Method for regulating and controlling chiral signal of peptidyl co-assembly by microfluidics

Abstract

The invention relates to a method for regulating and controlling chiral signals of a peptidyl co-assembly body through microfluidics. The method comprises the following steps: step 1, constructing a microfluidics laminar flow chip; step 2, preparing a first DMF mother solution of Fmoc-amino acid, and preparing a second DMF mother solution of 1-aminopyrene; 3, inputting the first DMF mother liquor and the second DMF mother liquor into a microfluidic laminar flow chip from a first inner inlet and a second inner inlet respectively, and inputting the aqueous solution into the microfluidic laminar flow chip from a first outer inlet and a second outer inlet respectively to obtain a peptidyl co-assembly; according to the method, the peptidyl circular polarization luminescent material with excellent luminescent property is developed; compared with the prior art, the peptidyl co-assembly obtained in the microfluidic laminar flow chip is more uniform in morphology, and shows circular dichroism and circular polarization light-emitting signals which are completely opposite to those of an assembly obtained in an open solution; the invention also provides an effective strategy for regulating and controlling the chiral optical signal of the co-assembly in a confined microfluidic environment.

Description

Technical Field

[0001] The present invention relates to the technical field of nanomaterials, and particularly to a method for microfluidic regulation of chiral signals of peptide-based co-assemblies. Background Art

[0002] The self-assembly of polypeptide molecules has always been an important research direction in the fields of chemistry, materials, and biology, especially in chiral optical properties and functional material design. Chirality is the core element in key processes such as molecular recognition, catalytic activity, and drug design. Whether chiral signals can be effectively regulated directly affects the applications of materials in biomedicine, chiral sensing, and chiral optical devices. Chiral signal inversion usually means that the main chirality of the assembly or molecule changes to the opposite direction, thus showing two completely different optical activities or functional characteristics in the same molecular system. For example, in chiral optical sensing, if controllable inversion of chiral signals can be achieved, different chiral environments can be detected in the same system or multi-mode sensing paths can be established; in drug synthesis and enantioselective catalysis, selective synthesis of different configuration drug precursors is expected by inverting chiral signals.

[0003] Traditional methods for chiral signal inversion mostly rely on chemical modification, changes in solution environment (such as pH, ionic strength, temperature, etc.), or application of external fields (such as light, ultrasound, electric field), etc. These methods not only have disadvantages such as high energy consumption, harsh reaction conditions, or poor reversibility, but also the obtained peptide-based co-assemblies usually have uneven morphologies. How to more simply, controllably, and at low cost regulate the chiral optical signals of peptide-based co-assemblies and obtain peptide-based co-assemblies with more uniform morphologies urgently needs to be solved. Summary of the Invention

[0004] The first aspect of the present invention aims to solve the above technical problems and provides a method for microfluidic regulation of chiral signals of peptide-based co-assemblies, which has the advantages of mild conditions, good controllability, strong repeatability, and significant cost reduction. The main idea is as follows:

[0005] A method for microfluidic regulation of chiral signals of peptide-based co-assemblies includes the following steps:

[0006] Step 1: Construct a microfluidic laminar flow chip. The microfluidic laminar flow chip includes an assembly channel, an inner channel, and an outer channel. Among them, the inner channel includes a first inner channel, a second inner channel, and a confluence channel. One end of the first inner channel and the second inner channel are respectively connected to one end of the confluence channel and form a Y-shaped structure; the other end of the confluence channel is connected to one end of the assembly channel, and the other ends of the first inner channel and the second inner channel respectively form a first inner inlet and a second inner inlet;

[0007] The outer channels include a first outer channel and a second outer channel. The first outer channel and the second outer channel are respectively arranged on both sides of the inner channel. One ends of the first outer channel and the second outer channel are respectively communicated with one end of the assembly channel, and the other ends of the first outer channel and the second outer channel respectively form a first outer entrance and a second outer entrance;

[0008] Step 2: Prepare a first DMF mother liquor of Fmoc-amino acid and a second DMF mother liquor of 1-aminopyrene;

[0009] Step 3: Input the first DMF mother liquor and the second DMF mother liquor into the microfluidic laminar flow chip from the first inner entrance and the second inner entrance respectively, and input the aqueous solution into the microfluidic laminar flow chip from the first outer entrance and the second outer entrance respectively. The first DMF mother liquor, the second DMF mother liquor and the aqueous solution form laminar flow and assemble in the assembly channel to obtain a peptide-based co-assembly;

[0010] Step 4: Age the assembly to obtain an assembly with an opposite chiral optical signal compared with the vial open system. The method provided by this solution uses a microfluidic laminar flow chip for multi-component self-assembly, and regulates the chiral optical signal of the peptide-based co-assembly in the microfluidic confined parallel laminar flow to obtain an assembly material with an opposite chiral signal direction in the vial open system; specifically, chiral Fmoc-amino acid and non-chiral luminescent molecule AP are used for co-assembly. On the one hand, the peptide-based co-assembly obtained in the microfluidic laminar flow chip exhibits an opposite chiral optical signal (CD and CPL) to the assembly obtained in the vial open system; on the other hand, the construction of circularly polarized luminescent materials can be realized. Compared with the prior art, this method not only has the advantages of mild conditions, good controllability, strong repeatability, and lower cost, but also lays an important foundation for the large-scale application in the fields of micro-nano devices, chiral sensors, and biomedical engineering in the future. At the same time, this method utilizes the multiple hydrogen bonds and π-π stacking interactions between chiral Fmoc-amino acid and 1-aminopyrene, and based on the solute conformation restriction and solvent ordered diffusion mechanism of the microfluidic laminar flow chip, successfully realizes the chiral transfer from chiral Fmoc-Ala to 1-aminopyrene in the co-assembly; the peptide-based co-assembly prepared by this method is a peptide-based circularly polarized luminescent material with excellent luminescent properties; compared with the peptide-based fibrous material assembled by the bottom-up method in the traditional open solution, the peptide-based co-assembly obtained in the microfluidic laminar flow chip of the present invention not only has a more uniform morphology, but also exhibits circular dichroism and circularly polarized luminescence signals that are completely opposite to the assembly obtained in the open solution, and proposes an effective strategy for regulating the chiral optical signal of the co-assembly in the confined microfluidic environment.

[0011] Preferably, the assembly channel is a straight channel; the confluence channel is a straight channel, and the confluence channel is docked with the assembly channel. This is more conducive to achieving laminar flow, thereby facilitating the improvement of the success rate of regulating the chiral optical signal of the peptide-based co-assembly.

