Method and equipment for carrying out synthetic reaction on reagent and filler

Abstract

The invention provides a method and equipment for performing synthetic reaction on a reagent and filler. The method comprises the following steps: S1, adding a reagent liquid with a preset volume into a synthesis container; s2, enabling a part of the reagent liquid with a preset volume to flow through the reaction filler to obtain a primary reaction liquid; s3, enabling one part of the primary reaction liquid with the preset volume to reversely flow through the reaction filler again to obtain secondary reaction liquid, and enabling the other part of the primary reaction liquid which does not reversely flow through the reaction filler to become discharge liquid separated from the reaction filler; and S4, repeating the steps S2 to S3 for multiple times, and discharging the residual reagent liquid in the synthesis container. According to the method, complete conversion processing of the residual liquid is realized, so that the purposes of reducing cost and protecting environment are realized, and meanwhile, the product purity can be remarkably improved. Compared with a conventional processing technology at the present stage, the method has the advantages that the controllability of the reaction process is high, conversion and synthesis are thorough, reagent waste is reduced, and the product quality can be improved.

Description

Technical Field

[0001] The invention relates to the technical field of synthesis processing, and in particular to a method and equipment for causing a reagent and a filler to undergo a synthesis reaction. Background Art

[0002] In the field of chemical synthesis and biotechnology, a variety of chemical reactions and processes are involved, in which a large number of chemical reagents and fillers (synthetic carriers) are used. These processes usually involve injecting specific chemical reagents into a reaction vessel through a liquid injection unit so as to react chemically with the fillers or other reactants in the container. However, existing technical solutions have certain limitations in the use of reagents and the treatment of residual liquid.

[0003] In current technology, the injection of chemical reagents and the discharge of residual liquid are usually non-selective. That is, during the chemical reaction, both the reagents that have participated in the reaction and the unreacted reagents will be discharged together. Figure 1 Taking nucleic acid synthesis technology as an example, the existing nucleic acid synthesis technology generally adopts the column synthesis method, see Figure 1 As shown, the method injects key chemical reagents such as A / G / C / T monomer reagents corresponding to four different bases and acetonitrile reagents into the synthesis container through the injection unit to promote the chemical reaction between the reagents and the connection sites on the solid phase carrier filler in the synthesis container, and gradually construct the target nucleic acid sequence. However, in this process, the chemical reagents injected into the synthesis container are directly discharged through the discharge pipe regardless of whether they are completely reacted.

[0004] This treatment method leads to several significant problems: First, the utilization rate of chemical reagents is generally not high, because the unreacted reagents are discharged together with the residual liquid and cannot be effectively utilized in the current or subsequent synthesis process; second, the purity of the synthetic product is affected, and the presence of these components reduces the purity of the final product; third, the overall synthesis cost is high. Due to the waste of reagents and product purity issues, more reagent inputs and subsequent purification steps are required in the synthesis process, which increases costs.

[0005] Among them, the reasons that affect the purity of the synthetic product are specifically as follows: each time any reagent among A / G / C / T is added to the synthesis container, each added reagent must be connected to the connection site on the filler inside the synthesis container through a synthesis reaction, and there are many connection sites (hereinafter referred to as units) on the filler inside the synthesis container; assuming that in the process of adding A monomer reagent, two units in the filler (assuming that they are named X unit and Y unit) are selected, wherein X unit reacts with A reagent, but Y unit does not react with A; then G reagent, C reagent, and T reagent are added, and X unit and Y unit react with G reagent, C reagent, and T reagent; at this time, two products are synthesized, that is, product 1 synthesized on X unit has a sequence of AGCT, and product 2 synthesized on Y unit has a sequence of GCT. Based on the above reasons, conventional synthesis technology reduces the utilization rate of chemical reagents and the purity of the final nucleic acid product, thereby to a certain extent limiting the application and development of synthesis technology in a wider range of fields.

[0006] Therefore, one of the technical problems to be solved by the present invention is how to improve the purity of the product, that is, how to enable each filler unit to synthesize a product with a desired base sequence. Summary of the invention

[0007] To this end, the technical problem to be solved by the present invention is to overcome the problems of low reagent utilization, low product purity and high synthesis cost in the prior art, and to provide a new method and equipment for synthesizing reagents and fillers.

