Peptide fragment collecting device after high-flux in-gel enzymolysis

Abstract

The utility model relates to the technical field of peptide fragment extraction, in particular to a high-flux in-gel post-enzymolysis peptide fragment collecting device, which comprises a reaction cylinder, a storage tray, a sealing ring, a base and supporting legs, the reaction cylinder is vertically arranged, a liquid inlet is arranged at the top of the reaction cylinder, and the liquid inlet is used for being communicated with a peristaltic pump; the storage tray is detachably arranged at the bottom of the inner side of the reaction cylinder and used for placing albumen glue, and a first liquid leakage hole is formed in the storage tray in the vertical direction in a penetrating mode; the liquid inlet is connected with the peristaltic pump, so that reaction liquid can be conveniently injected into the reaction cylinder, the reaction liquid and albumen glue are mixed and reacted, then solid-liquid separation is realized through the storage tray, the separated liquid flows downwards into the test tube, manual operation steps in the whole reaction and separation process are fewer, the labor capacity is effectively reduced, and the production efficiency is improved. Therefore, the risk of artificial contamination of the sample can be reduced, and the method is suitable for treating large-batch albumen glue samples.

Description

TECHNICAL FIELD

[0001] The utility model relates to the technical field of peptide segment extraction, especially to a high-throughput gel in enzymolysis peptide segment collection device. BACKGROUND

[0002] High-throughput gel refers to protein gel after protein electrophoresis, and the process of degrading the protein retained in the gel into peptide segments under the action of various proteases, the peptide segments obtained by enzymolysis are still preserved in the gel block, at this time, it is necessary to extract the peptide segments after the protein gel enzymolysis.

[0003] For example, the invention application with publication number CN117187328A discloses a method for high-pressure circulating enzyme extraction of soybean peptides, which comprises the following steps: uniformly mixing soybean protein isolate with a specific type of complex protease, adding water and stirring to wet, placing in a percolation tank, combining a temporary storage tank and a high-pressure pump to perform high-pressure circulating enzymolysis to obtain an enzymolysis filtrate, and performing activated carbon decolorization, nanofiltration membrane desalination, concentration, and drying on the enzymolysis filtrate to obtain soybean peptides with a peptide segment ratio of more than 80% with a molecular weight of more than 2000 Dal.

[0004] Currently, the extraction of peptide segments in the gel generally requires manual repeated dehydration and resolubilization operations, and when the number of samples to be processed increases, the amount of work to be performed increases exponentially, and the repeated operations increase the risk of human sample contamination. UTILITY MODEL CONTENT

[0005] Therefore, the utility model provides a high-throughput gel in enzymolysis peptide segment collection device, which is convenient for injecting a reaction liquid into the reaction cylinder to mix and react the reaction liquid with the protein gel through the setting of the liquid inlet and the peristaltic pump connection, and then realizes solid-liquid separation through the placement of the placement disc, the downward flow of the separated liquid into the test tube, fewer human operation steps in the whole reaction and separation process, effective reduction of labor, and further reduction of the risk of human sample contamination, which is suitable for the processing of a large number of protein gel samples.

[0006] The technical scheme of the utility model is as follows:

[0007] The utility model provides a high-throughput gel in enzymolysis peptide segment collection device, which comprises a reaction cylinder, and further comprises a placement disc, a sealing ring, a base and a supporting leg, wherein,

[0008] The reaction cylinder is vertically arranged, the top of the reaction cylinder is provided with a liquid inlet, and the liquid inlet is used for connecting a peristaltic pump;

[0009] The placing disc is detachably arranged at the inner bottom of the reaction cylinder and used for placing protein glue, and a first liquid leakage hole is vertically arranged through the placing disc;

[0010] The sealing ring is arranged between the placing disc and the reaction cylinder and sleeved on the placing disc;

[0011] The base is rotationally arranged at the lower end of the reaction cylinder, the top end of the base abuts against the bottom end of the placing disc, a second liquid leakage hole is vertically arranged through the base, and the base is used for driving the second liquid leakage hole to rotate so as to selectively communicate or dislocate the second liquid leakage hole and the first liquid leakage hole;

[0012] The supporting leg is detachably arranged at the lower end of the base.