[0012] Preferably, the assembly channel is a square channel, the width of the assembly channel is 300 μm, and the height of the assembly channel is 100 μm.

[0013] Preferably, the length of the assembly channel is 20 mm, which is conducive to achieving more stable laminar flow.

[0014] Preferably, the outer channel is a square channel, the width of the outer channel is 300 μm, and the height of the outer channel is 100 μm.

[0015] Preferably, the first outer channel and the second outer channel are symmetrically arranged on both sides of the inner channel.

[0016] In the second aspect of the present invention, the problem of how to improve the assembly efficiency is to be solved. Further, the first outer channel includes a straight section and an arc section, the straight section is connected to the arc section, the arc section is connected to the assembly channel, the straight section is parallel to the assembly channel, and the arc section adopts a circular arc structure; the second outer channel includes a straight section and an arc section, the straight section is connected to the arc section, the arc section is connected to the assembly channel, the straight section is parallel to the assembly channel, and the arc section adopts a circular arc structure; the first outer channel and the second outer channel are respectively vertically connected to the assembly channel. In this solution, through the combined design of pre-stabilizing the flow in the straight section, gently turning in the circular arc section, and vertically symmetric confluence, not only the directional transmission of the fluid with low resistance and low consumption in the microchannel system is realized, especially suitable for scenarios with high requirements for flow stability and energy efficiency in microfluidic laminar flow chips; but also it is conducive to the full collision of the solutions in the inner and outer channels, thereby accelerating the nucleation and growth of the assembly, and can effectively improve the assembly efficiency of the co-assembly.

[0017] Preferably, the confluence channel is a square channel, the width of the confluence channel is 300 μm, and the height of the confluence channel is 100 μm.

[0018] Preferably, the first inner channel and the second inner channel are symmetrically arranged.

[0019] In the third aspect of the present invention, the problem of efficient mixing or stable flow is to be solved. Further, the first inner channel is a square channel, and along the direction from the first inner inlet to the confluence channel, the width of the first inner channel gradually decreases; the second inner channel is a square channel, and along the direction from the second inner inlet to the confluence channel, the width of the second inner channel gradually decreases. In this solution, by gradually decreasing the widths of the two inner channels along the flow direction (while the height remains unchanged), the regulation of flow velocity and pressure is achieved by using the continuity equation and Bernoulli's principle in fluid mechanics; through the optimization of the geometric shape, the dynamic characteristics of the fluid are coordinated, which can not only achieve the effects of efficient mixing and stable flow, but also give full play to the confinement effect of laminar flow, thus being more conducive to changing the chiral optical signal of the peptide-based co-assembly.

[0020] Preferably, the height of the first inner channel is 100 μm, the width of the first inner channel at the first inner inlet is 300 μm, and the width of the first inner channel at the connection to the confluence channel is 220 μm; the second inner channel is a square channel, the height of the second inner channel is 100 μm, the width of the second inner channel at the second inner inlet is 300 μm, and the width of the second inner channel at the connection to the confluence channel is 220 μm. Such a design is not only conducive to the efficient mixing and assembly of the first DMF mother liquor and the second DMF mother liquor, but also conducive to achieving more stable laminar flow and giving full play to the confinement effect of laminar flow, thus being more conducive to changing the chiral optical signal of the peptide-based co-assembly.

[0021] Preferably, the length of the first inner channel is 2.91 mm; the length of the second inner channel is 2.91 mm.

[0022] Preferably, in step 2, the Fmoc-amino acid used is Fmoc-L-Ala or Fmoc-D-Ala.

[0023] Further, in step 2, the concentration of the first DMF mother liquor is 8 - 32 mM. A lower concentration will result in a poor confinement effect during the assembly in the microfluidic laminar flow chip, which is not conducive to regulating the chiral optical signal of the peptide-based co-assembly, while a higher concentration will cause the chip to be easily blocked, which is also not conducive to regulating the chiral optical signal of the peptide-based co-assembly. Controlling the concentration of the first DMF mother liquor to 8 - 32 mM will neither cause blockage nor can achieve a better confinement effect, and is more conducive to regulating the chiral optical signal of the peptide-based co-assembly.

[0024] Preferably, in step 2, the concentration of the first DMF mother liquor is 20 mM.

[0025] Further, in the step 2, the concentration of the second DMF mother liquor is 8-32 mM. A lower concentration will result in a poor confinement effect during the assembly in the microfluidic laminar flow chip, which is not conducive to regulating the chiral optical signal of the peptide-based co-assembly. While a higher concentration will cause the chip to be easily blocked, also not conducive to regulating the chiral optical signal of the peptide-based co-assembly. Controlling the concentration of the second DMF mother liquor to 8-32 mM will neither cause blockage nor result in a better confinement effect, which is more conducive to regulating the chiral optical signal of the peptide-based co-assembly.

[0026] Preferably, in the step 2, the concentration of the second DMF mother liquor is 20 mM.

[0027] In the fourth aspect of the present invention, the problem of controllable preparation of functional peptide-based materials is to be solved. Further, in the step (3), the input flow rate at the first inner inlet is controlled to be 1-4 μL / min; the input flow rate at the second inner inlet is controlled to be 1-4 μL / min; in the step (3), the input flow rate at the first outer inlet is controlled to be 4-16 μL / min; the input flow rate at the second outer inlet is controlled to be 4-16 μL / min. Within this flow rate range, a stable laminar flow can be formed, and the confinement effect advantage of the laminar flow can be fully exerted. It is not only more conducive to achieving the regulation of the chiral optical signal of the peptide-based co-assembly, but also a more uniform peptide-based co-assembly can be obtained. And for the first time, three key problems of maintaining laminar flow stability, precise assembly of nanostructures, and controllable chiral optical properties are synchronously solved in a dynamic microenvironment, providing a new technical path for the controllable preparation of functional peptide-based materials.

[0028] Preferably, in the step (3), the input flow rate at the first inner inlet is controlled to be 2 μL / min; the input flow rate at the second inner inlet is controlled to be 2 μL / min. Under this flow rate configuration, the laminar flow is more stable, and the obtained peptide-based assembly structure is more uniform.

[0029] Preferably, in the step (3), the input flow rate at the first outer inlet is controlled to be 8 μL / min; the input flow rate at the second outer inlet is controlled to be 8 μL / min. Under this flow rate configuration, the laminar flow is more stable, and the obtained peptide-based assembly structure is more uniform.

