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Murine original monoclonal antibody 3D8 identified hantaan virus glycoprotein neutralizing epitope peptide and application thereof

A technology of monoclonal antibody and Hantaan virus, which is applied in the direction of antiviral agents, viral antigen components, peptides, etc., can solve the problems of unclear and incompletely clear epitopes, and achieve good development and application prospects

Active Publication Date: 2012-10-17
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0007] However, the antigenic epitopes recognized by the 3G1 and 3D8 monoclonal antibodies and the mechanism of their neutralizing activity are still not completely clear. The phage display peptide library method was used to identify an epitope recognized by the 3G1 monoclonal antibody, while for the 3D8 monoclonal antibody Anti-recognized epitopes are unknown

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  • Murine original monoclonal antibody 3D8 identified hantaan virus glycoprotein neutralizing epitope peptide and application thereof
  • Murine original monoclonal antibody 3D8 identified hantaan virus glycoprotein neutralizing epitope peptide and application thereof
  • Murine original monoclonal antibody 3D8 identified hantaan virus glycoprotein neutralizing epitope peptide and application thereof

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Embodiment Construction

[0022] The peptide scanning technology used in the present invention has the advantage of being able to directly determine linear epitopes, which is achieved through the following technical solutions:

[0023]Synthesis of overlapping peptides: Synthesize overlapping peptides according to UniProtKB / Swiss-Prot database accession number: P08668-1 (Hantaan virus glycoprotein sequence, 1135aa). The length of each synthetic peptide is 15aa, and the adjacent peptides overlap by 11aa, totaling 281. Entrust Xi'an Lianmei Biotechnology Co., Ltd. to synthesize. The purity of the synthetic peptide is >85% (HPLC, MS), and the dry powder of the synthetic peptide is stored at -20°C.

[0024] The mouse-derived neutralizing antibody HTNV GP-specific monoclonal antibody 3D8 was provided by the applicant's Microbiology Department.

[0025] The B cell epitope peptides reactive with 3D8 monoclonal antibody were screened out by indirect ELISA method and Dot blot method for peptide scanning technol...

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Abstract

The invention discloses a neutralizing monoclonal antibody 3D8 identified HTNV-GP specific B cell epitope and key amino acid residue sequence of the epitope for treating hemorrhagic fever with renal syndrome (HFRS) caused by hantaan virus (HTNV) infection. The B cell epitope has an amino acid sequence of 882GFLCPEFPGSFRKKC896. The epitope peptide can be applied to study of mechanism of 3D8 neutralizing monoclonal antibody in treatment of HFRS, or preparation of drug for treating HFRS and preparation of novel vaccine strain aiming at hantaan virus 76-118, or development of novel diagnostic kitfor hantaan virus 76-118 as a target protein. The invention has good prospects of development and application in the field of HFRS specific immunotherapy.

Description

technical field [0001] The invention belongs to the technical field of prevention and treatment of Hantaan virus, and specifically relates to a specific glycoprotein (GP) epitope peptide recognized by a therapeutic monoclonal antibody 3D8 for hemorrhagic fever with renal syndrome (HFRS) and its application. Background technique [0002] Hantaan virus (HTNV) is an important virus among pathogenic microorganisms within the scope of verification of the Protocol on the Prohibition of Biological Weapons, belonging to the genus Hantavirus in the family Bunyaviridae. Hantaviruses are all single-stranded negative-sense RNA enveloped viruses. The genome includes three segments: large (L), medium (M), and small (S). The L segment encodes RNA polymerase, and the M segment encodes the envelope. Glycoprotein 1 and 2 (glycoprotein 1, G1 / Gn and glycoprotein 2, G2 / Gc), S segment encodes nucleocapsid protein (NP). The HTNV 76-118 strain first isolated by Li Haowang et al. in 1978 is the rep...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08G01N33/68A61K39/12A61P31/14A61P13/12
Inventor 张宇丝徐志凯闫果林庄然吴兴安张芳琳张赟徐竹蔚易静张春梅马樱刘蓓陈丽华杨琨宋朝君李琦方亮周幸春刘志佳金伯泉
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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