Full-automatic protease activity detection system

Abstract

The utility model provides a full-automatic protease activity detection system which comprises a full-automatic liquid treatment workstation which is respectively communicated with a multifunctional microplate reader, a constant-temperature and constant-humidity incubator, a centrifugal machine and a consumable rack, the full-automatic liquid treatment workstation, the multifunctional microplate reader, the constant-temperature and constant-humidity incubator, the centrifugal machine and the consumable rack are all connected with the main control system. The protease activity detection device not only can efficiently and accurately carry out large-batch protease activity detection and remarkably reduce the labor cost of experimental operation, but also can effectively avoid personal errors and ensure the accuracy of a detection result. By virtue of the characteristics of high flux and automation, the increasing requirements of modern biotechnology research and industrial production on protease activity detection are practically met.

Description

Technical Field

[0001] The utility model relates to the technical field of activity detection, in particular to a full-automatic protease activity detection system. Background Art

[0002] In the fields of biochemical research and industrial enzyme preparation production, detecting protease activity is a key link in analyzing product quality and its application performance. The current national standard GB / T 23527.1-2023 in China provides an authoritative and standardized method for the determination of protease activity in enzyme preparations, which plays a key role in ensuring the stability and consistency of enzyme preparation product quality and maintaining the normal development of the enzyme preparation industry. However, in the scenario of screening extracellular protease-producing strains, there are some limitations in traditional protease activity detection methods. The cumbersome manual operation steps not only increase the labor input in the laboratory and prolong the detection cycle, but also are easily interfered by human factors, reducing the repeatability and accuracy of detection results.

[0003] For example, the invention patent with the application number 202210072768.4, a large-volume full-automatic nucleic acid extraction method and device, discloses that the nucleic acid extraction method is to load reagents and consumables in designated positions; the pipetting robotic arm automatically picks up Tips heads and transfers samples and reagents from the sample area and reagent area to the deep well plate in sequence; the nucleic acid extraction device completes mixing, lysis, extraction, and purification; the pipetting robotic arm transfers the PCR reaction system construction reagents in the reagent area to the octuplet tube in the reaction construction system area and transfers the purified nucleic acid in the deep well plate to the octuplet tube; collect and process waste. This invention provides a device and method for full-automatic high-throughput large-volume nucleic acid extraction, which uses a pipetting workstation and a magnetic rod sleeve to optimize the division of labor method and structural layout to complete full-automatic high-throughput large-volume nucleic acid extraction. However, the device in this invention is based on a relatively large-volume reaction system, increasing the reagent consumption; increasing the detection cost, especially when multiple parallel tests need to be performed on each sample to ensure data reliability, this phenomenon is more prominent. Summary of the Utility Model

[0004] Aiming at the technical problems of high detection cost, poor repeatability, and low accuracy of the existing detection system, the utility model proposes a full-automatic protease activity detection system. The utility model aims to solve the problems encountered in large-scale and high-throughput protease activity detection according to the national standard method, and realize a high-precision, low-consumable, and automated large-scale protease activity detection method.

[0005] In order to achieve the above object, the technical solution of the utility model is realized as follows:

[0006] A fully automatic protease activity detection system, comprising a fully automatic liquid handling workstation, which is respectively connected to a multifunctional microplate reader, a constant temperature and humidity incubator, a centrifuge, and a consumable rack, and the fully automatic liquid handling workstation, the multifunctional microplate reader, the constant temperature and humidity incubator, the centrifuge, and the consumable rack are all connected to the main control system.

[0007] The fully automatic liquid handling workstation includes a workstation housing. Below the workstation housing, there is a workstation tabletop. A protease activity detection kit is placed on the workstation tabletop. An oscillator is installed beside the protease activity detection kit. The oscillator is connected to the main control system. There is a channel port a on the workstation tabletop beside the oscillator. The fully automatic liquid handling workstation is connected to the centrifuge through the channel port a. The centrifuge is located below the workstation tabletop.

[0008] Above the workstation housing, there are horizontal slide rails near the front and rear surfaces of the workstation housing respectively. A liquid transfer device and a plate transfer device are slidably arranged on the horizontal slide rails. The liquid transfer device cooperates with the protease activity detection kit to perform liquid addition and liquid extraction operations. The plate transfer device cooperates with the protease activity detection kit to perform consumable transfer operations.

