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Protein purification system and method

A protein purification and protein technology, which is applied in the field of protein purification, can solve problems such as inability to achieve continuous purification, and achieve the effect of substantially improving and improving purification efficiency

Active Publication Date: 2019-02-22
MABSPACE BIOSCIENCES (SUZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The first object of the present invention is to provide a protein purification system, which solves the problem that cannot be achieved between low temperature sterilization and normal temperature affinity chromatography by adding two filters and adding a heat exchanger between the two filters. The problem of continuous purification improves the purification efficiency

Method used

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  • Protein purification system and method

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Embodiment 1

[0050] A protein purification system such as figure 1As shown, the arrow in the figure indicates the flow direction of the protein solution, including the first sterilizing filter 1 (membrane pore size 0.22 μm), heat exchanger 2, second sterilizing filter 3 membrane (pore size 0.22 μm) and protein A affinity chromatography column 4.

[0051] Take Prism A (GE) packing material, Econoline 5 / 250 (Essential life) chromatography column with a diameter of 0.5cm, a column height of 20.5cm, and a column volume of 4ml as an example, and the operating flow rate is 300cm / h.

[0052] The source of the sample is the monoclonal antibody (Mab1) expressed by CHO cells. Pass through the first sterilizing filter, heat exchanger and second sterilizing filter in sequence, wherein the heat exchanger heats the protein solution to 25°C, and then enters the chromatographic column through sterilizing filtration.

[0053] The purification process in the chromatographic column is: wash the chromatogra...

Embodiment 2

[0055] A protein purification system such as figure 1 As shown, the arrow in the figure indicates the flow direction of the protein solution, including the first sterilizing filter 1 (membrane pore size 0.22 μm) connected in sequence), heat exchanger 2, the second sterilizing filter 3 (membrane pore size 0.22 μm) and protein A affinity Column 4

[0056] Taking Praesto Jetted A50 (Purolite) packing material, chromatography column Vantage 11 / 250 (Millipore), diameter 1.1cm, column height 20cm, column volume 20ml as an example, the operating flow rate is 300cm / h.

[0057] The source of the sample is the monoclonal antibody (Mab2) expressed by CHO cells. Pass through the first sterilizing filter, heat exchanger and second sterilizing filter in sequence, wherein the heat exchanger heats the protein solution to 25°C, and then enters the chromatographic column through sterilizing filtration.

[0058] The purification process in the chromatographic column is: wash the chromatographi...

Embodiment 3

[0072] MabSelect SuRe LX, chromatography column Vantage 11 / 250 (Millipore), diameter 1.1cm, column height 20cm, column volume 19.0mL, operating flow rate 220cm / h. The source of the sample is the monoclonal antibody expressed by CHO cells (No. Mab1). Rinse the chromatographic column with 0.2mol / L sodium hydroxide solution of 3CV, then use 5CV of equilibrium liquid (50mM Tris-HCl+150mM NaCl, pH7.4) to equilibrate the chromatographic column; the cell culture harvest liquid after clarification ( HCCF) is loaded onto the chromatographic column, and the loading capacity of the sample is 50g / L; the column bed is washed with 3CV of the equilibrium solution, and the intermediate cleaning buffer (50mM NaAc-HAc+1M NaCl, pH5.5, conductivity 88.5 mS / cm) to wash the column bed, then wash the column bed with 3CV equilibrium solution; then use 3.5CV of eluent (50mM NaAc-HAc, pH3.5) to elute and collect the eluate; finally use 3CV of 0.2mol Rinse the chromatography column with 1 / L sodium hydr...

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Abstract

The invention provides a protein purification system and method. A protein purification system comprises a first sterilizing filter, a heat exchanger, a second sterilizing filter and a protein A affinity column connected in series. The purification method includes that a protein solution is subjected to first filtration sterilization, heated, subjected to second filtration sterilization and then subjected to protein A affinity chromatography. The system and method solve the problem that continuous purification between low temperature sterilization and normal temperature affinity chromatographycannot be achieved by adding two filters and adding a heat exchanger between the two filters, thereby improving the purification efficiency.

Description

technical field [0001] The invention relates to the field of protein purification, in particular to a protein purification system and method. Background technique [0002] In recent years, the biopharmaceutical industry has flourished in our country. The low toxicity and side effects and high specific therapeutic efficacy of biological drugs have brought good news to the vast number of patients in our country. Nevertheless, the manufacturing process of biopharmaceuticals is still relatively backward, and the productivity is low. One of the factors limiting productivity is that batch production wastes a lot of effective production time. In contrast, continuous production can greatly improve production efficiency. Due to the adoption of new production processes often bring risks and increase investment in fixed assets. Therefore, on the basis of the existing production facilities and production methods, carrying out transformation to achieve continuous flow production can ...

Claims

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Application Information

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IPC IPC(8): C07K1/36C07K1/34C07K1/22C07K16/00
CPCC07K1/22C07K1/34C07K1/36C07K16/00
Inventor 徐志豪杨晓明汤炜张赶
Owner MABSPACE BIOSCIENCES (SUZHOU) CO LTD
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