[0008] In order to solve the above technical problems, the present invention provides a method for causing a reagent and a filler to undergo a synthesis reaction, characterized in that it comprises the following steps: step S1, providing a reagent liquid containing the reagent and a synthesis container loaded with a reaction filler, and adding a preset first volume of the reagent liquid into the synthesis container; step S2, allowing a portion of the reagent liquid with a preset second volume to flow through the reaction filler to obtain a primary reaction liquid; step S3, allowing a portion of the primary reaction liquid with a preset third volume to flow through the reaction filler again in reverse to obtain a secondary reaction liquid, and another portion of the primary reaction liquid that has not reversely flowed through the reaction filler becomes a discharge liquid that is separated from the reaction filler, and the fourth volume of the discharge liquid is the difference between the second volume and the third volume; step S4, after repeatedly repeating steps S2 to S3, the remaining reagent liquid in the synthesis container is discharged.

[0009] In one embodiment of the present invention, the first volume of the reaction liquid added to the synthesis container in step S1 is set to V1, and step S2 further includes: setting the second volume of the primary reaction liquid to V2, and the size of V2 is obtained by the following relationship: V2=X·V1 / n, wherein n is an integer greater than 1, and 1<X<n.

[0010] In one embodiment of the present invention, in step S3: step S3 further includes: setting the third volume of the secondary reaction liquid to V3, the size of V3 is obtained by the following relationship: V3 = (X-1)·V1 / n, setting the volume of the discharge liquid separated from the reaction filler to V4, then V4 = V2-V3 = V1 / n.

[0011] In one embodiment of the present invention, the number of repetitions of steps S2 to S3 in step S4 depends on the values ​​of n and X, and is positively correlated with the value of n and negatively correlated with the value of X.

[0012] In one embodiment of the present invention, in step S4, the number of repetitions of steps S2 to S3 is at most an integer obtained by rounding down the calculation result of (n-X+1).

[0013] In one embodiment of the present invention, in step S2, a portion of the reagent liquid flows through the reaction filler at a first speed Vx in a range of 5 to 100 μl / s; in step S3, a portion of the primary reaction liquid flows through the reaction filler again in the reverse direction at a second speed Vy in a range of 5 to 100 μl / s.

[0014] In one embodiment of the present invention, in step S2, a portion of the reagent liquid flows through the reaction filler and stands still for a first time T1 to obtain the primary reaction liquid; in step S3, a portion of the primary reaction liquid flows through the reaction filler again in the reverse direction and stands still for a second time T2 to obtain the secondary reaction liquid.

[0015] The present invention also provides a reagent filler synthesis processing device, comprising: a liquid injection unit, a synthesis container and a liquid driving unit, wherein the liquid injection unit is connected to a storage storing a reagent liquid containing the reagent, the synthesis container is loaded with a reaction filler, and the liquid injection unit is used to add the reagent liquid into the synthesis container; the liquid driving unit is connected to the synthesis container and is used to control the flow and flow direction of the reagent liquid in the synthesis container; the reagent filler synthesis processing device performs reagent filler synthesis processing by executing the following steps:

[0016] a. adding a preset first volume of the reagent liquid into the synthesis container through the liquid injection unit;

[0017] b. allowing a portion of the preset second volume of the reagent liquid to flow through the reaction filler through the liquid driving unit to obtain a primary reaction liquid;

[0018] c. using the liquid driving unit to reversely flow a portion of the first reaction liquid having a preset third volume through the reaction filler again to obtain a second reaction liquid, and another portion of the first reaction liquid that has not reversely flowed through the reaction filler becomes a discharge liquid separated from the reaction filler, and a fourth volume of the discharge liquid is a difference between the second volume and the third volume;

[0019] d. After the liquid driving unit repeats steps b to c for multiple times, the remaining reagent liquid in the synthesis container is discharged.

[0020] In one embodiment of the present invention, the reagent filler synthesis processing equipment includes a plurality of the injection units, a plurality of the synthesis containers and a plurality of the liquid driving units, wherein the plurality of the injection units and the plurality of the synthesis containers are arranged in a one-to-one correspondence, and any of the synthesis containers is connected to one of the liquid driving units.

[0021] In one embodiment of the present invention, the liquid driving unit includes a pump body and two docking nozzles, both of which are arranged on the pump body, one of which is connected to the synthesis container, and the other is connected to the discharge liquid collection device.

[0022] In one embodiment of the present invention, the reagent filler synthesis processing equipment further includes a control mechanism for controlling the actions of the liquid injection unit and the liquid driving unit in steps a to d.