[0009] There is a channel port b on the front surface of the workstation housing. The fully automatic liquid handling workstation is connected to the multifunctional microplate reader through the channel port b; there is a channel port c on the right surface of the workstation housing. The fully automatic liquid handling workstation is connected to the consumable rack through the channel port c; there is a channel port d on the rear surface of the workstation housing. The fully automatic liquid handling workstation is connected to the constant temperature and humidity incubator through the channel port d.

[0010] The protease activity detection kit includes a sample plate, multiple reaction plates, multiple reaction reagent tip boxes, a common tip box, and multiple reagent troughs. The sample plate, multiple reaction reagent tip boxes, the common tip box, and multiple reagent troughs are all arranged on the workstation tabletop. Multiple reaction plates and the sample plate are both placed on the workstation tabletop by the plate transfer device.

[0011] The pipetting device includes a multi-channel pipetting robotic arm. A slider Ⅰa is provided at the middle position of the multi-channel pipetting robotic arm. A gripper Ⅰ is provided at one end of the multi-channel pipetting robotic arm. A driving device Ⅰz is provided at the upper end of the control gripper Ⅰ. The driving device Ⅰz is connected to a ball screw Ⅰz through a coupling. The ball screw Ⅰz is slidably connected to the slider Ⅰa. The driving device Ⅰz drives the ball screw Ⅰz to rotate, thereby driving the gripper Ⅰ to move along the z-axis. A slider Ⅰb is provided below the slider Ⅰa. The multi-channel pipetting robotic arm is slidably arranged on a longitudinal slide rail Ⅰ through the slider Ⅰb. A driving device Ⅰy is provided at one end of the longitudinal slide rail Ⅰ. The driving device Ⅰy is connected to a ball screw Ⅰy through a coupling. The ball screw Ⅰy is connected to the slider Ⅰb. The driving device Ⅰy drives the ball screw Ⅰy to rotate, thereby driving the slider Ⅰb to move along the y-axis direction. Sliders Ⅰc are provided at both ends of the longitudinal slide rail Ⅰ. The longitudinal slide rail Ⅰ is slidably arranged on a transverse slide rail through the sliders Ⅰc. An X-axis driving device is provided at one end of the transverse slide rail. The X-axis driving device is connected to a ball screw Ⅰx through a coupling. The ball screw Ⅰx is connected to the slider Ⅰc. The X-axis driving device drives the ball screw Ⅰx to rotate, thereby driving the slider Ⅰc to move along the x-axis direction.

[0012] The plate moving device includes at least one plate moving robotic arm. The plate moving robotic arm is located on one side of the multi-channel pipetting robotic arm near the consumable rack direction. A slider Ⅱa is provided at the middle position of the plate moving robotic arm. A gripper Ⅱ is provided at one end of the plate moving robotic arm. A driving device Ⅱz is provided at the upper end of the gripper Ⅱ. The driving device Ⅱz is connected to a ball screw Ⅱz through a coupling. The ball screw Ⅱz is connected to the slider Ⅱa. The driving device Ⅱz drives the ball screw Ⅱz to rotate, thereby driving the gripper Ⅱ to move along the z-axis. A slider Ⅱb is provided below the slider Ⅱa. The plate moving robotic arm is slidably arranged on a longitudinal slide rail Ⅱ through the slider Ⅱb. A driving device Ⅱy is provided at one end of the longitudinal slide rail Ⅱ. The driving device Ⅱy is connected to a ball screw Ⅱy through a coupling. The ball screw Ⅱy is connected to the slider Ⅱb. The driving device Ⅱy drives the ball screw Ⅱy to rotate, thereby driving the slider Ⅱb to move along the y-axis direction. Sliders Ⅱc are provided at both ends of the longitudinal slide rail Ⅱ. The longitudinal slide rail Ⅱ is slidably arranged on a transverse slide rail through the sliders Ⅱc. An X-axis driving device is provided at one end of the transverse slide rail. The X-axis driving device is connected to a ball screw Ⅱx through a coupling. The ball screw Ⅱx is connected to the slider Ⅱc. When moving along the x-axis direction, the X-axis driving device drives the ball screw Ⅱx to rotate, thereby driving the slider Ⅱc to move along the x-axis direction.

[0013] A waste needle station is also provided at the edge of the workstation tabletop. A waste needle station outlet is provided below the waste needle station.

[0014] The workstation housing is a housing partially made of transparent material.