[0030] Further, in the step (3), the total flow rate in the microfluidic laminar flow chip is controlled to be 10-40 μL / min. It is beneficial to obtain a more uniform peptide-based co-assembly.

[0031] Preferably, in the step (3), the total flow rate in the microfluidic laminar flow chip is controlled to be 20 μL / min. A more uniform peptide-based co-assembly can be obtained.

[0032] Preferably, in the step (3), the aqueous solution used is distilled water.

[0033] Preferably, in the step 4, the aging time of the assembly is controlled to be 2 - 12 h.

[0034] Compared with the prior art, by using the method for microfluidically regulating the chiral signal of a peptide-based co-assembly provided by the present invention, the chiral transfer of Fmoc-amino acid to AP and the construction of the co-assembly are successfully achieved through multiple hydrogen bond and π-π stacking interactions between chiral Fmoc-amino acid and achiral luminescent compound AP. The peptide-based co-assembly obtained in the microfluidic laminar flow chip exhibits chiral optical signals (CD and CPL) opposite to those of the assembly obtained in the vial development system. Therefore, the chiral optical signal of the peptide-based co-assembly can be effectively regulated by this method. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as limiting the scope. For those of ordinary skill in the art, other related drawings can be obtained based on these drawings without creative efforts.

[0036] Figure 1 It is a schematic structural diagram of a microfluidic laminar flow chip prepared in the embodiment of the present invention.

[0037] Figure 2 It is a schematic structural diagram of another microfluidic laminar flow chip prepared in the embodiment of the present invention.

[0038] Figure 3 It is a structural diagram of Fmoc-L-Ala, Fmoc-D-Ala, and AP used in the embodiment of the present invention.

[0039] Figure 4 In it, a) is the CD signal diagram of the assembly prepared in Comparative Example 1 and Comparative Example 2; b) is the CD signal diagram of the assembly prepared in Example 3 and Example 4.

[0040] Figure 5 In it, a) is the CPL signal diagram of the assembly prepared in Comparative Example 1 and Comparative Example 2; b) is the CPL signal diagram of the assembly prepared in Example 3 and Example 4.

[0041] Figure 6 It is the circular dichroism spectrum diagram at different flow rates.

[0042] Figure 7 In it, a) is the SEM diagram of the assembly prepared in a vial (Comparative Example 1 or Comparative Example 2); b) is the SEM diagram of the assembly prepared in a microfluidic laminar flow chip (Example 3 or Example 4).

[0043] Figure 8 FTIR spectra of Fmoc-L-Ala and Fmoc-L-Ala / AP.

[0044] Figure 9 XRD patterns of AP and Fmoc-L-Ala / AP.

[0045] Figure 10 The obtained GIWAXS pattern.

[0046] Figure 11 In a), it is the structural diagram of the optimized dimer A; in b), it is the structural diagram of the optimized dimer B; in c), it is the ECD spectrum corresponding to dimer A and dimer B.

[0047] Marking explanations in the figure: inner channel 1, first inner inlet 11, first inner channel 12, second inner inlet 13, second inner channel 14, confluence channel 15; outer channel 2, first outer inlet 21, first outer channel 22, second outer inlet 23, second outer channel 24, straight segment 25, arc segment 26; assembly channel 3. Detailed implementation manners

[0048] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Usually, the components of the embodiments of the present invention described and shown in the accompanying drawings here can be arranged and designed in various different configurations. Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed present invention, but only represents the selected embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative efforts fall within the scope of protection of the present invention.

[0049] Embodiment 1

[0050] In this embodiment, a method for microfluidic regulation of the chiral signal of a peptide-based co-assembly is provided, including the following steps:

[0051] Step 1: Construct a microfluidic laminar flow chip. The microfluidic laminar flow chip includes an assembly channel 3, an inner channel 1, and an outer channel 2. Among them, as Figure 1 and Figure 2 shown, the inner channel 1 includes a first inner channel 12, a second inner channel 14, and a confluence channel 15. One end of the first inner channel 12 and the second inner channel 14 are respectively connected to one end of the confluence channel 15 and form a Y-shaped structure, as Figure 1As shown; the other end of the confluence channel 15 is connected to one end of the assembly channel 3, and the other ends of the first inner channel 12 and the second inner channel 14 respectively form a first inner inlet 11 and a second inner inlet 13, as Figure 1 shown; in implementation, the first inner channel 12 and the second inner channel 14 are preferably symmetrically arranged, which is more conducive to forming a more stable laminar flow.

[0052] As Figure 1 shown, the outer channel 2 includes a first outer channel 22 and a second outer channel 24. The first outer channel 22 and the second outer channel 24 are respectively arranged on both sides of the inner channel 1. One ends of the first outer channel 22 and the second outer channel 24 are respectively connected to one end of the assembly channel 3, and the other ends of the first outer channel 22 and the second outer channel 24 respectively form a first outer inlet 21 and a second outer inlet 23, as Figure 1 shown; in implementation, the first outer channel 22 and the second outer channel 24 are preferably symmetrically arranged on both sides of the inner channel 1, which is less likely to cause flow disturbance, and thus is more conducive to forming a more stable laminar flow.

[0053] In this embodiment, the assembly channel 3 adopts a straight channel, as Figure 1 shown; the confluence channel 15 also adopts a straight channel. The confluence channel 15 is docked with the assembly channel 3, and the internal dimensions of the confluence channel 15 and the assembly channel 3 are preferably constructed to be the same, which is more conducive to maintaining laminar flow and is beneficial to improving the success rate of regulating the chiral optical signal of the peptide-based co-assembly.

[0054] Since the first inner channel 12, the second inner channel 14 and the confluence channel 15 form a Y-shaped structure, as Figure 1 shown, in implementation, the angle between the first inner channel 12 and the second inner channel 14 can be determined according to actual needs. For example, in a preferred implementation manner, the angle between the first inner channel 12 and the second inner channel 14 can be 45-120 degrees. And for facilitating the full contact and mixing of the solutions in the first inner channel 12 and the second inner channel 14, in implementation, the angle between the first inner channel 12 and the second inner channel 14 can be preferably controlled to be 70-90 degrees. For example, in the preferred implementation manner provided in this embodiment, the angle between the first inner channel 12 and the second inner channel 14 can be preferably 77 degrees, 78 degrees, 79 degrees, 80 degrees, 81 degrees and 82 degrees, etc., which can not only achieve a better assembly effect but also be conducive to forming laminar flow.

