Dialysis concentration device in novel purification process of erythrocyte membrane monoclonal antibody

Abstract

The utility model relates to the technical field of dialysis concentration devices, in particular to a dialysis concentration device in a novel purification process of an erythrocyte membrane monoclonal antibody, which comprises a circulating device, a deep filtering device and a touch screen which are connected through a pipeline, the circulating device comprises a feeding barrel, a constant flow pump, a stirrer and a sample collecting barrel, and the deep filtering device is connected with the touch screen through a pipeline. Comprising a constant flow pump, a tangential flow ultrafiltration system and a sample collecting barrel, according to the utility model, a tangential flow filtering technology is adopted, a part of liquid is pressed through a filtering membrane by a transmembrane pressure difference generated when the liquid tangentially flows through the surface of the membrane, and the other part of liquid circularly flows along with an antibody intercepted in the system; and on the other hand, a buffer solution needing to be replaced is added into the circulating device, so that the aims of dialyzing, replacing and concentrating the antibody are fulfilled, the antibody can be stably filtered, and the antibody dialyzing and concentrating efficiency in the mass production process of the erythrocyte membrane monoclonal antibody is improved.

Description

Technical Field

[0001] The utility model relates to a dialysis concentration device, in particular to a dialysis concentration device in a new purification process of red blood cell membrane monoclonal antibody. Background Technique

[0002] Anti - red blood cell membrane antibody is the first autoantibody clarified in the human body and can cause autoimmune hemolytic anemia (AIHA). Anti - red blood cell antibody causes the destruction of red blood cells, reduces the number of red blood cells, destroys the human immune function, and has great harm to the human body, which has important clinical significance in medical practice. When the antibody is put into use, there are certain requirements for the concentration of the antibody and the preservation buffer solution, which requires us to further process the antibody during purification production.

[0003] The currently used dialysis fluid replacement method is to put the dialysis bag containing the antibody into the dialysis bucket for dialysis fluid replacement. This method utilizes the principle of semi - permeable membrane. The sample solution is placed inside the bag, and this dialysis bag is immersed in the buffer solution. The antibody in the sample solution is retained inside the bag, while salts and small - molecule substances continuously diffuse and dialyze outside the bag. Then, the buffer solution in the dialysis bucket is replaced multiple times until all the buffer solution inside the bag is replaced. For antibody concentration, we usually use ultrafiltration tube centrifugation or membrane package filtration to achieve the purpose of concentration. The current concentration and filtration methods need to be carried out separately in sequence, and the concentration rate during the concentration process is slow, which is time - consuming and labor - intensive for mass - produced antibodies. Summary of the Invention

[0004] The utility model provides a dialysis concentration device in a new purification process of red blood cell membrane monoclonal antibody to solve the problems existing in the above - mentioned prior art.

[0005] The technical solution adopted by the utility model is as follows:

[0006] A dialysis concentration device in a new purification process of red blood cell membrane monoclonal antibody, including a circulation device, a depth filtration device and a touch screen, which are connected by pipelines. The circulation device includes a feed bucket, a constant - flow pump I, a stirrer and a sample collection bucket I. The depth filtration device includes a constant - flow pump II, a tangential flow ultrafiltration system and a sample collection bucket I.

[0007] Furthermore, a long shaft passing through the top of the stirrer is connected inside the stirrer. The upper end of the long shaft is connected to a motor. There are two groups of stirring impellers connected to the long shaft inside the stirrer, and the connection part of the long shaft and the stirrer is sealed by a shaft seal.

[0008] Furthermore, a membrane package is provided inside the tangential flow ultrafiltration system, and a clamp is also provided on the upper part of the tangential flow ultrafiltration system. The clamp plays a role in fixing the membrane package, and the number and area size of the membrane packages can be adjusted according to the amount of antibody to be processed.

[0009] Further, the tangential flow ultrafiltration system is provided with a discharge port. On the pipeline connecting the circulation device and the tangential flow ultrafiltration system, a pressure gauge II and a conductivity meter are successively installed.

