Device for preparing in-situ synthesized peptide microarray and liquid supply system therefor
Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Filing Date
- 2025-12-31
- Publication Date
- 2026-08-13
Smart Images
Figure CN2025148309_13082026_PF_FP_ABST
Abstract
Description
An apparatus for preparing in-situ synthesized peptide chips and its liquid supply system
[0001] Priority information
[0002] This application claims priority to Chinese Patent Application No. 202510134504.0, filed on February 6, 2025, entitled "An apparatus for preparing in situ synthetic peptide chips and its liquid supply system", the entire contents of which are incorporated herein by reference. Technical Field
[0003] This invention relates to the field of in-situ synthetic peptide chip technology, and in particular to an apparatus for preparing in-situ synthetic peptide chips and its liquid supply system. Background Technology
[0004] Peptide microarrays are powerful bioinformatics analysis platforms and tools that can assist in disease diagnosis, new drug development, and biomarker screening. This is because a large number of functional peptide probe arrays are immobilized on the surface of peptide microarrays in a patterned manner.
[0005] The fabrication methods for peptide chips mainly fall into two categories: spotting and in-situ synthesis. With the continuous development of fabrication processes, peptide chips prepared by in-situ synthesis technology have the advantages of high throughput, high density, and small feature size.
[0006] Typical in-situ peptide synthesis methods are mainly divided into two types. One method uses inkjet printing technology to directly print monomers and other necessary reaction reagents onto the substrate surface. The other method is photoconductive patterning, such as using a semiconductor photolithography mask to precisely deprotect the N-terminal protecting groups of amino acid sequences at specific locations on the substrate, followed by coupling to extend the peptide chain, thereby achieving in-situ synthesis, such as in the fabrication of high-throughput peptide chips for health monitoring. In the photolithography method, the reagents for peptide chain extension are supplied, mixed, and pre-activated by the equipment's liquid supply system.
[0007] Photolithography can be used to pattern and deprotect peptide chips, allowing for the in-situ synthesis of different amino acids onto a substrate to create peptide chips containing thousands of different sequences. In this process, effectively transporting the amino acid monomer solution with protecting groups to the chip surface shares similarities with the liquid supply system of a peptide synthesizer, but also has its own unique characteristics. Peptides prepared by solid-phase peptide synthesizers often require purification to obtain high-purity target peptides. In-situ synthesized peptides cannot be purified, yet high purity is desired, thus requiring extremely high precision in the liquid supply, mixing, and pre-activation processes. Summary of the Invention
[0008] In the in-situ synthesis of peptide chips, a large number of peptide sequences are synthesized and directly immobilized on the substrate surface, making purification impossible. Therefore, the process is particularly sensitive to liquid path contamination, requiring more efficient cleaning steps to address this challenge.
[0009] This application aims to provide a new solution for in situ peptide synthesis technology that avoids liquid path contamination and eliminates the need for purification.
[0010] One aspect of this application discloses a liquid supply system for an in-situ peptide synthesis chip device, comprising: an amino acid coupling module for coupling amino acids to a target site, the amino acid coupling module including a first synthesis unit; an amino acid liquid supply module for supplying the amino acid liquid and coupling agent required by the synthesis unit to the amino acid coupling module, comprising: multiple amino acid liquid supply units, each amino acid liquid supply unit including an independent amino acid container and a corresponding output pipe and control valve, each amino acid container storing an amino acid monomer; the multiple amino acid liquid supply units supplying the corresponding amino acid to the first synthesis unit; and a coupling agent liquid supply unit including an independent... The system includes: a coupling agent container and an output pipeline; a cleaning agent supply unit, including an independent cleaning agent container, an output pipeline, and a control valve; a first manifold valve group, corresponding to the first synthesis unit; the first manifold valve group includes multiple input ports and one output port, the multiple input ports of the first manifold valve group being connected to the output pipelines of multiple amino acid supply units, coupling agent supply units, and cleaning agent supply units; a first mixing unit, corresponding to the first synthesis unit, connected to the output port of the first manifold valve group, used to mix the selected amino acid monomers and coupling agents in the first manifold valve group; the output port of the first mixing unit is connected to the first synthesis unit of the chip via a control valve.
