High-flux polypeptide synthesis equipment

Abstract

The invention discloses high-throughput polypeptide synthesis equipment. The device comprises a transverse plate, a vertical plate vertically and fixedly arranged on the top of the transverse plate, a storage tank cabinet arranged on one side of the transverse plate, a liquid storage tank arranged in the storage tank cabinet, four supporting plates arranged on the top of the transverse plate in a bilateral symmetry mode, a plurality of reaction kettles arranged on the tops of the supporting plates, and a transverse moving assembly arranged on the rear side wall of the vertical plate. The transverse moving assembly is arranged on the base, the front-back moving assembly is arranged on the transverse moving assembly, the electric telescopic rod is arranged on the front-back moving assembly, the mounting plate is arranged at the telescopic end of the electric telescopic rod, the liquid adding needles are arranged on the mounting plate, and each liquid adding needle is independently connected to the corresponding liquid storage tank through a pipeline.

Description

Technical Field

[0001] The invention relates to the technical field of laboratory automation equipment, and in particular to a high-throughput polypeptide synthesis device. Background Art

[0002] With the increase in investment in biotechnology and drug research and development, the demand for peptide synthesis is also growing. In particular, pharmaceutical companies attach great importance to the development of new drugs, making peptides more and more widely used as therapeutic drugs or diagnostic tools. The market size continues to expand and the growth momentum is good.

[0003] The mainstream method for peptide synthesis in the current market is solid phase peptide synthesis (SPPS), especially FMOC (9-fluorenylmethoxycarbonyl) protection strategy. This method allows efficient and automated synthesis of polypeptide chains and can be easily mass-produced. However, there are currently few types of automated equipment, and most of them are imported from foreign countries.

[0004] Existing peptide synthesis equipment has the following defects:

[0005] (1) Slow synthesis speed and low throughput: The current mainstream equipment uses single-channel pipetting, but there are many types of reagents, including 32 commonly used amino acids and 4-6 common reagents. Each time a reagent is added, the single channel is used. Currently, most of them are single reactors, and the efficiency of single peptide synthesis is too low to meet production needs.

[0006] (2) High price: Currently, the first-tier manufacturers are mainly Biotage, MultiSynTech GmbH, and Activotec. Automation equipment is usually expensive, with an average price of 1-2 million yuan, which may be a burden for small laboratories or research institutions with limited funds. This also increases the cost of the experiment.

[0007] (3) Waste of reagents: For single-channel pipetting, a large amount of DMF is required for pipe cleaning after adding each liquid, which greatly increases the reagent cost and the cost of hazardous chemical waste liquid treatment. In addition, DMF (N,N-dimethylformamide) is classified as a hazardous chemical, which is flammable, explosive and volatile, and has potential effects on the reproductive system and nervous system, which may cause pregnancy problems and neurodegeneration. Summary of the invention

[0008] In order to solve one or some technical problems existing in the prior art, the purpose of the present application is to provide a high-throughput polypeptide synthesis device to greatly improve the liquid addition efficiency.

[0009] In order to solve the above existing technical problems, one of the purposes of this application is achieved by adopting the following technical solution:

[0010] A high-throughput peptide synthesis device comprises a horizontal plate, a vertical plate fixedly arranged on the top of the horizontal plate, a storage tank cabinet arranged on one side of the horizontal plate, a liquid storage tank arranged in the storage tank cabinet, four support plates symmetrically arranged on the top of the horizontal plate, a plurality of reaction kettles arranged on the top of the support plate, a horizontal moving component arranged on the rear side wall of the vertical plate, a front-back moving component arranged on the horizontal moving component, an electric telescopic rod arranged on the front-back moving component, a mounting plate arranged at the telescopic end of the electric telescopic rod, and a plurality of liquid adding needles arranged on the mounting plate, each of the liquid adding needles being connected to the corresponding liquid storage tank by a separate pipeline.

