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Simple device for preparing plasma protein products through batch adsorption, and method thereof

A plasma protein, batch adsorption technology, applied in the field of medicine and biology, can solve the problems of limiting the application of the batch adsorption method, hindering the research on the preparation process of the batch adsorption method, and the lack of matching plasma proteins.

Inactive Publication Date: 2012-11-21
BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] With the in-depth understanding of the performance of existing media and the development of specific binding new media, batch adsorption has been applied to the preparation of various plasma proteins due to its simpler processing process. However, as far as laboratory research is concerned, batch adsorption preparation Plasma protein has no matching device and corresponding method so far. This situation has limited the application of batch adsorption method in the separation and extraction of plasma protein products, and also hindered the research on the preparation process of plasma protein products by batch adsorption method.

Method used

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  • Simple device for preparing plasma protein products through batch adsorption, and method thereof
  • Simple device for preparing plasma protein products through batch adsorption, and method thereof
  • Simple device for preparing plasma protein products through batch adsorption, and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation experiment of prothrombin complex

[0026] (1) medium balance

[0027] Place the filter cloth sheet 4 on the upper opening of the reaction bottle 2, seal the upper opening with the airtight cap 1, and turn the reaction bottle 2 upside down. Weigh 0.15g of DEAE-Sephedex A50 medium, measure 80ml of balance solution, and pour it into the reaction bottle 2 at the opening of the lower end in sequence. The balance solution is sodium citrate solution with pH 6.8, containing 0.05mol / L NaCl. After mixing evenly, balance the DEAE-Sephedex A50 medium at 4-8°C. After 24 hours, the airtight cover 3 seals the lower end opening, puts the reaction bottle 2 upright, replaces the upper end airtight cover 1 with an open cover 5, inverts the reaction bottle 2, pressurizes the middle part of the reaction bottle 2, and filters out the equilibrium liquid through the filter cloth sheet 4; Put the reaction bottle 2 upright, replace the open cap 5 with the upper closed c...

Embodiment 2

[0037] Example 2: Protein C separation and purification experiment

[0038] (1) medium balance

[0039] Weigh 0.3g of DEAE-Sephedex A50 medium, and take 300ml of balance solution respectively, the balance solution is sodium citrate solution with pH 7.0, containing 0.05mol / L NaCl. According to (1) in Example 1, the DEAE-Sephedex A50 medium was equilibrated twice.

[0040] (2) Adsorption of protein C

[0041] Take 100ml of healthy human mixed plasma; shaker temperature is 4°C, rotation speed is 250rpm, 50min. The adsorption of protein C was completed according to (2) in Example 1.

[0042] (3) Miscellaneous protein washing

[0043] Take 150ml of washing solution respectively, the washing solution is sodium citrate solution with pH 7.4, containing 0.1mol / L NaCl. According to the operation of (3) in Example 1, the miscellaneous protein was washed twice, each time for 30 min.

[0044] (4) Elution of protein C

[0045] Take 50ml of the eluent, which is a sodium citrate soluti...

Embodiment 3

[0049] Example 3: protein S separation and purification experiment

[0050] (1) medium balance

[0051]Weigh 0.2g of DEAE-Sephedex A50 medium, and take 120ml of balance solution respectively, the balance solution is sodium citrate solution with pH 7.0, containing 0.075mol / L NaCl. According to (1) in Example 1, the DEAE-Sephedex A50 medium was equilibrated twice.

[0052] (2) Adsorption of protein S

[0053] Take 100ml of healthy human mixed plasma; shaker temperature is 4°C, rotation speed is 230rpm, 40min. The adsorption of protein S was completed according to (2) in Example 1.

[0054] (3) Miscellaneous protein washing

[0055] Take 40ml of washing solution respectively, the washing solution is a sodium citrate solution with pH 7.0, containing 0.1mol / L NaCl. According to the operation of (3) in Example 1, the miscellaneous protein was washed twice, each time for 30 min.

[0056] (4) Elution of protein S

[0057] Take 25ml of eluent, which is a sodium citrate solution ...

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Abstract

The invention relates to the fields of the medical and biological technologies, and discloses a simple device for preparing plasma protein products through batch adsorption, and a method thereof. The device comprises an upper end sealing cover (1), a reaction bottle (2), a lower end sealing cover (3), a filter cloth sheet (4) and an openmouthed cover (5). The method is characterized in that two ends of the reaction bottle (2) are sealed through the upper end sealing cover (1), the filter cloth sheet (4) and the lower end sealing cover (3); the addition of a material in the reaction bottle (2) is realized through opening the lower end sealing cover (3); a liquid component in the reaction bottle (2) is filtered by the filter cloth sheet (4) through replacing the upper end sealing cover (5) by the openmouthed cover (5) and through pressurizing the middle part of the reaction bottle (2); and the whole batch adsorption preparation process comprising medium balancing, plasma protein adsorption, other protein washing and target plasma protein eluting is carried out in the reaction bottle (2). The device and the method simplify equipment and operations required by the preparation of the plasma proteins through the batch adsorption, and can simply realize the efficient separation of various plasma proteins. Same-batch experiments enable the screening of various conditions of the batch-adsorption preparation technology to be completed by utilizing a plurality of the devices.

Description

technical field [0001] The invention relates to the field of medical biotechnology, and relates to a simple device and method for preparing plasma protein products by batch adsorption. Background technique [0002] Batch adsorption is a commonly used name in the plasma protein industry. It is one of the methods for preparing plasma protein products. The specific process refers to: plasma or its derivative components are rich in various plasma proteins. Under certain conditions, plasma or its derivative components and The media with specific groups are fully mixed together, so that the target plasma protein is combined with the medium, and then the medium is filtered out, and then subsequent treatments such as washing and elution are performed to separate the target plasma protein. The typical application of this preparation method at home and abroad is for the preparation of prothrombin complex. [0003] With the in-depth understanding of the performance of existing media a...

Claims

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Application Information

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IPC IPC(8): C07K1/16C07K14/755C07K14/745C07K14/435
Inventor 曹海军王宗奎张学俊马莉刁戈邓锷林方昭李长清
Owner BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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