Inlet valve array, cleaning method and synthesis system based on inlet valve array

Abstract

The invention provides an inlet valve array, a cleaning method and a biosynthesis system based on the inlet valve array. A first connecting flow channel and a second connecting flow channel of the inlet valve array are connected with a cleaning main pipeline through pipelines; outlets of the first connecting flow channels of the two inlet valve arrays are connected with a first liquid inlet main pipeline through pipelines. A first connecting flow channel of one inlet valve array is connected with a second liquid inlet main pipeline through a pipeline, the first liquid inlet main pipeline and the second liquid inlet main pipeline are respectively connected with a column front input pipeline, and the column front input pipeline is connected with the input ends of a plurality of composite columns through a plurality of valves; an output end pipeline of each composite column is connected with an inlet of a post output pipeline through a corresponding valve; an outlet of the post-column output pipeline is connected with a discharge pipeline and a second liquid inlet main pipeline through valves; the device has the advantages that cross contamination generated when different materials are conveyed can be avoided, the synthesis purity is improved, and waste of the materials is reduced, so that the cost is reduced; and the pump can be switched without stopping.

Description

Technical Field

[0001] The present invention relates to the field of biosynthesis, in particular to an inlet valve array, a cleaning method and a biosynthesis system based on the inlet valve array. Background Art

[0002] Nucleic acid synthesizer is an automated device used to synthesize DNA or RNA using solid phase binding method. Nucleotide monomers are added according to the set program. Its synthesis method is the phosphoamidite method. The chemical steps of the phosphoamidite valve are: (1) Deprotection: Use trichloroacetic acid to remove the nucleotide protecting group attached to the solid phase support to obtain free 5 ， -hydroxyl group; (2) activation: mixing the phosphoramidite-protected nucleotide monomer with an activator to react and obtain an intermediate; (3) coupling: reacting the active intermediate with the deprotected nucleotide on the solid support; (4) capping: using a capping agent to block the unreacted 5 ， -hydroxyl groups, preventing extension in subsequent cycles and improving the purity of the product; (5) Oxidation: converting the phosphoramidite bond into a phosphotriester bond to obtain a stable nucleotide. Finally, repeat the above steps.

[0003] U.S. Patent Publication No. US20060280651A1, entitled "Industrial biosynthesizer system and method," primarily utilizes two delivery pumps to connect two inlet modules (for connecting phosphoramidite monomers and liquid reagents) to an external reaction vessel (synthesis column). The reagents are delivered to the reaction vessel in a specific sequence while controlling the flow rate. The inlet modules in this patent utilize multiple diaphragm valves and multi-port static diaphragm valves. This combination of diaphragm valves and static diaphragm valves presents a disadvantage: Residues remain after adding individual monomers, deprotecting agents, activators, capping agents, and oxidants, resulting in relatively low product purity.

[0004] The Chinese patent publication number is CN113000003B, and the invention is titled "System and Method for Nucleic Acid Synthesis." The invention involves the use of a rotary valve with alternating cleaning and material pipes, which helps prevent cross-contamination when adding the next material. The disadvantages are that the rotary valve is not cleaned thoroughly, leaving a certain amount of residue, and can only clean the common pipes. The rotary valve requires the pump to be stopped for switching, and can only meet laboratory-level production requirements. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the present invention provides an inlet valve array, a cleaning method, and a biosynthesis system based on the inlet valve array. These can avoid cross-contamination when transporting different materials, improve synthesis purity, reduce material waste, and thus reduce costs; and can be switched without stopping the pump.

[0006] To solve the above technical problems, the first aspect of the present disclosure provides an inlet valve array, comprising a valve body, a plurality of three-way valves and a plurality of two-way valves arranged on the valve body, a plurality of reagent delivery ports arranged on the valve body, each reagent delivery port being connected to a first inlet end of a corresponding three-way valve through an inlet channel, a second inlet end of the corresponding three-way valve being connected to an outlet end of a corresponding two-way valve through a channel on the valve body, a first connection channel and a second connection channel arranged on the valve body, an outlet end of each three-way valve being connected to the first connection channel through a corresponding channel, an inlet end of each two-way valve being connected to the second connection channel through a corresponding channel, and a two-way valve being arranged at the inlet of the first connection channel.

[0007] In some embodiments, each three-way valve comprises a lower fluid housing and an upper fluid housing, a fluid cavity being formed between the lower fluid housing and the upper fluid housing, a diaphragm being arranged between the lower fluid housing and the upper fluid housing, a first inlet end and a second inlet end being arranged on both sides of the lower fluid housing, the first inlet end and the second inlet end being connected to the fluid cavity formed between the lower fluid housing and the upper fluid housing, the fluid cavity formed between the lower fluid housing and the upper fluid housing being connected to a first outlet end arranged on the lower fluid housing, a piston being arranged on the upper fluid housing corresponding to the first inlet end and the second inlet end, the piston being connected to a valve rod, the valve rod being driven by a driving device in the valve housing, and the piston controlling the opening and closing of the first inlet end and the second inlet end to the fluid cavity.

