing pump 15 may be installed on the pipeline between the reagent container 7 and the four-way valve 14.

[0057] The degassing pump 15 can be a device for removing dissolved gases from a reagent, such as a vacuum degassing pump 15 or an ultrasonic degasser.

[0058] During storage and transportation, reagents may dissolve air bubbles. These air bubbles entering the detection flow path can affect flow stability and detection accuracy. The degassing pump 15 can remove air bubbles from the reagent before it enters the four-way valve 14 and the third pump 8, ensuring that the reagent entering the third pump 8 and subsequent flow paths is in a degassed state.

[0059] By setting up the degassing pump 15, the interference of air bubbles on the elution analysis process is reduced, and the stability of the flow path flow rate and the accuracy of the detection results are improved.

[0060] In some embodiments, a filter 16 may be provided on the pipeline between the sixth connection port 66 and the detector 9.

[0061] The filter 16 can be an in-line filter, such as a microporous filter membrane or sieve, used to filter particulate impurities in reagents or samples and prevent particles from entering the detector 9 and clogging the heating wire 91, synthesis column 92 or detection cell 93.

[0062] By setting up filter 16, the delicate internal components of detector 9 are protected, the risk of clogging is reduced, the service life of detector 9 is extended, and the cleanliness of samples and reagents entering detector 9 is ensured, thereby improving detection reliability.

[0063] In some embodiments, the detector 9 may include a heating wire 91, a synthesis column 92, and a detection cell 93 arranged sequentially from upstream to downstream.

[0064] The component through which the liquid flows first is called the upstream component, and the component through which it flows last is called the downstream component.

[0065] Heating wire 91 can be an electrothermal element wound or attached to the wall of the flow path tube, used to heat the liquid in the flow path to a suitable temperature (e.g., 37°C or the optimal temperature required by the method) to ensure that the sample reaction and elution separation are carried out under optimal temperature conditions.

[0066] The synthesis column 92 can be a chromatography column packed with a stationary phase, used to separate and elute different components in a sample. The detection cell 93 can be a colorimetric cell or a turbidimetric cell, used to measure the concentration of the complex solution by optical methods (e.g., turbidimetry) and obtain the concentration value of a specific protein (e.g., HbA1C) by conversion.

[0067] By integrating the heating wire 91, synthesis column 92, and detection cell 93 into a detector 9, the sample is heated, separated, and detected sequentially during transport, resulting in a compact process. The pre-positioning of the heating wire 91 ensures the sample reaches a suitable temperature before entering the synthesis column 92, improving separation efficiency and reaction rate. The synthesis column 92 effectively separates the sample, reducing interference between different components. The detection cell 93 directly detects the separated sample, enhancing detection accuracy and sensitivity.

[0068] Embodiments of this application also provide a protein analysis and detection method, applied to the protein analyzer described in the above embodiments. The protein analysis and detection method includes a sampling step, a mixing step, a sample injection step, and a detection step.

[0069] In the sampling process, refer to Figure 9 and Figure 10 The flow path switching device 6 switches the first connection port 61 and the second connection port 62 to connect, and the first valve port T1 and the second valve port T2 of the three-way valve 10 are connected. The sampling needle 1 draws blood samples through the first pump 3 and injects the drawn blood samples into the mixing pool 5.

[0070] For example, the first pump 3 can be a 0.5 mL syringe pump, and the blood sample volume drawn by the sampling needle 1 can be 3 μL.

[0071] In the mixing step, refer to Figure 11 The first port K1 and the fifth port K5 of the five-way valve 12 are connected. The hemolytic agent is injected into the mixing tank 5 through the second pump 4, and the hemolytic agent and blood sample in the mixing tank 5 are thoroughly mixed by repeated aspiration.

[0072] For example, the second pump 4 can be a 5mL syringe pump, and the volume of the hemolysing agent injected can be 300μL. Repeated aspiration can promote the mixing and reaction of the blood sample and the hemolysing agent to form a complex.

[0073] In the injection procedure, refer to Figure 12 The first pump 3 draws the mixed hemolysin and blood sample from the sampling needle 1; the flow path switching device 6 switches the first connection port 61 to the third connection port 63 and the fourth connection port 64 to the second connection port 62, and the first pump 3 injects the mixed hemolysin and blood sample drawn by the sampling needle 1 into the quantitative loop 2.

[0074] For example, the volume of mixed sample drawn by the first pump 3 can be greater than the volume of the metering loop 2. For example, if the volume of the metering loop 2 is 5 μL, the first pump 3 can draw 50 μL. The extra volume (e.g., 30 μL ± 5 μL) is used to rinse the residue at the inlet end of the metering loop 2 to ensure that the mixed sample inside the metering loop 2 is clean.

