Method for preparing high-purity rhNGF
A high-purity, cell culture technology, applied in the preparation methods of peptides, chemical instruments and methods, growth factors/inducing factors, etc., can solve the problems of high equipment requirements, unfavorable large-scale industrial amplification, and unfavorable large-scale industrial production, etc. achieve the effect of improving efficiency
Inactive Publication Date: 2018-07-03
XINTRUM PHARMA
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Problems solved by technology
The linear gradient elution method usually requires a dual-pump chromatography system, which requires high equipment and is not conducive to large-scale industrial production.
[0008] None of the above methods involve the removal of N-terminal truncation and abnormal variants, and the use of stage dynamic cleaning to remove precursor variants, and the chromatography step uses a linear gradient elution method, which is not conducive to large-scale industrial scale-up
[0009] Although each chromatography method in the prior art can remove some related impurities in rhNGF, a single method cannot satisfactorily remove all major impurities
Method used
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Embodiment 1
[0116] Example 1 Purification of rhNGF by Cation Exchange Chromatography and Hydrophobic Interaction Chromatography
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Abstract
The invention provides a method for preparing high purity rhNGF from CHO cell culture, which combines cation- exchange chromatography and hydrophobic interaction chromatography. The method is characterized in that a cleaning step is added before chromatography by the two methods. Experiments prove that by the purification method, rhNGF precursor molecular variants can be removed, the content of N-terminal truncated and abnormal variants in purified products can be greatly reduced. The product has high purity and can meet the requirement of clinical application.
Description
Technical field: [0001] The invention relates to a preparation method of high-purity rhNGF, in particular to a method for obtaining high-purity rhNGF from CHO cell culture. Background technique: [0002] RhNGF expressed by genetic engineering and produced in host cells usually contains a variety of impurities, including host cell proteins, nucleic acids, and variants of the expression product rhNGF, such as precursors, N-terminal truncations, and abnormalities; Impurities of various organic and inorganic components such as endotoxins, viral contaminants, and cell culture media components will be present. Among them, N-terminal truncation and abnormal variants and precursors are the most critical impurities that affect the quality of recombinant human nerve growth factor and must be removed. [0003] The physical and chemical properties of these impurities vary, and the most effective removal methods also vary. [0004] Currently, reports on the purification of rhNGF includ...
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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/48C07K1/20C07K1/18
CPCC07K14/48C07K1/18C07K1/20C07K1/36
Inventor 刘文超孙洪亮张怡
Owner XINTRUM PHARMA
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