[0023] In one embodiment of the present invention, the first volume of the reaction liquid added to the synthesis container in step a is set to V1, and step b further includes: setting the second volume of the primary reaction liquid to V2, and the size of V2 is obtained by the following relationship: V2=X·V1 / n, wherein n is an integer greater than 1, and 1<X<n; step c further includes: setting the third volume that will become the secondary reaction liquid to V3, and the size of V3 is obtained by the following relationship: V3=(X-1)·V1 / n, and setting the fourth volume of the discharge liquid separated from the reaction filler to V4, then V4=V2-V3=V1 / n, and the user can set the values ​​of V1, n, and X through the control mechanism.

[0024] The above technical solution of the present invention has the following advantages compared with the prior art:

[0025] The method and device for causing a synthetic reaction between a reagent and a filler of the present invention increase the number of contacts between the reaction liquid and the filler so that the two can fully contact and react, thereby achieving complete conversion processing of the residual liquid, which not only improves the utilization rate of the reagent, thereby achieving the purpose of cost reduction and environmental protection, but also can significantly improve the purity of the product. Compared with conventional processing technology at this stage, the present application has the advantages of high controllability of the reaction process, thorough mixed conversion effect, reduced reagent waste, and improved product quality, providing a new idea for the synthetic processing of reagents and fillers. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to make the contents of the present invention more clearly understood, the present invention is further described in detail below based on specific embodiments of the present invention in conjunction with the accompanying drawings.

[0027] Figure 1 It is a schematic diagram of the structure of an existing synthesis device;

[0028] Figure 2 It is a schematic diagram of the three-dimensional structure of the reagent filler synthesis processing equipment in the preferred embodiment of the present invention;

[0029] Figure 3 yes Figure 2 A schematic diagram of the structure of the reagent filler synthesis processing equipment when the mixed reaction liquid is separated from the reaction filler;

[0030] Figure 4 yes Figure 2 The schematic diagram of the structure of the reagent filler synthesis processing equipment is shown when part of the prepared discharge liquid contacts and reacts with the reaction filler again.

[0031] Explanation of the reference numerals in the specification: 100, liquid injection unit; 200, synthesis container; 210, filler; 220, main body; 230, connecting pipe; 240, Luer connector; 300, liquid drive unit; 310, pump body; 320, docking nozzle; 400, reagent liquid. DETAILED DESCRIPTION

[0032] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0033] Embodiment 1

[0034] This embodiment provides a method for causing a reagent and a filler to undergo a synthetic reaction, which is used for the synthesis of oligonucleotides or polypeptides, etc., and includes the following steps:

[0035] Step S1, providing a reagent liquid containing the reagent and a synthesis container loaded with a reaction filler, and adding a preset first volume of the reagent liquid into the synthesis container;

[0036] Furthermore, in this embodiment, the first volume V1 of the reagent liquid can be configured to be, for example, 150 μl, which can be used to perform a synthesis reaction in a synthesis container containing a filler.

[0037] Step S2, allowing a portion of the preset second volume of the reagent liquid to flow through the reaction filler to obtain a primary reaction liquid; further, in this embodiment, the reagent liquid inside the synthesis container flows out from the bottom thereof, so that part of the reagent liquid is separated from the reaction filler to become a primary reaction liquid.

[0038] Furthermore, in this embodiment, step S2 further includes:

[0039] The second volume of the primary reaction solution is set to V2, and the size of V2 is obtained by the following relationship:

[0040] V2=X·V1 / n, wherein n is an integer greater than 1, and 1<X<n. Based on this, the second volume of the primary reaction solution can be changed by adjusting the values ​​of n and X according to actual needs.

[0041] Specifically, n in this embodiment is set to 3, and X is preferably 2, so that the reaction process can take into account both reaction efficiency and synthesis effect. In different embodiments, the specific settings of n and X can be adaptively adjusted according to actual reaction requirements, and the present invention does not make specific restrictions on this, but in order to significantly improve product purity, X is preferably set between 1.5 and 2.5. Based on this, V2 in this embodiment = 100 μl, at this time, 50 μl of reagent liquid remains in the synthesis container after step S2.

[0042] Furthermore, the discharge rate Vx of the primary reaction liquid in this embodiment is 8 to 12 μl / s. It should be noted that, in this embodiment, the discharge rate Vx is obtained by the purity change law of the product obtained at different discharge rates. Specifically, in this embodiment, the purity of the corresponding synthesized product is detected in turn under multiple conditions of uniform gradient discharge rates, and the optimal discharge rate Vx is obtained after comparison, thereby making the product processing efficiency relatively faster and the purity relatively higher. For conventional synthetic processing processes, the range of Vx is generally configured to be 5 to 100 μl / s.

