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CN-121991204-A - Preparation method and application of monoclonal antibody of main capsid protein of mouse anti-infectious spleen and kidney necrosis virus

CN121991204ACN 121991204 ACN121991204 ACN 121991204ACN-121991204-A

Abstract

The application discloses a preparation method and application of a monoclonal antibody of main capsid protein of a mouse anti-infectious spleen and kidney necrosis virus, and relates to the technical field of monoclonal antibodies. The monoclonal antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region, the heavy chain variable region comprises complementarity determining regions CDR-H1, CDR-H2 and CDR-H3, the amino acid sequences of the heavy chain variable region are respectively shown as SEQ ID NO. 3-5, and the light chain comprises a light chain variable region, and the light chain variable region comprises complementarity determining regions CDR-L1, CDR-L2 and CDR-L3. The monoclonal antibody provided by the application has excellent affinity to the main capsid protein of the infectious spleen-kidney necrosis virus, has a titer of 1:320000, and can be used for detecting the infectious spleen-kidney necrosis virus or the main capsid protein of the infectious spleen-kidney necrosis virus.

Inventors

  • GUO CHANGJUN
  • YOU YANLIN
  • CAO XUEQIAN
  • PAN WEIQIANG
  • JIANG LAI
  • HE JIANGUO

Assignees

  • 中山大学

Dates

Publication Date
20260508
Application Date
20260403

Claims (10)

  1. 1. A monoclonal antibody of a main capsid protein of a mouse anti-infectious spleen and kidney necrosis virus is characterized in that the monoclonal antibody comprises a heavy chain and a light chain; The heavy chain comprises a heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2 and CDR-H3; the amino acid sequence of the CDR-H1 is shown as SEQ ID NO. 3; the amino acid sequence of the CDR-H2 is shown as SEQ ID NO. 4; the amino acid sequence of the CDR-H3 is shown in SEQ ID NO. 5; the light chain comprises a light chain variable region comprising complementarity determining regions CDR-L1, CDR-L2 and CDR-L3; the amino acid sequence of the CDR-L1 is shown in SEQ ID NO. 7; The amino acid sequence of the CDR-L2 is LVS; the amino acid sequence of the CDR-L3 is shown in SEQ ID NO. 8.
  2. 2. The monoclonal antibody according to claim 1, wherein the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 2.
  3. 3. The monoclonal antibody according to claim 1 or 2, wherein the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 6.
  4. 4. A bispecific antibody comprising a monoclonal antibody to the major capsid protein of the murine anti-infectious spleen-kidney necrosis virus of any one of claims 1 to 3.
  5. 5. A biological material associated with a monoclonal antibody to the main capsid protein of the murine anti-infectious spleen-kidney necrosis virus according to any one of claims 1 to 3, characterized in that said biological material comprises any one of A1) to A5); A1 A nucleic acid molecule encoding a monoclonal antibody of the major capsid protein of the murine anti-infectious spleen-kidney necrosis virus of any one of claims 1 to 3; A2 An expression cassette comprising A1) said nucleic acid molecule; a3 A) a recombinant vector comprising the nucleic acid molecule of A1) or the expression cassette of A2); A4 A) a recombinant microorganism comprising the nucleic acid molecule of A1) or the expression cassette of A2) or the recombinant vector of A3); a5 A) a recombinant cell comprising the nucleic acid molecule of A1) or the expression cassette of A2) or the recombinant vector of A3), said recombinant cell comprising no propagation material.
  6. 6. The biological material according to claim 5, wherein said nucleic acid molecule comprises a nucleic acid molecule encoding said heavy chain variable region and a nucleic acid molecule encoding said light chain variable region.
  7. 7. A method for preparing a monoclonal antibody against the main capsid protein of the infectious spleen and kidney necrosis virus according to any one of claims 1 to 3, wherein the monoclonal antibody is obtained by culturing the recombinant microorganism or recombinant cell according to claim 5 or 6, and isolating and purifying the recombinant microorganism or recombinant cell.
  8. 8. An antibody drug conjugate comprising a monoclonal antibody to the major capsid protein of the murine anti-infectious spleen and kidney necrosis virus of any one of claims 1 to 3.
  9. 9. A pharmaceutical composition is characterized in that the product comprises at least one of B1) to B3): b1 A monoclonal antibody to the major capsid protein of the murine anti-infectious spleen-kidney necrosis virus of any one of claims 1 to 3; b2 A biomaterial as claimed in claim 5 or 6; b3 An antibody drug conjugate according to claim 8.
  10. 10. Use of a monoclonal antibody of the main capsid protein of the murine anti-infectious spleen-kidney necrosis virus according to any one of claims 1 to 3 for the preparation of a reagent for the detection of infectious spleen-kidney necrosis virus or infectious spleen-kidney necrosis virus main capsid protein.