[0015] A lighting and sound-light alarm device is also provided above the workstation housing. The lighting and sound-light alarm device is connected to the main control system.

[0016] The beneficial effects of the present utility model:

[0017] It can not only measure the activity of a large number of proteases efficiently and accurately, significantly reducing the labor cost of experimental operations, but also effectively avoid human errors and ensure the accuracy of detection results. With the characteristics of high throughput and automation, this utility model effectively meets the growing demand for protease activity detection in modern biotechnology research and industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present utility model or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only some embodiments of the present utility model. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0019] Figure 1 It is a schematic diagram of the workbench surface of the full-automatic liquid handling workstation of the present utility model.

[0020] Figure 2 It is an overall external view of the full-automatic liquid handling workstation of the present utility model.

[0021] Figure 3 It is a schematic diagram of the structure of the multi-channel pipetting robotic arm of the present utility model.

[0022] Figure 4 It is a schematic diagram of the structure of the plate-transferring robotic arm of the present utility model.

[0023] In the figure, 1 is the workstation housing, 2 is the workstation tabletop, 3 is the longitudinal slide rail I, 4 is the transverse slide rail, 5 is the multi-channel pipetting robotic arm gripper I, 6 is the lighting and audible and visual alarm device, 7 is the longitudinal slide rail II, 8 is the slider IIa, 9 is the plate-transferring robotic arm gripper II, 10 is the reaction plate I for the enzyme solution to be measured, 11 is the Folin reagent reaction plate, 12 is the channel port a, 13 is the channel port b, 14 is the channel port c, and 15 is the channel port d. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0024] The following will clearly and completely describe the technical solutions in the embodiments of the present utility model with reference to the drawings in the embodiments of the present utility model. Obviously, the described embodiments are only some embodiments of the present utility model, rather than all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the protection scope of the present utility model.

[0025] As Figure 2As shown in the figure, a fully automatic protease activity detection system includes a fully automatic liquid handling workstation, which is used for experimental operations of protease activity detection; the fully automatic liquid handling workstation includes a workstation housing 1, and the workstation housing 1 is a housing partially made of transparent materials, which is convenient for observing the experimental detection process.

[0026] As Figure 1 shown, a workstation table 2 is provided below the workstation housing 1. A protease activity detection kit is placed on the workstation table 2. The protease activity detection kit includes multiple reaction plates, a sample plate, multiple reaction reagent tip boxes, a common tip box, and multiple reagent troughs. The sample plate, multiple reaction reagent tip boxes, the common tip box, and multiple reagent troughs are all arranged on the workstation table 2, and multiple reaction plates are placed on the workstation table 2 by a plate transfer device. In this embodiment, multiple reaction plates all adopt 96-well deep-well plates. In this embodiment, multiple reaction reagent tip boxes are arranged at the leftmost side of the workstation table 2. From the back to the front of the workstation table 2, there are a casein tip box, a trichloroacetic acid tip box, a sodium carbonate tip box, and a Folin reagent tip box in sequence. Multiple reagent troughs are arranged on the right side of multiple reaction reagent tip boxes. From the back to the front of the workstation table 2, there are a casein solution reagent trough, a trichloroacetic acid solution reagent trough, a sodium carbonate solution reagent trough, and a Folin reagent solution reagent trough in sequence. The common tip box is arranged on the right side of multiple reagent troughs and is located in the middle of the workstation table 2. An oscillator is provided on the right side of the common tip box, and the oscillator is connected to the main control system. The oscillator is used for performing timed and uniform oscillation operations on the well plates filled with the enzyme solution sample to be tested and reaction reagents, ensuring that the enzyme solution sample to be tested and reaction reagents are fully mixed; in front of the oscillator, a waste needle station is provided at the edge of the workstation table 2, and a waste needle station outlet is provided below the waste needle station. A garbage bag or trash can can be connected to the waste needle station outlet, which is convenient for the disposal of used pipette tips; a channel port a12 is provided on the workstation table 2 on the right side of the oscillator. The fully automatic liquid handling workstation is connected to a centrifuge through the channel port a12. The centrifuge is located below the workstation table 2 and is used to replace the slow qualitative filter paper filtration precipitation step in the national standard method; the sample plate is arranged on the right side of the channel port a12, and multiple reaction plates are arranged close to the sample plate. In this embodiment, multiple reaction plates include a 96-well microplate for final absorbance measurement, 4 reaction plates 10 for the enzyme solution to be tested, and 4 Folin reagent reaction plates 11. The final absorbance measurement uses a 96-well microplate, and other reaction plates are 96-well deep-well plates. The specific arrangement is as Figure 1 shown.

