CN-122011174-A - Monoclonal antibody NP8 of anti-influenza A virus NP protein and application thereof

Abstract

The invention discloses a monoclonal antibody NP8 of an anti-influenza A virus NP protein and application thereof, belonging to the technical field of monoclonal antibodies. The monoclonal antibody NP8 is obtained by screening through ELISA detection and sequencing analysis. NP8 comprises light chain CDR2 with the amino acid sequence LVS, light chain CDR1 and CDR3 with the amino acid sequences shown in SEQ ID NO.12-13 and heavy chain CDR1-CDR3 shown in SEQ ID NO. 16-18. The invention lays a foundation for preparing the influenza A virus detection kit.

Inventors

• ZHANG XIUWEN
• LIU PENG
• LIANG YAN

Assignees

• 保定国兰生物技术有限公司

Dates

Publication Date

20260512

Application Date

20250319

Claims (7)

1. 1. The monoclonal antibody NP8 of the anti-influenza A virus NP protein is characterized in that the light chain of the monoclonal antibody NP8 comprises a light chain CDR1 with an amino acid sequence shown as SEQ ID NO.12, a light chain CDR2 with an amino acid sequence of LVS and a light chain CDR3 with an amino acid sequence shown as SEQ ID NO. 13; The heavy chain of the monoclonal antibody NP8 comprises a heavy chain CDR1 with an amino acid sequence shown as SEQ ID NO.16, a heavy chain CDR2 with an amino acid sequence shown as SEQ ID NO.17 and a heavy chain CDR3 with an amino acid sequence shown as SEQ ID NO. 18.
2. 2. The monoclonal antibody NP8 of claim 1 wherein the amino acid sequence of the light chain variable region of the monoclonal antibody NP8 is shown in SEQ ID No.11 and the amino acid sequence of the heavy chain variable region of the monoclonal antibody NP8 is shown in SEQ ID No. 15.
3. 3. A hybridoma cell line (Mus museuus) NP8 secreting the monoclonal antibody NP8 of claim 1 or 2, wherein the hybridoma cell line NP8 is deposited with the China general microbiological culture collection center for which the preservation time is 2024, 7 months and 16 days, and the preservation address is North Star west line No. 1, no.3, and the preservation number is CGMCC No.46020 in the korean region of beijing.
4. 4. Use of the monoclonal antibody NP8 of claim 1 or 2 in the preparation of a product for detecting influenza a virus.
5. 5. The use of claim 4, wherein the product comprises an ELISA kit.
6. 6. An ELISA kit for detecting influenza a virus comprising the monoclonal antibody NP8 of claim 1 or 2.
7. 7. A method for detecting influenza a virus for non-diagnostic or therapeutic purposes, comprising the step of detecting a sample to be detected using the ELISA kit of claim 6.

Description

Monoclonal antibody NP8 of anti-influenza A virus NP protein and application thereof Technical Field The invention relates to the technical field of monoclonal antibodies, in particular to a monoclonal antibody NP8 of an anti-influenza A virus NP protein and application thereof. Background The 5 th segment of the genome of influenza A virus encodes the nucleoprotein (NP protein), which is the main internal structural protein of influenza A virus and the main component of viral nucleocapsid, and the NP protein is linked with viral RNA and viral polymerase (PB 1, PB2, PA) to form ribonucleocapsid complex (RNP), which plays an important role in the processes of viral replication, transcription, assembly and transportation. The NP protein is an important structural protein of the influenza A virus, has a relatively conserved sequence and high immunogenicity, and has important value for diagnosis and monitoring of the influenza A virus. Common Influenza A Virus (IAV) detection techniques include virus isolation and identification, serological methods, and molecular biological methods. The chicken embryo or cell is utilized for virus separation, is the most commonly used classical method for detecting IAV at home and abroad at present, is also the 'gold standard' for detecting influenza A, but the method has complicated operation and long time consumption, and is not suitable for rapid diagnosis of influenza A. The serological detection method mainly comprises Hemagglutination (HA) and hemagglutination inhibition experiment (HI), neutralization experiment, immunofluorescence experiment, enzyme-linked immunosorbent assay (ELISA) and colloidal gold immunochromatography. The molecular biological detection method of IAV mainly includes RT-PCR, real-time fluorescence quantitative PCR (qRT-PCR) and the like, and the detection method is relatively accurate, but the required condition is higher and the time is longer. The monoclonal antibody of NP protein is the most important raw material for detecting influenza A virus antigen, so that the preparation of the NP protein monoclonal antibody is needed urgently, and a foundation is laid for preparing an influenza A virus detection kit. Disclosure of Invention The invention aims to provide a monoclonal antibody NP8 of an anti-influenza A virus NP protein and application thereof, so as to solve the problems in the prior art. The monoclonal antibody NP8 provided by the invention has higher capability of combining with the NP protein of the influenza A virus. In order to achieve the above object, the present invention provides the following solutions: The invention provides a monoclonal antibody NP8 of an anti-influenza A virus NP protein, wherein the light chain of the monoclonal antibody NP8 comprises a light chain CDR1 with an amino acid sequence shown as SEQ ID NO.12, a light chain CDR2 with an amino acid sequence shown as LVS and a light chain CDR3 with an amino acid sequence shown as SEQ ID NO. 13; The heavy chain of the monoclonal antibody NP8 comprises a heavy chain CDR1 with an amino acid sequence shown as SEQ ID NO.16, a heavy chain CDR2 with an amino acid sequence shown as SEQ ID NO.17 and a heavy chain CDR3 with an amino acid sequence shown as SEQ ID NO. 18. Preferably, the amino acid sequence of the light chain variable region of the monoclonal antibody NP8 is shown as SEQ ID NO.11, and the amino acid sequence of the heavy chain variable region of the monoclonal antibody NP8 is shown as SEQ ID NO. 15. The invention also provides a hybridoma cell strain (Mus museuus) NP8 secreting the monoclonal antibody NP8, wherein the hybridoma cell strain NP8 is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) for 2024, for 7 months and 16 days, and the preservation address is number 3 of West Liu 1, games-support No. North Star in the Korean region of Beijing, and the preservation number is CGMCC No.46020. The invention also provides application of the monoclonal antibody NP8 in preparation of products for detecting influenza A virus. Preferably, the product comprises an ELISA kit. The invention also provides an ELISA kit for detecting influenza A virus, which comprises the monoclonal antibody NP8. The invention also provides a method for detecting influenza A virus for non-diagnosis or treatment purposes, which comprises the step of detecting a sample to be detected by using the ELISA kit. The invention discloses the following technical effects: According to the invention, 19 influenza A virus NP protein monoclonal antibodies are prepared and screened, the binding capacity of the monoclonal antibodies and the influenza A virus NP protein is detected by ELISA, and the monoclonal antibody NP8 with stronger binding capacity with the NP protein is selected and obtained, so that a foundation is laid for preparing an influenza A virus detection kit. Drawings In order to more clearly illustrate the embodi