[0008] In some embodiments, each two-way valve comprises a lower fluid housing and an upper fluid housing, a fluid cavity being formed between the lower fluid housing and the upper fluid housing, a diaphragm being arranged between the lower fluid housing and the upper fluid housing, a second outlet end being arranged on one side of the lower fluid housing, the outlet end being connected to the fluid cavity formed between the lower fluid housing and the upper fluid housing, a third inlet end being arranged on the lower fluid housing, the inlet end being connected to the fluid cavity, a piston being arranged on the upper fluid housing corresponding to the outlet end, the piston being connected to a valve rod, the valve rod being driven by a driving device in the second valve housing, and the piston controlling the opening and closing of the outlet end to the fluid cavity.

[0009] In some embodiments, the number of three-way valves is greater than or equal to eight, and the number of two-way valves is greater than or equal to nine.

[0010] To solve the above technical problems, the second aspect of the present disclosure provides a method for cleaning an inlet valve array, which utilizes the inlet valve array described above, the inlet end of the first connection channel of the valve array being connected to a cleaning main pipeline through a two-way valve, the second connection channel of the valve array being connected to the cleaning main pipeline, and the reagent delivery ports on the valve array being respectively connected to corresponding reagent delivery pipelines.

[0011] The cleaning method comprises the following steps:

[0012] Step 1) After the reagent solution enters from the first inlet end of the three-way valve through the corresponding reagent conveying port, it enters the first connecting flow channel through the outlet end of the three-way valve, and is output through the output port of the first connecting flow channel;

[0013] Step 2) Valve cleaning: the cleaning solvent enters the two-way valve of the corresponding three-way valve through the cleaning main pipeline, enters the three-way valve, and then enters the first connecting flow channel through the three-way valve, and is output through the output port of the first connecting flow channel;

[0014] Step 3) According to the above step 2), sequentially clean the three-way valve located in the input reagent and all three-way valves in the output port direction of the first connecting flow channel;

[0015] Step 4) Cleaning of the first connecting flow channel: the cleaning solvent enters the first connecting flow channel through the cleaning main pipeline, flows into the first connecting flow channel from the input end of the first connecting flow channel, and flows out from the output end of the first connecting flow channel.

[0016] In some embodiments, the first inlet end of the three-way valve is a normally closed end, and the second inlet end is a normally open end; when the first inlet end is opened, the second inlet end is closed; when the reagent enters, the first inlet end is opened; when the cleaning agent enters, the first inlet end is closed.

[0017] To solve the above technical problems, the third aspect of the present disclosure provides a biosynthesis system based on an inlet valve array, which comprises the above-mentioned inlet valve array, wherein the inlet valve array comprises three; the first connecting flow channel and the second connecting flow channel of each inlet valve array are respectively connected to a cleaning main pipeline through a pipeline; the outlet of the first connecting flow channel of two inlet valve arrays is connected to a first liquid inlet main pipeline through a pipeline; the first connecting flow channel of the other inlet valve array is connected to a second liquid inlet main pipeline through a pipeline, and a valve is arranged between the pipeline and the second liquid inlet main pipeline; the first liquid inlet main pipeline and the second liquid inlet main pipeline are respectively connected to a pre-column input pipeline, and the pre-column input pipeline is connected to the input ends of a plurality of synthesis columns through a plurality of valves; the output end pipeline of each synthesis column is connected to the inlet of a post-column output pipeline through a corresponding valve; the outlet of the post-column output pipeline is connected to a discharge pipeline and the second liquid inlet main pipeline through a valve; a pump is arranged on the first liquid inlet main pipeline and the second liquid inlet main pipeline respectively.

[0018] In some embodiments, the outlet of the post-column output conduit, the drain conduit, the second inlet valve array and the outlet conduit of the first connecting flow channel of the another inlet valve array are respectively connected with valves, the valves comprising a first position and a second position, in the first position, the outlet of the post-column output conduit is communicated with the inlet of the second inlet valve array, and the outlet conduit of the first connecting flow channel of the another inlet valve array is communicated with the drain conduit; in the second position, the outlet of the post-column output conduit is communicated with the drain conduit, and the outlet conduit of the first connecting flow channel of the another inlet valve array is communicated with the inlet of the second inlet valve array.

[0019] In some embodiments, the valve is a two-position four-way rotary valve.

[0020] In some embodiments, the pre-column input conduit is communicated with the post-column output conduit through a conduit.