[0011] Another aspect of this application provides an apparatus for preparing an in-situ synthesized peptide chip, comprising a liquid supply system according to any embodiment, and an exposure module for transferring deprotected sites onto the chip in a patterned manner using a mask.
[0012] By converting the independent supply of amino acid monomers into a single mixing module supplying several or all amino acid monomers, investment in mixing modules and heated spin coaters can be saved. Due to the risk of cross-contamination, cleaning is necessary. The mixing module is a common component, made of smooth glass and placed vertically. A spray nozzle is added to the top of the volumetric tube for easy cleaning. This solution is simple in structure, low in cost, quick to set up, and cost-effective. During the R&D phase, a direct-flow supply system can be quickly built to supply a single or several heated spin coaters, enabling process development or process optimization during pilot production. This solution can also be divided into multi-channel synthesis units using valve assemblies or selector valves. Valve assemblies prevent unused reagents from entering the process, while rotary selector valves simplify maintenance and allow for better liquid path layout. Attached Figure Description
[0013] The performance and advantages of the invention can be further understood by referring to the remainder of this specification and the accompanying drawings, in which the same reference numerals are used for the same component. In some cases, a sub-label is placed after a label followed by a hyphen to indicate one of many similar components. When a label is mentioned without specifically naming an existing sub-label, it refers to all of these similar components.
[0014] Figure 1A is a flowchart of a photolithography-based synthesis of polypeptide chips according to an embodiment of this application.
[0015] Figure 1B is a flowchart of the photolithography method for synthesizing polypeptide chips according to another embodiment of this application.
[0016] Figure 2A is a block diagram of the polypeptide chip in-situ synthesis system of the embodiment shown in Figure 1A.
[0017] Figure 2B is a block diagram of the polypeptide chip in-situ synthesis system of the embodiment shown in Figure 1B.
[0018] Figure 3 shows a liquid circuit scheme for an in-situ polypeptide chip synthesis system.
[0019] Figure 4 is a schematic diagram of the fluid path of the manifold section in the embodiment shown in Figure 3.
[0020] Figure 5 is a schematic diagram of a liquid circuit structure for the liquid circuit scheme shown in Figure 3; Embodiments of the present invention
[0021] The embodiments are described in more detail below with reference to the examples provided below. These examples are provided by way of illustration only and are not intended to be limiting.
[0022] This invention has many variations that can be anticipated by those skilled in the art, and can achieve the effects of this invention.
[0023] The use of terms to define space or location in this application, such as inside, outside, top, bottom, left, right, top, bottom, front, and back, is based on the placement of the object in its usage state.
[0024] In this application, the term "in-situ synthesis" refers to the process of directly synthesizing a polypeptide sequence at the desired application site on a matrix or substrate. In some exemplary embodiments, a linker with protecting groups is deposited on the substrate surface, which can be removed, followed by the addition of an amino acid layer, and polypeptide synthesis is achieved by chemically coupling amino acid monomers. In some exemplary embodiments, a specially designed photolithographic mask is used to protect the sites that do not need to be synthesized, while exposing the sites that need to be synthesized. Photoresist, under light, generates a deprotected substance, removing the protecting groups on amino groups, and the amino groups condense with the carboxyl groups of another amino acid molecule to synthesize the polypeptide.
[0025] In this application, the term "matrix or substrate" refers to a carrier in the process of biopeptide synthesis that has various chemical groups and can immobilize biomolecules such as proteins, antibodies, enzymes, peptides, DNA, etc.
[0026] In this application, the term "linker molecule" refers to a linker molecule laid on a treated glass slide in in situ synthesis, and amino acids can be synthesized into peptides based on linker molecules.
[0027] In this application, the term "protecting group" is used to protect the active side chain of amino acids. The protection of amino acids in peptide chemical synthesis is crucial and directly determines the success of the synthesis. These protecting groups are stable during the synthesis process, have no side reactions, and can be completely and quantitatively removed after the synthesis is completed.
[0028] In this application, the term "BOC protecting group" refers to tert-butyloxycarbonyl, an amino protecting group used in organic synthesis, particularly in peptide synthesis.
[0029] In this application, the term "chemical modification" refers to the process of attaching active groups or catalytic substances to the electrode surface by means of adsorption, coating, polymerization, chemical reaction, etc., to protect the electrode or improve the electrode's characteristic functions.