[0010] Further, an observation window is provided on the outer side of the stirrer.

[0011] Further, the data of the pressure gauge II, the conductivity meter and the constant flow pump II are transmitted to the touch screen, and the touch screen monitors the sample conductivity, pressure and flow rate during the operation of the whole device.

[0012] Compared with the prior art, the beneficial effects of the present utility model are as follows:

[0013] By adopting the tangential flow filtration technology, the transmembrane pressure difference generated by the tangential flow of the liquid across the membrane surface presses part of the liquid through the filter membrane, while the other part of the liquid circulates with the antibody retained in the system. While the circulating liquid is performing tangential flow filtration, on the other side, the buffer solution to be replaced is added to the circulation device, and finally the purpose of dialysis fluid replacement and concentration of the antibody is achieved. The circulating flow of the liquid throughout the process not only plays the role of ultrafiltration, but also can wash the filter membrane, preventing the antibody from aggregating on the membrane surface, ensuring the stable progress of antibody filtration, and improving the efficiency of antibody dialysis and concentration during the large-scale production of monoclonal antibodies against the erythrocyte membrane. Description of the Drawings

[0014] Figure 1 is the structural diagram of the dialysis and concentration device of the present utility model.

[0015] Figure 2 is the structural diagram of the depth filtration device.

[0016] Figure 3 is the structural diagram of the circulation device. Detailed Embodiments

[0017] Next, the technical solutions in the embodiments of the present utility model will be clearly and completely described in conjunction with the drawings in the embodiments of the present utility model. Obviously, the described embodiments are only a part of the embodiments of the present utility model, rather than all of the embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without making creative efforts shall fall within the protection scope of the present utility model. Embodiment

[0018] As Figures 1-3 shown, a dialysis and concentration device in a novel purification process of monoclonal antibodies against the erythrocyte membrane of the present utility model includes a touch screen, a depth filtration device and a circulation device.

[0019] The sample conductivity, pressure and flow rate during the operation of the whole device are monitored through the touch screen.

[0020] 1. Purification of Monoclonal Antibodies Against Red Blood Cell Membrane

[0021] 1.1 Preparation:

[0022] 1.1.1 Preparation of Raw Materials and Reagents:

[0023] Raw Materials: Ascites (note clone number and batch number)

[0024] Reagents: Equilibration buffer, washing buffer, dissociation buffer, dialysis buffer, and pH neutralization solution

[0025] 1.1.2 Preparation of Equipment and Consumables:

[0026] Equipment: Peristaltic pump, automated protein purification system or nucleic acid protein detector, magnetic stirrer, balance, pH meter

[0027] Consumables: Protein A column, beaker, graduated cylinder

[0028] 1.1.3 Environmental Preparation: General clean environment, environmental temperature 25°C

[0029] 1.2 Purification of Ascites (note clone number and batch number) on Protein A column

[0030] 2. Antibody Treatment:

[0031] 2.1 Dialysis and Concentration:

[0032] 2.1.1 Preparation: Sequentially pump 0.08 - 0.14M NaOH, pure water, and storage buffer 15 - 25mM Tris-NaCl (pH 7.5) or 8 - 12mM PBS (pH 7.4) into the circulation device to rinse the entire dialysis and concentration device;

[0033] 2.1.2 Sample Loading: After dissociation, the sample is collected in the feed bucket. The feed inlet of the circulation device is the pipe opening extending into the feed bucket. The sample solution is pumped into the circulation device from the feed bucket by peristaltic pump 1. The outlet at the bottom of the circulation device is opened, and the antibody is pumped into the depth filtration device by the power of peristaltic pump 2. Through the tangential flow ultrafiltration membrane in the depth filtration device, part of the liquid is displaced. This part of the permeate flows out from the outlet, and the retained reflux liquid returns to the circulation device for continuous circulation filtration;