[0021] In some embodiments, the pre-column input conduit is communicated with the plurality of synthesis columns through a one-in-multiple-out rotary valve, and the post-column output conduit is communicated with the plurality of synthesis columns through a multiple-in-one-out rotary valve; the pre-column input conduit is connected with the inlet end of the one-in-multiple-out rotary valve, the input end of each synthesis column is connected with the outlet end of the one-in-multiple-out rotary valve, the output end of each synthesis column is connected with the input end of the multiple-in-one-out rotary valve, and the output end of the multiple-in-one-out rotary valve is connected with the post-column output conduit; one of the outlet ends of the one-in-multiple-out rotary valve is connected with the input end of the multiple-in-one-out rotary valve through a conduit.

[0022] In some embodiments, a pre-column pressure sensor is arranged on the pre-column input conduit, and a post-column pressure sensor, an ultraviolet detector and a conductivity detector are arranged on the post-column output conduit.

[0023] In some embodiments, a check valve is arranged on the drain conduit.

[0024] Advantages of the present application:

[0025] 1. The inlet valve array adopted in the present application can meet the cleaning of the valve and the conduit when different kinds of monomers and reagents (deprotection agents, activation agents, oxidizing agents, capping agents, etc.) are added, so as to avoid pollution.

[0026] 2. The synthesis system of the present application can realize the flushing of the inlet valve array after the use of one reagent, clean the synthesis column and the conduit, and then input the next reagent, so as to avoid cross contamination and improve the synthesis purity. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 FIG. 1 is a sectional view of the inlet valve array of the present application.

[0028] Figure 2The structure schematic diagram of the inlet valve array of the present application.

[0029] Figure 3 The schematic diagram of the cleaning method of the inlet valve array of the present application.

[0030] Figure 4 The structure schematic diagram of the synthesis system of the present application. DETAILED DESCRIPTION

[0031] The present application will be further described below with reference to the drawings.

[0032] The technical content of the present application is described below through specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present application from the disclosure of the present application. The present application can also be implemented or applied through other different embodiments. Those skilled in the art can make various modifications and changes without departing from the spirit of the present application.

[0033] Before the specific embodiments of the present disclosure are described in detail, some terms used in the present disclosure are first explained.

[0034] Unless otherwise defined in the following, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. References made to technical terms used herein are intended to refer to the technical field as commonly understood by those skilled in the art, including those changes or substitutions of technology or replacement of equivalent technology that are obvious to those skilled in the art. Although the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present application. When a term appears herein, it is intended to refer to its corresponding term. All patents, published patent applications, and publications recited herein are incorporated by reference herein.

[0035] The terms "connected," "coupled," or "coupling," or similar terms as used herein are not limited to direct connections, but also include indirect connections.

[0036] The terms "on-line monitoring" or "real-time monitoring" as used herein refer to detecting certain parameters or properties of the buffer, reaction fluid, fluid flowing out of the flow reactor, such as pH value, pressure, flow rate, conductivity, etc. in real time during the use of the chromatography system. Unlike off-line detection or analysis, on-line monitoring or real-time monitoring can provide real-time feedback of the detection results.

[0037] The positional relationship words "upper", "lower", "left", "right", "front", "back", etc. as referred to herein are determined according to the layout direction of the drawings of the specification, and are only used to represent the relative positional relationship. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0038] Referring toFigure 1 As shown, the inlet valve array includes a valve body 1, one side of the valve body 1 is provided with a plurality of reagent delivery ports 2, such as Figure 2As shown, eight reagent delivery ports 2 are provided in total, and a single reagent delivery port 2 is connected to the first inlet end 4 of the corresponding three-way valve 3 through the first flow channel 7 in the valve body 1. The single three-way valve 3 comprises a lower fluid housing 31 and an upper fluid housing 32, and the lower fluid housing 31 and the upper fluid housing 32 form a fluid cavity, the first inlet end 4 of the three-way valve 3 is in communication with the fluid cavity, the second inlet end 5 of the three-way valve 3 is in communication with the fluid cavity, and the middle position of the fluid cavity is in communication with the first outlet end 6, and the first outlet end 6 is in communication with the vertical second flow channel 8 provided on the valve body 1. A diaphragm 33 is provided between the lower fluid housing 31 and the upper fluid housing 32. The upper fluid housing 32 is provided with a piston 34 corresponding to the positions of the first inlet end 4 and the second inlet end 5, respectively, the piston 34 is connected to a valve rod 35, the valve rod 35 is driven by a driving device in the valve shell 36, and the piston 34 controls the opening and closing of the first inlet end 4 and the second inlet end 5, respectively. The three-way valve 3 in the present application can adopt a normally closed electromagnetic valve on one side, for example, the first inlet end 4 is a normally closed end, when the electromagnetic valve is powered on, the first inlet end 4 is opened, and at this time, the second inlet end 5 is closed; when the electromagnetic valve is powered off, the first inlet end 4 is closed, and at this time, the second inlet end 5 is opened. The second inlet end 5 of the three-way valve 3 is in communication with the second outlet end 11 of the two-way valve 9 through the third flow channel 10 on the valve body 1. The two-way valve 9 comprises a lower fluid housing 91 and an upper fluid housing 92, and a fluid cavity is formed between the lower fluid housing 91 and the upper fluid housing 92, the other side position of the fluid cavity is in communication with the third inlet end 12, and the third inlet end 12 is in communication with the vertical fourth flow channel 13 on the valve body 1. The second outlet end 11 of the two-way valve 9 is in communication with the fluid cavity. A diaphragm 93 is provided between the lower fluid housing 91 and the upper fluid housing 92, and the upper fluid housing 92 is provided with a piston 94 corresponding to the positions of the second outlet end 11 and the third inlet end 12, respectively, the piston 94 is connected to a valve rod 95, the valve rod 95 is driven by a driving device in the valve shell 96, and the piston 94 controls the opening and closing of the second outlet end 11 and the third inlet end 12 with the fluid cavity. The two-way valve 9 can adopt an electromagnetic valve, when powered on, the second outlet end 11 is in communication with the fluid cavity, and when powered off, it is a normally closed end. The third inlet end 12 of the two-way valve 9 can be in a normally open state, that is, the opening and closing of the second outlet end 11 does not affect the third inlet end 12 to be in a normally open state. The second flow channel 8 on the valve body 1 is in communication with the first connection flow channel 14 on the valve body 1, and the first connection flow channel 14 is vertically arranged with the second flow channel 8 in space. The fourth flow channel 13 on the valve body 1 is in communication with the second connection flow channel 15, and the fourth flow channel 13 is vertically arranged with the second connection flow channel 15 in space. The first inlet end 4, the second inlet end 5, the first outlet end 6 of the three-way valve 9, and the second outlet end 11, the third inlet end 12 of the two-way valve 9 are respectively provided with a sealing ring 17 between the valve body 1.

