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Isoelectric protein purification device and method

A purification device and protein technology, applied in chemical instruments and methods, peptide preparation methods, organic chemistry, etc., can solve the problems of cumbersome steps and difficult control, and achieve simple control process, simple composition and structure, and large protein loading Effect

Pending Publication Date: 2017-10-13
赖兵
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This leads to cumbersome and difficult-to-control steps

Method used

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  • Isoelectric protein purification device and method
  • Isoelectric protein purification device and method
  • Isoelectric protein purification device and method

Examples

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Embodiment 1

[0057] Such as figure 1 , figure 2 As shown, this embodiment discloses a protein isoelectric purification device, which includes an electrophoresis tank 8, a power supply 7, a liquid storage tank 10 and a waste liquid tank 11; the electrophoresis tank 8 is a square straight tank made by splicing organic glass plates, The joints of the plexiglass plates are bonded and sealed with transparent silicone rubber.

[0058] Preferably, the power supply 7 is a 15V voltage regulating switching power supply.

[0059] Preferably, the thickness of the plexiglass plate is 3mm, and the overall size of the electrophoresis tank 8 is 120mm×22mm×30mm.

[0060] The inner surface of the electrophoresis tank 8 is longitudinally provided with two rows of chutes, and the chute is provided with a porous filter membrane 4 made of porous material; the porous filter membrane 4 divides the electrophoresis tank 8 into a sample pool 3, a positive pole electrode pool 101 and a negative pole Electrode cel...

Embodiment 2

[0082] This example discloses a protein isoelectric purification method, and the device used in this example is the same as that in Example 1.

[0083] When using this set of equipment to purify proteins, the specific methods are as follows:

[0084] Step 1: Dissolve bovine hemoglobin in Na 2 HPO 4 / NaH 2 PO 4 (50mM, pH6.8) buffer solution to prepare a 5mg / mL solution, and add it to the sample cell 3;

[0085] Step 2: Add Na to the positive electrode pool 101 and the negative electrode pool 1 respectively 2 HPO 4 / NaH 2 PO 4 (50mM, pH6.8) buffer;

[0086] Step 3: Turn on the power supply 7 for 10 minutes of electrophoresis purification;

[0087] In this embodiment, the isoelectric point of bovine hemoglobin is about 6.8, and bovine hemoglobin is almost uncharged at this pH. There was no obvious accumulation of bovine hemoglobin in the positive and negative electrode cells 1 at each time point.

Embodiment 3

[0089] This example discloses a protein isoelectric purification method, and the device used in this example is the same as that in Example 1.

[0090] When using this set of equipment to purify proteins, the specific methods are as follows:

[0091] Step 1: Dissolve bovine hemoglobin in TrisHCl (50mM, pH8.5) buffer solution to prepare a 5mg / mL solution, and add it to the sample pool 3;

[0092] Step 2: adding TrisHCl (50mM, pH8.5) buffer solution to the positive electrode pool 101 and the negative electrode pool 1 respectively;

[0093] Step 3: Turn on the power supply 7 for 10 minutes of electrophoresis purification;

[0094] In this embodiment, the isoelectric point of bovine hemoglobin is about 6.8, and bovine hemoglobin is negatively charged in TrisHCl buffer solution with pH 8.5, and it migrates toward the positive electrode in electrophoresis.

[0095] The result of the electrophoretic purification of the positive electrode pool 101 under the environment of pH 5.0 is:...

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Abstract

The invention discloses a method and device for carrying out protein purification on the basis of isoelectric point difference of proteins. A principle of the method and the device is as follows: during electrophoresis, a target protein hardly swims in a buffer solution, of which isoelectric points are the same as those of the target protein. Impurity proteins swim to the positive pole or negative pole of an electrophoresis bath due to negative charges or positive charges and are concentrated on electrode bath areas, so as to be separated from the target protein. A sample is isolated from positive and negative pole areas of the electrophoresis bath by adopting a microporous material, so that the target protein can be reserved in a sample pool after electrophoresis. Meanwhile, the buffer solution in an electrode bath is uninterruptedly updated during electrophoresis, so that the condition that the swimming of the impurity proteins stops and the impurity proteins and polarized accumulated ions are taken away due to pH polarization and the concentration polarization of various charged ions is avoided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein isoelectric purification device and method. Background technique [0002] Proteins are macromolecular substances that realize the functions of living bodies, including enzymes, antibodies, cytokines, structural proteins, membrane proteins, and many other types. Because proteins have a series of activities from catalytic reactions to signal transduction, their applications in scientific research, medicine, food, industry, etc. are becoming more and more extensive. [0003] One protein is often mixed with other proteins. In order to obtain pure protein for application purposes, it is often necessary to purify the protein. Taking antibodies used in pharmaceuticals as an example, their purity usually needs to be over 99%. Protein purification methods include salting out, various chromatography, ultrafiltration, and electrophoresis. Among them, salting out is a very rough me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/28
CPCC07K1/28
Inventor 周洲赖兵陈娇杨莉黄迎春翟祎星
Owner 赖兵
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