[0013] On the basis of the above technical scheme, preferably, the reaction cylinder comprises a transparent cylinder body, an end cover and a liquid inlet pipe, wherein,

[0014] The inner side of the lower end of the transparent cylinder body is provided with a sink, the upper end of the base is rotationally connected in the sink, the sealing ring is arranged in the sink and abuts against the groove wall of the sink, the upper end of the placing disc is arranged in the interior of the reaction cylinder, and the outer diameter of the upper end of the placing disc is less than or equal to the inner diameter of the reaction cylinder;

[0015] The end cover is screwed at the upper end of the transparent cylinder body;

[0016] The liquid inlet pipe is fixedly connected and vertically penetrates the end cover, and the liquid inlet is arranged at the upper end of the liquid inlet pipe.

[0017] On the basis of the above technical scheme, preferably, the outer side surface of the lower end of the transparent cylinder body and the outer side surface of the lower end of the base, and the outer side surface of the end cover are all provided with anti-skid lines.

[0018] On the basis of the above technical scheme, preferably, the side of the placing disc is provided with a ring groove, wherein,

[0019] The sealing ring is sleeved on the ring groove.

[0020] On the basis of the above technical scheme, preferably, the bottom of the base is provided with three threaded holes, wherein,

[0021] The three threaded holes are annularly arranged along the circumference of the base;

[0022] The supporting leg is provided with three, and one supporting leg is screwed on each threaded hole.

[0023] On the basis of the above technical scheme, preferably, the supporting leg comprises a supporting rod and an anti-skid ball, wherein,

[0024] One end of the support rod is screwed in the corresponding threaded hole, and the other end is fixedly connected with the anti-skid ball.

[0025] On the basis of the above technical scheme, preferably, a hemispherical protrusion is fixedly arranged on the side of the upper end of the base, a first sliding groove is vertically arranged on the inner side wall of the lower end of the transparent cylinder, and a second sliding groove is circumferentially arranged on the inner side wall of the lower end of the transparent cylinder, wherein,

[0026] The first sliding groove and the second sliding groove are arranged in a T shape and are in communication;

[0027] The protrusion selectively slides in the first sliding groove or in the second sliding groove.

[0028] On the basis of the above technical scheme, preferably, a funnel is further included, wherein,

[0029] The funnel is screwed at the bottom of the base, and the interior of the funnel is in communication with the second liquid leakage hole.

[0030] On the basis of the above technical scheme, preferably, an anti-skid protrusion is arranged on the outer surface of the funnel.

[0031] On the basis of the above technical scheme, preferably, a test tube rack is further included, wherein,

[0032] The test tube rack is placed directly below the funnel;

[0033] A test tube insertion hole is arranged at the top of the test tube rack.

[0034] The high-flux gel enzymolysis post-peptide segment collection device of the utility model has the following beneficial effects relative to the prior art:

[0035] (1) By setting the liquid inlet and the peristaltic pump connection, it is convenient to inject the reaction liquid into the reaction cylinder through the peristaltic pump, so that the reaction liquid is mixed and reacted with the protein glue in the reaction cylinder. After the reaction, solid-liquid separation is realized through the first floor liquid leakage hole and the second liquid leakage hole on the placement disc and the base. After separation, the protein glue is left on the placement disc, waiting for the next reaction. The reaction liquid flows downward into the test tube after passing through the first floor liquid leakage hole and the second liquid leakage hole, realizing the collection of the peptide segment solution. The artificial operation steps of the whole reaction and separation process are less, effectively reducing the labor amount, and further reducing the risk of sample human pollution, suitable for the processing of large quantities of protein glue samples.

[0036] (2) By setting the base for driving the second liquid leakage hole to rotate, so that the second liquid leakage hole and the first liquid leakage hole are selectively communicated or dislocated, it is convenient to control the on-off of the second liquid leakage hole and the first liquid leakage hole, and further convenient to provide sufficient reaction time for the reaction liquid and the protein glue, and improve the extraction purity of the peptide segment.

[0037] (3) A sink is provided on the inner side of the lower end of the transparent cylinder, the upper end of the base is rotatably connected to the sink, and the storage tray is detachably provided on the inner bottom of the reaction cylinder. At the same time, the end cover is screwed on the upper end of the transparent cylinder, so that the device can be disassembled and disassembled, which is convenient for later cleaning.

[0038] (4) By setting up a funnel, it is convenient to output the reaction liquid in a directional manner. At the same time, by setting up a test tube rack, it is convenient to position the test tube under the funnel, so that the reaction liquid can be injected into the test tube more accurately and is not easy to spill.