[0075] In the detection steps, refer to Figure 13The flow path switching device 6 switches the fifth connection port 65 to the third connection port 63, and the fourth connection port 64 to the sixth connection port 66. The reagent in the reagent container 7 and the hemolysin mixed in the quantitative loop 2 and the blood sample are delivered to the detector 9 by the third pump 8 for elution analysis.

[0076] The third pump 8 can be a plunger pump, which provides high-pressure driving force to heat the reagent through the heating wire 91, elute and separate it through the synthesis column 92, and then deliver it to the detection cell 93 for turbidimetric detection.

[0077] For example, through the coordination of the above steps, the sampling, mixing, injection and detection steps are closely linked in sequence. The first pump 3 realizes sampling and injection, the second pump 4 independently realizes hemolytic agent injection and mixing, and the third pump 8 independently realizes reagent elution. Each step does not interfere with each other, which improves the coordination and overall efficiency of the detection process, while ensuring detection accuracy and repeatability.

[0078] In some embodiments, the protein analysis detection method may further include a pre-sampling solution filling step. This pre-sampling solution filling step may include multiple sub-steps for completing flow path filling, air bubble removal, and component cleaning before formal detection.

[0079] In the first sub-step, refer to Figure 2 The flow path switching device 6 switches the fifth connection port 65 to the second connection port 62, and the first valve port T1 of the three-way valve 10 is connected to the third valve port T3. The reagent in the reagent container 7 is filled into the first filling and cleaning flow path by the third pump 8 and the fourth pump 11.

[0080] This step can be used to prefill the flow path between the third pump 8 and the detector 9 with reagents (e.g., the first reagent) during the startup preparation stage or before detection, to remove air bubbles in the flow path, and to achieve pre-pump filling and liquid replacement.

[0081] In the second sub-step, refer to Figure 3 The first port K1 of the five-way valve 12 is connected to the second port K2, and the hemolytic agent in the hemolytic agent container 18 is drawn into the second pump 4 through the second pump 4. For example, the second pump 4 can draw 5 mL of hemolytic agent to fill the chamber of the second pump 4 with hemolytic agent.

[0082] In the third sub-step, refer to Figure 4 The first port K1 of the five-way valve 12 is connected to the fourth port K4. The hemolytic agent inside is injected into the sampling needle 1 and the mixing tank 5 by the second pump 4, and flows to the waste liquid outlet through the outlet of the mixing tank 5. For example, the second pump 4 can push 2 mL of hemolytic agent through the mixing tank 5 to achieve the initial cleaning of the mixing tank 5.

[0083] In the fourth sub-step, refer to Figure 5The flow path switching device 6 connects the seventh connection port 67 to the second connection port 62, the second port K2 of the five-way valve 12 connects to the third port K3, and the first valve port T1 and the second valve port T2 of the three-way valve 10 connect. The hemolytic agent in the hemolytic agent container 18 is drawn into the first pump 3. For example, the first pump 3 can draw 0.5 mL of hemolytic agent.

[0084] In the fifth sub-step, refer to Figure 6 The flow path switching device 6 switches the connection between the first connection port 61 and the second connection port 62, and the hemolysing agent inside is pushed out by the first pump 3 to clean the sampling needle 1 and the mixing tank 5, and flows to the waste liquid port through the outlet of the mixing tank 5. For example, the first pump 3 can push 1.5 mL of hemolysing agent through the inner wall of the sampling needle 1 to clean the inner wall of the sampling needle 1.

[0085] In the sixth sub-step, refer to Figure 7 and Figure 8 The first port K1 of the five-way valve 12 is connected to the fifth port K5. The hemolysing agent inside is pushed into the mixing tank 5 by the second pump 4 to clean the mixing tank 5. After cleaning, the hemolysing agent in the mixing tank 5 is withdrawn by the second pump 4. For example, the second pump 4 can push 2 mL of hemolysing agent through the mixing tank 5 and then withdraw it to empty the mixing tank 5.

[0086] Through the aforementioned pre-sampling solution filling steps, the filling and venting of the first pump 3, the second pump 4, the third pump 8, and related flow paths, as well as the cleaning of the inner and outer walls of the sampling needle 1 and the mixing tank 5, were completed before the formal testing. This ensured the cleanliness of the initial testing state and the absence of air bubbles in the flow paths, reducing the risk of sample contamination and air bubble interference, and providing a fundamental guarantee for the accuracy and repeatability of subsequent tests. Simultaneously, each filling and cleaning step was carried out in an orderly manner through the switching of the flow path switching device 6, the three-way valve 10, and the five-way valve 12, without interfering with the main testing process. This allowed for parallel preparation during the previous sample testing process, shortening the overall experimental interval time.