[0039] Refer to Figure 2The schematic diagram of the structure of the inlet valve array includes multiple three-way valves 3 and multiple two-way valves 9. As shown in the figure, there are eight three-way valves 3 and eight two-way valves 9 connected in pairs. The reagent delivery ports 2 on the valve array 1 are respectively connected to corresponding reagent delivery pipelines 18; the first outlet ends 6 of the eight three-way valves 3 are respectively connected to the first connecting flow channel 14, and a two-way valve 9 is provided at the inlet end of the first connecting flow channel 14. The second outlet end 11 and the third inlet end 12 of the two-way valve 9 are respectively connected to the first connecting flow channel 14; the second connecting flow channel 15 is connected to the third inlet end 12 of the eight two-way valves 9. The inlet ends of the first connecting flow channel 14 and the second connecting flow channel 15 are connected to the main cleaning pipeline 16.

[0040] When using, Figure 3 As shown, the reagent is input from the reagent delivery port 2 at the top of the inlet valve array. At this time, the first inlet end 4 of the three-way valve 3 is opened, and then the reagent enters the first connecting channel 14 through the first outlet end 6 of the three-way valve 3, and then flows out along the outlet end of the first connecting channel 14. During cleaning, the three-way valve 3 for inputting the reagent is disconnected, the first inlet end 4 of the three-way valve 3 is closed, and the second inlet end 5 of the three-way valve 3 is opened. At the same time, the second outlet end 11 of the two-way valve 9 connected to the three-way valve 3 is opened, and the cleaning liquid enters the two-way valve 9 through the cleaning main pipeline 16, and enters the second inlet end 5 of the three-way valve 3 through the second outlet end 11 of the two-way valve 9, achieving the purpose of cleaning the second inlet end 5 of the three-way valve 3. Finally, the cleaning liquid flows out through the first connecting channel 14. Similarly, the second outlet end 11 of the two-way valve 9 is opened in sequence from top to bottom, and the cleaning liquid cleans the second inlet end 5 of the three-way valve 3 respectively until the second inlet end 5 of the three-way valve 3 at the outlet position of the first connecting channel 14 is cleaned. Finally, open the second outlet end 11 of the two-way valve 9 at the inlet position of the first connecting flow channel 14. At this time, the first connecting flow channel 14 is connected to the two-way valve 9, and the cleaning liquid enters from the third inlet end 12 of the two-way valve 9, flows out from the second outlet end 11 of the two-way valve 9, and flows out from top to bottom along the first connecting flow channel 14, thereby achieving the purpose of cleaning the common channel of the three-way valve.

