CN-116333104-B - Human-mouse chimeric antibody against Dabiebanda virus Np protein and application thereof

Abstract

An antibody of human mouse chimeric anti-Dabiedada virus Np protein and application thereof, wherein the light chain variable region of the antibody has a nucleotide sequence of SEQ ID NO. 1 and an amino acid sequence of SEQ ID NO. 2, the light chain hypervariable region sequence has a nucleotide sequence of SEQ ID NO. 3, a nucleotide sequence of SEQ ID NO. 4 and an amino acid sequence of SEQ ID NO. 5, the heavy chain variable region has a nucleotide sequence of SEQ ID NO. 6 and an amino acid sequence of SEQ ID NO. 7, and the heavy chain hypervariable region has a nucleotide sequence of SEQ ID NO. 8, a nucleotide sequence of SEQ ID NO. 9 and an amino acid sequence of SEQ ID NO. 10. The antibody can specifically bind to the large allophana virus antigen (Np). The immunochromatography reagent based on the CDs/SiO 2 fluorescent nanospheres has the advantages of simplicity, rapidness, high sensitivity, strong specificity and the like, and is suitable for on-site rapid detection.

Inventors

• ZHU JIN
• DING SHOUNIAN
• WANG CHUNHUI
• ZHOU TINGTING
• LIN HONG
• YANG ZHAN

Assignees

• 中国人民解放军东部战区疾病预防控制中心

Dates

Publication Date

20260505

Application Date

20230404

Claims (4)

1. 1. The human-mouse chimeric antibody against the Np protein of the Dabiebanda virus is characterized in that the light chain variable region of the antibody has an amino acid sequence shown as SEQ ID NO. 2, and the heavy chain variable region has an amino acid sequence shown as SEQ ID NO. 7.
2. 2. Use of the antibody of claim 1 for the preparation of a medicament for diagnosing DBV virus infection.
3. 3. Use of the antibody of claim 1 for the preparation of a medicament for diagnosing fever with thrombocytopenia syndrome.
4. 4. The antibody of claim 1 modified with CDs/SiO 2 fluorescent nanospheres.

Description

Human-mouse chimeric antibody against Dabiebanda virus Np protein and application thereof Technical Field The invention belongs to the technical field of antibodies, and particularly relates to a human-mouse chimeric antibody against a Dabiebanda virus Np protein and application thereof. Background SFTS is an acute natural epidemic disease caused by Dabieda virus (Dabie bandavirus; once known as the "fever with thrombocytopenia syndrome bunyavirus (SEVERE FEVER WITH thrombocytopenia syndrome bunyavirus, DBV)", the clinical manifestations of SFTS, especially severe and fatal cases of SFTS, include viral hemorrhagic fever (viral hemorrhagic fever, VHF) with systemic symptoms including thrombocytopenia and/or leukopenia, systemic inflammatory response syndrome (systemic inflammatory response syndrome, SIRS), gastrointestinal symptoms, liver dysfunction, disseminated intravascular coagulation and multiple organ failure. Currently, no vaccine and specific drugs approved by the U.S. food and drug administration or the national food and drug administration for clinical use are available, and the main treatment means for DBV is symptomatic support treatment and treatment of complications. Ribavirin, although recommended for clinical use in china, was found to have no significant efficacy in altering clinical outcome. Inflammatory factor storm and impaired immune responses such as innate immunity, cellular immunity, humoral immunity and the like play an important role in the pathogenic process of DBV. The pathogenic mechanisms of DBV infection have not been fully understood so far, which place a great limitation on the clinical treatment of SFTS. Therefore, research on early diagnosis of DBV infection and therapeutic drugs is carried out, and the method has theoretical guiding significance for the treatment of severe SFTS. After immunization of mice with anti-large banda virus antigen, it was found that monoclonal antibodies have an immunoprotection effect on mice when challenged with virus. The result suggests that the monoclonal antibody can be used as a convenient, rapid and effective treatment means for DBV infection, and provides strong evidence for developing monoclonal antibodies for treating SFTS in the future. Therefore, for better application in clinical diagnosis and treatment, development of human-mouse chimeric anti-Dabiedada virus antibodies is extremely important. Disclosure of Invention The invention provides a human-mouse chimeric antibody against a Dabiedada virus antigen Np and application thereof, wherein monoclonal antibodies with high affinity to the DBV antigen Np are screened by monoclonal antibody technology, amino acid sequences and nucleotide sequences of heavy chain and light chain variable regions endowed with different characteristics of the antibodies are obtained, human-mouse chimeric transformation is carried out on heavy chain and light chain constant regions of the antibodies, and specific antigen binding is finally obtained. Based on CDs/SiO 2 fluorescent nanospheres, the preparation of the fluorescent-labeled anti-DBV specific labeled antibody is used for researching and developing a virus immunodetection reagent, has the advantages of simplicity, rapidness, high sensitivity, strong specificity and the like, and is suitable for on-site rapid detection. The technical scheme is that the human-mouse chimeric antibody against the Dabiebanda virus Np protein has the amino acid sequences shown in SEQ ID NO. 3, SEQ ID NO. 4 and SEQ ID NO. 5 in the light chain hypervariable region and the amino acid sequences shown in SEQ ID NO. 8, SEQ ID NO. 9 and SEQ ID NO. 10 in the heavy chain hypervariable region. The human mouse chimeric antibody against the Dabiedada virus Np protein has the amino acid sequence shown in SEQ ID NO. 2 in the light chain variable region and the amino acid sequence shown in SEQ ID NO. 7 in the heavy chain variable region. The vector is an antibody IgG1 subtype secretion eukaryotic expression vector. The application of the antibody in preparing medicines for diagnosing and treating DBV virus infection. The application of the antibody in preparing medicines for diagnosing and treating fever with thrombocytopenia syndrome. The antibody is modified by CDs/SiO 2 fluorescent nanospheres. The invention provides the human-mouse chimeric monoclonal antibody hmNP with high protection, high specificity and high affinity for Np. NP is a protein expressed by the Dabiedada virus, so the monoclonal antibody hmNP of the present invention can be used in the diagnosis, treatment and prevention research of plague. The invention prepares the murine monoclonal antibody 6B5 by taking DBV antigen Np as a target molecule, and prepares the human-mouse chimeric antibody hm NP105 on the basis of NP105. And (3) identifying the working capacity of the prepared murine monoclonal antibody NP105 and the human chimeric antibody hm NP105. Immunological detection shows that the mouse monoclonal antibod