[0027] As Figure 2As shown in the figure, the fully automatic liquid handling workstation is connected to the multifunctional microplate reader through the channel port b13 on the front right side of the workstation housing 1. The multifunctional microplate reader is located in front of the fully automatic liquid handling workstation and is used for detecting the absorbance value of the reaction solution. The fully automatic liquid handling workstation is connected to the consumable rack through the channel port c14 on the right side of the workstation housing 1. The consumable rack is located on the right side of the fully automatic liquid handling workstation and is used for storing and managing various consumables required for experiments, realizing the automatic replenishment of consumables. The consumable rack, as an important facility in the experimental process, automatically tracks and manages the usage status of experimental consumables. When a certain protease activity detection step is completed, the main control system controls the plate transfer robotic arm through pre-programmed instructions, moves the used reaction plate to the dedicated recycling area of the consumable rack, and synchronously takes out a new reaction plate from the consumable rack and places it at the predetermined position on the workstation table 2, ensuring the continuity and efficiency of the entire detection process.

[0028] The fully automatic liquid handling workstation is connected to the thermostatic and humidostatic incubator through the channel port d15 on the rear right side of the workstation housing 1. The thermostatic and humidostatic incubator is located behind the fully automatic liquid handling workstation and is used to maintain a stable temperature, providing suitable temperature conditions for the proteolysis reaction and the Folin reagent reaction. The fully automatic liquid handling workstation, the multifunctional microplate reader, the thermostatic and humidostatic incubator, the centrifuge, and the consumable rack are all connected to the main control system.

[0029] The main control system is a computer, and the Freedom EVOware software is used to implement the control process of the detection. The main control system realizes signal interaction and data transmission with devices such as the fully automatic liquid handling workstation, the multifunctional microplate reader, the thermostatic and humidostatic incubator, the centrifuge, and the consumable rack through the USB interface, comprehensively coordinating and controlling the entire detection process. The main control system has a powerful program programming function, allowing users to customize and execute a series of experimental processes, including but not limited to variable setting, logical judgment, loop control, and timed waiting and other diversified experimental processes.

[0030] Near the upper part inside the workstation housing 1, transverse sliding rails 4 are respectively provided close to the front and rear surfaces of the workstation housing 1. A liquid transfer device and a plate transfer device are slidably arranged on the transverse sliding rails 4. The liquid transfer device cooperates with the protease activity detection kit to perform liquid addition and liquid extraction operations. The plate transfer device cooperates with the protease activity detection kit to perform consumable transfer operations, and the consumable is a reaction plate. The liquid transfer device and the plate transfer device together form an automated material transfer system, which can accurately and efficiently complete a series of operations such as consumable transfer, sample transfer, and reaction solution distribution according to the preset program.

[0031] As Figure 3As shown in the figure, the pipetting device includes a multi-channel pipetting robotic arm. A slider Ⅰa is provided at the middle position of the multi-channel pipetting robotic arm. A gripper Ⅰ5 is provided at one end of the multi-channel pipetting robotic arm. A driving device Ⅰz is provided at the upper end of the control gripper Ⅰ5. The driving device Ⅰz is connected to a ball screw Ⅰz through a coupling. The ball screw Ⅰz is connected to the slider Ⅰa through a nut. When moving along the z-axis, the slider Ⅰa remains stationary, and the driving device Ⅰz drives the ball screw Ⅰz to rotate, thereby driving the gripper Ⅰ5 to move along the z-axis. A slider Ⅰb is provided below the slider Ⅰa. The multi-channel pipetting robotic arm is slidably arranged on a longitudinal slide rail Ⅰ3 through the slider Ⅰb. A driving device Ⅰy is provided at one end of the longitudinal slide rail Ⅰ3. The driving device Ⅰy is connected to a ball screw Ⅰy through a coupling. The ball screw Ⅰy is connected to the slider Ⅰb through a nut. When moving along the y-axis, the driving device Ⅰy drives the ball screw Ⅰy to rotate, thereby driving the slider Ⅰb to move along the y-axis, and further driving the multi-channel pipetting robotic arm to move along the y-axis. The rotation of the ball screw Ⅰz and the ball screw Ⅰy does not affect each other. Sliders Ⅰc are provided at both ends of the longitudinal slide rail Ⅰ3. The longitudinal slide rail Ⅰ3 is slidably arranged on a transverse slide rail 4 through the slider Ⅰc. An X-axis driving device is provided at one end of the transverse slide rail 4. The X-axis driving device is connected to a ball screw Ⅰx through a coupling. The ball screw Ⅰx is connected to the slider Ⅰc through a nut. When moving along the x-axis, the X-axis driving device drives the ball screw Ⅰx to rotate, thereby driving the slider Ⅰc to move along the x-axis, and further driving the longitudinal slide rail Ⅰ3 to move along the x-axis. The X-axis driving device, the driving device Ⅰy, and the driving device Ⅰz cooperate together to control the multi-channel pipetting robotic arm to perform X-axis, Y-axis, and Z-axis movements for liquid aspiration and liquid addition operations. In this embodiment, the multi-channel pipetting robotic arm adopts a 96-channel pipetting robotic arm. The driving device Ⅰx, the driving device Ⅰy, and the x-axis driving device are not shown in the figure.