[0041] Reference Figure 4As shown, based on the inlet valve array-based biosynthesis system, the system uses five synthesis columns. The system comprises a first inlet valve array 20, a second inlet valve array 21 and a third inlet valve array 22, and the first connecting flow channel and the second connecting flow channel of the first inlet valve array 20, the second inlet valve array 21 and the third inlet valve array 22 are communicated with the cleaning main pipeline 16, the first connecting flow channel of the first inlet valve array 20 and the second inlet valve array 21 is connected with the first liquid inlet main pipeline 23 through a pipeline, and the first liquid inlet main pipeline 23 is provided with a pump one 24; the outlet end of the first liquid inlet main pipeline 23 is connected with the inlet end of the pre-column input pipeline 25, the outlet end of the pre-column input pipeline 25 is communicated with each synthesis column 26 through a pre-column six-position seven-way rotary valve 27, the inlet end of the pre-column input pipeline 25 is communicated with the pre-column six-position seven-way rotary valve 27, and the five outlet ends of the pre-column six-position seven-way rotary valve 27 are connected with the inlet ends of the five synthesis columns 26 through pipelines. The pipeline of the outlet end of the five synthesis columns 26 is connected with a post-column output pipeline 29 through a post-column six-position seven-way rotary valve 28, the pipeline of the outlet end of the five synthesis columns 26 is connected with the five inlet ends of the post-column six-position seven-way rotary valve 28, and the outlet end of the post-column six-position seven-way rotary valve 28 is connected with the post-column output pipeline 29; wherein one outlet end of the pre-column six-position seven-way rotary valve 27 is connected with one inlet end of the post-column six-position seven-way rotary valve 28 through a pipeline, so that the direct communication between the pre-column input pipeline 25 and the post-column output pipeline 29 can be realized without passing through the synthesis column 26.

[0042] The system further comprises a second liquid inlet main pipeline 201 and a discharge pipeline 202; the outlet end of the post-column output pipeline 29, the pipeline of the first connecting flow channel of the third inlet valve array 22, the second liquid inlet main pipeline 201 and the discharge pipeline 202 are respectively connected with a valve 203, the valve 203 has two positions, when being in the first position, the second liquid inlet main pipeline 201 is communicated with the outlet end of the post-column output pipeline 29, and the pipeline of the first connecting flow channel of the third inlet valve array 22 is communicated with the discharge pipeline 202; when being in the second position, the outlet end of the post-column output pipeline 29 is communicated with the discharge pipeline 202, and the pipeline of the first connecting flow channel of the third inlet valve array 22 is communicated with the second liquid inlet main pipeline 201. The valve 203 can be a two-position four-way rotary valve. The second liquid inlet main pipeline 201 is provided with a pump two 204, and the outlet end of the second liquid inlet main pipeline 201 is connected with the inlet end of the pre-column input pipeline 25.

[0043] The pre-column input pipeline 25 is provided with a first pressure sensor 205, the post-column output pipeline 29 is provided with a second pressure sensor 206, an ultraviolet detector 207 and a conductivity detector 208. The discharge pipeline 202 is provided with a check valve 209.

[0044] In use, the following steps are performed:

[0045] (1) Synthesis column cleaning

[0046] Open the two-way valve of the first connection channel inlet of the first inlet valve array 20 or the second inlet valve array 21 and the third inlet valve array 22, and the cleaning liquid enters the first connection channel of the first inlet valve array 20 or the second inlet valve array 21 and the third inlet valve array 22, respectively; at this time, the valve 203 is in the second position, and the cleaning liquid flows into one of the synthesis columns 26 through the first liquid inlet main pipeline 23, the second liquid inlet main pipeline 201, and the pre-column input pipeline 25; then the cleaning liquid flows out of the synthesis column 26, passes through the post-column output pipeline 29 and the discharge pipeline 202, and is discharged.

[0047] (2) Deprotection

[0048] Open one of the three-way valves in the third valve array 22, for example, the fifth three-way valve from top to bottom, at this time, the deprotection agent is input from the reagent delivery port, and flows into the first connection channel of the third valve array 22 through the three-way valve; at this time, the valve 203 is in the second position, and the deprotection agent enters the synthesis column 26 cleaned in step (1) through the second liquid inlet main pipeline 201 and the pre-column input pipeline 25, and after the reaction, the deprotection agent is discharged and collected through the post-column output pipeline 29 and the discharge pipeline 202.

[0049] When cleaning, the synthesis column 26 needs to be cleaned, and the third inlet valve array 22 also needs to be cleaned: when cleaning the third inlet valve array 22, the cleaning liquid is cleaned in the order of the above-mentioned inlet valve array, that is, the cleaning liquid sequentially cleans the three-way valve inputting the deprotection agent, then sequentially cleans the three-way valves downward along the three-way valve, and finally opens the two-way valve of the first connection channel inlet of the third inlet valve array 22, and the cleaning liquid cleans from the first connection channel inlet to the outlet of the third inlet valve array 22; each time cleaning, the valve 203 is in the second position, the cleaning liquid flows into the first liquid inlet main pipeline 23, the second liquid inlet main pipeline 201, and the pre-column input pipeline 25, and enters the synthesis column 26 of the deprotection reaction in step (2), and after cleaning, the cleaning liquid is discharged and collected through the post-column output pipeline 29 and the discharge pipeline 202. At the same time, open the two-way valve of the first connection channel inlet of the first inlet valve array 20 or the second inlet valve array 21, and the cleaning liquid enters the first connection channel of the first inlet valve array 20 or the second inlet valve array 21, respectively; the cleaning liquid flows into the synthesis column 26 of the deprotection reaction in step (2) through the first liquid inlet main pipeline 23 and the pre-column input pipeline 25, and then the cleaning liquid flows out of the synthesis column 26, passes through the post-column output pipeline 29 and the discharge pipeline 202, and is discharged and collected.

