Liquid phase separation equipment for separating and purifying active protein

Abstract

The utility model provides liquid phase separation equipment for separating and purifying active protein, and relates to the technical field of active protein separation and purification. The feeding assembly is composed of a peristaltic pump, a mounting frame and a stepping motor, the peristaltic pump is arranged at the top of the mounting frame, the stepping motor is arranged at the bottom of the mounting frame, firstly, under the action of the first separation assembly, different components are separated out according to the size of protein molecules, target active protein enters the transfer storage tank to be temporarily stored, and the low temperature is kept; furthermore, the solution in the transfer storage tank is conveyed into the ultrafiltration mechanism to be subjected to concentration and impurity removal operation, the solution subjected to ultrafiltration enters the second separation assembly, high-purity separation of the active protein is achieved through affinity interaction, and compared with a traditional protein separation device, the device has the advantage that the purity of the separated active protein is improved.

Description

TECHNICAL FIELD

[0001] The utility model relates to active protein separation and purification technical field especially, it is a kind of liquid separation equipment for separating and purifying active protein. BACKGROUND

[0002] In the biotechnology field, the separation and purification of active protein is crucial, active protein plays a key role in biopharmaceutical research and development, disease diagnosis reagent production and other aspects, and its separation and purification process directly affects protein quality and activity, accurate and efficient separation can provide reliable basis for subsequent research, help to promote the continuous development of biotechnology, unlock more life mystery;

[0003] Traditional separation and purification method often has many drawbacks, its operation process is complicated, lead to low efficiency, it is difficult to quickly obtain a large number of high-purity active protein, extreme conditions in the separation process, such as too high temperature, pressure or unsuitable chemical reagent, easy to change protein structure, lead to protein inactivation, thereby greatly limit its in-depth application and development in the biotechnology field;

[0004] However, single chromatographic separation technology is difficult to meet the separation requirements of high purity and high activity at one time, and the processing capacity is limited, and some simple filtration or precipitation method cannot accurately separate the protein with specific activity;

[0005] Therefore, the utility model provides a kind of liquid separation equipment for separating and purifying active protein. INVENTION CONTENTS

[0006] The utility model aims at solving the shortcomings in prior art, and provides a kind of liquid separation equipment for separating and purifying active protein.

[0007] In order to realize the above-mentioned purpose, the utility model adopts the following technical scheme: a kind of liquid separation equipment for separating and purifying active protein, comprising;

[0008] Feed assembly, feed assembly is made of peristaltic pump, mounting bracket and stepper motor, the peristaltic pump is arranged at the top of mounting bracket, and the stepper motor is arranged at the bottom of mounting bracket;

[0009] First separation assembly, the first separation assembly is made of first chromatographic separation column, first cover plate and first pump ball, the first cover plate is arranged at the top opening of first chromatographic separation column, and first pump tube is arranged between the first cover plate and first pump ball;

[0010] Transfer tank, the top of transfer tank is fixedly communicated with the bottom of first chromatographic separation column, and the bottom of transfer tank is provided with ultrafiltration mechanism;

[0011] The second separation assembly is composed of a second chromatographic separation column, a second cover plate arranged at the top opening of the second chromatographic separation column, a second pump ball, and a second pump pipe arranged between the second cover plate and the second pump ball.

[0012] As a preferred embodiment, the output end of the stepper motor is connected with the peristaltic pump through the mounting frame, and the output end of the peristaltic pump is fixedly communicated with the first chromatographic separation column.

[0013] The beneficial effect of the further scheme is that the peristaltic pump is powered under the action of the stepper motor, and the flow adjustment range of the peristaltic pump is controlled at 0.1-100 ml / min, so that the solution can be stably passed into the first separation assembly through the first silica gel pipe.

[0014] As a preferred embodiment, the transfer tank is arranged outside the first fixed frame, and the first fixed frame is arranged with a first connecting frame connected with the top of the transfer tank, and the first connecting frame is arranged with three first connecting frames arranged in a ring shape.

[0015] The beneficial effect of the further scheme is that the first chromatographic separation column is positioned under the action of the first fixed frame, so that the first chromatographic separation column is prevented from tilting or shifting during use, and the first fixed frame is fixedly connected with the top of the transfer tank under the action of the first connecting frame, so that the stability of the device during use is further improved.

[0016] As a preferred embodiment, the top of the ultrafiltration mechanism is communicated with the transfer tank, the bottom of the ultrafiltration mechanism is arranged with a micro water pump, and the micro water pump is fixedly communicated with a second silica gel pipe between the second chromatographic separation column.