[0039] (5) By arranging the first slide groove and the second slide groove in a T-shape and connecting them, the protrusion selectively slides in the first slide groove or in the second slide groove, which facilitates limiting the rotatable angle of the base through the second slide groove and facilitates the alignment and misalignment operations of the second leakage hole and the first leakage hole. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0041] Figure 1 This is a stereoscopic diagram of a high-throughput in-gel enzymatic hydrolysis peptide collection device of the present invention;

[0042] Figure 2 It is a cross-sectional view of the reaction tube of the present invention;

[0043] Figure 3 A three-dimensional diagram of the base of the present invention;

[0044] Figure 4 A perspective view of a reaction tube according to the present invention;

[0045] Figure 5 A three-dimensional diagram of the storage tray of the present invention;

[0046] In the figure: 1. Reaction cylinder; 2. Storage tray; 3. Sealing ring; 4. Base; 5. Support leg; 6. Funnel; 7. Tube rack; 11. Transparent cylinder; 12. End cover; 13. Infusion tube; 51. Support rod; 52. Anti-slip ball; 101. Liquid inlet; 102. Sink; 103. First slide groove; 104. Second slide groove; 201. First leakage hole; 202. Ring groove; 401. Second leakage hole; 402. Threaded hole; 403. Protrusion; 701. Test tube jack. DETAILED DESCRIPTION

[0047] The following will be combined with the specific embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0048] like Figure 1 As shown, a high-throughput in-gel enzymatic hydrolysis peptide collection device of the present invention includes a reaction cylinder 1, a storage tray 2, a sealing ring 3, a base 4 and support legs 5.

[0049] The reaction cylinder 1 is vertically arranged, and a liquid inlet 101 is provided on the top of the reaction cylinder 1 . The liquid inlet 101 is used to connect to a peristaltic pump, and the peristaltic pump is used to pump the reaction liquid into the reaction cylinder 1 .

[0050] The storage tray 2 is detachably arranged at the inner bottom of the reaction cylinder 1 and is used to place the protein glue. The reaction liquid pumped into the reaction cylinder 1 is mixed and reacted with the protein glue placed on the storage tray 2. A first leakage hole 201 is vertically penetrated on the storage tray 2. The first leakage hole 201 is used to transport the reaction liquid in the reaction cylinder 1 downward.

[0051] The sealing ring 3 is provided between the storage tray 2 and the reaction cylinder 1 and is sleeved on the storage tray 2 to achieve a seal between the side of the storage tray 2 and the side of the reaction cylinder 1 to prevent leakage. Specifically, Figure 5 As shown, a ring groove 202 is provided on the side of the storage tray 2 , and the sealing ring 3 is sleeved on the ring groove 202 .

[0052] The base 4 is rotatably arranged at the lower end of the reaction cylinder 1, and the top of the base 4 is in contact with the bottom of the storage tray 2. A second leakage hole 401 is vertically provided on the base 4. The base 4 is used to drive the second leakage hole 401 to rotate so that the second leakage hole 401 and the first leakage hole 201 are selectively connected or misaligned; when drainage is required, a test tube is placed under the base 4, and the base 4 is rotated to connect the second leakage hole 401 and the first leakage hole 201 up and down. At this time, the liquid can pass through the second leakage hole 401 and the first leakage hole 201 and fall downward into the test tube to realize the collection of the peptide solution. When reaction is required, the base 4 is rotated in the opposite direction to interlace the second leakage hole 401 and the first leakage hole 201. Even if the second leakage hole 401 and the first leakage hole 201 are misaligned up and down and not connected, the reaction liquid cannot be discharged into the test tube from the second leakage hole 401.

[0053] The support legs 5 are detachably provided at the lower end of the base 4 to support the reaction cylinder 1 and facilitate the placement of test tubes below the reaction cylinder 1. Figure 3As shown, the bottom of the base 4 is provided with three threaded holes 402, wherein the three threaded holes 402 are arranged in a circumferential annular array along the base 4; as shown Figure 1 As shown, the support leg 5 is provided with three, each threaded hole 402 on the screw joint of a support leg 5; the support leg 5 comprises a support rod 51 and an anti-skid ball 52, one end of the support rod 51 is screwed into the corresponding threaded hole 402, and the other end is fixedly connected with the anti-skid ball 52; the reaction cylinder 1 is supported by three support legs 5.

[0054] In the above structure, the reaction cylinder 1 comprises a transparent cylinder 11, an end cover 12 and a liquid inlet pipe 13.