[0032] As Figure 4As shown in the figure, the plate transfer device includes at least one plate transfer robotic arm, which is located on one side of the multi-channel pipetting robotic arm close to the consumable rack. A slider IIa 8 is provided at the middle position of the plate transfer robotic arm. A direction-adjustable gripper II 9 is provided at one end of the plate transfer robotic arm. A driving device IIz is provided at the upper end of the gripper II 9. The driving device IIz is connected to the ball screw IIz through a coupling. The ball screw IIz is connected to the slider IIa 8 through a nut. When moving along the z-axis, the slider IIa 8 remains stationary, and the driving device IIz drives the ball screw IIz to rotate, thereby driving the gripper II 9 to move along the z-axis. A slider IIb is provided below the slider IIa 8. The plate transfer robotic arm is slidably arranged on the longitudinal slide rail II 7 through the slider IIb. A driving device IIy is provided at one end of the longitudinal slide rail II 7. The driving device IIy is connected to the ball screw IIy through a coupling. The ball screw IIy is connected to the slider IIb through a nut. When moving along the y-axis, the driving device IIy drives the ball screw IIy to rotate, thereby driving the slider IIb to move along the y-axis, and further driving the plate transfer robotic arm to move along the y-axis. The rotations of the ball screw IIz and the ball screw IIy do not affect each other. Sliders IIc are provided at both ends of the longitudinal slide rail II 7. The longitudinal slide rail II 7 is slidably arranged on the transverse slide rail 4 through the slider IIc. An X-axis driving device is provided at one end of the transverse slide rail 4. The X-axis driving device is connected to the ball screw IIx through a coupling. The ball screw IIx is connected to the slider IIc through a nut. When moving along the x-axis, the X-axis driving device drives the ball screw IIx to rotate, thereby driving the slider IIc to move along the x-axis, and further driving the longitudinal slide rail II 7 to move along the x-axis. The X-axis driving device, the driving device IIy, and the driving device IIz cooperate to control the plate transfer robotic arm to perform X-axis, Y-axis, and Z-axis movements for the transfer operation of the reaction plate. The driving device IIx and the driving device IIy are not shown in the figure.

[0033] Above the workstation housing 1, there is also a lighting and audible-visual alarm device 6 to ensure the visibility of operations and timely warning of potential abnormal situations. The lighting and audible-visual alarm device 6 is connected to the main control system.

[0034] Using the full-automatic protease activity detection system of the present utility model, taking 96 samples as an example, the high-throughput and fully automatic detection of neutral protease activity is carried out as follows:

[0035] S1. Preparation work: As Figure 1 shown, add 96 samples to be tested into the sample plate (96-well deep-well plate). Pre-arrange the prepared casein, trichloroacetic acid, sodium carbonate, Folin reagent, and multiple 96-well deep-well plates, etc., according to the layout of the full-automatic liquid handling workstation table 2; and set the thermostatic and humidostatic incubator to a specified temperature.