[0017] The beneficial effect of the further scheme is that the solution stored in the ultrafiltration mechanism can be pumped into the second chromatographic separation column for secondary separation through the second silica gel pipe under the action of the micro water pump.

[0018] As a preferred embodiment, the collection tank is arranged outside the second chromatographic separation column and is arranged with a second fixed frame, and the outer side of the second cover plate is arranged with a second connecting frame connected with the collection tank, and the second connecting frame is arranged with three second connecting frames arranged in a ring shape.

[0019] The beneficial effect of the further scheme is that the second chromatographic separation column is fixed under the action of the second fixing frame, so that the second chromatographic separation column is prevented from tilting or displacement during use, which weakens the separation effect, and the second fixing frame is connected and fixed with the collection box through the second connecting frame, so that the stability of the device during use is improved.

[0020] As a preferred embodiment, the first pump ball is in communication with the inside of the first chromatographic separation column through the first pump pipe and the first cover plate.

[0021] The beneficial effect of the further scheme is that under the action of the first pump ball, the inside of the first chromatographic separation column is pressurized by pressing the first pump ball through the first pump pipe, so that the separation speed of the solution in the first chromatographic separation column is accelerated.

[0022] As a preferred embodiment, the second pump ball is in communication with the inside of the second chromatographic separation column through the second pump pipe and the second cover plate.

[0023] The beneficial effect of the further scheme is that under the action of the second pump ball, the inside of the second chromatographic separation column is pressurized by pressing the second pump ball through the second pump pipe, so that the separation speed of the solution in the second chromatographic separation column is accelerated.

[0024] Compared with the prior art, the device has the advantages and positive effects that:

[0025] 1. In the device, different components are separated according to the size of protein molecules under the action of the first separation assembly, and the target active protein is temporarily stored in the transfer tank and kept at low temperature, and then the solution in the transfer tank is transported to the ultrafiltration mechanism for concentration and impurity removal operation, and the solution after ultrafiltration is introduced into the second separation assembly to realize high-purity separation of the active protein through affinity, so that the purity of the active protein after separation is improved.

[0026] 2. In the device, the first pump ball or the second pump ball is pressed to pressurize the first chromatographic separation column or the second chromatographic separation column corresponding to the first pump ball or the second pump ball, so that the separation speed of the solution in the first chromatographic separation column or the second chromatographic separation column is effectively accelerated when the pressure in the first chromatographic separation column or the second chromatographic separation column is increased, thereby improving the working efficiency of the device. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 It is a front view of the liquid phase separation equipment for separating and purifying active protein according to the present application;

[0028] Figure 2 It is an exploded view of the liquid phase separation equipment for separating and purifying active protein according to the present application;

[0029] Figure 3 This is an exploded view of the feeding component in a liquid phase separation device for separating and purifying active proteins according to the present invention.

[0030] Figure 4 This is an exploded view of the first separation component in a liquid phase separation device for separating and purifying active proteins according to the present invention.

[0031] Figure 5 This is an exploded view of the second separation component in a liquid phase separation device for separating and purifying active proteins according to the present invention.

[0032] Figure Labels

[0033] 1. Feeding assembly; 11. Peristaltic pump; 12. Mounting bracket; 13. Stepper motor;

[0034] 2. First separation component; 21. First chromatographic separation column; 22. First cover plate; 23. First pump tube; 24. First pump ball;

[0035] 3. Transfer storage tank; 31. First connecting frame; 32. First fixing frame;

[0036] 4. Ultrafiltration mechanism; 41. Miniature water pump;

[0037] 5. Second separation component; 51. Second chromatographic separation column; 52. Second cover plate; 53. Second pump tube; 54. Second pump ball;

[0038] 6. Collection box; 61. Second connecting frame; 62. Second fixing frame;

[0039] 7. First silicone tube; 71. Second silicone tube. Detailed Implementation

[0040] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0041] Example 1

[0042] like Figures 1-5 As shown, this utility model provides a technical solution: a liquid phase separation device for separating and purifying active proteins, comprising;

[0043] The feeding assembly 1 is composed of a peristaltic pump 11, a mounting frame 12 and a stepping motor 13, the peristaltic pump 11 is arranged at the top of the mounting frame 12, and the stepping motor 13 is arranged at the bottom of the mounting frame 12;

[0044] The first separation assembly 2 is composed of a first chromatographic separation column 21, a first cover plate 22 and a first pump ball 24, the first cover plate 22 is arranged at the top opening of the first chromatographic separation column 21, and the first pump ball 24 is arranged between the first cover plate 22 and the first pump pipe 23;