[0055] In implementation, the cross-sectional shapes of the assembly channel 3, the inner channel 1, and the outer channel 2 can be determined according to actual requirements. However, based on previous experiments, in this embodiment, the cross-sections of the assembly channel 3, the inner channel 1, and the outer channel 2 all adopt rectangles, which is more convenient for forming laminar flow and is also more conducive to maintaining the laminar flow state. For example, in this embodiment, the assembly channel 3, the outer channel 2, the inner channel 1, and the confluence channel 15 are all respectively constructed as square channels, and the dimensions of the channels need to be strictly controlled. If the dimensions of the channels are too small, the chip is likely to be blocked, while if the dimensions of the channels are too large, it is similar to the existing vial open system and cannot produce the corresponding technical effects. Therefore, the dimensions of the channels need to be strictly controlled to better exert the confinement effect of laminar flow while forming laminar flow. Specifically, in this embodiment, the first inner channel 12 adopts a square channel, the height of the first inner channel 12 is 100 μm, and the width of the first inner channel 12 is 300 μm. At the same time, the second inner channel 14 adopts a square channel, the height of the second inner channel 14 is 100 μm, and the width of the second inner channel 14 is 300 μm; the height of the confluence channel 15 is 100 μm, and the width of the confluence channel 15 is also 300 μm; correspondingly, the height of the assembly channel 3 is also constructed as 100 μm, and the width of the assembly channel 3 is also constructed as 300 μm. At the same time, since the outer channel 2 is directly connected to the assembly channel 3, in implementation, the dimensions of the outer channel 2 are preferably the same as those of the assembly channel 3. For example, in this embodiment, the height of the outer channel 2 is also constructed as 100 μm, and the width of the outer channel 2 is also constructed as 300 μm, which is more conducive to exerting the confinement effect of laminar flow.

[0056] In implementation, the first outer channel 22, the second outer channel 24, and the assembly channel 3 can also form a Y-shaped structure. However, in the preferred implementation provided in this embodiment, the end of the first outer channel 22 and the end of the second outer channel 24 are respectively vertically connected to the assembly channel 3, as Figure 1 shown, and the first outer channel 22 includes a straight segment 25 and an arc segment 26. The straight segment 25 is connected to the arc segment 26, the arc segment 26 is connected to the assembly channel 3, the straight segment 25 is parallel to the assembly channel 3, and the arc segment 26 adopts a circular arc structure, as Figure 1 shown; at the same time, the second outer channel 24 includes a straight segment 25 and an arc segment 26. The straight segment 25 is connected to the arc segment 26, the arc segment 26 is connected to the assembly channel 3, the straight segment 25 is parallel to the assembly channel 3, and the arc segment 26 adopts a circular arc structure, as Figure 1As shown. More specifically, during implementation, the length of the straight segment 25 can be greater than or equal to 2 mm. For example, in this embodiment, the length of the straight segment 25 is 3 mm. At the same time, the outer diameter of the arc segment 26 can be greater than or equal to 4 mm. For example, in this embodiment, the outer diameter of the arc segment 26 is 9.26 mm. Through the combined design of pre-stabilizing the flow by the straight segment 25, gently turning by the arc segment, and vertically symmetric confluence, not only is the directional transmission of the fluid in this microchannel system with low resistance and low consumption achieved, which is especially suitable for scenarios with high requirements for flow stability and energy efficiency in microfluidic laminar flow chips; but also it is beneficial for the sufficient collision of the solutions inside and outside the channels, thereby accelerating the nucleation and growth of the assembly and improving the assembly efficiency of the co-assembly.

[0057] It can be understood that during implementation, the length of the assembly channel 3 can be determined according to actual needs. In this embodiment, the length of the assembly channel 3 is 20 mm, which is relatively long and is beneficial for achieving more stable laminar flow. Similarly, during implementation, the lengths of the first inner channel 12 and the second inner channel 14 can be determined according to actual needs. By way of example, in this embodiment, the length of the first inner channel 12 is 2.91 mm; the length of the second inner channel 14 is 2.91 mm.

[0058] During implementation, other structures of the microfluidic laminar flow chip are the same as those in the prior art and will not be elaborated here.

[0059] Step 2: Calculate the concentration of the first DMF mother liquor based on the concentration of the peptide-based co-assembly prepared in the vial open system, and prepare the first DMF mother liquor of Fmoc-amino acid. Similarly, calculate the concentration of the second DMF mother liquor based on the concentration of the peptide-based co-assembly prepared in the vial open system, and prepare the second DMF mother liquor of 1-aminopyrene (AP). Among them, Fmoc-amino acid has chirality; Fmoc-alanine (Fmoc-Ala) includes two isomers, Fmoc-L-Ala and Fmoc-D-Ala. These two isomers are a pair of enantiomers with a mirror-symmetric spatial structure. In this embodiment, Fmoc-amino acid preferably uses Fmoc-L-Ala or Fmoc-D-Ala.

[0060] In specific implementation, the Fmoc-amino acid is dissolved in N,N-dimethylformamide (DMF) to form a first DMF stock solution. Since the concentration of the peptide-based co-assembly prepared by this method is the same as that of the peptide-based co-assembly (abbreviated as assembly) prepared in the corresponding vial open system, the concentration of the first DMF stock solution can be calculated according to the concentration of the first DMF stock solution used in the vial open system or the concentration of the assembly prepared in the vial open system. Similarly, 1-aminopyrene is dissolved in N,N-dimethylformamide (DMF) to form a second DMF stock solution. Since the concentration of the peptide-based co-assembly prepared by this method is the same as that of the peptide-based co-assembly (abbreviated as assembly) prepared in the corresponding vial open system, the concentration of the second DMF stock solution can be calculated according to the concentration of the second DMF stock solution used in the vial open system or the concentration of the assembly prepared in the vial open system. In specific implementation, a lower concentration will result in a poor confinement effect during the assembly in the microfluidic laminar flow chip, which is not conducive to regulating the chiral optical signal of the peptide-based co-assembly, while a higher concentration will cause the chip to be easily blocked, which is also not conducive to regulating the chiral optical signal of the peptide-based co-assembly. Therefore, it is necessary to precisely and strictly control the concentrations of the first DMF stock solution and the second DMF stock solution. Correspondingly, the concentration of the peptide-based co-assembly prepared in the vial open system is also limited. According to previous experiments, the concentration of the first DMF stock solution used in the vial open system can be preferably controlled at 20-80 mM. At the same time, the concentration of the second DMF stock solution used in the vial open system can be preferably controlled at 20-80 mM, which is more conducive to implementing this method. Based on this, in implementation, the concentration of the first DMF stock solution can preferably be 8-32 mM. For example, the concentration of the first DMF stock solution can preferably be 18 mM, 19 mM, 20 mM, 21 mM, etc. At the same time, the concentration of the second DMF stock solution can preferably be 8-32 mM. For example, the concentration of the second DMF stock solution can preferably be 18 mM, 19 mM, 20 mM, 21 mM, etc. As an example, in this embodiment, the concentrations of both the first DMF stock solution and the second DMF stock solution are 20 mM.