[0030] In this application, the term "activation" refers to the activation reaction. In protein biosynthesis, various proteins must be activated before being incorporated into the peptide chain. Then, their specific tRNAs carry them to the ribosomes, where they can condense into peptide chains using mRNA as a template.
[0031] In this application, the term "valve assembly" or "multi-channel distribution valve assembly" refers to a valve having multiple channels, including a one-inlet-multiple-outlet valve assembly and a multi-inlet-one-outlet valve assembly.
[0032] In this application, the terms "rotary selector valve" and "multi-channel distribution valve assembly" include multiple channels, typically used to divert fluid from a single source to multiple destinations; or to converge multiple flows into a single flow.
[0033] In this application, the term "manifold" refers to a valve assembly that brings together fluid from multiple channels to a single outlet.
[0034] In this application, the term "comprising" means including, but not limited to, the following elements, that is, it does not exclude other elements.
[0035] In this application, the terms "about" and "approximately" represent a range of accuracy that a person skilled in the art would understand while still ensuring the accuracy of the technical effects of the features in question. This term typically indicates a deviation of 10% from the numerical values shown, preferably 5%.
[0036] To address the challenges of liquid supply systems in in-situ peptide synthesis systems, this application proposes a liquid supply system for an in-situ peptide synthesis chip device, comprising: an amino acid coupling module for coupling amino acids to a target location, the amino acid coupling module including a first synthesis unit; and an amino acid supply module for supplying the amino acid liquid and coupling agent required by the synthesis unit to the amino acid coupling module, comprising: multiple amino acid supply units, each amino acid supply unit including an independent amino acid container and a corresponding output pipe and control valve, each amino acid container storing one amino acid monomer; the multiple amino acid supply units supplying the corresponding amino acid to the first synthesis unit; and coupling... The coupling agent supply unit includes an independent coupling agent container and an output pipeline; the cleaning agent supply unit includes an independent cleaning agent container, an output pipeline, and a control valve; a first manifold valve group corresponds to the first synthesis unit; the first manifold valve group includes multiple input ports and one output port, and the multiple input ports of the first manifold valve group are connected to the output pipelines of multiple amino acid supply units, coupling agent supply units, and cleaning agent supply units; a first mixing unit corresponds to the first synthesis unit and is connected to the output port of the first manifold valve group for mixing the selected amino acid monomers and coupling agents in the first manifold valve group; the output port of the first mixing unit is connected to the first synthesis unit of the chip through a control valve.
[0037] In at least one embodiment, the liquid supply system further includes a nitrogen supply unit for filling the connected container or pipeline with nitrogen gas, causing the liquid in the container or pipeline to be output along the pipeline. In at least one embodiment, the nitrogen supply unit is connected to the input port of the first to Nth mixing units. In at least one embodiment, the nitrogen supply unit is connected to the input port of the amino acid liquid supply unit. In at least one embodiment, the nitrogen supply unit is connected to the input port of the first to Nth synthesis units.
[0038] In at least one embodiment, the amino acid coupling module further includes second to Nth synthesis units, and the amino acid supply module further includes second to Nth manifold valve groups and second to Nth mixing units connected one-to-one with the second to Nth synthesis units; each amino acid supply unit's output pipe is provided with a multi-channel distribution valve group, so that the amino acid monomers of each amino acid supply unit are distributed to different branches, and each branch corresponds to one of the first to Nth manifold valve groups; the number of channels in the multi-channel distribution valve group corresponds to the number of synthesis units.
[0039] In at least one embodiment, the cleaning agent supply unit includes a first output branch pipe and a second output branch pipe;
[0040] The first output branch pipe is configured with multiple branches, the number of which corresponds to the number of synthesis units; the branches of the first output branch pipe are fluidly connected to the corresponding manifold valve group.
[0041] The second output branch pipe is also configured with multiple branches, and the branches of the second output branch pipe are connected to the corresponding mixing unit.
[0042] In at least one embodiment, the amino acid supply module further includes a coupling agent / excipient supply unit, which includes an output pipe connected to the input port of the manifold valve assembly.
[0043] In at least one embodiment, the liquid supply system further includes a waste liquid collection unit for collecting waste liquid in the pipeline after cleaning the liquid path.
[0044] In at least one embodiment, the amino acid supply unit is positioned horizontally in the vertical direction lower than the position of the mixing unit, so that after the amino acid supply unit completes the supply of liquid to the corresponding mixing unit, the liquid is returned by gravity.
