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Binding molecule for resisting WNV infection

A technology that combines molecules and nucleic acid molecules, which is applied in the field of molecular immunology and can solve the problem of no specific drugs for WNV infection.

Active Publication Date: 2021-01-01
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no specific drug for the treatment of WNV infection, and early detection is the main measure to control the spread of WNV

Method used

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  • Binding molecule for resisting WNV infection
  • Binding molecule for resisting WNV infection
  • Binding molecule for resisting WNV infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Preparation of WN-XH2 monoclonal antibody

[0077] 1. Experimental materials

[0078] PRT / KIgG1 carrier (see patent literature: A monoclonal antibody that specifically binds to PD-1, application number: 201810273628.7)

[0079] 2. Steps

[0080] (1) Using the method of gene synthesis to synthesize the antibody heavy chain variable region sequence (sequence shown in SEQ ID NO: 4), light chain variable region sequence (sequence shown in SEQ ID NO: 8), and use them Methods of Molecular Cloning Fragments were cloned into PRT / KIgG1.

[0081] (2) Transfect the antibody expression recombinant vector constructed in step 1 into 293T cells in the logarithmic growth phase, change the fresh medium 6-8 hours after transfection, and incubate at 37°C 8% CO 2 Incubate for 96 hours in an incubator. The transfection supernatant was collected, centrifuged at 4000 rpm for 1 hour, and purified by Protein A affinity chromatography. SDS-PAGE and Western Blot experiments were us...

Embodiment 2

[0084] Example 2 Determination of binding activity and neutralizing activity of WN-XH2 monoclonal antibody

[0085] 1. Determination of binding activity of monoclonal antibody

[0086] Dilute the EDIII antigen of WNV to 1 μg / mL in the coating solution, add 100 μL per well to the enzyme-linked plate, and place it in a humid chamber at 4°C overnight. Wash the enzyme-linked plate 3 times with a plate washer, block with 1.5% casein, 200 μL per well, and block for 1 hour at 37°C in a humid box. Dilute the antibody to different concentrations with 1xPBS, add 100μL per well to the enzyme-linked plate, react in a wet box at 37°C for 1h, wash the enzyme-linked plate 3 times, add goat anti-human (Fab') 2-HRP secondary antibody and react at room temperature for 45min , wash the enzyme-linked plate 5 times and add 100 μL TMB substrate for color development, react for 3 minutes and use 100 μL 2N H 2 SO 4 The reaction was terminated, and the enzyme-linked immunosorbent assay was read at ...

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Abstract

The invention discloses a binding molecule for resisting WNV infection. The binding molecule is a monoclonal antibody aiming at WNV E protein. In-vitro experiments prove that the monoclonal antibody can be specifically combined with WNV E protein, has relatively high affinity activity and can neutralize WNV viruses. On one hand, the research result provides a new method for clinical diagnosis, andon the other hand, a candidate drug is provided for clinical WNV infection resistance.

Description

technical field [0001] The invention belongs to the field of molecular immunology and relates to a combination molecule for resisting WNV infection. Background technique [0002] West Nile virus (WNV) infection can cause West Nile fever, West Nile virus encephalitis and meningitis, and is a zoonotic, natural foci, acute infectious disease. Since WNV broke out in New York, USA in 1999, the virus has spread rapidly to many countries and regions in the world, becoming a viral disease that seriously threatens human health, and has attracted the attention of the global public health community. WNV can infect a variety of mosquitoes and birds, and spread the virus along the migration path of birds through the alternate infection of mosquitoes and birds; mosquitoes can also transmit WNV to a variety of mammals by biting WNV-infected birds , such as humans, horses, dogs, cats and poultry such as chickens, geese, etc.; the virus can also be transmitted vertically through blood trans...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13G01N33/577G01N33/569A61K39/42A61P31/14
CPCC07K16/1081G01N33/577G01N33/56983A61P31/14C07K2317/565C07K2317/56C07K2317/92C07K2317/76C07K2317/35G01N2333/185G01N2469/10A61K2039/505Y02A50/30
Inventor 肖鹤冯健男沈倍奋卢星陈国江王晶乔春霞罗龙龙李新颖刘成华
Owner ACADEMY OF MILITARY MEDICAL SCI
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