[0061] Step 3: Input the first DMF mother liquor and the second DMF mother liquor into the microfluidic laminar flow chip from the first inner inlet 11 and the second inner inlet 13 respectively, and input the aqueous solution into the microfluidic laminar flow chip from the first outer inlet 21 and the second outer inlet 23 respectively. The first DMF mother liquor, the second DMF mother liquor and the aqueous solution form a laminar flow and assemble in the assembly channel 3 to obtain a peptide co-assembly (or called a nanosheet assembly). In this embodiment, it is necessary to comprehensively consider the liquid holding capacity of the microfluidic laminar flow chip and whether a stable laminar flow state can be maintained in combination with the structure and flow rate of the microfluidic laminar flow chip. If the flow rate is too small, a stable laminar flow may not be formed, and if the flow rate is too large, the laminar flow will turn into a turbulent flow; in addition, when the flow rate is too large, the advantage of the laminar flow, that is, the uniform and rapid diffusion and collision between the molecules forming the assembly, will also disappear, which is not conducive to obtaining a peptide co-assembly with a more uniform structure. Therefore, the flow rate needs to be strictly controlled. According to previous experiments, in specific implementation, the input flow rate at the first inner inlet 11 can be preferably controlled at 1-4 μL / min, and the input flow rate at the second inner inlet 13 can be preferably controlled at 1-4 μL / min. Correspondingly, the input flow rate at the first outer inlet 21 can be preferably controlled at 4-16 μL / min, and the input flow rate at the second outer inlet 23 can be preferably controlled at 4-16 μL / min; so that the total flow rate in the microfluidic laminar flow chip is preferably controlled at 10-40 μL / min. According to previous experiments, within this flow rate range, a stable laminar flow can be formed, and the confinement effect advantage of the laminar flow can be fully exerted. Not only can a peptide co-assembly with a more uniform structure be obtained, but also it is more conducive to achieving the purpose of regulating the chiral optical signal of the peptide co-assembly. That is, through the systematic optimization of hydrodynamic parameters, this solution synchronously solves the three key problems of maintaining laminar flow stability, precisely assembling nanostructures, and controlling chiral optical properties in a dynamic microenvironment for the first time, providing a new technical path for the controllable preparation of functional peptide-based materials.

[0062] In implementation, the input flow rate at the first inner inlet 11 can be preferably controlled at 2 μL / min or 3 μL / min. At the same time, the input flow rate at the second inner inlet 13 can be preferably controlled at 2 μL / min or 3 μL / min, and the input flow rate at the first outer inlet 21 can be preferably controlled at 7 μL / min or 8 μL / min; the input flow rate at the second outer inlet 23 can be preferably controlled at 7 μL / min or 8 μL / min, so that the total flow rate of the microfluidic laminar flow chip is 18-22 μL / min, preferably controlled at 20 μL / min. Under this flow rate configuration, the laminar flow is more stable, and the obtained peptide assembly structure is more uniform.

[0063] In implementation, the aqueous solution can preferably use distilled water.

[0064] Step 4: Age the peptide co-assembly to obtain a peptide co-assembly with an opposite chiral optical signal compared to the vial open system. During implementation, the aging time of the peptide co-assembly can preferably be controlled to be 2 - 12 h.

[0065] Example 2

[0066] The main difference between this Example 2 and the above examples lies in that in the method for microfluidic regulation of the chiral signal of the peptide co-assembly provided in this example, there are differences in the structure of the microfluidic laminar flow chip. Specifically, in this example, along the direction from the first inner inlet 11 to the confluence channel 15, the height of the first inner channel 12 remains unchanged, but the width of the first inner channel 12 gradually decreases, as Figure 2 shown; at the same time, along the direction from the second inner inlet 13 to the confluence channel 15, the height of the second inner channel 14 remains unchanged, but the width of the second inner channel 14 gradually decreases, as Figure 2 shown. By gradually decreasing the widths of the two inner channels along the flow direction (while the height remains unchanged), the regulation of the flow velocity and pressure is achieved by using the continuity equation and Bernoulli's principle in hydrodynamics. This design coordinates the fluid dynamic characteristics through the optimization of the geometry, and finally achieves the technical goal of efficient mixing or stable flow.

[0067] As an example, as Figure 2 shown, the first inner channel 12 is a square channel, the height of the first inner channel 12 is 100 μm, the width of the first inner channel 12 at the first inner inlet 11 is 300 μm, and the width of the first inner channel 12 at the connection with the confluence channel 15 is 220 μm; at the same time, as Figure 1 shown, the second inner channel 14 is a square channel, the height of the second inner channel 14 is 100 μm, the width of the second inner channel 14 at the second inner inlet 13 is 300 μm, and the width of the second inner channel 14 at the connection with the confluence channel 15 is 220 μm. Such a design not only facilitates the efficient mixing and assembly of the first DMF mother liquor and the second DMF mother liquor, but also facilitates the realization of more stable laminar flow and gives full play to the confinement effect of laminar flow, thereby being more conducive to changing the chiral optical signal of the peptide co-assembly.

[0068] During implementation, the lengths of the first inner channel 12 and the second inner channel 14 can be determined according to actual needs. In this example, the length of the first inner channel 12 is 2.91 mm; the length of the second inner channel 14 is 2.91 mm, as Figure 2 shown.