[0055] Among them, the transparent cylinder 11 is made of transparent glass, which is convenient for observing the reactants in the reaction cylinder 1, the inner side of the lower end of the transparent cylinder 11 is provided with a sink 102, the upper end of the base 4 is rotatably connected in the sink 102, the sealing ring 3 is in the sink 102, and the sealing ring 3 abuts against the groove wall of the sink 102, the upper end of the object tray 2 is arranged in the inside of the reaction cylinder 1, and the outer diameter of the upper end of the object tray 2 is less than or equal to the inner diameter of the reaction cylinder 1; in this mechanism, it is convenient to disassemble the base 4 and the object tray 2, and then it is convenient to clean, at the same time, the sink 102 is used to limit the sealing ring 3, and then the installation position of the object tray 2 is determined, when the base 4 rotates, the object tray 2 is not easy to displace, the top surface of the base 4 and the bottom surface of the object tray 2 are kept in abutment, and liquid leakage is prevented.

[0056] The end cover 12 is screwed on the upper end of the transparent cylinder 11, which is convenient to disassemble the end cover 12, and then it is convenient to clean.

[0057] The liquid inlet pipe 13 penetrates the end cover 12 along the vertical direction and is fixedly connected, the liquid inlet 101 is arranged at the upper end of the liquid inlet pipe 13, and the upper end of the liquid inlet pipe 13 is connected with the peristaltic pump through the pipeline.

[0058] In order to facilitate disassembly, the outer side surface of the lower end of the transparent cylinder 11 and the outer side surface of the lower end of the base 4, and the outer side surface of the end cover 12 are all provided with anti-skid lines, which are convenient for screwing operation and facilitate disassembly of each part.

[0059] In addition, in order to facilitate the control of the rotation angle of the base 4, as shown Figure 3 As shown, the side of the upper end of the base 4 is fixedly provided with a hemispherical protrusion 403, as shown Figure 4 As shown, a first sliding groove 103 is vertically arranged on the inner side wall of the lower end of the transparent cylinder 11, and a second sliding groove 104 is circumferentially arranged on the inner side wall of the lower end of the transparent cylinder 11, wherein the first sliding groove 103 and the second sliding groove 104 are arranged in T shape and are communicated, and as shown Figure 2As shown, the protrusion 403 selectively slides in the first sliding groove 103 or in the second sliding groove 104, when the base 4 needs to be disassembled or assembled, the protrusion 403 slides in the first sliding groove 103, when the base 4 needs to be rotated, the protrusion 403 slides in the second sliding groove 104, the second sliding groove 104 is used to limit the rotatable angle of the base 4, facilitating the alignment and misalignment operation of the second liquid leakage hole 401 and the first liquid leakage hole 201.

[0060] In addition, in order to facilitate the smooth introduction of the reaction solution into the test tube, the high-throughput in-gel enzymatic digestion peptide collection device further comprises a funnel 6 and a test tube rack 7.

[0061] The funnel 6 is screwed at the bottom of the base 4, and the inside of the funnel 6 is communicated with the second liquid leakage hole 401, and the outer surface of the funnel 6 is provided with anti-skid protrusions, facilitating the disassembly of the funnel 6 and facilitating subsequent cleaning.

[0062] The test tube rack 7 is placed directly below the funnel 6; the top of the test tube rack 7 is provided with a test tube insertion hole 701, when the test tube needs to be used, as shown, the test tube rack 7 is placed directly below the funnel 6, then the test tube end is inserted into the test tube insertion hole 701, and the tube opening of the test tube is aligned with the funnel 6. Figure 1

[0063] The use method of the high-throughput in-gel enzymatic digestion peptide collection device of the utility model is as follows:

[0064] When the peptide is extracted, the protein glue is placed on the storage tray 2, then the end cover 12 is installed, the infusion tube 13 is communicated with the peristaltic pump through the pipeline; the test tube rack 7 is placed directly below the funnel 6, then the test tube end is inserted into the test tube insertion hole 701, and the tube opening of the test tube is aligned with the funnel 6; the reaction solution is pumped into the reaction cylinder 1 through the peristaltic pump, after the reaction solution reacts with the protein glue, the first liquid leakage hole 201 and the second liquid leakage hole 401 are aligned up and down by rotating the base 4 or rotating the reaction cylinder 1 (the two can be relatively rotated), after the reaction solution passes through the second liquid leakage hole 401 and the first liquid leakage hole 201, it falls into the test tube, realizing the collection of the peptide solution; the reaction solution is repeatedly transported through the peristaltic pump, and the liquid discharge operation is repeated, which can realize the repeated dehydration and resolubilization of the protein glue, realizing multiple extraction.

