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Bifunctional phage and uses thereof

A bacteriophage, functional technology, applied in the field of nanomedicine, can solve the problems of displaying targeted functional peptides at the same time, untried and other problems, and achieve the effect of overcoming the lack of targeting and inhibiting the formation of new blood vessels

Inactive Publication Date: 2017-06-30
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many current studies only use its targeting, but have not attempted to display both targeting functional peptides and therapeutic functional peptides on a phage

Method used

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  • Bifunctional phage and uses thereof
  • Bifunctional phage and uses thereof
  • Bifunctional phage and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1. Biological panning:

[0016] 1.1. Use 0.1M NaHCO3 coating solution to dilute VEGF protein into four gradient concentrations of 20g / mL, 4g / mL, 0.8g / mL, and 0.16g / mL.

[0017] 1.2. Incubate 400 μL of protein solution with a concentration of 20 g / mL in a 24-well cell culture plate overnight at 4°C.

[0018] 1.3. Discard the protein solution, add 500 μL of 5 mg / mL BSA solution, block at 37°C for 2 hours; discard the blocking solution.

[0019] 1.4. Add 500 μL of 5% Tween-20 in PBST solution and incubate for 6 minutes, discard and repeat 6 times.

[0020] 1.5, will contain 10 10 400 μL PBS of the pfu phage library was added as input, blocked for 1 hour at 37°C, and the supernatant was discarded.

[0021] 1.6. Add 500 μL of 5% Tween-20 in PBST solution and incubate for 6 minutes, discard and repeat 6 times to wash away unbound and non-specifically bound phages.

[0022] 1.7. Add 400 μL of 0.1M HCl with pH=2.2 and incubate for 5-10 minutes to elute the specifically boun...

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Abstract

The present invention discloses a bifunctional phage and uses thereof. According to the present invention, a WR polypeptide capable of specifically recognizing vascular endothelial growth factor (VEGF) is successfully obtained through phage biopanning, and the WR polypeptide and a tumor-targeting recognition peptide are simultaneously displayed on filamentous phage to form a bifunctional phage, wherein the phage can specifically recognize tumor and can also block the entering of the VEGF secreted by tumor cells into vascular endothelial cells around tumor so as to inhibit the formation of new blood vessels around the tumor; with the technology, the tumor cells cannot proliferate due to insufficient nutrients and cannot successfully pass through the capillaries, such that the tumor metastasis cannot be achieved; and the bifunctional phage overcomes the disadvantages of deficiency in targeting, high cost, complex preparation and the like of the traditional targeting drugs.

Description

technical field [0001] The invention belongs to the field of nanomedicine and relates to a bifunctional phage and its application in inhibiting tumor growth. Background technique [0002] In 1979, Dr. Folkman of Harvard University proposed the famous tumor angiogenesis hypothesis, which was accepted after nearly 30 years of research. The hypothesis holds that tumors highly express a series of endogenous angiogenic factors under conditions such as hypoxia, and quiescent endothelial cells are activated by these cytokines and undergo cell proliferation, migration and arrangement into a luminal structure. Tumor volume growth and metastasis need to rely on these new blood vessels to provide nutrients, remove metabolites and provide pathways. This hypothesis has been proved by many experiments: the growth of tumors on chick embryonic chorion (CAM) is limited in the avascular stage, but grows rapidly after vascularization; the volume of subcutaneously transplanted tumors in mice i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01A61K47/69A61P35/00
CPCC12N7/00C12N2795/14121C12N2795/14142C12N2795/14145
Inventor 毛传斌李龑杨明英
Owner ZHEJIANG UNIV
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