Fully human monoclonal antibody for neutralizing enterovirus 71 (EV71) and application of fully human monoclonal antibody

A monoclonal antibody and enterovirus technology, applied in the field of biological immunology, can solve problems such as adverse reactions, lack of anti-EV71 drugs, inability to prevent EV71 virus subtypes, etc., and achieve good biocompatibility

Active Publication Date: 2019-12-20
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the first half of 2016, the vaccine against EV71 virus has been approved for marketing. This is currently the only vaccine that can be used to prevent hand, foot and mouth disease. It can curb the incidence of hand, foot and mouth disease caused by EV71 virus to a certain extent, but there are some limitations : People over the age of 5, patients with fever, patients with acute and chronic disease attacks, and those with allergies should not receive the vaccine; those who are allergic to the active substance of the vaccine and kanamycin sulfate should not be vaccinated; the vaccination has individual differences and can Some adverse effects; does not protect against all genotypes of EV71 virus, not effective for patients already infec

Method used

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  • Fully human monoclonal antibody for neutralizing enterovirus 71 (EV71) and application of fully human monoclonal antibody
  • Fully human monoclonal antibody for neutralizing enterovirus 71 (EV71) and application of fully human monoclonal antibody
  • Fully human monoclonal antibody for neutralizing enterovirus 71 (EV71) and application of fully human monoclonal antibody

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Embodiment 1

[0053] Example 1: Preparation of a fully human monoclonal antibody neutralizing enterovirus 71

[0054] 1. Sorting of memory B cells

[0055] 1.1 Acquisition of peripheral blood mononuclear cells

[0056] 3.5ml of fresh blood was drawn from volunteers infected with enterovirus 71 and placed in an anticoagulant tube. After diluting the fresh anticoagulated whole blood with PBS in an equal volume, slowly add it to the lymphocyte separation medium (domestic Solarbio, Cat.NO.P8610), take it out carefully after centrifugation, and absorb the second layer of cloudy monocytes The layer was washed in a centrifuge tube filled with PBS, the supernatant was discarded by centrifugation, and then washed once with PBS, and the collected cells were peripheral blood mononuclear cells (PBMC).

[0057] 1.2 Memory B cell sorting

[0058] Centrifuge the isolated PBMCs from volunteers to remove the supernatant thoroughly, every 10 7 Resuspend the cell pellet with 40 μL buffer. Add 10μL Biotin...

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Abstract

The invention belongs to the field of biological immunology, and more particularly relates to a fully human monoclonal antibody for neutralizing enterovirus 71 (EV71) and application of the fully human monoclonal antibody. An amino acid sequence of a heavy chain variable region is shown as any one of SEQ ID NO:14-20, and an amino acid sequence of a light chain variable region is shown as any one of SEQ ID NO:21-26. The invention further provides a nucleic acid molecule encoding the antibody, an expression vector, application and a composition. Binding molecules can prevent the EV71 from infecting susceptible cells and are fully human derived, compared with other animal derived (such as rat-derived) anti-EV71 molecules, immunogenicity is low, affinity is good, the treating effect is good, and side effects are low.

Description

technical field [0001] The invention relates to the field of biological immunology, in particular to a fully human monoclonal antibody neutralizing enterovirus 71 and its use. Background technique [0002] Hand foot mouth disease (HFMD) is a common acute infectious disease of infants and young children caused by enterovirus. Small herpes or small ulcers appear in other parts, most of the children will heal by themselves in about 1 week, and a few children may cause complications such as myocarditis, pulmonary edema, aseptic meningitis, etc., and the rapid development of severe children may lead to death. Currently, there is no effective treatment. medicine. In recent decades, HFMD has broken out and become popular in many parts of the world. In 2008, my country included hand, foot and mouth disease as a category C infectious disease under statutory report management, requiring it to be reported to the Ministry of Health within 24 hours. From 2009 to 2015, my country's heal...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/13G01N33/569G01N33/577A61K39/42A61P31/14
CPCA61K2039/505A61P31/14C07K16/1009C07K2317/24C07K2317/56C07K2317/76G01N33/56983G01N33/577
Inventor 辛洪波胡萍魏强刘苏俊
Owner NANCHANG UNIV
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