[0036] S2. The main control system controls the 96-channel pipetting robotic arm to obtain 96 new pipette tips from the common tip box, controls the 96-channel pipetting robotic arm to aspirate 96 × 400 uL of the enzyme solution sample to be tested from the sample plate, and adds the enzyme solution sample to be tested to the reaction plate for the enzyme solution to be tested. The above operations are repeated until the aliquoting of 4 reaction plates for the enzyme solution to be tested (the reaction plate for the enzyme solution to be tested is a 96-well deep well plate) is completed. As Figure 1 shown, the reaction plate for the enzyme solution to be tested is a blank plate for the enzyme solution to be tested or experimental plate 1 for the enzyme solution to be tested or experimental plate 2 for the enzyme solution to be tested or experimental plate 3 for the enzyme solution to be tested;

[0037] S3. The main control system controls the 96-channel pipetting robotic arm to discard 96 pipette tips to the waste needle station, re-obtain 96 pipette tips from the trichloroacetic acid tip box, and aspirate 96 × 800 uL of trichloroacetic acid solution from the trichloroacetic acid solution reagent tank and inject it into the blank plate for the enzyme solution to be tested. After completion, the main control system controls the 96-channel pipetting robotic arm to return the 96 pipette tips to the trichloroacetic acid tip box to achieve the recycling of the tips and reduce the waste of consumables;

[0038] S4. The main control system controls the 96-channel pipetting robotic arm to obtain 96 pipette tips from the casein tip box, and aspirate 96 × 400 uL of casein solution from the casein solution reagent tank and inject it into experimental plate 1 for the enzyme solution to be tested, experimental plate 2 for the enzyme solution to be tested, and experimental plate 3 for the enzyme solution to be tested respectively. After completion, the main control system controls the 96-channel pipetting robotic arm to put the 96 pipette tips back into the casein tip box for recycling;

[0039] S5. The plate transfer robotic arm, according to the preset instructions in the main control system, sequentially transfers the 4 reaction plates for the enzyme solution to be tested to the oscillator for thorough mixing, and then transports them to the thermostatic and humidity-controlled incubator preheated to 30 degrees Celsius;

[0040] S6. Start timing for 10 minutes;

[0041] S7. After the timing ends, the main control system controls the plate transfer robotic arm to return the 4 reaction plates for the enzyme solution to be tested to the starting positions of the 4 reaction plates for the enzyme solution to be tested on the tabletop 2 of the fully automatic liquid handling workstation in sequence;

[0042] S8. The main control system controls the 96-channel pipetting robotic arm. After obtaining 96 corresponding pipette tips from the casein tip box, aspirate 96 × 400 uL of casein solution from the casein solution reagent tank and inject it into the blank plate for the enzyme solution to be tested. Then, control the 96-channel pipetting robotic arm to return the 96 pipette tips to the casein tip box;

[0043] S9. The main control system controls the 96-channel pipetting robotic arm to obtain 96 pipette tips from the trichloroacetic acid tip box, and separately aspirate 96×800 uL of trichloroacetic acid solution from the trichloroacetic acid solution reagent tank into the experimental plates 1, 2, and 3 of the enzyme solution to be tested. After that, it controls the 96-channel pipetting robotic arm to return the 96 pipette tips to the trichloroacetic acid tip box;

[0044] S10. The main control system controls the plate transfer robotic arm to send 4 experimental plates of the enzyme solution to be tested to the oscillator for mixing, and after mixing, returns the 4 experimental plates of the enzyme solution to their original positions;

[0045] S11. Start the 10-minute timing again;

[0046] S12. After the timing ends, transfer the 4 experimental plates of the enzyme solution to be tested to the centrifuge for centrifugation in sequence using the plate transfer robotic arm. Set the centrifugation conditions as 3000 rpm and the centrifugation time as 5 minutes;

[0047] S13. After centrifugation, the main control system controls the plate transfer robotic arm to transfer the 4 centrifuged experimental plates of the enzyme solution to be tested back to the workstation table 2 in the original order;