[0045] The transfer tank 3 is fixedly communicated with the bottom of the first chromatographic separation column 21, and the bottom of the transfer tank 3 is provided with the ultrafiltration mechanism 4;

[0046] The second separation assembly 5 is composed of a second chromatographic separation column 51 and a second cover plate 52 and a second pump ball 54, the second cover plate 52 is arranged at the top opening of the second chromatographic separation column 51, the second pump ball 54 is arranged between the second cover plate 52 and the second pump pipe 53, and the bottom of the second chromatographic separation column 51 is provided with the collection box 6, the output end of the stepping motor 13 penetrates through the mounting frame 12 and is connected with the peristaltic pump 11, the output end of the peristaltic pump 11 is fixedly communicated with the first chromatographic separation column 21, the top of the ultrafiltration mechanism 4 is communicated with the transfer tank 3, the bottom of the ultrafiltration mechanism 4 is provided with a micro water pump 41, the micro water pump 41 is fixedly communicated with the second chromatographic separation column 51 through the second silica gel pipe 71, first, different components are separated according to the size of protein molecules under the action of the first separation assembly 2, the target active protein enters the transfer tank 3 for temporary storage and low-temperature preservation, further, the solution in the transfer tank 3 is transported into the ultrafiltration mechanism 4 for concentration and impurity removal operation, the solution after ultrafiltration enters the second separation assembly 5 to realize high-purity separation of the active protein through affinity, and the purity of the active protein after separation is improved.

[0047] Further, as shown in Figures 4-5 The first pump ball 24 is communicated with the inside of the first chromatographic separation column 21 through the first pump pipe 23 and the first cover plate 22, the second pump ball 54 is communicated with the inside of the second chromatographic separation column 51 through the second pump pipe 53 and the second cover plate 52, and under the action of the first pump ball 24 and the second pump ball 54, the first pump ball 24 or the second pump ball 54 can be pressed according to the needs during use to pressurize the first chromatographic separation column 21 or the second chromatographic separation column 51 corresponding thereto, at this time, when the pressure in the first chromatographic separation column 21 or the second chromatographic separation column 51 rises, the separation speed of the internal solution can be effectively accelerated, thereby improving the working efficiency of the device.

[0048] In the above scheme, there is also a problem that the first chromatographic separation column 21 tilts during use,Figure 4 As shown in the figure: in this scheme, the transfer tank 3 is located outside the first fixed frame 32, which is provided with the first fixed frame 32, and the first connecting frame 31 connected with the top of the transfer tank 3 is arranged on the first fixed frame 32. The first connecting frame 31 is provided with three, and the three first connecting frames 31 are annularly distributed. Under the action of the first fixed frame 32, the first chromatographic separation column 21 is limited, so that the first chromatographic separation column 21 does not tilt or displace during use. Under the action of the first connecting frame 31, the first fixed frame 32 is connected and fixed with the top of the transfer tank 3, further improving the stability of the device during use.

[0049] In the above scheme, the second chromatographic separation column 51 also has the problem of tilting during use, such as Figure 5 As shown in the figure: in this scheme, the collection box 6 is located outside the second chromatographic separation column 51 and is provided with the second fixed frame 62, and the outer side of the second cover plate 52 is provided with the second connecting frame 61 connected with the collection box 6. The second connecting frame 61 is provided with three, and the three second connecting frames 61 are annularly distributed. Under the action of the second fixed frame 62, the second chromatographic separation column 51 is fixed, so that the second chromatographic separation column 51 does not tilt or displace during use, which reduces the separation effect. Further, the second fixed frame 62 is connected and fixed with the collection box 6 through the second connecting frame 61, improving the stability of the device during use.

[0050] Example 2

[0051] In the utility model, the peristaltic pump 11 can accurately control the feeding speed of the active protein solution, the flow regulation range is 0.1-100ml / min, and the solution is ensured to pass through the first silica gel pipe 7 into the first chromatographic separation column 21 stably.

[0052] In the utility model, the first chromatographic separation column 21 selects a gel filtration chromatographic column, the filler is agarose gel with high resolution, the column length is 50-100cm, and the inner diameter is 2-5cm, so that the active protein can be preliminarily separated according to the size of protein molecules.

[0053] In the utility model, the transfer tank 3 adopts stainless steel material, the volume is 5-20L, has constant temperature control function, can maintain the temperature of the temporarily stored protein solution at 4-8 DEG C, and prevents protein degradation.

[0054] In the utility model, the ultrafiltration mechanism 4 has an ultrafiltration membrane molecular weight cut-off of 10-100kDa, can concentrate and further remove impurities of the protein solution, and the operating pressure is 0.1-0.5MPa.