[0043] Further, in order to ensure that the reaction reagent can fully contact the filler and undergo a synthesis reaction, a portion of the reagent liquid in this embodiment flows through the reaction filler and stands for a first time T1 to obtain the primary reaction liquid. In different embodiments, the value of the preset first time T1 can be adjusted accordingly according to experimental tests in combination with different chemical reactions, and the present invention does not impose specific limitations on this.

[0044] Step S3, making a part of the preset third volume of the primary reaction liquid flow through the reaction filler again in reverse to obtain a secondary reaction liquid, and another part of the primary reaction liquid that has not flowed through the reaction filler in reverse becomes a discharge liquid separated from the reaction filler, and the fourth volume of the discharge liquid is the difference between the second volume and the third volume.

[0045] Furthermore, in this embodiment, step S3 further includes: setting the third volume of the secondary reaction liquid to V3, the size of V3 is obtained by the following relationship: V3 = (X-1) · V1 / n, setting the fourth volume of the discharge liquid separated from the reaction filler to V4, then V4 = V2-V3 = V1 / n. Thus, the purpose of synchronously carrying out the residual liquid repeated reaction and the discharge liquid collection is achieved. In this article, the term residual liquid refers to the reagent liquid that has been in contact with the reaction filler and reacted.

[0046] Specifically, in this embodiment, V3 and V4 are both 50 μl. At this time, 50 μl of discharge liquid is obtained after step S3, and the liquid volume in the synthesis container is 100 μl, which is specifically composed of the remaining 50 μl reagent liquid in the synthesis container in step S2 and the secondary reaction liquid returned to the synthesis container in step S3.

[0047] Furthermore, the reflux rate Vy of the portion of the primary reaction liquid that flows through the reaction filler again in the reverse direction is 6-10 μl / s. Similarly, for conventional synthesis processing, the range of Vy is generally configured to be 5-100 μl / s.

[0048] Further, in step S3, a portion of the primary reaction liquid flows through the reaction filler again in the reverse direction and is left to stand for a second time T2 to obtain the secondary reaction liquid, thereby increasing the contact time between the reagent in the primary reaction liquid and the filler to achieve the purpose of improving the actual utilization rate. In different embodiments, the preset second time T2 can be adjusted accordingly according to the type of actual reagent and environmental parameters, and the present invention does not make specific limitations on this.

[0049] Step S4, after repeating steps S2 to S3 for multiple times, the remaining reagent liquid in the synthesis container is discharged.

[0050] According to the method of the present invention, step S2 and step S3 can be regarded as a cycle. Those skilled in the art can imagine that when the cycle consisting of step S2 and step S3 is repeated multiple times, according to different set values ​​of n and X, part of the reagent liquid may contact and react with the filler more than twice. Therefore, the terms "primary reaction liquid" and "secondary reaction liquid" herein mean that within one cycle, part of the reagent liquid contacts and reacts with the filler for the first time, and contacts and reacts with the filler for the second time, and does not limit the actual number of contacts and reactions across multiple cycles.

[0051] Furthermore, in this embodiment, the number of repetitions of step S2 to step S3 depends on the values ​​of n and X, is positively correlated with the value of n, and is negatively correlated with the value of X. The specific parameter values ​​can be freely set according to actual processing requirements.

[0052] Specifically, since the monomer reagents used in the coupling stage of chemical synthesis are expensive, when used in the coupling stage, the number of repetitions (cycles) of step S2 to step S3 can be increased by appropriately increasing n and decreasing X, thereby improving the utilization rate of the monomer reagents and reducing the experimental cost; while the price of reagents such as acetonitrile required for the deprotection, oxidation, capping and other stages is relatively low, and the number of repetitions (cycles) of step S2 to step S3 can be reduced by appropriately decreasing n and increasing X, and even no cycle is required.

[0053] Furthermore, in this embodiment, in step S4, an upper limit may be set for the number of repetitions of step S2 to step S3, and the number of repetitions of step S2 to step S3 is at most an integer obtained by rounding down the calculation result of (n-X+1):

[0054] In the present embodiment, after repeating step S2~step S3 for 3 times, the reagent liquid of 50 μ l remaining in the described synthesis container is discharged completely, so far completes the processing of a complete reagent and filler building-up reaction.It can be seen from this that after repeating step S2~step S3 for 3 times, the reagent liquid of 50 μ l remaining in the synthesis container is less than the second volume V2-100 μ l in step 2 at this moment, if step 2 to step 3 is carried out again at this moment, the reagent liquid in the synthesis container will be emptied completely in step 2, and filler is still exposed to air after step 3, and this is undesirable, because this is an invalid circulation step.For this reason, as mentioned above, can be set upper limit for step S2~step S3 repetition number.In the present embodiment, because n is 3 and X is 2, the repetition number of step S2~step S3 is at most the result of calculation of (n-X+1) rounded down obtained integer=3.