[0011] Furthermore, the lateral movement component includes a horizontal box fixedly arranged on the rear side wall of the vertical plate, an adjusting motor fixedly arranged on the left side wall of the horizontal box, an opening opened on the rear side wall of the horizontal box, a threaded rod penetrating the left side wall of the horizontal box and rotating forward and reversely through the adjusting motor, a connecting block penetrating the opening and threadedly connected to the threaded rod, and a moving block fixedly arranged on the rear side wall of the connecting block, and the front-back moving component is arranged on the moving block.

[0012] Furthermore, the forward and backward moving component includes a horizontal box 2 fixedly arranged on the moving block 1, an adjusting motor 2 fixedly arranged on the rear side wall of the horizontal box 2, an opening 2 opened on the outer side wall of the horizontal box 2, a threaded rod 2 penetrating the rear side wall of the horizontal box 2 and rotating forward and reversely by the adjusting motor 2, a connecting block 2 penetrating the opening 2 and threadedly connected to the threaded rod 2, a moving block 2 fixedly arranged on the connecting block 2, and the electric telescopic rod is arranged at the bottom of the moving block 2.

[0013] Furthermore, the plurality of liquid adding needles are arranged in a 7 by 7 square array.

[0014] Furthermore, the plurality of reactors are arranged in a 6 by 8 arrangement.

[0015] Furthermore, a cover box is provided between the vertical plate and the horizontal plate.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] (1) During use, the dosing needle is driven to move forward and backward by operating the lateral moving component. The dosing needle is driven to move forward and backward by operating the forward and backward moving component, so that several dosing needles can be displaced in parallel in the area above the reactor. The pipeline controlled by the solenoid valve at the rear end is directly connected to the liquid storage tank. Each reagent has a separate pipeline and is added by a rotary plunger pump, which greatly improves the efficiency of dosing. Compared with the existing technology, it solves the problems of slow synthesis speed and low flux: the current mainstream equipment uses single-channel pipetting, but there are many types of reagents, there are 32 commonly used amino acids, and there are 4-6 common reagents. Each reagent is added in a single channel. At present, most reactors are single, and the efficiency of single polypeptide synthesis is too low to meet the technical problem of production needs. Since each dosing needle only adds fixed reagents, it can be added directly without pipeline cleaning, which greatly reduces the time and cost of the cleaning step, and avoids a large amount of hazardous waste liquid pollution and treatment costs. Compared with the existing technology, it solves the problem of waste reagents: the single-channel pipetting solution requires a large amount of DMF for pipeline cleaning after adding a liquid, which greatly increases the reagent cost and the cost of hazardous waste liquid treatment. DMF (N,N-dimethylformamide) is classified as a hazardous chemical. It is flammable, explosive and volatile, and has potential effects on the reproductive and nervous systems, which may lead to technical problems such as pregnancy problems and neurodegeneration. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a schematic diagram of the overall structure of this application;

[0019] Figure 2 It is a schematic diagram of the structure of the lateral moving component and the front-back moving component in this application;

[0020] Figure 3 It is a structural schematic diagram of the threaded rod 1 in this application;

[0021] Figure 4 This is a schematic diagram of the structure of the threaded rod 2 in this application;

[0022] In the figure: 1. Storage tank cabinet; 2. Cover box; 3. Vertical plate; 4. Horizontal plate; 5. Horizontal box one; 6. Adjustment motor one; 7. Horizontal box two; 8. Opening one; 9. Adjustment motor two; 10. Moving block one; 11. Moving block two; 12. Electric telescopic rod; 13. Mounting plate; 14. Dosing needle; 15. Support plate; 16. Reactor; 17. Connection block one; 18. Connection block two; 19. Opening two; 20. Threaded rod one; 21. Threaded rod two; 22. Lateral moving assembly; 23. Forward and backward moving assembly. DETAILED DESCRIPTION

[0023] Below, the present application is further described in conjunction with the accompanying drawings and specific implementation methods. It should be noted that, under the premise of no conflict, the various embodiments or technical features described below can be arbitrarily combined to form a new embodiment.