[0050] (3) Activation + coupling

[0051] The corresponding input monomer needs to be activated. At this time, one of the three-way valves in the second inlet valve array 21 is opened, for example, the seventh three-way valve from top to bottom, and the phosphoramidite-protected nucleotide monomer is input from the reagent delivery port, flows into the first connecting channel of the second inlet valve array 21 through the three-way valve, and the monomer enters the first liquid inlet main pipe 23 and the pre-column input pipe 25 from the first connecting channel of the second inlet valve array 21; at the same time, the valve 203 is in the second position, and the input of the activator, at this time, one of the three-way valves in the third inlet valve array 22 is opened, for example, the sixth three-way valve from top to bottom, and the activator is input from the reagent delivery port, flows into the first connecting channel of the third inlet valve array 22 through the three-way valve, and the activator enters the second liquid inlet main pipe 201 and the pre-column input pipe 25 from the first connecting channel of the third inlet valve array 22. The two reagents enter the above-mentioned synthesis column 26 through the pre-column input pipe 25, and at the same time, the valve 203 is in the first position, and the intermediate is circulated and reacted between the second liquid inlet main pipe 201, the pre-column input pipe 25, the synthesis column 26, and the post-column output pipe 29, so that the active intermediate is fully coupled with the deprotected nucleotide on the solid support. After a period of coupling, the valve 203 is in the second position, and finally the reagents flow out of the synthesis column 26, pass through the post-column output pipe 29 and the discharge pipe 202, and are collected.

[0052] Further cleaning: The three-way valves of the second inlet valve array 21 and the third inlet valve array 22 need to be cleaned according to the above steps. The seventh three-way valve and the following three-way valves of the second inlet valve array 21 are cleaned; the sixth three-way valve and the following three-way valves of the third inlet valve array 22 are cleaned. Each time the valve 203 is in the second position, the cleaning liquid flows into the first liquid inlet main pipe 23, the second liquid inlet main pipe 201, and the pre-column input pipe 25, and enters the above-mentioned synthesis column 26. After cleaning, the cleaning liquid flows out of the synthesis column 26, passes through the post-column output pipe 29 and the discharge pipe 202, and is collected. Finally, the two-way valves on the first connecting channel entrances of the first inlet valve array 20, the second inlet valve array 21, and the third inlet valve array 22 are opened, and the cleaning liquid enters the first connecting channel of the first inlet valve array 20, the second inlet valve array 21, and the third inlet valve array 22, respectively. The cleaning liquid flows into the above-mentioned synthesis column 26 through the first liquid inlet main pipe 23 and the pre-column input pipe 25, and then the cleaning liquid flows out of the synthesis column 26, passes through the post-column output pipe 29 and the discharge pipe 202, and is collected.

[0053] (4) Oxidation

[0054] Open one of the three-way valves in the third valve array 22, for example, the third three-way valve from the top, at this time, the oxidizing agent is input from the reagent delivery port, flows into the first connecting flow channel of the third valve array 22 through the three-way valve; at this time, the valve 203 is in the second position, the oxidizing agent enters the synthetic column 26 cleaned in step (1) through the second liquid inlet main pipeline 201, the pre-column input pipeline 25, and after the reaction, the deprotection agent is discharged through the post-column output pipeline 29, the discharge pipeline 202.

[0055] When cleaning, the synthetic column 26 needs to be cleaned, and the third inlet valve array 22 needs to be cleaned at the same time: when cleaning the third inlet valve array 22, the cleaning liquid is cleaned in the order of the above-mentioned cleaning sequence of the inlet valve array, that is, the cleaning liquid sequentially cleans the three-way valve through which the oxidizing agent is input, then sequentially cleans the three-way valves downward along the three-way valve, and finally opens the two-way valve at the first connecting flow channel inlet of the third inlet valve array 22, and the cleaning liquid cleans from the first connecting flow channel inlet to the outlet of the third inlet valve array 22; each time, the valve 203 is in the second position, the cleaning liquid flows into the first liquid inlet main pipeline 23, the second liquid inlet main pipeline 201, and the pre-column input pipeline 25, and enters the above-mentioned synthetic column 26, and after cleaning, the cleaning liquid is discharged through the post-column output pipeline 29 and the discharge pipeline 202. At the same time, open the two-way valve at the first connecting flow channel inlet of the first inlet valve array 20 or the second inlet valve array 21, and the cleaning liquid enters the first connecting flow channel of the first inlet valve array 20 or the second inlet valve array 21, respectively, and then the cleaning liquid flows into the above-mentioned synthetic column 26 through the first liquid inlet main pipeline 23 and the pre-column input pipeline 25, and then flows out of the synthetic column 26, and is discharged through the post-column output pipeline 29 and the discharge pipeline 202.