[0065] The above only describes the preferred embodiment of the utility model, and does not limit the utility model, any modification, equivalent replacement, improvement, etc. within the spirit and principle of the utility model should be included in the protection scope of the utility model.​

Claims

1. A high-throughput in-gel post-digestion peptide collection device comprising a reaction cartridge (1), characterized in that: It also includes a storage tray (2), a sealing ring (3), a base (4) and a support leg (5), wherein, The reaction cylinder (1) is vertically arranged, and a top of the reaction cylinder (1) is provided with a liquid inlet (101) for communicating with a peristaltic pump; The storage tray (2) is detachably arranged at an inner bottom of the reaction cylinder (1) and used for placing protein glue, and a first liquid leakage hole (201) is vertically arranged on the storage tray (2); The sealing ring (3) is arranged between the storage tray (2) and the reaction cylinder (1) and is sleeved on the storage tray (2); The base (4) is rotatably arranged at a lower end of the reaction cylinder (1), a top end of the base (4) abuts against a bottom end of the storage tray (2), a second liquid leakage hole (401) is vertically arranged on the base (4), and the base (4) is used for driving the second liquid leakage hole (401) to rotate, so that the second liquid leakage hole (401) and the first liquid leakage hole (201) selectively communicate or are misaligned; The support leg (5) is detachably arranged at a lower end of the base (4).

2. A high-throughput in-gel post-digestion peptide collection device as claimed in claim 1, wherein: The reaction cylinder (1) comprises a transparent cylinder body (11), an end cover (12) and a transfusion pipe (13), wherein, An inner side of a lower end of the transparent cylinder body (11) is provided with a sink (102), an upper end of the base (4) is rotatably connected in the sink (102), the sealing ring (3) is in the sink (102) and abuts against a groove wall of the sink (102), an upper end of the storage tray (2) is arranged in an interior of the reaction cylinder (1), and an outer diameter of the upper end of the storage tray (2) is less than or equal to an inner diameter of the reaction cylinder (1); The end cover (12) is screwed at an upper end of the transparent cylinder body (11); The transfusion pipe (13) penetrates the end cover (12) in a vertical direction and is fixedly connected, and the liquid inlet (101) is arranged at an upper end of the transfusion pipe (13).

3. A high-throughput in-gel post-digestion peptide collection device as claimed in claim 2, wherein: Anti-skid lines are arranged on an outer side surface of the lower end of the transparent cylinder body (11), an outer side surface of the lower end of the base (4) and an outer side surface of the end cover (12).

4. A high-throughput in-gel post-digestion peptide collection device as defined in claim 2, characterized in that: A side of the storage tray (2) is provided with a ring groove (202), wherein, The sealing ring (3) is sleeved on the ring groove (202).

5. A high-throughput in-gel post-digestion peptide collection device as defined in claim 3, characterized in that: A bottom of the base (4) is provided with three threaded holes (402), wherein, The three threaded holes (402) are arranged in a circumferential annular array along the base (4); The support leg (5) is provided with three, and one support leg (5) is screwed on each threaded hole (402).

6. A high-throughput in-gel post-digestion peptide collection device as defined in claim 5, characterized in that: The support leg (5) comprises a support rod (51) and an anti-skid ball (52), wherein, One end of the support rod (51) is screwed in the corresponding threaded hole (402), and the other end is fixedly connected with the anti-skid ball (52).

7. A high-throughput in-gel post-digestion peptide collection device as defined in claim 2, characterized in that: A side of an upper end of the base (4) is fixedly provided with a hemispherical protrusion (403), a first sliding groove (103) is vertically arranged on an inner side wall of a lower end of the transparent cylinder body (11), and a second sliding groove (104) is circumferentially arranged on the inner side wall of the lower end of the transparent cylinder body (11), wherein, The first chute (103) and the second chute (104) are arranged in T shape and communicate with each other; The protrusion (403) selectively slides in the first chute (103) or the second chute (104).

8. A high-throughput in-gel post-digestion peptide collection device as defined in claim 1, characterized in that: Further comprising a funnel (6), wherein, The funnel (6) is screwed at the bottom of the base (4), and the inner part of the funnel (6) communicates with the second liquid leakage hole (301).

9. A high-throughput in-gel post-digestion peptide collection device as defined in claim 8, characterized in that: The outer surface of the funnel (6) is provided with anti-skid protrusions.

10. The high-throughput in-gel post-digestion peptide collection device according to claim 8, characterized in that: Further comprising a test tube rack (7), wherein, The test tube rack (7) is placed directly below the funnel (6); the top of the test tube rack (7) is provided with a test tube insertion hole (701).

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