[0024] In the description of the present application, it should be understood that the terms "up", "down", "left", "right", etc., indicating directions or positional relationships are based on the directions or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore should not be understood as a limitation on the present application.

[0025] The terms "first", "second", etc. in this application are used to distinguish similar objects, and are not used to describe a specific order or sequence. It should be understood that the data used in this way can be interchanged where appropriate, so that the embodiments of the present application can be implemented in an order other than those illustrated or described herein, and the objects distinguished by "first", "second", etc. are generally of one type, and the number of objects is not limited. For example, the first object can be one or more. In addition, "and / or" in the specification and claims represents at least one of the connected objects, and the character " / " generally indicates that the objects associated with each other are in an "or" relationship of a high-throughput peptide synthesis device.

[0026] Embodiment 1:

[0027] like Figure 1-Figure 4 As shown, the present embodiment provides a high-throughput polypeptide synthesis device, which includes a horizontal plate 4, a vertical plate 3 vertically fixed on the top of the horizontal plate 4, a storage tank cabinet 1 arranged on one side of the horizontal plate 4, a liquid storage tank arranged in the storage tank cabinet 1, four support plates 15 symmetrically arranged on the top of the horizontal plate 4, a plurality of reaction kettles 16 arranged on the top of the support plate 15, a lateral moving component 22 arranged on the rear side wall of the vertical plate 3, a front-rear moving component 23 arranged on the lateral moving component 22, an electric telescopic rod 12 arranged on the front-rear moving component 23, a mounting plate 13 arranged at the telescopic end of the electric telescopic rod 12, and a plurality of liquid adding needles 14 arranged on the mounting plate 13, each of which is connected to the corresponding liquid storage tank by a separate pipeline.

[0028] A high-throughput peptide synthesis device includes a horizontal plate 4, a vertical plate 3 is vertically fixed on the top of the horizontal plate 4, a storage tank cabinet 1 is arranged on one side of the horizontal plate 4, a liquid storage tank is arranged in the storage tank cabinet 1, four support plates 15 are symmetrically arranged on the top of the horizontal plate 4, a plurality of reaction kettles 16 are arranged on the top of the support plate 15, a horizontal moving component 22 is arranged on the rear side wall of the vertical plate 3, a front-rear moving component 23 is arranged on the horizontal moving component 22, an electric telescopic rod 12 is arranged on the front-rear moving component 23, a mounting plate 13 is arranged at the telescopic end of the electric telescopic rod 12, a plurality of liquid adding needles 14 are arranged on the mounting plate 13, and each liquid adding needle 14 is connected to the corresponding liquid storage tank by a separate pipeline. When in use, the liquid adding needle 14 is driven to move forward and backward by operating the lateral moving component 22, and the liquid adding needle 14 is driven to move forward and backward by operating the forward and backward moving component 23, so that several liquid adding needles 14 can be parallelly displaced in the area above the reactor 16. The pipeline controlled by the solenoid valve at the rear end is directly connected to the liquid storage tank. Each reagent has a separate pipeline and is added by a rotary plunger pump, which greatly improves the efficiency of adding liquid. Compared with the existing technology, it solves the problems of slow synthesis speed and low flux: the current mainstream equipment uses single-channel pipetting, but there are many types of reagents, there are 32 commonly used amino acids, and there are 4-6 common reagents. Each reagent is added in a single channel. At present, most of them are single reactors 16, and the efficiency of single polypeptide synthesis is too low to meet the technical problems of production needs. Since each dosing needle only adds fixed reagents, it can add liquid directly without pipeline cleaning, which greatly reduces the time and cost of the cleaning step, and avoids a large amount of hazardous waste liquid pollution and treatment costs. Compared with the existing technology, it solves the problem of reagent waste: the single-channel pipetting solution requires a large amount of DMF for pipeline cleaning after adding a liquid, which greatly increases the reagent cost and the cost of hazardous waste liquid treatment. And DMF (N, N-dimethylformamide) is classified as a hazardous chemical, which is flammable, explosive and volatile, and has potential effects on the reproductive system and nervous system, which may cause pregnancy problems and technical problems of neurodegeneration.