[0087] In some embodiments, the protein analysis detection method may further include a post-detection cleaning step. The post-detection cleaning step can be performed after each detection to clean the sampling needle 1 and the mixing chamber 5, preventing sample residue from causing cross-contamination in subsequent detections.

[0088] In the first cleaning sub-step, refer to Figure 14 The first port K1 of the five-way valve 12 is connected to the fourth port K4. The hemolysing agent inside is injected into the sampling needle 1 and the mixing tank 5 by the second pump 4. The hemolysing agent in the mixing tank 5 flows to the waste liquid port. This step can be used to clean the outer wall of the sampling needle 1 and the mixing tank 5. For example, the second pump 4 can push 900 μL of hemolysing agent through the outer wall of the sampling needle 1.

[0089] In the second cleaning sub-step, refer to Figure 15 The flow path switching device 6 switches the connection between the first connecting port 61 and the second connecting port 62, and the hemolysing agent inside is pushed out by the first pump 3 to clean the sampling needle 1 and the mixing tank 5, and flows to the waste liquid port through the outlet of the mixing tank 5. This step can be used to clean the inner wall of the sampling needle 1 and the mixing tank 5. For example, the first pump 3 can push 100 μL of hemolysing agent through the inner wall of the sampling needle 1.

[0090] In the third cleaning sub-step, refer to Figure 16 The first port K1 of the five-way valve 12 is connected to the fifth port K5. The hemolytic agent inside is injected into the sampling needle 1 and the mixing tank 5 by the second pump 4, and flows to the waste liquid outlet through the outlet of the mixing tank 5. This step can further clean the mixing tank 5, for example, the second pump 4 can push 2 mL of hemolytic agent through the mixing tank 5.

[0091] In the fourth cleaning sub-step, refer to Figure 17 The hemolysin in the mixing tank 5 after cleaning is removed by the second pump 4, thus emptying the mixing tank 5.

[0092] Through the above-mentioned post-test cleaning steps, the inner and outer walls of the sampling needle 1 and the mixing pool 5 are cleaned and emptied in multiple steps after each test, effectively removing residual samples and reagents, reducing the risk of cross-contamination, and ensuring the accuracy of the next test.

[0093] In some embodiments, after completing a full detection and cleaning step, the protein analyzer can repeat the pre-sampling solution filling step, sampling step, mixing step, injection step, detection step, and post-detection cleaning step to perform the next protein analysis detection.

[0094] In the repeated detection steps, the third pump 8 can be connected to the reagent container 7 containing the second reagent or the reagent container 7 containing the third reagent by switching the four-way valve 14. This allows the third pump 8 to deliver the second reagent or the third reagent, which is different from the first reagent, to the detector 9 through the fifth connection port 65, the third connection port 63, the quantitative loop 2, the fourth connection port 64, and the sixth connection port 66. After being heated by the heating wire 91 and eluted by the synthesis column 92, the reagent is then subjected to turbidimetric detection in the detection cell 93.

[0095] The second and third reagents can be elution reagents with different components or concentrations than the first reagent, and are respectively contained in different reagent containers 7, for elution analysis of different components in the sample or for gradient elution.

[0096] By repeating the above steps, the protein analyzer can continuously test multiple samples, or perform multi-step elution analysis using different reagents on the same sample, thus improving throughput and analytical capabilities. Furthermore, because a post-test cleaning step is performed after each test, and the flow path is refilled with a pre-sampling solution before each test, the cleanliness of the flow path and the accuracy of the test results are ensured during continuous testing.

[0097] Although embodiments of the invention have been described in conjunction with the accompanying drawings, those skilled in the art can make various modifications and variations without departing from the spirit and scope of the invention, and such modifications and variations all fall within the scope defined by the appended claims.