[0055] In this invention, the second chromatographic separation column 51 is an affinity chromatographic column. The specific ligand is selected according to the target active protein. For example, for a certain enzyme protein, its specific substrate analog can be used as the ligand. The column length is 30-80cm and the inner diameter is 1-3cm, so as to achieve high-purity separation of active proteins.

[0056] In this invention, the collection box 6 is made of glass, and its inner wall is treated with silanization to reduce protein adsorption. The box is equipped with a stirring device with a rotation speed of 50-200 rpm to ensure the collected protein solution is uniform.

[0057] Working principle:

[0058] like Figures 1-5 As shown, the active protein solution is pumped by a peristaltic pump 11 through the first silicone tube 7 into the first chromatographic separation column 21 at a set flow rate. Under the action of the first chromatographic separation column 21, different components are separated according to the size of the protein molecules. The target active protein component is temporarily stored in the transfer tank 3 and kept at a low temperature. Then, the solution in the transfer tank 3 is transported to the ultrafiltration unit 4 for concentration and impurity removal. The ultrafiltered solution enters the second chromatographic separation column 51, where high-purity separation of the active protein is achieved through affinity. Finally, it is collected in the collection box 6. During the collection process, the stirring device runs continuously to ensure the homogeneity of the protein solution.

[0059] The above description is merely a preferred embodiment of the present utility model and is not intended to limit the present utility model in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments for application in other fields. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present utility model without departing from the technical solution of the present utility model shall still fall within the protection scope of the technical solution of the present utility model.

Claims

1. A liquid separation apparatus for the isolation and purification of active proteins, characterized in that, Comprise; The feeding assembly (1) is composed of a peristaltic pump (11), a mounting frame (12) and a stepping motor (13), the peristaltic pump (11) is arranged at the top of the mounting frame (12), and the stepping motor (13) is arranged at the bottom of the mounting frame (12); The first separation assembly (2) is composed of a first chromatographic separation column (21), a first cover plate (22) and a first pump ball (24), the first cover plate (22) is arranged at the top opening of the first chromatographic separation column (21), and a first pump pipe (23) is arranged between the first cover plate (22) and the first pump ball (24); The top of the transfer tank (3) is fixedly communicated with the bottom of the first chromatographic separation column (21), and the bottom of the transfer tank (3) is provided with an ultrafiltration mechanism (4); The second separation assembly (5) is composed of a second chromatographic separation column (51) and a second cover plate (52), a second pump ball (54), the second cover plate (52) is arranged at the top opening of the second chromatographic separation column (51), a second pump pipe (53) is arranged between the second cover plate (52) and the second pump ball (54), and a collection box (6) is arranged at the bottom of the second chromatographic separation column (51).

2. The liquid separation apparatus for separating and purifying active proteins according to claim 1, characterized in that: The output end of the stepping motor (13) penetrates through the mounting frame (12) and is connected with the peristaltic pump (11), and the output end of the peristaltic pump (11) is fixedly communicated with the first chromatographic separation column (21).

3. The liquid separation device for separating and purifying active protein according to claim 1, characterized in that: The first fixed frame (32) is arranged outside the transfer tank (3), the first fixed frame (32) is provided with a first connecting frame (31) connected with the top of the transfer tank (3), the first connecting frame (31) is provided with three, and the three first connecting frames (31) are arranged in a ring shape.

4. The liquid separation device for separating and purifying active protein according to claim 1, characterized in that: The top of the ultrafiltration mechanism (4) is communicated with the transfer tank (3), the bottom of the ultrafiltration mechanism (4) is provided with a micro water pump (41), and the micro water pump (41) is fixedly communicated with the second chromatographic separation column (51) through a second silica gel pipe (71).

5. The liquid separation device for separating and purifying active protein according to claim 1, characterized in that: The second fixed frame (62) is arranged outside the second chromatographic separation column (51), the second connecting frame (61) is arranged outside the second cover plate (52) and connected with the collection box (6), the second connecting frame (61) is provided with three, and the three second connecting frames (61) are arranged in a ring shape.

6. The liquid separation device for separating and purifying active protein according to claim 1, characterized in that: The first pump ball (24) is communicated with the inside of the first chromatographic separation column (21) through the first pump pipe (23) and the first cover plate (22).

7. The liquid separation device for separating and purifying active protein according to claim 1, characterized in that: The second pump ball (54) is communicated with the inside of the second chromatographic separation column (51) through the second pump pipe (53) and the second cover plate (52).

Images