[0056] (5) capping

[0057] The capping agent A and the capping agent B are input through the second valve array 21 and the third valve array 22, respectively, for example, the first inlet end of the eighth three-way valve from the top of the second valve array 21 and the third valve array 22 is connected to the reagent delivery pipeline of the capping agent A and the capping agent B, respectively. The capping agent A is delivered to the pre-column input pipeline 25 through the first liquid inlet main pipeline 23, and then enters the above-mentioned synthetic column 26. After the reaction, the reagent solution is discharged through the post-column output pipeline 29 and the discharge pipeline 202; similarly, the capping agent B is delivered to the pre-column input pipeline 25 through the second liquid inlet main pipeline 201, and then enters the above-mentioned synthetic column 26. After the reaction, the reagent solution is discharged through the post-column output pipeline 29 and the discharge pipeline 202.

[0058] When cleaning, the three-way valves of the second inlet valve array 21 and the third inlet valve array 22 need to be cleaned according to the above steps. Since the delivery end of the capping agent is at the lowermost three-way valve, the corresponding three-way valve needs to be cleaned. During each cleaning, the valve 203 is in the second position, the cleaning liquid flows into the first liquid inlet main pipeline 23, the second liquid inlet main pipeline 201, the pre-column input pipeline 25, and enters the above-mentioned synthesis column 26, after cleaning, the cleaning liquid is discharged through the post-column output pipeline 29 and the discharge pipeline 202. Finally, open the two-way valves of the first connection flow channel inlet of the first inlet valve array 20, the second inlet valve array 21, and the third inlet valve array 22, and the cleaning liquid enters the first inlet valve array 20, the second inlet valve array 21, and the third inlet valve array 22, respectively. The first connection flow channel cleaning liquid flows into the first liquid inlet main pipeline 23, the pre-column input pipeline 25, and enters the above-mentioned synthesis column 26, and then the cleaning liquid flows out of the synthesis column 26, passes through the post-column output pipeline 29 and the discharge pipeline 202, and is collected.

Claims

1. Inlet valve array, characterized in that, It includes a valve body, on which are arranged a plurality of three-way valves and a plurality of two-way valves; a plurality of reagent delivery ports are arranged on the valve body, each reagent delivery port is connected to the first inlet end of the corresponding three-way valve through the inlet flow channel; the second inlet end of the corresponding three-way valve is connected to the outlet end of the corresponding two-way valve through the flow channel on the valve body; a first connecting flow channel and a second connecting flow channel are arranged on the valve body; the outlet end of each of the three-way valves is respectively connected to the first connecting flow channel through the corresponding flow channel; the inlet end of each of the two-way valves is respectively connected to the second connecting flow channel through the corresponding flow channel; a two-way valve is arranged at the inlet of the first connecting flow channel.

2. The inlet valve array according to claim 1, characterized in that: Each of the three-way valves includes a lower fluid housing and an upper fluid housing, a fluid cavity is formed between the lower fluid housing and the upper fluid housing; a diaphragm is provided between the lower fluid housing and the upper fluid housing; a first inlet end and a second inlet end are provided on both sides of the lower fluid housing, the first inlet end and the second inlet end are respectively connected to the fluid cavity formed between the lower fluid housing and the upper fluid housing; a fluid cavity formed between the lower fluid housing and the upper fluid housing is connected to the first outlet end provided on the lower fluid housing; a piston is respectively provided on the upper fluid housing corresponding to the first inlet end and the second inlet end, the piston is connected to a valve stem, and the valve stem is driven by a driving device in the valve housing; the piston controls the opening and closing of the first inlet end and the second inlet and the fluid cavity.

3. The inlet valve array according to claim 1, characterized in that: Each of the two-way valves has a lower fluid housing 2 and an upper fluid housing 2, and a fluid cavity is formed between the lower fluid housing 2 and the upper fluid housing 2; a diaphragm 2 is provided between the lower fluid housing 2 and the upper fluid housing 2; a second outlet is provided on one side of the lower fluid housing 2, and the outlet is connected to the fluid cavity formed between the lower fluid housing 2 and the upper fluid housing 2; a third inlet is provided on the lower fluid housing 2, and the inlet is connected to the fluid cavity; a piston 2 is provided at a position corresponding to the outlet on the upper fluid housing 2, and the piston 2 is connected to the valve stem 2, and the valve stem 2 is driven by a driving device in the valve housing 2; the piston 2 controls the connection and disconnection between the outlet and the fluid cavity.

4. The inlet valve array according to claim 1, characterized in that: The number of the three-way valves is greater than or equal to eight; the number of the two-way valves is greater than or equal to nine.