[0069] Example 3

[0070] In this example, the peptide-based co-assembly is prepared by using the method for microfluidically regulating the chiral signal of the peptide-based co-assembly provided in Example 1 or Example 2. Among them, the microfluidic laminar flow chip is constructed according to Step 1; in Step 2, the Fmoc-amino acid used is commercially available Fmoc-L-Ala (L-Ala), as Figure 3 shown, and it is dissolved in N,N-dimethylformamide (DMF) to obtain a first DMF mother liquor with a concentration of 20 mM; at the same time, 1-aminopyrene is dissolved in N,N-dimethylformamide (DMF) to obtain a second DMF mother liquor with a concentration of 20 mM; then, according to the method of Step 3, the first DMF mother liquor and the second DMF mother liquor are respectively input into the microfluidic laminar flow chip from the first inner inlet 11 and the second inner inlet 13, and distilled water aqueous solutions are respectively input into the microfluidic laminar flow chip from the first outer inlet 21 and the second outer inlet 23; among them, the flow rates of the first inner channel 12 and the second inner channel 14 are respectively 2 μL / min; the flow rates of the first outer channel 22 and the second outer channel 24 are respectively 8 μL / min; the total flow rate of the microfluidic laminar flow chip is 20 μL / min; the first DMF mother liquor, the second DMF mother liquor, and the distilled water aqueous solutions form a laminar flow and assemble in the assembly channel 3, and finally the Fmoc-L-Ala / AP peptide-based co-assembly is obtained. Then, according to the method of Step 4, the prepared peptide-based co-assembly is aged for at least 8 h, and finally the required Fmoc-L-Ala / AP peptide-based co-assembly is obtained.

[0071] Example 4

[0072] In this example, the peptide-based co-assembly is prepared by using the method for microfluidically regulating the chiral signal of the peptide-based co-assembly provided in Example 1 or Example 2. Among them, the microfluidic laminar flow chip is constructed according to Step 1; in Step 2, the Fmoc-amino acid used is commercially available Fmoc-D-Ala (D-Ala), as Figure 3As shown, it was dissolved in N,N-dimethylformamide (DMF) to obtain a first DMF mother liquor with a concentration of 20 mM; meanwhile, 1-aminopyrene was dissolved in N,N-dimethylformamide (DMF) to obtain a second DMF mother liquor with a concentration of 20 mM; then, according to the method in step 3, the first DMF mother liquor and the second DMF mother liquor were respectively input into the microfluidic laminar flow chip from the first inner inlet 11 and the second inner inlet 13, and the distilled water aqueous solution was respectively input into the microfluidic laminar flow chip from the first outer inlet 21 and the second outer inlet 23; wherein, the flow rates of the first inner channel 12 and the second inner channel 14 were respectively 2 μL / min; the flow rates of the first outer channel 22 and the second outer channel 24 were respectively 8 μL / min; the total flow rate of the microfluidic laminar flow chip was 20 μL / min; the first DMF mother liquor, the second DMF mother liquor, and the distilled water aqueous solution formed a laminar flow and assembled in the assembly channel 3, and finally, the Fmoc-D-Ala / AP peptide co-assembly was obtained. Then, according to the method in step 4, the prepared peptide co-assembly was aged for at least 8 h, and finally, the desired Fmoc-D-Ala / AP peptide co-assembly was obtained.

[0073] Comparative Example 1

[0074] The Fmoc-L-Ala / AP assembly was prepared in a traditional vial open system, and the specific method was as follows:

[0075] In the vial open system, the Fmoc-amino acid used was commercially available Fmoc-L-Ala (as Figure 3 shown), and it was dissolved in N,N-dimethylformamide (DMF solution) to obtain a first DMF mother liquor, and the concentration of the first DMF mother liquor could be 50 mM; meanwhile, 1-aminopyrene (AP) was dissolved in N,N-dimethylformamide (DMF solution) to obtain a second DMF mother liquor, and the concentration of the second DMF mother liquor could also be 50 mM. Then, 40 μL of the prepared first DMF mother liquor was taken with a pipette into a 5 mL vial (such as a conical flask or a beaker), and 40 μL of the prepared second DMF mother liquor was taken with a pipette into the 5 mL vial; then, 120 μL of DMF solution was added to the vial and gently shaken, and finally, 800 μL of distilled water aqueous solution was added to the vial for assembly to obtain an assembly, and then, according to the same method as in Example 3, the assembly was aged (such as static aging) for 8 h to obtain the desired Fmoc-L-Ala / AP assembly.

[0076] In implementation, it is necessary to accurately calculate the concentration of the first DMF mother liquor and the concentration of the second DMF mother liquor in the vial open system, so that the concentration of the peptide-based co-assembly prepared in this example is the same as the concentration of the peptide-based co-assembly prepared in Example 3, in order to obtain a co-assembly with opposite chiral optical signals at the same concentration. That is to say, in implementation, in some cases, the concentration of the first DMF mother liquor and the concentration of the second DMF mother liquor when preparing the peptide-based co-assembly by the method described in Example 3 can be calculated according to the concentration of the peptide-based co-assembly prepared in the vial open system. In some cases, the concentration of the first DMF mother liquor and the concentration of the second DMF mother liquor in the vial open system can be calculated according to the concentration of the peptide-based co-assembly prepared by the method described in Example 3. By comparison, the ratio of the concentration of the first DMF mother liquor used in the method described in Example 3 to the concentration of the first DMF mother liquor used in the corresponding vial open system (i.e., this example) is 2:5; at the same time, the ratio of the concentration of the second DMF mother liquor used in the method described in Example 3 to the concentration of the second DMF mother liquor used in the corresponding vial open system (i.e., this example) is 2:5. It is not only possible to obtain peptide-based co-assemblies with the same concentration, but also conducive to more stably obtaining two co-assemblies with opposite chiral optical signals.

[0077] In addition, other test conditions of Comparative Example 1 are the same as those of Example 3.

[0078] Comparative Example 2

[0079] Prepare the Fmoc-D-Ala / AP assembly in a traditional vial open system. The specific method is as follows:

[0080] In the vial open system, the Fmoc-amino acid used is commercially available Fmoc-D-Ala (as Figure 3 shown), and it is dissolved in N,N-dimethylformamide (DMF solution) to obtain the first DMF mother liquor, and the concentration of the first DMF mother liquor can be 50 mM; at the same time, 1-aminopyrene (AP) is dissolved in N,N-dimethylformamide (DMF solution) to obtain the second DMF mother liquor, and the concentration of the second DMF mother liquor can also be 50 mM. Then, 40 μL of the prepared first DMF mother liquor is taken with a pipette into a 5 mL vial (such as an Erlenmeyer flask or a beaker), and 40 μL of the prepared second DMF mother liquor is taken with a pipette into the 5 mL vial; then 120 μL of DMF solution is added to the vial and shaken gently, and finally 800 μL of distilled water aqueous solution is added to the vial for assembly to obtain an assembly, and then the assembly is aged in the same method as in Example 1 for 8 h to obtain the required Fmoc-D-Ala / AP assembly.

[0081] It is understandable that the concentration of the assembly prepared in this embodiment is the same as that of the peptide-based co-assembly prepared in Example 4. In addition, the other test conditions of Comparative Example 2 are the same as those of Example 4 and Comparative Example 1.