[0055] Furthermore, the method disclosed in this embodiment for causing a synthetic reaction between a reagent and a filler can be applied, for example, to nucleic acid synthesis processing, and can be specifically used in the various stages of coupling, deprotection, oxidation, and capping of nucleic acid synthesis, and is particularly suitable for the coupling stage of nucleic acid synthesis. Different types of reagents are used in the coupling, deprotection, oxidation, and capping stages of nucleic acid synthesis, respectively, because the ATCG monomer reagent required in the coupling stage is expensive, and the price of reagents such as acetonitrile required for the deprotection, oxidation, and capping stages is relatively low.

[0056] Furthermore, based on the above process, this embodiment can improve the efficiency of nucleic acid synthesis and processing, greatly reduce reagent loss and processing costs, and has practical significance for nucleic acid synthesis and processing. In addition, the above process can be applied to expensive chemical reagent synthesis reactions, and it can also be applied to deprotection, oxidation, capping and other stages, thereby achieving the purpose of reducing costs and increasing efficiency.

[0057] Embodiment 2

[0058] See also Figure 2 As shown, this embodiment provides a reagent filler synthesis reaction device, which includes a liquid injection unit 100, a synthesis container 200 and a liquid driving unit 300, wherein the liquid injection unit 100 is connected to a storage device storing a reagent liquid containing the reagent, and is used to inject a reagent liquid 400 into the synthesis container 200, and the liquid injection unit 100 can be connected to a reagent container storing a reagent required for the synthesis reaction; the synthesis container 200 is loaded with a reaction filler 210, and a synthesis reaction space is provided for the filler 210 and the reagent liquid 400, and the filler 210 is arranged on the synthesis container 200. The body 220 of the synthesis container 200 is inside; the liquid driving unit 300 is connected with the synthesis container 200, and is used to control the flow and flow direction of the reagent liquid in the synthesis container 200 to achieve a circulating reaction or discharge of the liquid. The liquid driving unit 300 includes a pump body 310 and two docking nozzles 320, and the docking nozzles 320 are all arranged on the pump body 310, one of the docking nozzles 320 is connected with the outlet end of the synthesis container 200, and the other docking nozzle 320 is connected with a discharge liquid collecting device not shown in the figure.

[0059] The specific steps for carrying out the reagent filler 210 synthesis reaction process using the above reagent filler synthesis reaction equipment are as follows:

[0060] Step a: adding a preset first volume of the reagent liquid 400 into the synthesis container 200 through the liquid injection unit 100 .

[0061] Further, see Figure 3 and Figure 4As shown, in this embodiment, the reagent filler 210 synthesis processing equipment may include a plurality of the liquid injection units 100, a plurality of the synthesis containers 200 and a plurality of the liquid driving units 300, wherein the plurality of the liquid injection units 100 and the plurality of the synthesis containers 200 are arranged in a one-to-one correspondence, and any of the synthesis containers 200 is connected to one of the liquid driving units 300, thereby multiple synthesis reactions can be carried out simultaneously to multiply the synthesis processing efficiency of the reagent filler 210.

[0062] Furthermore, the synthesis container 200 in this embodiment is configured as a cylindrical structure. In different implementations, its specific shape can be adjusted according to actual usage requirements. At the same time, in addition to the injection unit 100, the present application can also use other devices to add the reagents required for the synthesis reaction from the reagent container to the synthesis container 200, and the present invention does not impose specific limitations on this.

[0063] Specifically, in this embodiment, the first volume V1 of the reagent liquid 400 added to the corresponding synthesis container by any injection unit 100 is configured to be, for example, 150 μl, that is, 150 μl of the reagent liquid 400 enters the corresponding synthesis container 200 through multiple injection units 100 .

[0064] Step b: using the liquid driving unit 300 to allow a portion of the preset second volume of the reagent liquid 400 to flow through the reaction filler 210 to obtain a primary reaction liquid.