[0045] In at least one embodiment, the manifold valve assembly is also connected to a waste liquid collection unit.
[0046] Another aspect of this application provides an apparatus for preparing in-situ synthesized peptide chips, comprising a liquid supply system according to any embodiment, and an exposure module for transferring deprotection sites onto the chip in a patterned manner using a mask. In one embodiment, the apparatus further includes a photoresist spin-coating and pre-baking module for photoresist spin-coating and pre-baking; and a heating amplification deprotection module for amplifying the protection of the N-terminus of acid / alkali-generating deprotection sites on the chip surface after exposure excitation.
[0047] In-situ synthesis of peptide chips using photolithography involves the parallel chemical synthesis of peptide probes via photoconductive mapping on a substrate surface. A mask is used to pattern the peptide probes. Based on the deprotection method of the N-terminal protecting group of amino acid monomers, two types are used: one where the N-terminal protecting group is removed with acid or alkali, requiring photoresist (as shown in Figure 1A); and another where the N-terminal protecting group is photosensitive, allowing direct deprotection by light of a specific wavelength (as shown in Figure 1B). These two methods also determine whether spin-coating with photoresist is necessary. Probe synthesis follows a cycle of "deprotection-washing-coupling-washing" in solid-phase peptide synthesis.
[0048] This application addresses the characteristics of in-situ synthesis of peptide chips using photolithography, combining semiconductor industry technology with solid-state synthesis technology. In at least one embodiment, an automated system for in-situ synthesis of peptide chips using photolithography is provided, as shown in Figure 2A. The system shown in Figure 2A consists of five subsystems: a photoresist spin coating and pre-baking module 300, an exposure module 400, a heating amplification module 500, a heating spin coating amino acid coupling module 200, and an amino acid supply module 100. The photoresist spin coating and pre-baking module 300 is responsible for photoresist spin coating and pre-baking, and consists of several spin coaters and hot plates. The exposure module 400 uses a mask to transfer the deprotected sites onto the chip in a patterned form, mainly consisting of a photolithography machine. The heating amplification module 500 further amplifies the sites on the chip surface excited by exposure to produce acid / alkali and remove the protection at the N-terminus of the sites, and consists of hot plates. The heating spin coating amino acid coupling module 200 cleans the photoresist and removed protective groups from the chip surface, and performs amino acid coupling and post-coupling cleaning, and consists of several heating spin coaters. The amino acid supply module 100 is responsible for the delivery and mixing of amino acid coupling reagents and is a supply system.
[0049] Compared to the system shown in Figure 2A, the system shown in Figure 2B only includes an amino acid supply module 100, an exposure module 400, and a heated spin-coating amino acid coupling module 200. It does not require the participation of a photoresist spin-coating and post-baking module or a heated amplification and deprotection module. The working process of the heated spin-coating amino acid coupling module 200 and the amino acid supply module 100 is consistent with the working process of the corresponding modules in the system shown in Figure 2A. The amino acid monomer independent supply liquid circuit system also follows the one-to-one correspondence of AAn-Hn.
[0050] Figure 3 is a schematic diagram of the liquid circuit in another embodiment. In the figure, the cleaning agent supply unit 102 outputs cleaning agent R1 through two output pipes. The cleaning agent liquid in the first output branch pipe is connected to the manifold valve group 301 for cleaning the manifold valve group 301 and its liquid circuit. The cleaning agent in the second output branch pipe of the cleaning agent supply unit 102 flows to the mixing unit 104 for cleaning the mixing unit 104 and its liquid circuit. At the same time, the coupling agent output from the coupling agent supply unit 101 also enters the mixing unit 104 after passing through the distribution valve group 302. In the mixing unit 104, the amino acid monomer and the coupling agent are mixed and activated before being sent to the synthesis unit 201. The waste liquid after the liquid circuit cleaning is collected by the waste liquid collection unit 202.
