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CN-120795134-B - A-stream virus N protein monoclonal antibody 19C7 and application thereof

CN120795134BCN 120795134 BCN120795134 BCN 120795134BCN-120795134-B

Abstract

The invention provides a A-flow virus N protein monoclonal antibody 19C7 and application thereof, wherein the amino acid sequence of a light chain CDR1 sequence, a CDR2 sequence and a CDR3 sequence of the A-flow virus N protein monoclonal antibody 19C7 is shown as SEQ ID NO. 1-3, and the amino acid sequence of a heavy chain CDR1 sequence, a CDR2 sequence and a CDR3 sequence is shown as SEQ ID NO. 4-6. The antibody has high specificity and wide coverage rate.

Inventors

  • ZHAO DIPING
  • YANG PINGPING
  • Fei Jiayao
  • ZHOU BIN
  • ZHANG YONGLEI
  • CHEN YARU

Assignees

  • 厦门康基生物科技有限公司
  • 杭州宝太生物科技有限公司

Dates

Publication Date
20260505
Application Date
20250812

Claims (9)

  1. 1. A monoclonal antibody 19C7 of a influenza A virus N protein is characterized in that the amino acid sequences of a light chain CDR1 sequence, a CDR2 sequence and a CDR3 sequence are respectively shown in SEQ ID NO. 1-3, the amino acid sequences of a heavy chain CDR1 sequence, a CDR2 sequence and a CDR3 sequence are respectively shown in SEQ ID NO. 4-6, and the monoclonal antibody 19C7 can identify NP proteins of the influenza A virus H1N 1.
  2. 2. The monoclonal antibody 19C7 of claim 1, wherein the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 7 and the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 8.
  3. 3. The monoclonal antibody 19C7 of the N protein of the alphavirus according to claim 1, wherein the amino acid sequence of the light chain is shown as SEQ ID NO. 9, and the amino acid sequence of the heavy chain is shown as SEQ ID NO. 10.
  4. 4. A reagent for detecting the protein of alphavirus N, comprising the monoclonal antibody 19C7 of the protein of alphavirus N according to any one of claims 1 to 3.
  5. 5. A kit for detecting the protein a of alphavirus N, comprising the monoclonal antibody 19C7 of the protein a of alphavirus N according to any one of claims 1 to 3.
  6. 6. Use of the monoclonal antibody 19C7 to the N protein of the influenza a virus according to any one of claims 1 to 3 for the preparation of a reagent for diagnosing influenza a.
  7. 7. A nucleic acid molecule encoding the heavy and light chain variable regions of the alphavirus N protein monoclonal antibody 19C7 of any one of claims 1 to 3.
  8. 8. An expression vector comprising the nucleic acid molecule of claim 7.
  9. 9. A host cell comprising the expression vector of claim 8.

Description

A-stream virus N protein monoclonal antibody 19C7 and application thereof Technical Field The invention relates to a monoclonal antibody 19C7 of an N protein of a influenza A virus and application thereof, belonging to the technical field of antibodies. Background Influenza is a very contagious respiratory disease caused by influenza virus. Common symptoms include fatigue, fever, aversion to cold, dry cough and body pain. These symptoms typically self-alleviate within 1-2 weeks, but complications may occur and cause life-threatening secondary infections. Influenza viruses infected by humans are largely classified into three major classes, type a, type B and type C, and these viruses are further subdivided into different subtypes and strains. New influenza strains occur annually, on the one hand because of the ability of influenza viruses to slowly mutate, and on the other hand, to benefit from their rapid recombination mechanisms. Slow variation causes seasonal fluctuations throughout the year, while rapid recombination may induce new strains that induce global pandemics. Influenza a virus (Flu a) is particularly susceptible to genetic recombination due to its broad host range (covering animals such as humans, dogs, birds, pigs, horses, whales, seals, etc.). In particular, the genome of influenza a virus consists of 8 loosely connected segments, each carrying at least one key gene. These genes are responsible for directing the expression of major viral proteins such as Hemagglutinin (HA), neuraminidase (NA), nucleoprotein (NP) and the like. Currently, diagnostic assays for influenza include rapid influenza diagnostic assays (immunodiagnostics), direct fluorescent antibody staining, virus culture, and molecular detection. These assays are generally capable of distinguishing influenza a from influenza b, and also of identifying different subtypes of influenza a. Immunodiagnostic reagents have been put into routine use since their FDA approval was obtained 1999. The reagent has simple and convenient operation and low cost, and can rapidly obtain results within 10-30 minutes, thereby enabling doctors to timely prescribe medicines within the effective period of antiviral medicines (1-2 days after symptoms appear). Most immunodiagnostic reagents are implemented by detecting Nucleoprotein (NP), a highly conserved protein in influenza virus that is not prone to mutation. The main limitation of the current mainstream methods is the low sensitivity and instability. To actually improve the detection sensitivity, it is important to develop monoclonal antibodies capable of recognizing existing and new strains. Disclosure of Invention The invention provides a monoclonal antibody 19C7 of an N protein of a influenza A virus and application thereof, which can effectively solve the problems. A monoclonal antibody 19C7 of the N protein of the A-flow virus has the amino acid sequences of a light chain CDR1 sequence, a CDR2 sequence and a CDR3 sequence shown in SEQ ID NO. 1-3, and the amino acid sequences of a heavy chain CDR1 sequence, a CDR2 sequence and a CDR3 sequence shown in SEQ ID NO. 4-6. In some embodiments, the amino acid sequence of the light chain variable region of the A-stream virus N protein monoclonal antibody 19C7 is shown in SEQ ID NO. 7, and the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 8. In some embodiments, the amino acid sequence of the light chain of the A-flow virus N protein monoclonal antibody 19C7 is shown as SEQ ID NO. 9, and the amino acid sequence of the heavy chain is shown as SEQ ID NO. 10. A reagent for detecting the protein of the alphavirus N, which comprises the monoclonal antibody 19C7 of the protein of the alphavirus N. A kit for detecting a protein of a alphavirus N, comprising said monoclonal antibody 19C7 of a protein of a alphavirus N. Use of the monoclonal antibody 19C7 against the N protein of the influenza a virus in the preparation of a reagent for diagnosing, preventing or treating influenza a. A nucleic acid molecule encoding the heavy chain variable region and the light chain variable region of the alphavirus N protein monoclonal antibody 19C7. An expression vector comprising said nucleic acid molecule. A host cell comprising said expression vector. The preparation method of the A-flow virus N protein monoclonal antibody 19C7 is characterized by comprising the following steps: S1, recombinant expression of H1N1-NP and H1N1-NP-C proteins; s2, immunizing an animal by taking H1N1-NP protein as an immunogen, separating spleen cells and electrically fusing the spleen cells and myeloma cells; S3, taking the H1N1-NP-C protein as a detecting antigen, and screening hybridoma cell strains for identifying H1N1-NP-C fragments; S4, extracting mRNA of the hybridoma cell, cloning heavy chain and light chain variable region genes, constructing a recombinant expression vector, and carrying out recombinant expression and purification to obtain the recombinant a