[0065] Furthermore, the body 220 of the synthesis container 200 in this embodiment is connected to the docking nozzle 320 via a connecting pipe 230 , and a Luer connector 240 is provided on the body 220 to connect the outlet end of the synthesis container 200 to the connecting pipe 230 .

[0066] In other embodiments, in addition to the liquid driving unit 300, other types of pumps or double-acting cylinders may be used to control the flow and flow direction of the reagent liquid 400, and the present invention does not impose any specific limitation on this.

[0067] In this embodiment, the volume of the primary reaction liquid is set to V2, wherein the size of V2 is obtained by the following relationship:

[0068] V2=X·V1 / n, wherein n is an integer greater than 1, and 1<X<n. Based on this, the second volume of the primary reaction solution can be changed by adjusting the values ​​of n and X according to actual needs.

[0069] Specifically, n in this embodiment is set to 3, and X is preferably 2, so that the reaction process can take into account both reaction efficiency and synthesis effect. In different embodiments, the specific settings of n and X can be adaptively adjusted according to actual reaction requirements, and the present invention does not make specific restrictions on this, but in order to significantly improve product purity, X is preferably set between 1.5 and 2.5. Based on this, V2 in this embodiment = 100 μl, at this time, 50 μl of reagent liquid 400 remains in the synthesis container 200.

[0070] Furthermore, the discharge rate Vx of the primary reaction liquid in this embodiment is 8 to 12 μl / s. It should be noted that, in this embodiment, the discharge rate Vx is obtained by the purity change law of the product obtained at different discharge rates. Specifically, in this embodiment, the purity of the corresponding synthesized product is detected in turn under multiple conditions of uniform gradient discharge rates, and the optimal discharge rate Vx is obtained after comparison, thereby making the product processing efficiency relatively faster and the purity relatively higher. For conventional synthetic processing processes, the range of Vx is generally configured to be 5 to 100 μl / s.

[0071] Further, in order to ensure that the reaction reagent can fully contact the filler 210 and undergo a synthesis reaction, a portion of the reagent liquid 400 in this embodiment flows through the reaction filler 210 and stands for a first time T1 to obtain the primary reaction liquid. In different embodiments, the value of the preset first time T1 can be adjusted accordingly according to experimental tests in combination with different chemical reactions, and the present invention does not impose specific limitations on this.

[0072] Step c, using the liquid driving unit 300 to reversely flow a portion of the first reaction liquid of the preset third volume through the reaction filler 210 in the synthesis container 200 again to obtain a second reaction liquid, and at the same time, using the liquid driving unit 300 to reversely flow another portion of the first reaction liquid that has not reversely flowed through the reaction filler 210 to become a discharge liquid separated from the reaction filler 210, and a fourth volume of the discharge liquid is the difference between the second volume and the third volume;

[0073] Furthermore, in this embodiment, step c further includes: setting the third volume of the secondary reaction liquid to V3, the size of V3 is obtained by the following relationship: V3 = (X-1) V1 / n, setting the fourth volume of the discharge liquid separated from the reaction filler 210 to V4, then V4 = V2-V3 = V1 / n. Thus, the purpose of performing the repeated reaction of the residual liquid and the collection of the product simultaneously is achieved.

[0074] Specifically, in this embodiment, V3 and V4 are both 50 μl. At this time, 50 μl of discharge liquid is obtained after step c, and the liquid volume in the synthesis container 200 is 100 μl, which is specifically composed of the remaining 50 μl reagent liquid 400 in the synthesis container 200 in step b and the secondary reaction liquid returned to the synthesis container 200 in step c.

[0075] Furthermore, the secondary reaction liquid flows through the liquid driving unit again through the reaction filler 210 in the synthesis container 300 at a reflux rate Vy of 6-10 μl / s. Similarly, for conventional synthesis processes, the range of Vy is generally configured to be 5-100 μl / s.

[0076] Further, in step c, a portion of the primary reaction liquid flows through the reaction filler 210 again in the reverse direction and is allowed to stand for a time T2 to obtain the secondary reaction liquid, thereby increasing the contact time between the reagent in the primary reaction liquid and the filler 210, so as to achieve the purpose of improving the actual utilization rate. In different embodiments, the preset second time T2 is adjusted accordingly according to the type of actual reagent and environmental parameters, and the present invention does not make specific limitations on this.

[0077] Step d: After the liquid driving unit 300 repeats steps b to c for multiple times, the remaining reagent liquid 400 in the synthesis container 200 is discharged.