Description

Preparation method and application of monoclonal antibody of main capsid protein of mouse anti-infectious spleen and kidney necrosis virus Technical Field The application relates to the technical field of monoclonal antibodies, in particular to a preparation method and application of a monoclonal antibody of main capsid protein of a mouse anti-infectious spleen-kidney necrosis virus. Background Along with the continuous increase of the aquaculture density, various diseases frequently occur, and the healthy and sustainable development of the aquaculture industry is seriously hindered, wherein the viral diseases cause quick spread, large harm and difficult prevention and control, and the ingestion of the cultured fishes is reduced, the behaviors are abnormal, the death rate is increased, and huge economic loss is caused. Iridovirus is one of the most popular and most pathogenic viral pathogens in freshwater fish, especially Infectious Spleen and Kidney Necrosis Virus (ISKNV) of the genus megaly, and has a serious influence on the health and sustainable development of aquaculture industry. At present, in detection and research of ISKNV, polyclonal antibodies based on whole virus antigens and corresponding ELISA methods have been developed, but the limitations of strong cross reactivity, unstable sensitivity and the like still exist, and the popularization of the polyclonal antibodies in practical application is limited. Monoclonal antibody technology has been widely used in the field of virus diagnosis and control because of its advantages such as high specificity, good uniformity, sustainable production, etc. However, monoclonal antibodies against the ISKNV major capsid protein (Major capsid protein, MCP) are still freshly reported, and lack of monoclonal antibodies capable of accurately recognizing ISKNV-MCP epitopes and suitable for low-cost, high-throughput detection in the related art has hindered development of the ISKNV virus rapid detection kit and in-depth research of infection mechanism. Based on the above, a preparation method and application of a high-specificity murine monoclonal antibody aiming at ISKNV main capsid protein still need to be developed. Disclosure of Invention The embodiment of the application provides a preparation method and application of a monoclonal antibody of main capsid protein of a mouse anti-infectious spleen and kidney necrosis virus. Specifically, the application expresses and purifies recombinant MCP protein through a prokaryotic or eukaryotic system, immunizes a BALB/c mouse, and combines a hybridoma technology to screen a cell strain capable of stably secreting anti-MCP monoclonal antibody, thereby obtaining the monoclonal antibody with high affinity and strong specificity. To achieve the above object, a first aspect of embodiments of the present application provides a monoclonal antibody against a major capsid protein of infectious spleen and kidney necrosis virus in mice, wherein the monoclonal antibody comprises a heavy chain and a light chain; The heavy chain comprises a heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2 and CDR-H3; the amino acid sequence of the CDR-H1 is shown as SEQ ID NO. 3; the amino acid sequence of the CDR-H2 is shown as SEQ ID NO. 4; the amino acid sequence of the CDR-H3 is shown in SEQ ID NO. 5; the light chain comprises a light chain variable region comprising complementarity determining regions CDR-L1, CDR-L2 and CDR-L3; the amino acid sequence of the CDR-L1 is shown in SEQ ID NO. 7; The amino acid sequence of the CDR-L2 is LVS; the amino acid sequence of the CDR-L3 is shown in SEQ ID NO. 8. In some embodiments, the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2. In some embodiments, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 6. In a second aspect, embodiments of the present application provide a bispecific antibody comprising a monoclonal antibody comprising the above-described major capsid protein of murine anti-infectious spleen and kidney necrosis virus. A third aspect of embodiments of the present application provides a biomaterial associated with a monoclonal antibody to a major capsid protein of the murine anti-infectious spleen-kidney necrosis virus of the above first aspect, comprising any one of A1) to A5); A1 Nucleic acid molecules encoding monoclonal antibodies to the major capsid proteins of the murine anti-infectious spleen-kidney necrosis virus described above; A2 An expression cassette comprising A1) said nucleic acid molecule; a3 A) a recombinant vector comprising the nucleic acid molecule of A1) or the expression cassette of A2); A4 A) a recombinant microorganism comprising the nucleic acid molecule of A1) or the expression cassette of A2) or the recombinant vector of A3); a5 A) a recombinant cell comprising the nucleic acid molecule of A1) or the expression cassette of A2) or the recombinant vector of