[0048] S14. The main control system controls the 96-channel pipetting robotic arm to obtain 96 pipette tips from the common tip box, aspirate 200 uL of the supernatant of the centrifuged experimental plates of the enzyme solution to be tested, and place it into the corresponding Folin reagent reaction plate as shown. Control the 96-channel pipetting robotic arm to discard the 96 pipette tips obtained from the common tip box. Control the 96-channel pipetting robotic arm to obtain 96 pipette tips from the sodium carbonate tip box, and aspirate 96×1000 uL of sodium carbonate solution from the sodium carbonate solution reagent tank and place it into the corresponding Folin reagent reaction plate. Control the 96-channel pipetting robotic arm to return the 96 pipette tips obtained from the sodium carbonate tip box. Control the 96-channel pipetting robotic arm to obtain 96 pipette tips from the Folin reagent tip box, and aspirate 96×200 uL of Folin reagent from the Folin reagent solution reagent tank and place it into the corresponding Folin reagent reaction plate. Control the 96-channel pipetting robotic arm to return the 96 pipette tips obtained from the Folin reagent tip box, and control the plate transfer robotic arm to send the corresponding Folin reagent reaction plate to the oscillator. After mixing the mixed solution in the corresponding Folin reagent reaction plate, send it to the constant temperature and humidity incubator; Figure 1 Repeat the above operations 4 times until all 4 Folin reagent reaction plates are processed. The Folin reagent reaction plate (the Folin reagent reaction plate is a 96-well deep plate) is a Folin reagent blank plate or Folin reagent experimental plate 1 or Folin reagent experimental plate 2 or Folin reagent experimental plate 3;

[0049] Repeat the above operations 4 times until all 4 Folin reagent reaction plates are processed. The Folin reagent reaction plate (the Folin reagent reaction plate is a 96-well deep plate) is a Folin reagent blank plate or Folin reagent experimental plate 1 or Folin reagent experimental plate 2 or Folin reagent experimental plate 3;

[0050] S15. Start the third 20-minute timing;

[0051] S16. After the timing ends, the main control system controls the plate transfer robotic arm to successively take out the 4 Folin reagent reaction plates that have completed the Folin reaction from the thermostatic and humid incubator and place them back in the original position on the workstation table 2.

[0052] S17. The main control system controls the plate transfer robotic arm to fetch a new 96-deep well plate from the consumable rack to the workstation table 2, controls the 96-channel pipetting robotic arm to obtain 96 pipette tips from the common pipette tip box, transfers the 96×200uL reaction solution in the Folin reagent reaction plate to the 96-well microplate, controls the 96-channel pipetting robotic arm to discard the 96 pipette tips obtained from the common pipette tip box, and controls the plate transfer robotic arm to send the 96-well microplate to the multifunctional microplate reader to measure the absorbance value at a wavelength of 680nm. This step is also carried out four times in a cycle until the detection of the 4 Folin reagent reaction plates is completed.

[0053] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. An automatic protease activity detection system, characterized in that, It includes a fully automatic liquid handling workstation, which is respectively connected to a multi-functional microplate reader, a constant temperature and humidity incubator, a centrifuge, and a consumable rack. The fully automatic liquid handling workstation, the multi-functional microplate reader, the constant temperature and humidity incubator, the centrifuge, and the consumable rack are all connected to the main control system.

2. The fully automatic protease activity detection system according to claim 1, wherein The fully automatic liquid handling workstation includes a workstation housing (1). Below the workstation housing (1), there is a workstation tabletop (2). A protease activity detection kit is placed on the workstation tabletop (2). An oscillator is installed beside the protease activity detection kit. The oscillator is connected to the main control system. There is a channel port a (12) on the workstation tabletop (2) beside the oscillator. The fully automatic liquid handling workstation is connected to the centrifuge through the channel port a (12). The centrifuge is located below the workstation tabletop (2).

3. The fully automatic protease activity detection system according to claim 2, wherein Above the workstation housing (1), horizontal slide rails (4) are respectively provided near the front and rear surfaces of the workstation housing (1). A liquid transfer device and a plate transfer device are slidably arranged on the horizontal slide rails (4). The liquid transfer device cooperates with the protease activity detection kit for liquid addition and liquid extraction operations. The plate transfer device cooperates with the protease activity detection kit for consumable transfer operations.

4. The fully automatic protease activity detection system according to claim 3, characterized in that, A channel port b (13) is provided on the front surface of the workstation housing (1). The fully automatic liquid handling workstation is connected to the multi-functional microplate reader through the channel port b (13); a channel port c (14) is provided on the right surface of the workstation housing (1). The fully automatic liquid handling workstation is connected to the consumable rack through the channel port c (14); a channel port d (15) is provided on the rear surface of the workstation housing (1). The fully automatic liquid handling workstation is connected to the constant temperature and humidity incubator through the channel port d (15).

5. The fully automatic protease activity detection system according to claim 4, characterized in that, The protease activity detection kit includes a sample plate, multiple reaction plates, multiple reaction reagent tip boxes, a common tip box, and multiple reagent troughs. The sample plate, multiple reaction reagent tip boxes, the common tip box, and multiple reagent troughs are all arranged on the workstation tabletop (2). Multiple reaction plates and the sample plate are both placed on the workstation tabletop (2) by the plate transfer device.

