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Long-acting interferon and preparation method thereof

An interferon, long-acting technology, applied in the field of biotechnology research, can solve the problems of biological activity and half-life, and achieve the effect of simple preparation method and prolonged half-life

Inactive Publication Date: 2017-08-22
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this strategy is conceivable in theory, it faces the following specific questions when designing: Can this fusion protein be expressed efficiently in E. coli? What are the best conditions for its separation, purification and tag removal? Can IFN-C2 fusion interferon bind to IgG? What is its biological activity and half-life?

Method used

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Embodiment Construction

[0023] biomaterials:

[0024] 1. pET-ELP vector: constructed in our laboratory (document: Wen-Jun Liu, Qian Wu, Bi Xu, Xin-YuZhang, Xiao-Li Xia, Huai-Chang Sun. Single-step purification of recombinant proteins using elastin-like peptide -mediated inverse transition cycling and self-processing module from Neisseria meningitides FrpC. Protein Expression and Purification, 2014, 98:18-24).

[0025] 2. Escherichia coli DH5α: imported from BD Biosciences Clontech, USA, and preserved in our laboratory.

[0026] 3. BL21(DE3) Escherichia coli: imported from Novagen, USA, and preserved in our laboratory.

[0027] 4. MDBK cells: imported from ATCC in the United States and preserved in our laboratory.

[0028] 5. Vesicular stomatitis virus (VSV): preserved by our laboratory (document: Xu Yaoxian, Zhou Xiaofeng, Liu Lide. Molecular Virology. Wuhan: Hubei Science and Technology Press, 2000.300-302).

[0029] 6. pUC57 vector: purchased from GenScript Biotechnology (Nanjing) Co., Ltd.

[...

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Abstract

The invention belongs to the field of biotechnological research, and particularly relates to long-acting interferon and a preparation method thereof. The long-acting interferon is composed of interferon and streptococcal G protein C2 structural domain, and the preparation method includes the steps of fusion expression vector, fusion interferon expression and purification, purification tag removal and fusion interferon recovery. Compared with the existing recombinant interferon, the long-acting interferon has the advantages of being simple in preparation process, low in cost, long in half-life period and the like, and can be used for the preparation of long-acting interferon for human and animals.

Description

technical field [0001] The invention relates to the field of biotechnology research, in particular to a long-acting interferon and a preparation method thereof. Background technique [0002] Interferon is an effective therapeutic drug for viral diseases and an adjuvant therapeutic drug for tumors. Most of the existing recombinant interferons are histidine-tagged fusion proteins expressed in Escherichia coli, which need to be purified by affinity columns. Affinity columns are not only expensive, but also difficult to prepare on a large scale. Elastin-like polypeptide (ELP) is a polymer molecule synthesized based on the repeating sequence of elastin. It has temperature-sensitive reversible phase transition characteristics. The temperature solution is in a condensed state. Therefore, the ELP fusion protein can not only be separated and purified by simple centrifugation at a specific temperature, but also has a purification efficiency comparable to that of affinity chromatogr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/70
CPCC07K14/555C07K2319/31C12N15/62C12N15/70C12N2800/22
Inventor 孙怀昌谭笑孙永越夏文龙张鑫宇夏晓莉
Owner YANGZHOU UNIV
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