Metal affinity fusion protein tag and its application

A fusion protein and affinity technology, applied in the field of bioengineering, can solve the problems of affecting protein activity, affecting protein refolding stability, solubility, activity reduction, etc.

Active Publication Date: 2019-06-25
INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

[0003] It has been found that the fusion of His-tag at the amino or carboxyl terminal of the protein will affect the renaturation[1], stability[2] or solubility[3] of the protein; His-tag may also affect the a

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  • Metal affinity fusion protein tag and its application
  • Metal affinity fusion protein tag and its application
  • Metal affinity fusion protein tag and its application

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Embodiment Construction

[0028] This patent provides a new short peptide sequence to replace His-tag. The sequence is selected from the amino-terminal 1-25 amino acid residues of the natural human copper ion transporter (MDHSHHMGMSYMDSNSTMQPSHHHP, SEQ ID No: 2), referred to as MCT-tag, which contains the ATCUN binding site and a methionine-rich region , and a stretch of three consecutive histidine residues. MCT-tag can be chelated with metal ions such as nickel and copper ions, so as to replace His-tag and fuse with the target protein, which is convenient for the purification of recombinant protein. MCT-tag is derived from human natural protein sequence, which can be retained on the target protein without removing MCT-tag, avoiding the complicated process of subsequent enzyme digestion and purification, improving production efficiency and reducing production costs. Unexpectedly, after MCT-tag fusion, the expression of the target protein is usually promoted, and the effect on the folding of the target...

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Abstract

The invention discloses a novel metal affinity fusion protein tag, which is derived from an extracellular amino terminal fragment of a natural human copper ion transporter, which can be used as the metal affinity tag instead of His-tag, and can be used for purifying a recombinant fusion protein; the invention also discloses a novel fusion protein fused to the tag, and the novel fusion protein hasparticular physiological and pharmacological activities.

Description

technical field [0001] The invention belongs to the field of bioengineering, and specifically relates to a novel metal-affinity fusion protein tag having affinity for metal ions, and a fusion protein thereof. Background technique [0002] Metal chelate affinity chromatography, also known as immobilized metal ion affinity chromatography. This method utilizes the metal ion binding properties of some amino acids on the protein surface, such as histidine, tryptophan and other residues, to separate and purify proteins through the adsorption of solid-phase substrates. Histidine tag (His-Tag), as a synthetic sequence, can be used for the fusion expression of foreign proteins to facilitate the purification of recombinant proteins. A peptide composed of continuous histidine residues (usually 6-10 histidines) is fused to the amino-terminal, carboxy-terminal or middle of the target protein, which can be expressed in various expression systems (bacteria, yeast and mammalian cells) suc...

Claims

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Application Information

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IPC IPC(8): C07K14/47C07K19/00C07K1/22C12N15/12C12N15/62
Inventor 王楠王燕潘微彤雷琼沈苍颉宋晓涵
Owner INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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