6. The fully automatic protease activity detection system according to claim 4 or 5, characterized in that, The pipetting device includes a multi-channel pipetting robotic arm. A slider Ⅰa is provided at the middle position of the multi-channel pipetting robotic arm. A gripper Ⅰ(5) is provided at one end of the multi-channel pipetting robotic arm. A driving device Ⅰz is provided at the upper end of the control gripper Ⅰ(5). The driving device Ⅰz is connected to a ball screw Ⅰz through a coupling. The ball screw Ⅰz is slidably connected to the slider Ⅰa. The driving device Ⅰz drives the ball screw Ⅰz to rotate, thereby driving the gripper Ⅰ(5) to move along the z-axis. A slider Ⅰb is provided below the slider Ⅰa. The multi-channel pipetting robotic arm is slidably arranged on a longitudinal slide rail Ⅰ(3) through the slider Ⅰb. A driving device Ⅰy is provided at one end of the longitudinal slide rail Ⅰ(3). The driving device Ⅰy is connected to a ball screw Ⅰy through a coupling. The ball screw Ⅰy is connected to the slider Ⅰb. The driving device Ⅰy drives the ball screw Ⅰy to rotate, thereby driving the slider Ⅰb to move along the y-axis direction. Sliders Ⅰc are provided at both ends of the longitudinal slide rail Ⅰ(3). The longitudinal slide rail Ⅰ(3) is slidably arranged on a transverse slide rail (4) through the sliders Ⅰc. An X-axis driving device is provided at one end of the transverse slide rail (4). The X-axis driving device is connected to a ball screw Ⅰx through a coupling. The ball screw Ⅰx is connected to the slider Ⅰc. The X-axis driving device drives the ball screw Ⅰx to rotate, thereby driving the slider Ⅰc to move along the x-axis direction.

7. The fully automatic protease activity detection system according to claim 6, wherein The plate moving device includes at least one plate moving robotic arm. The plate moving robotic arm is located on one side of the multi-channel pipetting robotic arm close to the consumable rack direction. A slider Ⅱa(8) is provided at the middle position of the plate moving robotic arm. A gripper Ⅱ(9) is provided at one end of the plate moving robotic arm. A driving device ⅠⅡz is provided at the upper end of the gripper Ⅱ(9). The driving device Ⅱz is connected to a ball screw Ⅱz through a coupling. The ball screw Ⅱz is connected to the slider Ⅱa(8). The driving device Ⅱz drives the ball screw Ⅱz to rotate, thereby driving the gripper Ⅱ(9) to move along the z-axis. A slider Ⅱb is provided below the slider Ⅱa(8). The plate moving robotic arm is slidably arranged on a longitudinal slide rail Ⅱ(7) through the slider Ⅱb. A driving device Ⅱy is provided at one end of the longitudinal slide rail Ⅱ(7). The driving device Ⅱy is connected to a ball screw Ⅱy through a coupling. The ball screw Ⅱy is connected to the slider Ⅱb. The driving device Ⅱy drives the ball screw Ⅱy to rotate, thereby driving the slider Ⅱb to move along the y-axis direction. Sliders Ⅱc are provided at both ends of the longitudinal slide rail Ⅱ(7). The longitudinal slide rail Ⅱ(7) is slidably arranged on the transverse slide rail (4) through the sliders Ⅱc. An X-axis driving device is provided at one end of the transverse slide rail (4). The X-axis driving device is connected to a ball screw Ⅱx through a coupling. The ball screw Ⅱx is connected to the slider Ⅱc. When moving along the x-axis direction, the X-axis driving device drives the ball screw Ⅱx to rotate, thereby driving the slider Ⅱc to move along the x-axis direction.

8. The fully automatic protease activity detection system according to claim 2, wherein A waste needle station is also provided at the edge of the workstation tabletop (2), and a waste needle station outlet is provided below the waste needle station.

9. The fully automatic protease activity detection system according to claim 2, wherein The workstation housing (1) is a housing partially made of transparent material.

10. The fully automatic protease activity detection system according to claim 2, wherein A lighting and audible and visual alarm device (6) is also provided above the workstation housing (1), and the lighting and audible and visual alarm device (6) is connected to the main control system.

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