Claims

1. A protein analyzer, characterized in that, include: Sampling needle (1), quantitative loop (2), first pump (3), second pump (4), mixing cell (5), flow path switching device (6), at least one reagent container (7), third pump (8) and detector (9); The sampling needle (1) is used to draw blood samples and inject them into the mixing pool (5). The second pump (4) is connected to the mixing pool (5) and is used to inject a hemolytic agent into the mixing pool (5). The flow path switching device (6) includes a first connection port (61), a second connection port (62), a third connection port (63), a fourth connection port (64), a fifth connection port (65), and a sixth connection port (66). The sampling needle (1) is connected to the first connecting port (61), and the first pump (3) is connected to the second connecting port (62); when the flow path switching device (6) switches the connection between the first connecting port (61) and the second connecting port (62), the sampling needle (1), the first connecting port (61), the second connecting port (62) and the first pump (3) constitute the first detection flow path; One end of the quantitative loop (2) is connected to the third connecting port (63), and the other end of the quantitative loop (2) is connected to the fourth connecting port (64); when the flow path switching device (6) switches the first connecting port (61) to the third connecting port (63) and the fourth connecting port (64) to the second connecting port (62), the sampling needle (1), the first connecting port (61), the third connecting port (63), the quantitative loop (2), the fourth connecting port (64), the second connecting port (62) and the first pump (3) constitute the second detection flow path; The reagent container (7) is connected to the fifth connection port (65), the inlet of the detector (9) is connected to the sixth connection port (66), the third pump (8) is disposed on the pipeline between the reagent container (7) and the fifth connection port (65), the flow path switching device (6) switches the fifth connection port (65) to the third connection port (63), and the fourth connection port (64) to the sixth connection port (66), so that the reagent container (7), the third pump (8), the fifth connection port (65), the third connection port (63), the quantitative loop (2), the fourth connection port (64), the sixth connection port (66) and the detector (9) constitute the third detection flow path.

2. The protein analyzer according to claim 1, characterized in that, The protein analyzer also includes a three-way valve (10) and a fourth pump (11). The first valve port (T1) of the three-way valve (10) is connected to the second connecting port (62) through a pipeline. The second valve port (T2) of the three-way valve (10) is connected to the sampling needle (1) through a pipeline. The third valve port (T3) of the three-way valve (10) is connected to the inlet of the fourth pump (11) through a pipeline. The outlet of the fourth pump (11) is used to connect to the waste liquid outlet. When the first valve port (T1) and the second valve port (T2) of the three-way valve (10) are connected, the sampling needle (1) is connected to the second connecting port (62). The flow path switching device (6) switches the fifth connection port (65) to the second connection port (62); when the first valve port (T1) and the third valve port (T3) of the three-way valve (10) are connected, the reagent container (7), the third pump (8), the fifth connection port (65), the second connection port (62), the three-way valve (10), and the fourth pump (11) constitute the first filling and cleaning flow path.

3. The protein analyzer according to claim 2, characterized in that, The protein analyzer also includes a hemolysin container (18). The flow path switching device (6) further includes a seventh connection port (67), and the hemolytic agent container (18) is connected to the seventh connection port (67); when the flow path switching device (6) switches the seventh connection port (67) to the second connection port (62), and the first valve port (T1) of the three-way valve (10) is connected to the second valve port (T2), the hemolytic agent container (18), the seventh connection port (67), the second connection port (62), the three-way valve (10) and the first pump (3) constitute a second filling and cleaning flow path.

4. The protein analyzer according to claim 3, characterized in that, The protein analyzer also includes a five-way valve (12). The second pump (4) is connected to the first port (K1) of the five-way valve (12). The hemolytic agent container (18) is connected to the second port (K2) of the five-way valve (12) through a pipeline. The seventh port (67) is connected to the third port (K3) of the five-way valve (12). The fourth port (K4) of the five-way valve (12) is connected to the sampling needle (1) through a pipeline. The fifth port (K5) of the five-way valve (12) is connected to the mixing tank (5) through a pipeline. When the second port (K2) and the third port (K3) of the five-way valve (12) are connected, the seventh port (67) is connected to the hemolytic agent container (18). When the first port (K1) and the second port (K2) of the five-way valve (12) are connected, the hemolytic agent container (18), the second port (K2), the first port (K1) and the second pump (4) constitute a third filling and cleaning flow path; When the first port (K1) and the fourth port (K4) of the five-way valve (12) are connected, the second pump (4), the first port (K1), the fourth port (K4) and the sampling needle (1) constitute the fourth filling cleaning flow path; When the first port (K1) of the five-way valve (12) is connected to the fifth port (K5), the second pump (4), the first port (K1), the fifth port (K5), and the mixing tank (5) constitute the fifth filling and cleaning flow path.

5. The protein analyzer according to claim 4, characterized in that, A mixer (13) is installed on the pipeline between the fifth interface (K5) and the mixing pool (5).

6. The protein analyzer according to claim 1, characterized in that, The reagent container (7) is provided in three parts. The protein analyzer also includes a four-way valve (14). The three reagent containers (7) are respectively connected to the three valve ports of the four-way valve (14) through pipelines. The other valve port of the four-way valve (14) is connected to the inlet of the third pump (8).