5. A method for cleaning an inlet valve array, characterized in that: The inlet valve array comprises the inlet valve array according to claim 1-4, wherein the inlet end of the first connecting flow channel of the valve array is connected to the cleaning main pipeline through a two-way valve, and the second connecting flow channel of the valve array is connected to the cleaning main pipeline; the reagent delivery ports on the valve array are respectively connected to corresponding reagent delivery pipelines; The cleaning method comprises the following steps: Step 1) The reagent solution enters the first inlet of the three-way valve through the corresponding reagent delivery port, enters the first connecting channel through the outlet of the three-way valve, and is output through the output port of the first connecting channel; Step 2) Valve cleaning: The cleaning solvent enters the two-way valve corresponding to the three-way valve through the cleaning main pipeline, then enters the three-way valve, enters the first connecting flow channel through the three-way valve, and is output through the output port of the first connecting flow channel; Step 3) Cleaning the three-way valve located at the input reagent and all three-way valves in the direction of the output end of the first connecting flow channel in sequence according to the above step 2); Step 4) Cleaning the first connecting flow channel: The cleaning solvent enters the first connecting flow channel through the cleaning main pipeline, flows from the input end of the first connecting flow channel into the output end of the first connecting flow channel and flows out.

6. The method for cleaning the inlet valve array according to claim 5, characterized in that: The first inlet end of the three-way valve is a normally closed end, and the second inlet end is a normally open end; when the first inlet end is opened, the second inlet end is closed; when the reagent enters, the first inlet end is opened; when the cleaning agent enters, the first inlet end is closed.

7. A biosynthetic system based on an inlet valve array, characterized in that: It comprises the inlet valve arrays as described in claims 1-4, wherein three inlet valve arrays are used; the first connecting flow channel and the second connecting flow channel of each inlet valve array are respectively connected to the cleaning main pipeline through pipelines; the outlets of the first connecting flow channels of two of the inlet valve arrays are connected to the first liquid inlet main pipeline through pipelines; the first connecting flow channel of another inlet valve array is connected to the second liquid inlet main pipeline through a pipeline, and an on-off valve is provided between the pipeline and the second liquid inlet main pipeline; the first liquid inlet main pipeline and the second liquid inlet main pipeline are respectively connected to the pre-column input pipeline, and the pre-column input pipeline is connected to the input ends of multiple synthesis columns through multiple valves; the output end pipeline of each synthesis column is connected to the inlet of the post-column output pipeline through a corresponding valve; the outlet of the post-column output pipeline is connected to the discharge pipeline and the second liquid inlet main pipeline through a valve; pumps are respectively provided on the first liquid inlet main pipeline and the second liquid inlet main pipeline.

8. The biosynthesis system based on the inlet valve array according to claim 7, characterized in that: The outlet of the post-column output pipeline, the discharge pipeline, the second liquid inlet main pipeline and the output pipeline of the first connecting flow channel of another inlet valve array are respectively connected to valves, and the valve includes a first position and a second position. In the first position, the outlet of the post-column output pipeline is connected to the inlet of the second liquid inlet main pipeline, and the output pipeline of the first connecting flow channel of another inlet valve array is connected to the discharge pipeline; in the second position, the outlet of the post-column output pipeline is connected to the discharge pipeline, and the output pipeline of the first connecting flow channel of another inlet valve array is connected to the inlet of the second liquid inlet main pipeline.

9. The biosynthesis system based on the inlet valve array according to claim 8, characterized in that: The valve is a two-position four-way rotary valve.

10. The biosynthesis system based on an inlet valve array according to claim 7, characterized in that: The front-column input pipeline and the post-column output pipeline are connected through a pipeline.

11. The biosynthesis system based on an inlet valve array according to claim 7, characterized in that: The pre-column input pipeline is connected to the multiple synthesis columns through a one-inlet and multiple-outlet rotary valve, and the post-column output pipeline is connected to the multiple synthesis columns through a multiple-inlet and single-outlet rotary valve; the pre-column input pipeline is connected to the inlet end of the one-inlet and multiple-outlet rotary valve, and the input end of each synthesis column is connected to the output end of the one-inlet and multiple-outlet rotary valve; the output end of each synthesis column is connected to the input end of the multiple-inlet and single-outlet rotary valve, and the output end of the multiple-inlet and single-outlet rotary valve is connected to the post-column output pipeline; one of the output ends of the one-inlet and multiple-outlet rotary valve is connected to the input end of the multiple-inlet and single-outlet rotary valve through a pipeline.

12. The biosynthesis system based on an inlet valve array according to claim 7, characterized in that: The pre-column input pipeline is provided with a pre-column pressure sensor; the post-column output pipeline is provided with a post-column pressure sensor, an ultraviolet detector and a conductivity detector.

13. The biosynthesis system based on an inlet valve array according to claim 7, characterized in that: A check valve is provided on the discharge pipe.

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