[0034] 2.1.3 Buffer Exchange: While loading the sample, store buffer 15 - 25mM Tris-NaCl (pH 7.5) or 8 - 12mM PBS (pH 7.4) is stored in the circulation device, and the circulating liquid enters the circulation, finally playing the role of buffer exchange;

[0035] 2.1.4 Sample collection: Collect a small amount of sample from the sample collection port of the circulation device to detect its concentration. After the concentration meets the requirements, sample collection can be carried out. After the sample collection is completed, wash the entire system with preservation buffer 15 - 25 mM Tris-NaCl (pH 7.5) or 8 - 12 mM PBS (pH 7.4) to recover the antibodies remaining in the system; Example

[0036] Under the condition that other devices remain unchanged, the tangential flow filtration membrane module in the depth filtration device can be replaced with others with different pore sizes according to the usage purpose and the particle size of the treatment object. The filter membrane with a pore size between 0.1 - 10 microns is suitable for filtering microorganisms, viruses, proteins and other fine particles; the filter membrane with a pore size between 0.001 - 0.1 microns is suitable for filtering high molecular compounds, colloids, pigments, impurities, etc.; the filter membrane with a pore size between 0.001 - 0.01 microns is suitable for filtering small molecule organic compounds, ions, high polymers, etc.

[0037] For those skilled in the art, it is obvious that the present utility model is not limited to the details of the above exemplary embodiments, and can be implemented in other specific forms without departing from the spirit or basic characteristics of the present utility model. Therefore, from any point of view, the embodiments should be regarded as exemplary and non-limiting. The scope of the present utility model is defined by the appended claims rather than the above description. Therefore, all changes falling within the meaning and scope of the equivalent elements of the claims are intended to be included in the present utility model, and any reference signs in the claims should not be regarded as limiting the claims involved.

Claims

1. A dialysis concentration device in a novel purification process of erythrocyte membrane monoclonal antibody, characterized in that: The invention comprises a circulation device (1), a deep filter device (2) and a touch screen (3), which are connected by pipelines. The circulation device (1) comprises a feed barrel (8), a constant flow pump (19), a stirrer (4) and a sample collection barrel (9). The deep filter device (2) comprises a constant flow pump (22), a tangential flow ultrafiltration system (21) and a sample collection barrel (13).

2. The dialysis concentration device in the novel purification process of red blood cell membrane monoclonal antibody according to claim 1, characterized in that: The circulation device (1) is provided with a discharge port and a feed port, and a pressure gauge (20) is installed on the pipeline connected to the feed port.

3. The dialysis concentration device in the novel purification process of erythrocyte membrane monoclonal antibody according to claim 1, characterized in that: The stirrer (4) is connected to a long shaft (17) passing through the top of the stirrer (4), the upper end of the long shaft is connected to a motor (15), the stirrer (4) has two sets of stirring impellers (16) connected to the long shaft (17), and the connection between the long shaft (17) and the stirrer (4) is sealed by a shaft seal (28).

4. The dialysis concentration device in the novel purification process of red blood cell membrane monoclonal antibody according to claim 1, characterized in that: A membrane package (27) is provided inside the tangential flow ultrafiltration system (21), and a clamp (26) is also provided on the upper part of the tangential flow ultrafiltration system (21).

5. The dialysis concentration device in the novel purification process of red blood cell membrane monoclonal antibody according to claim 1, characterized in that: The tangential flow ultrafiltration system (21) is provided with a discharge port and a feed port. A second pressure gauge (23) and a conductivity meter (25) are installed in sequence on the pipeline connecting the circulation device (1) and the tangential flow ultrafiltration system (21).

6. The dialysis concentration device in the novel purification process of erythrocyte membrane monoclonal antibody according to claim 1, characterized in that: An observation window (18) is provided on the outer side of the stirrer (4).

7. The dialysis concentration device in the novel purification process of erythrocyte membrane monoclonal antibody according to claim 1, characterized in that: The data of the second pressure gauge (23), the conductivity meter (25) and the second constant flow pump (22) are transmitted to the touch screen (3).

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