[0029] Furthermore, the lateral moving component 22 includes a horizontal box 5 fixedly arranged on the rear side wall of the vertical plate 3, an adjusting motor 6 fixedly arranged on the left side wall of the horizontal box 5, an opening 8 opened on the rear side wall of the horizontal box 5, a threaded rod 20 penetrating through the left side wall of the horizontal box 5 and rotating forward and reversely through the adjusting motor 6, a connecting block 17 penetrating through the opening 8 and threadedly connected to the threaded rod 20, a moving block 10 fixedly arranged on the rear side wall of the connecting block 17, and the front and rear moving component 23 is arranged on the moving block 10.

[0030] The horizontal moving assembly 22 includes a horizontal box 15 fixedly arranged on the rear side wall of the vertical plate 3, an adjusting motor 16 fixedly arranged on the left side wall of the horizontal box 15, an opening 18 provided on the rear side wall of the horizontal box 15, a threaded rod 20 which is rotated forward and reversely by the adjusting motor 6 is provided through the left side wall of the horizontal box 5, a connecting block 17 which is threadedly connected to the threaded rod 20 is provided through the opening 8, a moving block 10 is fixedly arranged on the rear side wall of the connecting block 17, and the front-back moving assembly 23 is arranged on the moving block 10. When in use, the adjusting motor 6 is operated, and the front-back moving assembly 23 is moved forward and backward by the forward and reverse rotation of the adjusting motor 6, the adjusting motor 6 drives the threaded rod 20 to rotate, the threaded rod 20 drives the connecting fast one to move, the connecting fast one drives the moving block 10 to drive, and the moving block 10 drives the front-back moving assembly 23 to move, thereby realizing the front-back moving assembly 23.

[0031] Furthermore, the front-rear moving assembly 23 includes a horizontal box 27 fixedly arranged on the moving block 10, an adjusting motor 29 fixedly arranged on the rear side wall of the horizontal box 27, an opening 21 opened on the outer side wall of the horizontal box 27, a threaded rod 21 penetrating through the rear side wall of the horizontal box 27 and rotated forward and reversely by the adjusting motor 29, a connecting block 218 penetrating through the opening 219 and threadedly connected to the threaded rod 21, a moving block 11 fixedly arranged on the connecting block 218, and an electric telescopic rod 12 is arranged at the bottom of the moving block 11.

[0032] The forward and backward moving assembly 23 includes a horizontal box 27 fixedly arranged on the moving block 10, an adjusting motor 29 fixedly arranged on the rear side wall of the horizontal box 27, an opening 219 provided on the outer side wall of the horizontal box 27, a threaded rod 21 penetrating through the rear side wall of the horizontal box 27 for forward and reverse rotation by the adjusting motor 29, a connecting block 28 threadedly connected to the threaded rod 21 penetrating through the opening 219, a moving block 211 fixedly arranged on the connecting block 218, and the electric telescopic rod 12 arranged at the bottom of the moving block 21. When in use, the adjusting motor 29 is started, and the forward and reverse rotation of the adjusting motor 29 is used to realize the forward and backward movement of the liquid adding needle 14, the adjusting motor 29 drives the threaded rod 21 to rotate, the threaded rod 21 drives the moving block 21 to move, the moving block 11 drives the connecting block 28 to move, and the connecting block 2 drives the liquid adding needle 14 to move, thereby realizing the forward and backward movement of the liquid adding needle 14.

[0033] Furthermore, the plurality of liquid adding needles 14 are arranged in a 7 by 7 square array, forty-nine in total, which greatly improves the liquid adding efficiency.

[0034] Furthermore, a plurality of reaction kettles 16 are arranged on the support plate 15 in a 6 by 8 arrangement, and up to 192 reaction kettles 16 can be synthesized simultaneously, thereby improving the liquid addition efficiency.