[0078] Furthermore, in this embodiment, the number of repetitions of step S2 to step S3 depends on the values ​​of n and X, is positively correlated with the value of n, and is negatively correlated with the value of X. The body parameters can be set according to actual processing requirements.

[0079] Specifically, since the monomer reagent used in the coupling stage is expensive, when used in the coupling stage, the number of repetitions (cycles) of step S2 to step S3 can be increased by appropriately increasing n and reducing X, thereby improving the utilization rate of the monomer reagent and reducing the experimental cost; while the price of reagents such as acetonitrile required for the deprotection, oxidation, capping and other stages is relatively low, and the number of repetitions (cycles) of step S2 to step S3 can be reduced by appropriately reducing n and increasing X, and even no circulation is required and the reagents can be directly discharged through a peristaltic pump.

[0080] Furthermore, in this embodiment, in step d, an upper limit may be set for the number of repetitions of step b to step c, and the number of repetitions of step b to step c is at most an integer obtained by rounding down the calculation result of (n-X+1):

[0081] In this embodiment, after repeating step b to step c three times, the remaining 50 μl of the reagent liquid 400 in the synthesis container 200 is completely discharged, thus completing a complete processing process of the synthesis reaction of the reagent and the filler 210.

[0082] Furthermore, in this embodiment, the reagent filler 210 synthesis processing equipment may also include a control mechanism for controlling the actions of the injection unit 100 and the liquid driving unit 300 in the steps a to d. Specifically, the action of the injection unit 100 adding reagent liquid into the synthesis container 200 in step a, the action of the liquid driving unit 300 allowing a portion of the reagent liquid 400 to flow through the reaction filler 210 in step b, the action of the liquid driving unit 300 allowing a portion of the primary reaction liquid to flow through the reaction filler 210 again in reverse in step c, and the action of the liquid driving unit 300 repeating steps b to c multiple times and discharging the remaining reagent liquid 400 in the synthesis container 200 in step d are all performed under the control of the control mechanism.

[0083] Furthermore, the control mechanism stores a program for executing the above-mentioned reagent filler 210 synthesis reaction method, and the injection unit 100, the synthesis container 200 and the liquid drive unit 300 are respectively connected to the control mechanism. In the actual production and processing process, the operator can control the above-mentioned structure in real time through the control mechanism, thereby improving the flexibility of the use of the equipment, and the parameters can also be preset through the control mechanism, thereby improving the degree of automation of the equipment.

[0084] Furthermore, the user can set the values ​​of V1, n, X, etc. through the control mechanism. The specific preset parameters of the control mechanism include n and X that limit the number of cycles, a first speed Vx at which a portion of the reagent liquid flows through the reaction filler, a second speed Vy at which a portion of the primary reaction liquid flows through the reaction filler 210 again in the opposite direction, a first time T1 at which a portion of the reagent liquid flows through the reaction filler 210 and remains still, a second time T2 at which a portion of the primary reaction liquid flows through the reaction filler 210 again in the opposite direction, and a first volume V1 of the reaction liquid added to the synthesis container 200, etc.

[0085] In summary, the present invention achieves complete conversion of the residual liquid by enhancing the contact between the reaction liquid and the filler 210 to ensure sufficient interaction between the two. This technology not only significantly improves the use efficiency of the reagents and achieves the purpose of cost saving, but also effectively improves the purity of the final product.

[0086] Obviously, the above embodiments are merely examples for clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the protection scope of the invention.

Claims

1. A method for causing a reagent and a filler to undergo a synthetic reaction, characterized in that: The steps include: Step S1, providing a reagent liquid containing the reagent and a synthesis container loaded with a reaction filler, and adding a preset first volume of the reagent liquid into the synthesis container; Step S2, allowing a portion of the preset second volume of the reagent liquid to flow through the reaction filler to obtain a primary reaction liquid; Step S3, making a part of the first reaction liquid with a preset third volume flow through the reaction filler again in reverse to obtain a second reaction liquid, and another part of the first reaction liquid that does not flow through the reaction filler in reverse becomes a discharge liquid separated from the reaction filler, and a fourth volume of the discharge liquid is a difference between the second volume and the third volume; Step S4, after repeating steps S2 to S3 for multiple times, the remaining reagent liquid in the synthesis container is discharged.

2. The method for causing a reagent and a filler to undergo a synthetic reaction according to claim 1, wherein: The first volume of the reaction liquid added to the synthesis container in step S1 is set to V1, Step S2 further includes: setting the second volume of the primary reaction liquid to V2, where the value of V2 is obtained by the following relationship: V2=X·V1 / n, wherein n is an integer greater than 1, and 1<X<n.

