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BTNL2 recombinant protein, preparation method thereof and application of BTNL2 recombinant protein in preparation of medicine for treating inflammatory bowel disease

A recombinant protein and drug technology, applied in the field of biomedicine, can solve the problems of persistent disease, lack of treatment methods, and increased risk of colorectal cancer, and achieve the effects of reducing enteritis, reducing mortality, and reducing weight loss in mice

Pending Publication Date: 2022-05-06
SICHUAN ACADEMY OF MEDICAL SCI SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is currently no specific and effective treatment for inflammatory bowel disease, and the condition is often protracted, and chronic inflammatory bowel disease greatly increases the risk of colorectal cancer in patients1

Method used

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  • BTNL2 recombinant protein, preparation method thereof and application of BTNL2 recombinant protein in preparation of medicine for treating inflammatory bowel disease
  • BTNL2 recombinant protein, preparation method thereof and application of BTNL2 recombinant protein in preparation of medicine for treating inflammatory bowel disease
  • BTNL2 recombinant protein, preparation method thereof and application of BTNL2 recombinant protein in preparation of medicine for treating inflammatory bowel disease

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Experimental program
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Effect test

Embodiment 1

[0042] Embodiment 1, the preparation of recombinant protein BTNL2-Fc

[0043] 1. Construction of expression vector

[0044]The full cDNA sequence of mouse BTNL2-Fc was obtained by synthesis, wherein the nucleotide sequence of the extracellular region of mouse BTNL2 (27-452) is shown in SEQ ID NO.6, and the nucleotide sequence of the recombinant protein BTNL2-Fc is shown in SEQ ID NO. Shown in .1; Utilize EcoRI and NheI to digest the pFUSE-hIgG2-Fc vector (purchased from Invivogen), excise the 720bp fragment, and reclaim the 3424bp vector fragment; synthesize the BTNL2-Fc containing EcoRI and NheI restriction sites The entire cDNA sequence was digested with EcoRI and NheI and cloned into the recovered vector fragment by T4 DNA ligase, and the formed vector was named pINFUSE-mBTNL2-hIgG2-Fc.

[0045] 2. Transfection, expression and purification

[0046] The constructed expression vector was transfected into 293F cells (1L cultured cells, 1×10 6 / ml), at 37°C, 5% CO 2 , 130rp...

Embodiment 2

[0048] Embodiment 2, the preparation of recombinant protein BTNL2-his

[0049] 1. Construction of expression vector

[0050] Obtain the full cDNA sequence of human BTNL2 by synthesis, wherein the amino acid encoded by the extracellular region is the 24th lysine to the 453rd serine of human BTNL2, the nucleotide sequence of the human BTNL2 extracellular region nucleic acid sequence (24-453) As shown in SEQ ID NO.5;

[0051] The PET28a vector was digested with BamHI and XhoI, and the complete cDNA sequence of human BTNL2 was synthesized as a template, and primers F-5'-CGGGATCCATGAAGCAGTCAGAAGACTTTAG-3'(SEQ ID NO.3), R-5'-CCGCTCGAGT CTAATGGTGATGGTGATGATGTGAGAGAGAAAAAGTTG-3'( SEQ ID NO.4) was performed to obtain the full cDNA sequence of human BTNL2-His by PCR, which was digested with BamHI and XhoI and cloned into the vector fragment by T4DNA ligase, and the formed vector was named PET28a-hBTNL2-His. The nucleotide sequence of the recombinant protein BTNL2-his is shown in SEQ I...

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Abstract

The invention provides a BTNL2 recombinant protein, a preparation method thereof and an application of the BTNL2 recombinant protein in preparation of drugs for treating inflammatory bowel disease, the BTNL2 recombinant protein comprises BTNL2-Fc which is a fusion polypeptide coded by a BTNL2 gene cell membrane extracellular region of a mouse and a heavy chain Fc structural domain of immunoglobulin IgG, the amino acid encoded in the extracellular region of the BTNL2 gene cell membrane of the mouse is from the 27th aspartic acid to the 452th serine of the BTNL2 of the mouse; or the recombinant BTNL2-his is a fusion polypeptide coded by a human BTNL2 gene cell membrane extracellular region and a histidine tag structural domain, and amino acids coded by the human BTNL2 gene cell membrane extracellular region are from 24th lysine to 453th serine of human BTNL2. The recombinant protein can greatly reduce the severity of mouse enteritis and reduce the death rate, which indicates that the recombinant protein can be used as a medicine for treating the inflammatory bowel disease.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a BTNL2 recombinant protein, its preparation method and its application in the preparation of medicines for treating inflammatory bowel disease. Background technique [0002] Inflammatory bowel disease (Inflammatory bowel disease, IBD) is a chronic autoimmune disease involving the whole colon, mainly including ulcerative colitis (Ulcerative colitis, UC) and Crohn's disease (Crohn's disease, CD). In recent years, the incidence of inflammatory bowel disease in my country has been increasing year by year, and it has become an important cause of common diseases of the digestive system and chronic diarrhea, and most of the patients are young and middle-aged, so it has been paid more and more attention. Currently, there is no specific and effective treatment for inflammatory bowel disease, and the condition is often protracted, and chronic inflammatory bowel disease greatly increas...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/64C12N15/85A61K38/17A61K47/68A61K48/00A61P1/00A61P29/00A61P1/04
CPCC07K14/47C07K16/18C12N15/64C12N15/85A61K47/6811A61K48/005A61K48/0008A61P1/00A61P29/00A61P1/04C07K2319/30C07K2319/21C07K2317/52C12N2800/107A61K38/00
Inventor 王晨辉彭倩文潘婷杜艳芸
Owner SICHUAN ACADEMY OF MEDICAL SCI SICHUAN PROVINCIAL PEOPLES HOSPITAL
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