[0035] Furthermore, a cover box 2 is provided between the vertical plate 3 and the horizontal plate 4. The cover box 2 is provided to protect and cover the reaction kettle 16.

[0036] Furthermore, the peptide synthesis steps are as follows:

[0037] 1. Swelling: Use DCM / DMF to swell the resin joint for 30 minutes, drain the solution, add DMF to wash once, 3 minutes / time, and drain the solution;

[0038] 2. Deprotection: add piperidine for the first deprotection, react for 5 min, and drain the solution; add piperidine for the second time, react for 15 min, and drain the solution; wash with DMF 5 times, 3 min / time, and drain the solution;

[0039] 3. Coupling: Add amino acid, DIC and Oxyma, react for at least 1 hour, drain the solution, wash three times with DMF, 3 minutes each time, and drain the solution;

[0040] 4. Repeat steps 2-3.

[0041] The above-mentioned implementation modes are only preferred implementation modes of the present application and cannot be used to limit the scope of protection of the present application. Any non-substantial changes and substitutions made by technicians in this field on the basis of the present application shall fall within the scope of protection required by the present application.

Claims

1. A high-throughput peptide synthesis device, characterized in that: The invention comprises a horizontal plate (4), a vertical plate (3) vertically fixed on the top of the horizontal plate (4), a storage tank cabinet (1) arranged on one side of the horizontal plate (4), a liquid storage tank arranged in the storage tank cabinet (1), four support plates (15) symmetrically arranged on the top of the horizontal plate (4), a plurality of reaction kettles (16) arranged on the top of the support plate (15), a horizontal moving component (22) arranged on the rear side wall of the vertical plate (3), a front-back moving component (23) arranged on the horizontal moving component (22), an electric telescopic rod (12) arranged on the front-back moving component (23), a mounting plate (13) arranged at the telescopic end of the electric telescopic rod (12), and a plurality of liquid adding needles (14) arranged on the mounting plate (13), wherein each of the liquid adding needles (14) is connected to the corresponding liquid storage tank by a separate pipeline.

2. A high-throughput peptide synthesis device according to claim 1, characterized in that: The lateral moving assembly (22) comprises a horizontal box (5) fixedly arranged on the rear side wall of the vertical plate (3), an adjusting motor (6) fixedly arranged on the left side wall of the horizontal box (5), an opening (8) opened on the rear side wall of the horizontal box (5), a threaded rod (20) penetrating the left side wall of the horizontal box (5) and rotating forward and reversely through the adjusting motor (6), a connecting block (17) penetrating the opening (8) and threadedly connected to the threaded rod (20), and a moving block (10) fixedly arranged on the rear side wall of the connecting block (17), and the front-back moving assembly (23) is arranged on the moving block (10).

3. A high-throughput peptide synthesis device according to claim 2, characterized in that: The front-rear moving assembly (23) comprises a second horizontal box (7) fixedly arranged on the first moving block (10), a second adjusting motor (9) fixedly arranged on the rear side wall of the second horizontal box (7), a second opening (19) provided on the outer side wall of the second horizontal box (7), a second threaded rod (21) penetrating the rear side wall of the second horizontal box (7) and being rotated forward and reversely by the second adjusting motor (9), a second connecting block (18) penetrating the second opening (19) and being threadedly connected to the second threaded rod (21), and a second moving block (11) fixedly arranged on the second connecting block (18), wherein the electric telescopic rod (12) is arranged at the bottom of the second moving block (11).

4. A high-throughput peptide synthesis device according to claim 3, characterized in that: The plurality of liquid adding needles (14) are arranged in a 7 by 7 square array.

5. A high-throughput peptide synthesis device according to claim 4, characterized in that: The plurality of reaction kettles (16) are arranged in a 6 by 8 arrangement.

6. A high-throughput peptide synthesis device according to claim 5, characterized in that: A cover box (2) is provided between the vertical plate (3) and the horizontal plate (4).

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