[0051] As shown in the embodiment in Figure 4, the amino acids are split into multiple channels and then converged into a common pipeline of a multi-inlet / one-outlet manifold valve group or rotary selector valve. This multi-channel reuse improves efficiency and allows for the holding of more amino acids. The valve group has multi-stage valves, reducing the risk of reagents entering the mixing unit unused in the process. The rotary selector valve is smaller, reducing the risk of cross-contamination. A backflow valve is added before the amino acid enters the manifold valve group connector. When the amino acid supply ends, the solution is returned to the backflow valve, providing a buffer between the amino acid monomer solution and the manifold valve group, reducing the risk of contamination. The manifold valve group is equipped with a bypass, and the common flow channel of the valve group can be cleaned online, reducing the risk of cross-contamination in the common pipeline of the manifold valve group. It also enables parallel synthesis of multi-channel units, allowing for verification of batch-to-batch and batch-to-batch differences when multiple substrate batches are processed during the R&D phase, achieving process optimization and scale-up. It can also serve as a platform for small-batch peptide chip synthesis, used for developing post-processing techniques and application research.
[0052] Figure 5 shows one embodiment, which adopts the liquid path scheme shown in Figure 3, illustrated by a valve assembly. The liquid path after the amino acid monomer reagent enters the valve assembly is shown in Figure 4. Twenty natural amino acid monomers are delivered, with R1 being DMF, R2 being HATU, and R3 being DIEA. The liquid path is constructed according to Figure 5. The in-situ synthetic peptide chip automated system is set up with 8 synthesis units. After the module construction and debugging are completed, the automated program is set up to execute small-batch synthesis of the peptide chip for application research, using 8 pre-treated substrates for 60 synthesis cycles.
[0053] In Figure 5, V1-V3 are the control valves for R1-R3. R1-R3 are connected to the nitrogen control unit, which controls the liquid extrusion rate. R1 is divided into two branch pipes. The first output branch pipe's R1 is controlled by the V4 control valve and is accompanied by nitrogen extrusion. Then, the first output branch pipe's R1 is divided into multiple branches corresponding to the number of synthesis units. Each branch is connected to the manifold valve group VB. 01 -VB m VB manifold 01 -VB mThe quantity corresponds one-to-one with the synthesis unit; R1 of the second output branch pipe is V012-V m2 Controlled by multiple branch control valves, the amino acid monomer AA is divided into multiple branches, with each branch corresponding to the number of synthesis units. Each branch is then connected to a manifold valve group VB. 01 -VB m The branch control valves for R2 are V014-Vm4; the branch control valves for R3 are V015-Vm5; AV01-AVm are volumetric valves corresponding to the number of synthesis units; MV01-MVn are mixing units corresponding to the synthesis units; the manifold valve group VB 01 -VB m The corresponding output branches are connected to the corresponding branches of the second path R1, R2, and R3, respectively, to the corresponding volumetric liquid dispensers AV01-AVm; V016-Vm6 are the output control valves of volumetric liquid dispensers AV01-AVm; V017-Vm7 are the nitrogen output control valves at volumetric liquid dispensers AV01-AVm; V019-Vm9 are the waste liquid collection unit control valves at mixing units MV01-MVn; V018-Vm8 are the output control valves of mixing units V019-Vm9, accompanying nitrogen extrusion supply. Sm01-Smn are back suction valves; Vm01-Vmn are the manifold valve group VB. m The control valve is located in the middle. A single cycle process is as follows:
[0054] 1. Spin-coating photoresist onto substrate and pre-baking
[0055] 1) The substrate enters the photoresist spin coating and pre-baking module from the boat box. After cleaning the surface on the spin coater, a certain thickness of photoresist is spin coated.
[0056] 2) Place the product on the pre-baking hot plate for pre-baking. Once the baking time is up, transfer it to the cold plate to cool to room temperature.
[0057] 3) The substrate is transferred to the exposure module, and the activation of the coupled amino acid monomers is carried out simultaneously.
[0058] 2. Exposure Patterning
[0059] 1) The substrate is placed into the exposure unit with the mask containing the current cycle pattern. After alignment, the shutter is opened for exposure.
[0060] 2) The substrate is transferred from the exposure machine to the heating, amplification, and deprotection module.
[0061] 3. Heating amplification protection module
[0062] 1) The exposed substrate is transferred to the hot plate of the heating amplification and deprotection module to generate acid, which removes the protective substrate of the patterned sites, and then transferred to the cold plate to return to room temperature;
[0063] 2) The substrate is removed and transferred to a heated spin coater for amino acid coupling.
