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Staphylococcus aureus fusion protein and protein expression vector and purification method thereof

A fusion protein, Staphylococcus aureus technology, applied in the field of biomedicine, can solve the problems of limiting the application and development of CD47-SIRPα immune checkpoint blocking technology, short antibody half-life, and high production cost

Inactive Publication Date: 2019-06-28
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the disadvantages of antibody therapy, such as antibody-dependent ADCC effect, short antibody half-life, and high preparation costs, the application and development of CD47-SIRPα immune checkpoint blockade technology in anti-tumor therapy are severely limited.

Method used

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  • Staphylococcus aureus fusion protein and protein expression vector and purification method thereof
  • Staphylococcus aureus fusion protein and protein expression vector and purification method thereof
  • Staphylococcus aureus fusion protein and protein expression vector and purification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0041] The following examples are only representative to clarify and understand the essence of the present invention, but do not limit the scope of the present invention in any way.

[0042] 1. Materials and Reagents

[0043] (1) Strains and plasmids

[0044] Escherichia coli DH5α, BL21 strain, pET28a and pET15b plasmids are preserved by our laboratory.

[0045] (2) Main reagents

[0046]

[0047]

[0048] (3) Main instruments and equipment

[0049]

[0050]

[0051] (4) Preparation of main solutions and reagents

[0052] 1. LB liquid culture medium

[0053] Weigh 10g tryptone, 5g yeast powder, 10g NaCl, add 800ml ddH 2 O dissolved. Adjust the pH value to 7.4 with NaOH, add ddH 2 O was adjusted to 1000ml, sterilized by high pressure steam for 20min, and stored at 4°C for later use.

[0054] 2. LB solid medium

[0055] On the basis of LB liquid medium, add agar powder with a final concentration of 15g / L, sterilize by high-pressure steam for 20min, dump the ...

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Abstract

The invention discloses fusion protein containing staphylococcus aureus SSL6 protein and a mouse granulocyte-macrophage colony stimulating factor (GM-CSF) and an expression vector and purification method thereof. The fusion protein is composed of the SSL6 (aglucon of CD47) and the GM-CSF. On one hand, it is hopeful to achieve the anti-tumor immunity regulating effect of the SSL6; on the other hand, it is hopeful to achieve the immunity optimization effect of a GM-CSF-mediated tumor microenvironment. Maturation of dentritic cells is induced, a Th1 immune response and a CD8+CTL killing effect are promoted, and thus the anti-tumor effect is synergistically achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a fusion protein comprising Staphylococcus aureus protein SSL6 and granulocyte-macrophage colony-stimulating factor GM-CSF, an expression carrier thereof, and a purification method. technical background [0002] Malignant tumor is one of the most important malignant diseases that endanger human life and health, and its morbidity and mortality are high in my country and even in the world. At present, tumor treatment is mainly through surgical resection, radiotherapy and chemotherapy. However, due to the characteristics of tumor cells that are prone to metastasis, diffusion and insensitivity to radiotherapy and chemotherapy drugs, it often leads to the failure of tumor treatment. On the basis of traditional treatment methods, emerging tumor treatment methods have emerged as the times require, such as tumor immunotherapy, which has become one of the most active field...

Claims

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Application Information

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IPC IPC(8): C07K19/00C07K1/22C12N15/62C12N15/70C12N1/21C12R1/19
Inventor 李咏生余华张潇熊浚智
Owner THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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