[0082] Tests: (1) Circular dichroism test.

[0083] After the assemblies obtained in the aforementioned Comparative Example 1 and Comparative Example 2 were allowed to stand and age, the chiral properties of the assemblies were characterized by circular dichroism (CD).

[0084] The circular dichroism test method is as follows: First, take the aged assembly into a quartz cuvette with a pipette, and then put it into a circular dichroism instrument for testing.

[0085] As Figure 4 shown in a) of the figure, the Fmoc-L-Ala / AP assembly prepared in Comparative Example 1 and the Fmoc-D-Ala / AP assembly prepared in Comparative Example 2, these two chiral enantiomers showed obviously symmetric mirror-image CD signals after co-assembly; the peaks at 425 nm, 375 nm and 350 nm in the figure correspond to the absorption region of the pyrene group, indicating the effective chiral transfer from Fmoc-Ala to AP.

[0086] Similarly, after the peptide-based co-assemblies obtained in the aforementioned Example 3 and Example 4 were allowed to stand and age, the chiral properties of the peptide-based co-assemblies were characterized by circular dichroism (CD). Specifically, the CD signal was tested in the same way. Unexpectedly, for the peptide-based co-assembly constructed by the microfluidic laminar flow chip, the dominant positive CD signal peak at 418 nm became a negative CD signal, indicating that the chiral arrangement of the peptide-based co-assembly may have been reversed, as Figure 4 shown in b) of the figure, that is, Fmoc-Ala / AP showed opposite CD signals in the vial and the microfluidic laminar flow chip. Such results indicate that the aforementioned microfluidic laminar flow chip can effectively regulate the chiral optical signal of the peptide-based co-assembly and has great application prospects in controlling the chiral signal of the peptide-based co-assembly.

[0087] In addition, when this method is adopted, the circular dichroism signal intensities at different flow rates are as Figure 6 shown. If the flow rate is too small, a stable laminar flow may not be formed, and if the flow rate is too large, the laminar flow will turn into a turbulent flow, both of which will lead to the weakening of the circular dichroism signal.

[0088] (2) Circularly polarized luminescence property test.

[0089] Since it was found in the present invention that the CD signal of the peptide-based co-assembly obtained in the microfluidic laminar flow chip and vial system was reversed, it is possible to achieve the regulation of the circularly polarized luminescence (CPL) property, that is, the present method can also be used to regulate the circularly polarized luminescence (CPL) property.

[0090] For verification, the CPL sample was prepared as follows: The aged peptide-based co-assembly was centrifuged first, and then evenly coated on a quartz wafer.

[0091] As Figure 5 shown in a) of -3 , it was found through testing that the Fmoc-L-Ala / AP obtained in Comparative Example 1 and the Fmoc-D-Ala / AP assembly obtained in Comparative Example 2 showed negative CPL signals and positive CPL signals at 550 nm, respectively, indicating that the pyrene group dimers were successfully formed in the solid state. The luminescence dissymmetry factors (glum) of Fmoc-L-Ala / AP and Fmoc-D-Ala / AP were -4.7×10 -3 and 5.0×10 Figure 5 , respectively, as shown in a) of Figure 5 . More importantly, the chiral signal inversion also occurred in the excited state for the assemblies constructed by different methods. As shown in b) of

[0092] , the assemblies Fmoc-L-Ala / AP and Fmoc-D-Ala / AP obtained in Experimental Example 3 and Example 4 showed positive CPL signals and negative CPL signals at 550 nm, respectively, thus demonstrating that the method provided by the present invention successfully achieved the regulation of the CPL signal based on different assembly environments.

[0093] The peptide-based co-assembly flowing out of the microfluidic laminar flow chip in the aforementioned Example 3 and Example 4 was collected in a vial. After standing and aging for 8 h, the morphology of the assembly was characterized by scanning electron microscopy (SEM).

[0094] The preparation method of the SEM sample was as follows: First, place the silicon wafer on the filter paper, take 20 μL of the aged Fmoc-L-Ala / AP or Fmoc-D-Ala / AP assembly and drop it on the silicon wafer. After standing for 1 minute, suck it dry with the filter paper.

[0095] In the co-assembly obtained in the vial, the co-assembly formed a disordered linear fibrous structure, as shown in a) of Figure 7 , because the different components were unevenly distributed and assembled without restraint in the assembly environment of the vial; while the peptide-based co-assembly obtained in the microfluidic laminar flow chip showed a more ordered nano-sheet structure, as shown in Figure 7As shown in b) of [reference], it is because the diffusion and mass transfer performance of the molecules in the confined microfluidic environment are enhanced.

[0096] (4) Experiment on the assembly mechanism of the assembly

[0097] In order to study the assembly mechanism of the assembly in the vial and the microfluidic laminar flow chip, the inventors carried out characterizations of Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD).

[0098] The sample preparation for Fourier transform infrared spectroscopy is as follows: The aged assembly obtained in the vial or the microfluidic laminar flow chip is centrifuged, and after drying, it is tested.

[0099] The results are as Figure 8 shown. In the spectrum of Fmoc-L-Ala, the carbonyl groups of the amide and carboxyl groups are shown at 1702 and 1718 cm -1 , and the stretching vibrations in the amide I and II regions are at 1618 and 1529 cm -1 . In the Fmoc-L-Ala / AP assembly constructed in the microfluidic laminar flow chip, the carbonyl groups of the amide and carboxyl groups disappear, and the amide I and amide II regions shift, indicating that the amide and carboxyl groups of Fmoc-L-Ala may participate in the co-assembly with AP in the microfluidic laminar flow chip. At the same time, similar results were obtained for the assembly in the open system of the vial.

[0100] The sample preparation for X-ray diffraction is as follows: The aged assembly obtained in the vial or the microfluidic laminar flow chip is centrifuged, and then coated on a silicon wafer for testing to determine the change in the molecular arrangement before and after co-assembly.

[0101] The results of X-ray diffraction are as Figure 9 shown. The self-assembly of AP in the microfluidic laminar flow chip shows diffraction peaks at 10-15°. After co-assembly with L-Ala, a series of new peaks appear at 10.12, 15.24, 17.72, 20.36, 22.70, 25.54, 27.72, and 30.78°, and the corresponding lattice spacing ratios are 1:√2:√3:2:√5:√6:√7:3, which can be attributed to the typical body-centered cubic (BCC) phase.