[0064] 4. Preparation of Coupled Amino Acids
[0065] 1) Cleaning
[0066] a) During the previous coupling process, after the amino acid mixture is mixed and ready for spraying, Vm3 opens the waste liquid path, closes the AV path, V4 opens the DMF path, Vm1 valve opens, the DMF flush valve assembly is flushed for 20 seconds, V4 switches to the nitrogen path to purge the pipeline. Repeat twice, and all valves return to normal.
[0067] b) Open valve Vm6, open the DMF line on V4, open valve Vm1, flush valve assembly VBm and volumetric flask to MV bottle, open the nitrogen line on V4 to vent the nitrogen line, return V4 to normal, close valve Vm6, open valve Vm7 to vent nitrogen from MV, close valve Vm7. Repeat once, all valves are restored;
[0068] c) When valve Vm2 opens, DMF enters the volumetric AV and the level reaches level 3. Then, valve Vm2 closes, valve Vm6 opens, and DMF is placed into MV. Valve Vm6 closes, and this process is repeated once. When valve Vm7 opens, nitrogen gas forces the DMF out of MV, and all valves return to normal.
[0069] d) Open valves Vm6 and Vm2 in sequence to rinse the DMF volumetric flask, then close valves Vm2 and Vm6 in sequence. Repeat once, then open valve Vm7 to expel the DMF from the MV cavity with nitrogen gas, and all valves return to normal.
[0070] e) Open valves Vm6 and Vm2, add a certain amount of DMF to MV, close valves Vm2 and Vm6 in sequence, open valve Vm8 to the liquid injection passage, nitrogen gas will squeeze out the DMF to clean the liquid injection pipeline, MV will be empty, all valves will be restored, repeat once.
[0071] f) Multiple units can be operated simultaneously.
[0072] 2) Mixing and Activation
[0073] a) When V00n opens the passage inside the AAn bottle, the Vmn valve opens, and nitrogen pushes the AAn solution to the volumetric AV. When the liquid level in the volumetric AV reaches the set level, Vmn closes, the back suction valve Smn draws back, V00n restores the gas pressure balance state, the Vm6 valve opens, and the AAn solution is put into MV. The Vm6 valve closes.
[0074] b) V2 opens the passage inside the HATU bottle, Vm4 valve opens, nitrogen pushes the HATU solution to AV, the liquid level in the volumetric AV reaches the set level, Vm4 valve closes, V2 restores the gas pressure balance state, Vm6 valve opens, the HATU solution is put into MV, Vm6 valve closes, and DIEA enters in the same process.
[0075] c) Open the nitrogen line to MV through valves Vm8 and Vm9, bubble and stir, then close valves Vm8 and Vm9 and shut off the nitrogen.
[0076] d) When the Vm9 valve is turned on to the spray passage, the Vm7 valve is turned on to push the mixture into the spray pipe. When the Vm7 valve is turned off, the activation solution is ready (at this time, cleaning step 1 can begin).
[0077] 5. Amino acid coupling reaction
[0078] a) When the substrate enters the heated spin coater, the surface photoresist is first cleaned and rinsed. After rinsing, the Vm7 valve is opened to spray the liquid, and the spraying stops when the Vm7 valve is closed.
[0079] b) After amino acid coupling is complete, Vm7 will empty the pipeline and restore all valves to normal.
[0080] c) Clean the surface of the substrate after the reaction, spin dry, and return it to the boat box. This cycle is now complete, and we are ready for the next cycle.
[0081] After the synthesis process was completed, the substrate was post-processed, and the quality inspection sample was pre-processed by mass spectrometry and data was collected. The spectrum showed that there was almost no cross-contamination in the liquid path, and the small batch preparation was qualified.
[0082] In this application, amino acid monomers share a common mixing unit and synthesis unit, i.e., AA1-n to H1~m (n>m), balancing initial investment and flexibility. The development of process flow and parameters during the R&D phase can be achieved by rapidly building an optimized liquid circuit system in conjunction with a few heated spin coaters, offering both economic efficiency and timeliness. Furthermore, the optimized solution can be used for small-batch pilot production and scale-up, optimizing the process flow and parameters during the pilot production phase to meet the needs of the small-scale to pilot-scale stages.
