A fusion protein of endothelial cell growth factor and its application in regenerative medicine

A technology of fusion protein and growth factor, applied in the field of genetic engineering, can solve the problems of loss of inherent biological activity of growth factor, the biological activity of VEGF needs to be improved, and the immobilization and release behavior of VEGF cannot be quantitatively regulated, so as to improve the anticoagulant performance. , Improve angiogenesis-inducing ability, and promote the effect of rapid endothelialization

Inactive Publication Date: 2011-11-30
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, using this method, the growth factor still exists in a soluble state and its half-life is only 4 hours, and the biological activity of VEGF still needs to be improved.

Method used

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  • A fusion protein of endothelial cell growth factor and its application in regenerative medicine
  • A fusion protein of endothelial cell growth factor and its application in regenerative medicine
  • A fusion protein of endothelial cell growth factor and its application in regenerative medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 2

[0036] The plasmid pcDNA3.1-VEGF-Fc containing the target gene constructed in Example 1 was transfected into 293F cells, the cell supernatant was collected, and the VEGF-Fc fusion protein was purified using rProtein A FF column. The purified fusion protein was detected by immunoblotting with an anti-VEGF antibody, and there were no other non-specific exposure bands in the figure, only one band similar to the expected molecular weight (46KD). This result shows that the present invention has prepared VEGF-Fc fusion protein ( image 3 ). The resulting fusion protein is used to modify the surface of the hydrophobic material to form an endothelial cell growth factor fusion protein matrix for endothelial cell culture, as shown in the schematic diagram ( Figure 4 ) shown.

Embodiment 2

[0038] Example 3

[0039] The invention detects the adhesion of endothelial cells on the surface of differently modified polystyrene culture dishes by MTT method, and then investigates the effect of VEGF-Fc fusion protein matrix on the adhesion of endothelial cells.

Embodiment 3

[0041]

[0042] Example 4

[0043] In the present invention, it is further shown by cell proliferation experiments that compared with the addition of VEGF in the solution, the addition of VEGF-Fc fusion protein in the solution can effectively promote the proliferation of endothelial cells and the VEGF-Fc fusion protein can better maintain the cells with the prolongation of culture time. proliferative activity ( Figure 6 ). We can conclude that VEGF-Fc fusion protein can effectively prolong the biological activity of VEGF in vitro, and VEGF-Fc fusion protein matrix can promote endothelial cell proliferation persistently.

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Abstract

The invention utilizes gene recombination technology to construct a fusion protein (VEGF-Fc) of endothelial cell growth factor VEGF and Fc domain of immunoglobulin G. On the one hand, the fusion protein can better maintain the biological activity of VEGF and prolong its half-life; The specific affinity of endothelial cells can promote the adhesion and proliferation of endothelial cells and induce the directional differentiation of stem cells to endothelial cells, and improve the biological activity of anticoagulation and induction of angiogenesis of materials.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to the preparation of a fusion protein (VEGF-Fc) of endothelial cell growth factor (VEGF) and immunoglobulin Fc (Fc) by gene recombination technology, and the fusion protein can be used as stromal endothelial cell growth Factors are used in tissue engineering, regenerative medicine-related large-scale cell expansion and endothelial / vascular induced repair and other applied research. [0002] Background technique [0003] Anticoagulation and vascularization of materials have always been one of the bottlenecks in tissue engineering and regenerative medicine research, especially for regenerative tissues with a size of more than 200 μm, insufficient vascularization will lead to insufficient supply of oxygen and nutrients to cells lead to apoptosis in tissue [1]. In order to solve this problem, some experts proposed that on the one hand, blood vessels can be transplanted in v...

Claims

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Application Information

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IPC IPC(8): C12N15/79C07K19/00C12M3/00C12N5/071
Inventor 杨军于美华杜凤移
Owner NANKAI UNIV
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