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Purification method for recombinant human interleukin-12

A technology of interleukin and purification method, which is applied in the field of purification of recombinant human interleukin-12, which can solve the problems of incomplete renaturation, reduced titer, and loss of biological activity of the protein, and achieves low production cost and good stability , highly active effect

Inactive Publication Date: 2016-09-07
KANGLITAI BIOMEDICAL (QINGDAO) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because this method uses ammonium sulfate precipitation, it is necessary to refold the precipitated denatured rhIL-12 protein. During this process, some proteins cannot be completely refolded, or lose their biological activity, resulting in a decrease in titer.

Method used

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  • Purification method for recombinant human interleukin-12
  • Purification method for recombinant human interleukin-12
  • Purification method for recombinant human interleukin-12

Examples

Experimental program
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Effect test

Embodiment 1

[0067] Sample collection and clarification filtration

[0068] 1 ready

[0069] 1.1 Stainless steel barrels: Prepare three 20L stainless steel barrels, wash them with MQ water three times, and sterilize them at 180°C for 4-6 hours.

[0070] 1.2 Silicone tubing (silicone tubing) L / s 15: 5-7 pieces of silicone tubing, soaked in 0.2M NaOH for more than 4 hours, rinsed with tap water, and washed 3 times with MQ water. Sterilize at 121°C for 30 minutes.

[0071] 1.3 Filter: ①Millistak + HC Pod Depth Filter,D0HC media series 0.054m 2 , the pore size is 0.5-10μm, and the pressure is 0.2Mpa.

[0072] ②Millistak+HC Pod Depth Filter, A1HC media series 0.027m 2 , Pore diameter <0.5μm, withstand pressure 0.2Mpa.

[0073] 1.4 PBS: Prepare 10L of PBS, pH 7.2, and filter to sterilize.

[0074] 2 Sample collection and clarification and filtration

[0075] 2.1 Rinse the filter: Connect the inlet, outlet and exhaust port of the filter with silicone tubes, and the outlet port of the D0HC...

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Abstract

The invention relates to a purification method for recombinant human interleukin-12. The method includes the steps that cell culture fluid containing recombinant human interleukin-12 is clarified and filtered through a deep filtering system; 2, Q Sepharose HP purification is conducted; 3, Phenyl Sepharose 6FF Highsub purification is conducted; 4, SP Sepharose HP purification is conducted; 5, Superdex 200 purification is conducted. The method has the advantages of being simple in process, good in stability, uniform in product, low in production cost, environmentally friendly and high in activity and is suitable for industrial production.

Description

technical field [0001] The invention relates to a purification method of recombinant human interleukin-12 (rhIL-12). It belongs to the field of biopharmaceuticals. Background technique [0002] Interleukin 12 (IL-12), also known as cellular mycotic lymphocyte maturation factor (CLMF) or natural killer cell stimulating factor (NKSF), is a heterodimeric protein mainly produced by macrophages and B cells, etc. Somatic cytokines can improve the killing activity of NK cells and T cells, promote the response ability of specific CTL cells, promote the proliferation of activated NK cells and T cells, induce the production of cytokines such as IFN-γ, and promote the development of Th1 cells , up-regulate the expression of various cell surface molecules, and inhibit the synthesis of IgE. IL-12 will play an important role in the immunotherapy of viral diseases and parasitic diseases in terms of anti-tumor, acute radiation sickness, improving immunity, anti-tumor, tumor radiotherapy a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/54C07K1/36C07K1/34C07K1/16
CPCC07K14/5434
Inventor 赵毅姜会春黄冰
Owner KANGLITAI BIOMEDICAL (QINGDAO) CO LTD
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