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Chromatographic method for improving recovery rate of target protein

A technology of target protein and recovery rate, applied in the field of biopharmaceuticals, can solve the problems of low recovery rate and optimization of target protein recovery rate, and achieve the effect of improving recovery rate, easy commercialization and large-scale application, and shortening man-hours.

Active Publication Date: 2021-03-02
YICHANG HEC CHANGJIANG PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main purpose of the above method is to remove impurities or improve resolution, and it is not optimized for the recovery rate of the target protein, so the recovery rate is low

Method used

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  • Chromatographic method for improving recovery rate of target protein
  • Chromatographic method for improving recovery rate of target protein
  • Chromatographic method for improving recovery rate of target protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A chromatographic method for improving the recovery rate of target protein, said method comprising the following steps:

[0037] S1: Sample solution treatment: use the solution containing the target protein as the raw material, add salts, adjust the pH value, and obtain the processed sample solution;

[0038] S2: Carry out adsorption chromatography: take the chromatography medium and fill the chromatography column, the filling height is 28cm;

[0039] S3: controlling the temperature of the sample solution after the treatment, and performing a chromatographic operation.

[0040] The target protein in the step S1 is insulin glargine precursor.

[0041] Preferably, the chromatography medium in the step S2 is an affinity chromatography filler.

[0042] Preferably, the salts in the step S1 are magnesium sulfate with different concentrations.

[0043] Preferably, in the step S1, the pH value is adjusted to 4.8, and the alkali used for adjustment is 4 mol / L sodium hydroxide...

Embodiment 2

[0056] A chromatographic method for improving the recovery rate of target protein, said method comprising the following steps:

[0057]S1: Sample solution treatment: use the solution containing the target protein as the raw material, add salts, adjust the pH value, and obtain the processed sample solution;

[0058] S2: Carry out adsorption chromatography: take the chromatography medium and fill the chromatography column, the filling height is 28cm;

[0059] S3: controlling the temperature of the sample solution after the treatment, and performing a chromatographic operation.

[0060] The target protein in the step S1 is insulin glargine precursor.

[0061] Preferably, the chromatography medium in the step S2 is an affinity chromatography filler.

[0062] Preferably, the salt in the step S1 is magnesium sulfate, and the added amount is 0.15 mol / L.

[0063] Preferably, in the step S1, the pH value is adjusted to 4.8, and the alkali used for adjustment is 4 mol / L sodium hydrox...

Embodiment 3

[0076] A chromatographic method for improving the recovery rate of target protein, said method comprising the following steps:

[0077] S1: Sample solution treatment: use the solution containing the target protein as the raw material, add salts, adjust the pH value, and obtain the processed sample solution;

[0078] S2: Carry out adsorption chromatography: take the chromatography medium and fill the chromatography column, the filling height is 28cm;

[0079] S3: controlling the temperature of the sample solution after the treatment, and performing a chromatographic operation.

[0080] The target protein in the step S1 is insulin glargine precursor.

[0081] Preferably, the chromatography medium in the step S2 is an affinity chromatography filler.

[0082] Preferably, the salt in the step S1 is magnesium sulfate, and the added amount is 0.15 mol / L.

[0083] Preferably, in the step S1, different pH values ​​of the sample solution are adjusted.

[0084] Preferably, in the ste...

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Abstract

The invention relates to a chromatographic method for improving the recovery rate of a target protein. The method comprises the following steps: using a solution containing a target protein as a raw material, adding salts, regulating the pH value to obtain a treated sample solution, filling a chromatographic column with a chromatographic medium, controlling the sample loading temperature of the treated sample solution, and carrying out chromatographic operation. According to the method, the salt is added into the sample, and the pH value and temperature of the sample solution are adjusted, sothat the binding environment of the target protein and the chromatography medium is improved, the capture efficiency of the sample is improved, and the recovery rate is up to 95% or above.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and relates to a chromatography method for improving the recovery rate of a target protein. Background technique [0002] Protein drugs are one of the important development directions in the field of biopharmaceuticals. To study or prepare protein drugs requires a suitable protein separation and purification technology to recover the target protein from the solution containing the target protein. Protein separation and purification is to use the inherent similarity and difference between different proteins to remove protein and non-protein impurities. Chromatography based on adsorption mechanism is one of the most commonly used protein separation and purification methods. [0003] Adsorption chromatography is a purification technique that separates samples through adsorption and desorption between the stationary phase and the mobile phase. The adsorption force usually includes: the e...

Claims

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Application Information

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IPC IPC(8): C07K1/22C07K1/18C07K1/20C07K14/62C12N9/76
CPCC07K1/22C07K1/18C07K1/20C07K14/62C12N9/6427C12Y304/21004
Inventor 李爽贾永峰苏周刘晔戴祖冰
Owner YICHANG HEC CHANGJIANG PHARMA CO LTD