[0083] The methods provided in the exemplary embodiments in this specification are merely examples, and an example of one method does not limit an example of another method. Apparatus / methods discussed in one drawing may be added to or interchanged with apparatus / methods in other drawings. Furthermore, specific numerical data values (e.g., specific quantities, numbers, categories, etc.) or other specific information are used only for discussing exemplary embodiments and are not intended to limit the exemplary embodiments.
Claims
1. A liquid supply system for an in-situ peptide chip device, comprising: An amino acid coupling module for coupling amino acids to a target position, the amino acid coupling module comprising a first synthetic unit; An amino acid supply module, used to provide the amino acid liquid and coupling agent required by the synthesis unit to the amino acid coupling module, includes: Multiple amino acid supply units are provided, each of which includes an independent amino acid container and a corresponding output pipe and control valve. Each amino acid container stores one type of amino acid monomer. The multiple amino acid supply units supply the corresponding amino acid to the first synthesis unit. The coupling agent supply unit includes a separate coupling agent container and an output pipe; The cleaning agent supply unit includes a separate cleaning agent container, output pipeline, and control valve; The first manifold valve group corresponds to the first synthesis unit; the first manifold valve group includes multiple input ports and one output port, and the multiple input ports of the first manifold valve group are connected to the output pipes of the multiple amino acid supply units, coupling agent supply units, and cleaning agent supply units. The first mixing unit, corresponding to the first synthesis unit, is connected to the output port of the first manifold valve group and is used to mix the selected amino acid monomers and coupling agents in the first manifold valve group. The output port of the first mixing unit is connected to the first synthesis unit of the chip via a control valve.
2. The liquid supply system according to claim 1, characterized in that: The liquid supply system also includes a nitrogen supply unit, which is used to fill the connected container or pipeline with nitrogen so that the liquid in the container or pipeline is output along the pipeline.
3. The liquid supply system according to claim 2, characterized in that: The amino acid coupling module further includes second to Nth synthesis units, and the amino acid supply module further includes second to Nth manifold valve groups and second to Nth mixing units connected one-to-one with the second to Nth synthesis units; each amino acid supply unit's output pipe is provided with a multi-channel distribution valve group, so that the amino acid monomers of each amino acid supply unit are distributed to different branches, and each branch corresponds to one of the first to Nth manifold valve groups; the number of channels of the multi-channel distribution valve group corresponds to the number of synthesis units.
4. The liquid supply system according to claim 3, characterized in that: The cleaning agent supply unit includes a first output branch pipe and a second output branch pipe; The first output branch pipe is configured with multiple branches, the number of which corresponds to the number of synthesis units; the branches of the first output branch pipe are fluidly connected to the corresponding manifold valve group. The second output branch pipe is also configured with multiple branches, and the branches of the second output branch pipe are connected to the corresponding mixing unit.
5. The liquid supply system according to claim 3, characterized in that: The nitrogen supply unit is connected to the input port of the first to Nth mixing units.
6. The liquid supply system according to claim 3, characterized in that: The nitrogen supply unit is connected to the input port of the amino acid supply unit.
7. The liquid supply system according to claim 3, characterized in that: The nitrogen supply unit is connected to the input ports of the first to Nth synthesis units.
8. The liquid supply system according to claim 1, characterized in that: The amino acid supply module also includes a coupling agent / excipient supply unit, which includes an output pipe connected to the input port of the manifold valve assembly.
9. The liquid supply system according to claim 2, characterized in that: It also includes a waste liquid collection unit for collecting waste liquid in the pipeline after the cleaning fluid path.
10. The liquid supply system according to claim 3, characterized in that: The amino acid supply unit is positioned horizontally below the mixing unit in the vertical direction, allowing the amino acid supply unit to return the liquid to the corresponding mixing unit by gravity after supplying the liquid.
11. The liquid supply system according to claim 9, characterized in that: The manifold valve assembly is also connected to a waste liquid collection unit.
12. An apparatus for preparing in-situ synthesized polypeptide chips, comprising the liquid supply system of claim 1, and The exposure module is used to transfer deprotected sites onto the chip in a patterned manner using a mask.
13. The device according to claim 12, characterized in that: The equipment also includes a photoresist spin coating and pre-baking module, which is used for photoresist spin coating and pre-baking; and The heating amplification deprotection module is used to amplify the protection of the N-terminus of the acid / alkali removal site at the site excited by exposure on the chip surface.