A kind of monoclonal antibody e44 against Rift Valley fever virus and its application

A monoclonal antibody, Rift Valley fever technology, applied in the field of microbiology and immunology, to achieve good stability, good neutralization protection effect

Active Publication Date: 2022-08-02
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004]Currently, no Rift Valley fever vaccines and neutralizing monoclonal antibodies at home and abroad have been approved for marketing

Method used

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  • A kind of monoclonal antibody e44 against Rift Valley fever virus and its application
  • A kind of monoclonal antibody e44 against Rift Valley fever virus and its application
  • A kind of monoclonal antibody e44 against Rift Valley fever virus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Screening and preparation of anti-RVF monoclonal antibodies

[0038] 1.1 Packaging of human recombinant adenovirus type 5 (HAdV5-GnGcopt) expressing Rift Valley fever virus GnGc protein

[0039] The recombinant GnGc protein-encoding gene (GenBank: DQ380208.1) was cloned into the expression vector pDC316, and then co-transfected with the backbone plasmid (pBHGlox_E1, Cre) of the AdMax adenovirus system into HEK293 cells. After inoculation into 293F cells for propagation, the virus titer was measured and stored at -80°C for use (see CN105483140A for the construction method).

[0040] 1.2 Immunization of rhesus monkeys

[0041] First, 2 mL of rhesus monkey blood was collected intravenously before immunization and the serum was separated as a negative serum control. At the same time, 1×10 8Rhesus monkeys were immunized with IFUs of HAdV5-GnGcopt by intramuscular injection. Rhesus monkeys were immunized again with the same dose of HAdV5-GnGcopt on the 28th day a...

Embodiment 2

[0151] Example 2. Pharmacodynamic studies

[0152] 2.1 Determination of Neutralizing Antibody Affinity

[0153] Protein A chip was used to detect the affinity of E44 and Gn protein by Biacore T200 instrument. The antibody was first diluted to 0.5 μg / mL, and the antibody was captured by the Protein A chip at a flow rate of 10 μL / min. The Gn protein was then diluted to 320nM, 160nM, 80nM, 40nM and 20nM and passed through the E44-captured Protein A chip at a flow rate of 30μL / min and bound for 120s to determine the binding rate (K a ), and then dissociated for 600 s to determine the dissociation rate (K d ). Finally, the affinity of the antibody to be tested is calculated by the ratio of Ka and Kd (K D ). Finally, the affinity between E44 and Gn protein was measured to be 2..58nM. The results are shown in Figure 13 As shown in Table 16, it can be seen from the graph that the binding speed of E44 and Gn protein is faster and the dissociation speed is slower, so the affinity...

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Abstract

The invention discloses a monoclonal antibody against Rift Valley fever virus. The antibody is obtained by screening with flow sorting-single cell PCR technology and has a unique CDR partition. The invention also discloses that the antibody is used in the preparation of Rift Valley fever. Application of heat therapy drugs. The monoclonal antibody disclosed in the invention has high-efficiency and specific anti-RVF activity, and also has the characteristics of high expression, high degree of humanization and good stability, and is suitable for industrial production.

Description

technical field [0001] The invention discloses an antibody, which belongs to the fields of microbiology and immunology. Background technique [0002] Rift Valley fever is a zoonotic mosquito-borne infectious disease caused by the Rift Valley fever virus, named after it was first isolated from the Great Rift Valley in East Africa. It is mainly prevalent in the African region, and imported cases have been reported in Asia. In 2016, my country discovered the first imported case of Rift Valley fever. The potential risk of Rift Valley fever spreading across regions also poses challenges to my country's biosecurity. In 2015, 2016 and 2018, WHO released the list of viruses most in need of research, and Rift Valley fever virus is among them. In July 1997, the 7th meeting of the Ad Hoc Group of Governmental Experts of Biological Weapons States Parties included Rift Valley Fever together with Ebola and Marburg as a virus-like warfare agent against humans. The clinical symptoms of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C12N15/13C12N15/85C12N5/10A61K39/42A61P31/14
CPCC07K16/10C12N15/85A61P31/14C07K2317/56C07K2317/565C07K2317/52C07K2317/76C07K2317/92C12N2800/107A61K2039/505
Inventor 陈薇李建民郝勐于长明侯利华卞婷陈旖房婷刘树玲
Owner ACADEMY OF MILITARY MEDICAL SCI
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