7. The protein analyzer according to claim 6, characterized in that, A degassing pump (15) is installed on the pipeline between the reagent container (7) and the four-way valve (14).

8. The protein analyzer according to claim 1, characterized in that, A filter (16) is installed on the pipeline between the sixth connection port (66) and the detector (9).

9. The protein analyzer according to claim 1, characterized in that, The detector (9) includes a heating wire (91), a synthesis column (92), and a detection cell (93) arranged sequentially from upstream to downstream.

10. A protein analysis and detection method, characterized in that, The protein analysis detection method, applied to the protein analyzer as described in any one of claims 1-9, comprises: Sampling steps: The flow path switching device (6) switches the first connection port (61) and the second connection port (62) to connect, the first valve port (T1) and the second valve port (T2) of the three-way valve (10) are connected, the sampling needle (1) is drawn by the first pump (3), and the drawn blood sample is injected into the mixing pool (5). Mixing steps: The first port (K1) and the fifth port (K5) of the five-way valve (12) are connected, and the hemolytic agent is injected into the mixing pool (5) through the second pump (4), and the hemolytic agent and blood sample in the mixing pool (5) are thoroughly mixed by repeated aspiration; Sample injection steps: The sampling needle (1) is used to extract the mixed hemolysin and blood sample by the first pump (3); the flow path switching device (6) switches the first connection port (61) to the third connection port (63) and the fourth connection port (64) to the second connection port (62), and the mixed hemolysin and blood sample extracted by the sampling needle (1) is injected into the quantitative loop (2) by the first pump (3); Detection steps: The flow path switching device (6) switches the fifth connection port (65) to the third connection port (63), and the fourth connection port (64) to the sixth connection port (66). The reagent in the reagent container (7) and the hemolysin mixed in the quantitative loop (2) and the blood sample are transported together to the detector (9) through the third pump (8) for elution analysis.

11. The protein analysis and detection method according to claim 1, characterized in that, The protein analysis and detection method further includes: Pre-sampling solution filling step: The flow path switching device (6) switches the fifth connection port (65) to the second connection port (62), and the first valve port (T1) of the three-way valve (10) is connected to the third valve port (T3). The reagent in the reagent container (7) is filled into the first filling and cleaning flow path by the third pump (8) and the fourth pump (11). The first port (K1) of the five-way valve (12) is connected to the second port (K2), and the hemolytic agent in the hemolytic agent container (18) is drawn into the second pump (4) through the second pump (4); The first port (K1) of the five-way valve (12) is connected to the fourth port (K4). The hemolytic agent inside is injected into the sampling needle (1) and the mixing tank (5) through the second pump (4), and flows to the waste liquid port (17) through the outlet of the mixing tank (5). The flow path switching device (6) switches the seventh connection port (67) to the second connection port (62), the second port (K2) of the five-way valve (12) to the third port (K3), and the first valve port (T1) and the second valve port (T2) of the three-way valve (10) to be connected. The hemolytic agent in the container (18) is drawn into the first pump (3) through the first pump (3). The flow path switching device (6) switches the first connection port (61) and the second connection port (62) to connect, and pushes out the hemolytic agent inside through the first pump (3) to clean the sampling needle (1) and the mixing tank (5), and flows to the waste liquid port (17) through the outlet of the mixing tank (5). The first port (K1) of the five-way valve (12) is connected to the fifth port (K5). The hemolysin inside is pushed into the mixing tank (5) by the second pump (4) to clean the mixing tank (5). After cleaning, the hemolysin inside the mixing tank (5) is removed by the second pump (4).

12. The protein analysis and detection method according to claim 10, characterized in that, The protein analysis and detection method further includes: Post-detection cleaning steps: The first port (K1) of the five-way valve (12) is connected to the fourth port (K4). The hemolysing agent inside is injected into the sampling needle (1) and the mixing tank (5) through the second pump (4). The hemolysing agent in the mixing tank (5) flows to the waste liquid port (17). The flow path switching device (6) switches the first connection port (61) and the second connection port (62) to connect, and pushes out the hemolytic agent inside through the first pump (3) to clean the sampling needle (1) and the mixing tank (5), and flows to the waste liquid port (17) through the outlet of the mixing tank (5). The first port (K1) of the five-way valve (12) is connected to the fifth port (K5). The hemolytic agent inside is injected into the sampling needle (1) and the mixing tank (5) by the second pump (4), and flows to the waste liquid port (17) through the outlet of the mixing tank (5). The hemolysing agent in the mixing tank (5) after cleaning is removed by the second pump (4).

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