[0102] (5) Research experiment on the mechanism of chiral signal inversion

[0103] In order to study the possible mechanism for the assembly to obtain opposite chiral signals in the vial and the microfluidic laminar flow chip, further characterizations and calculations of grazing incidence wide-angle scattering (GIWAXS) and time-dependent density functional theory (TDDFT) were carried out.

[0104] The sample preparation for grazing incidence wide-angle scattering is similar to the sample preparation process for X-ray diffraction; along qz The L-Ala / AP copolymer aggregates of the axis exhibit a series of strong scattering bands at 0.437, 0.640, 0.749, 0.863, 1.003 and as shown. The ratios of these scattering peaks are 1:√2:√3:2:√5:√6, corresponding to typical BCC packing, which is consistent with the results of the XRD pattern. In addition, two bands located at 1.710 and Figure 10 are observed in the q xy direction, with distances of 3.70 and respectively. These two distances can be expressed as the π-π stacking distances of pyrene dimers in the co-assembly. Based on this, it is hypothesized that the inversion of the chiral signal of the assembly may be caused by different arrangement angles of the AP dimers. To verify this hypothesis, AP dimers A and B with different stacking angles are constructed; the structures of these dimers are optimized using the b3lyp / 6-311g(d) basis set to obtain the optimized structures. The π-π stacking distance and angle in dimer A are

[0105] and 13°, as shown in a) of ; the distance and angle of dimer B are Figure 11 and 14°, as shown in b) of Figure 11 . Through electronic circular dichroism (ECD) calculations based on time-dependent density functional theory (TDDFT), the structural basis for the inversion of the chiral signal of the co-assemblies formed in the vial system and the microfluidic laminar flow chip is further understood, as shown in c) of Figure 11 . Combining the above experiments and theoretical calculations, it can be concluded that the inversion of the chiral optical signal may be due to the opposite angles of the AP dimers formed in the vial and the microfluidic laminar flow chip.

[0106] As described above, it is only the specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention can easily think of changes or substitutions, which should all be covered within the protection scope of the present invention.​

Claims

1. A method for regulating the chiral signal of a peptide-based co-assembly by microfluidics, characterized in that, It includes the following steps: Step 1: Construct a microfluidic laminar flow chip, which includes an assembly channel, an inner channel, and an outer channel. Among them, the inner channel includes a first inner channel, a second inner channel, and a confluence channel. One end of the first inner channel and the second inner channel are respectively connected to one end of the confluence channel and form a Y-shaped structure; the other end of the confluence channel is connected to one end of the assembly channel, and the other ends of the first inner channel and the second inner channel respectively form a first inner inlet and a second inner inlet; The outer channel includes a first outer channel and a second outer channel. The first outer channel and the second outer channel are respectively arranged on both sides of the inner channel. One end of the first outer channel and the second outer channel are respectively connected to one end of the assembly channel, and the other ends of the first outer channel and the second outer channel respectively form a first outer inlet and a second outer inlet; Step 2: Prepare a first DMF mother liquor of Fmoc-amino acid and a second DMF mother liquor of 1-aminopyrene; Step 3: Input the first DMF mother liquor and the second DMF mother liquor into the microfluidic laminar flow chip from the first inner inlet and the second inner inlet respectively, and input the aqueous solution into the microfluidic laminar flow chip from the first outer inlet and the second outer inlet respectively. The first DMF mother liquor, the second DMF mother liquor, and the aqueous solution form a laminar flow and assemble in the assembly channel to obtain a peptide co-assembly; Step 4: Age the peptide co-assembly to obtain an assembly with an opposite chiral optical signal compared with the vial open system.

2. The method for regulating the chiral signal of the peptide-based co-assembly by microfluidics according to claim 1, wherein The assembly channel is a straight channel; the confluence channel is a straight channel, and the confluence channel is docked with the assembly channel.

3. The method for regulating the chiral signal of the peptide-based co-assembly by microfluidics according to claim 1, wherein The assembly channel is a square channel, the width of the assembly channel is 300 μm, and the height of the assembly channel is 100 μm; The outer channel is a square channel, the width of the outer channel is 300 μm, and the height of the outer channel is 100 μm; The confluence channel is a square channel, the width of the confluence channel is 300 μm, and the height of the confluence channel is 100 μm.

4. The method for regulating the chiral signal of the peptide-based co-assembly by microfluidics according to claim 1, wherein The first outer channel and the second outer channel are symmetrically arranged on both sides of the inner channel; And / or, the first inner channel and the second inner channel are symmetrically arranged.

5. The method for regulating the chiral signal of the peptide-based co-assembly by microfluidics according to claim 1, wherein, The first outer channel includes a straight segment and an arc segment. The straight segment is connected to the arc segment, and the arc segment is connected to the assembly channel. The straight segment is parallel to the assembly channel, and the arc segment adopts a circular arc structure; The second outer channel includes a straight segment and an arc segment. The straight segment is connected to the arc segment, and the arc segment is connected to the assembly channel. The straight segment is parallel to the assembly channel, and the arc segment adopts a circular arc structure; The first outer channel and the second outer channel are respectively vertically connected to the assembly channel.

6. The method for regulating the chiral signal of the peptide-based co-assembly according to claim 1, wherein In the step 2, the Fmoc-amino acid is Fmoc-L-Ala or Fmoc-D-Ala.

7. The method for regulating the chiral signal of the microfluidic-regulated peptide-based co-assembly according to claim 1, wherein, In the step 2, the concentration of the first DMF mother liquor is 8-32 mM; the concentration of the second DMF mother liquor is 8-32 mM.

8. The method for regulating the chiral signal of the peptide-based co-assembly by microfluidics according to claim 7, wherein In the step 2, the concentration of the first DMF mother liquor is 20 mM, and the concentration of the second DMF mother liquor is 20 mM.

9. The method for regulating the chiral signal of the microfluidic-regulated peptide-based co-assembly according to any one of claims 1-8, characterized in that, In step (3), the input flow rate at the first inner inlet is controlled to be 1 - 4 μL / min; the input flow rate at the second inner inlet is controlled to be 1 - 4 μL / min; The input flow rate at the first outer inlet is controlled to be 4 - 16 μL / min; the input flow rate at the second outer inlet is controlled to be 4 - 16 μL / min.

10. The method for regulating the chiral signal of the peptide-based co-assembly by microfluidics according to claim 9, characterized in that, In step (3), the input flow rate at the first inner inlet is controlled to be 2 μL / min; the input flow rate at the second inner inlet is controlled to be 2 μL / min; the input flow rate at the first outer inlet is controlled to be 8 μL / min; the input flow rate at the second outer inlet is controlled to be 8 μL / min.

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