3. The method for causing a reagent and a filler to undergo a synthetic reaction according to claim 2, wherein: Step S3 further comprises: The third volume of the secondary reaction liquid is set to V3, and the size of V3 is obtained by the following relationship: V3=(X-1)·V1 / n, Assuming that the fourth volume of the discharge liquid leaving the reaction packing is V4, V4=V2-V3=V1 / n.

4. The method for causing a reagent and a filler to undergo a synthetic reaction according to claim 3, wherein: The number of repetitions of step S2 to step S3 in step S4 depends on the values ​​of n and X, and is positively correlated with the value of n and negatively correlated with the value of X.

5. The method for causing a reagent and a filler to undergo a synthetic reaction according to claim 3, wherein: In step S4, the number of repetitions of steps S2 to S3 is at most an integer obtained by rounding down the calculation result of (n-X+1).

6. The method for causing a reagent and a filler to undergo a synthetic reaction according to claim 1, wherein: In step S2, a portion of the reagent liquid flows through the reaction filler at a first speed Vx ranging from 5 to 100 μl / s; in step S3, a portion of the primary reaction liquid flows through the reaction filler again in the reverse direction at a second speed Vy ranging from 5 to 100 μl / s.

7. The method for causing a reagent and a filler to undergo a synthetic reaction according to claim 1, wherein: In step S2, a portion of the reagent liquid flows through the reaction filler and stands for a first time T1 to obtain the primary reaction liquid; in step S3, a portion of the primary reaction liquid flows through the reaction filler again in the reverse direction and stands for a second time T2 to obtain the secondary reaction liquid.

8. A reagent filler synthesis processing equipment, characterized in that: include: A liquid injection unit, a synthesis container and a liquid drive unit, wherein: The injection unit is connected to a storage device storing a reagent liquid containing the reagent, the synthesis container is loaded with a reactive filler, and the injection unit is used to add the reagent liquid into the synthesis container; The liquid driving unit is in communication with the synthesis container and is used to control the flow and flow direction of the reagent liquid in the synthesis container; The reagent filler synthesis processing equipment performs reagent filler synthesis processing by executing the following steps: Step a, adding a preset first volume of the reagent liquid into the synthesis container through the liquid injection unit; Step b, allowing a portion of the preset second volume of the reagent liquid to flow through the reaction filler through the liquid driving unit to obtain a primary reaction liquid; Step c, using the liquid driving unit to reversely flow a portion of the first reaction liquid with a preset third volume through the reaction filler again to obtain a second reaction liquid, and another portion of the first reaction liquid that has not reversely flowed through the reaction filler becomes a discharge liquid separated from the reaction filler, and a fourth volume of the discharge liquid is a difference between the second volume and the third volume; Step d: After the liquid driving unit repeats steps b to c for multiple times, the remaining reagent liquid in the synthesis container is discharged.

9. The reagent filler synthesis processing equipment according to claim 8, characterized in that: The reagent filler synthesis processing equipment comprises a plurality of the injection units, a plurality of the synthesis containers and a plurality of the liquid driving units, wherein the plurality of the injection units and the plurality of the synthesis containers are arranged in a one-to-one correspondence, and any of the synthesis containers is connected to one of the liquid driving units.

10. The reagent filler synthesis processing equipment according to claim 8, characterized in that: The liquid driving unit comprises a pump body and two docking nozzles, both of which are arranged on the pump body, one of which is connected to the synthesis container, and the other is connected to the discharge liquid collecting device.

11. The reagent filler synthesis processing equipment according to claim 8, characterized in that: The reagent filler synthesis processing equipment also includes a control mechanism for controlling the actions of the liquid injection unit and the liquid driving unit in steps a to d.

12. The reagent filler synthesis processing equipment according to claim 11, characterized in that: The first volume of the reaction liquid added to the synthesis container in step a is set to V1, Step b further comprises: setting the second volume of the primary reaction liquid to V2, wherein the size of V2 is obtained by the following relationship: V2=X·V1 / n, wherein n is an integer greater than 1, and 1<X<n; Step c further includes: setting the third volume of the secondary reaction liquid to V3, the size of V3 is obtained by the following relationship: V3 = (X-1) · V1 / n, setting the fourth volume of the discharge liquid separated from the reaction filler to V4, then V4 = V2-V3 = V1 / n, The user can set the values ​​of V1, n, and X through the control mechanism.

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