CN-122011169-A - Monoclonal antibody of anti-rabies virus G protein, detection reagent and application thereof

Abstract

The invention discloses a monoclonal antibody for resisting rabies virus G protein, a detection reagent and application thereof. A monoclonal antibody of rabies virus G protein comprises a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown as SEQ ID No.1, and the amino acid sequence of the light chain variable region is shown as SEQ ID No. 2. The monoclonal antibody with higher affinity and detection sensitivity for rabies virus G protein lays a foundation for the research and development and popularization of colloidal gold test strips and fluorescent immune test strips.

Inventors

• YUAN TINGTING
• ZHAO RONGMAO
• MA MENGZHEN
• SU SHOUFENG
• WEI DANPING
• ZHAO FANGYUAN
• CHEN JUAN
• GONG YUJIE

Assignees

• 北京纳百生物科技有限公司

Dates

Publication Date

20260512

Application Date

20260415

Claims (5)

1. 1. A monoclonal antibody against rabies virus G protein, wherein the monoclonal antibody against rabies virus G protein comprises a heavy chain variable region and a light chain variable region; The amino acid sequence of the heavy chain variable region of the monoclonal antibody is shown as SEQ ID No. 1; The amino acid sequence of the light chain variable region of the monoclonal antibody is shown as SEQ ID No. 2; The heavy and light chain variable regions are each comprised of a complementarity determining region and a framework region, the complementarity determining regions being comprised of CDR1, CDR2 and CDR 3; the amino acid sequence of CDR1 of the heavy chain variable region of the monoclonal antibody is shown as SEQ ID No. 5; the amino acid sequence of CDR2 of the heavy chain variable region of the monoclonal antibody is shown as SEQ ID No. 6; the amino acid sequence of CDR3 of the heavy chain variable region of the monoclonal antibody is shown as SEQ ID No. 7; the amino acid sequence of CDR1 of the light chain variable region of the monoclonal antibody is shown as SEQ ID No. 8; The amino acid sequence of CDR2 of the light chain variable region of the monoclonal antibody is shown as SEQ ID No. 9; the amino acid sequence of CDR3 of the light chain variable region of the monoclonal antibody is shown as SEQ ID No. 10.
2. 2. The monoclonal antibody against rabies virus G protein according to claim 1, characterized in that: the nucleotide sequence of the heavy chain variable region of the monoclonal antibody is shown as SEQ ID No. 3; The nucleotide sequence of the light chain variable region of the monoclonal antibody is shown as SEQ ID No. 4.
3. 3. An expression vector comprising the nucleotide sequence of claim 2.
4. 4. A host cell comprising the nucleotide sequence of claim 2 or the expression vector of claim 3.
5. 5. A test strip comprising the monoclonal antibody of claim 1.

Description

Monoclonal antibody of anti-rabies virus G protein, detection reagent and application thereof Technical Field The invention relates to the technical field of rapid biological detection of animal epidemic diseases, in particular to a monoclonal antibody of anti-rabies virus G protein, a detection reagent and application thereof. Background Rabies virus is a pathogen causing rabies, and is an acute central nervous system infectious disease of human and animal co-patients. Rabies virus (Rabies virus, RABV), belonging to the genus rabies (Lyssavirus) of the order Mononegavirales (Mononegavirales) rhabdoviridae (Rhabdoviridae), is a bullet-shaped single-strand negative-strand RNA virus with a lipid envelope, whose genome is about 11.6 kb in full length, encodes five structural proteins, nucleoprotein (N), matrix protein (M), glycoprotein (G), matrix protein (L) and phosphoprotein (P). The rabies virus N protein is used as a core component of a nucleocapsid, forms a spiral ribonucleoprotein complex (RNP) by wrapping a virus single-strand negative-strand RNA genome, protects nucleic acid from degradation by host nuclease, and is used as a template support for transcription and replication, and the phosphorylation modification state of the rabies virus N protein further regulates and controls the space-time regulation of virus gene expression. The M protein is positioned between the envelope and the RNP, has the membrane binding and nucleocapsid targeting capability, mediates virus assembly and budding through bridging, maintains the characteristic bullet-shaped morphology and negatively regulates the transcription activity of the virus. The G protein is used as the main component of the fiber on the surface of the envelope, is a key antigen for inducing the organism to generate neutralizing antibodies and cellular immune responses, and is also a core target point for the design of the existing inactivated vaccine and genetic engineering vaccine. The L protein is taken as RNA-dependent RNA polymerase (RdRp), is the largest structural protein of the virus, is responsible for the whole processes of mRNA transcription, methylation, genome replication and the like, and has the catalytic function dependent on an active complex formed by the L protein and the P protein, so that the synthesis and expression of the viral genome are completed together. While P protein realizes immune escape by interfering host interferon signal path (IRF-3/STAT 1 path) and inhibiting apoptosis and other mechanisms. The death rate of rabies is nearly 100%, the active immune prevention and control is mainly dependent on vaccination, and the level of neutralizing antibody of rabies virus is more than or equal to 0.5 IU/mL, which is generally considered to have protective power, and is the standard widely adopted by World Health Organization (WHO) and various national health departments. The method for monitoring the antibody level of the vaccinated animal comprises a neutralization test, ELISA and immunochromatography, wherein rabies virus is needed in the neutralization test, the operation risk is high, the general laboratory does not have the condition, and the immunochromatography (colloidal gold and fluorescent microsphere test strip) is simple and quick to operate compared with the ELISA method, the result can be judged in 10-15 minutes, and the cost is low. The fluorescent microsphere test strip is used for quantitatively analyzing the result by means of a simple reader, so that the fluorescent microsphere test strip is more suitable for detecting rabies virus antibodies. The method has respective advantages and disadvantages, and in order to meet clinical application, the rabies virus monoclonal antibody with high specificity and sensitivity is prepared on the basis of the existing G protein antigen, and the G protein antigen and the antibody are used as raw materials to develop a fluorescent microsphere immunochromatography detection method, so that the virus neutralization test has better correlation, and has important significance for clinical veterinary application. Disclosure of Invention Therefore, the invention provides a monoclonal antibody for resisting rabies virus G protein, a detection reagent and application thereof. In order to achieve the above object, the present invention provides the following technical solutions: According to a first aspect of embodiments of the present invention, there is provided an anti-rabies virus G protein monoclonal antibody comprising a heavy chain variable region and a light chain variable region; The amino acid sequence of the heavy chain variable region of the monoclonal antibody is shown as SEQ ID No. 1; The amino acid sequence of the light chain variable region of the monoclonal antibody is shown as SEQ ID No. 2; The heavy and light chain variable regions are each comprised of a complementarity determining region and a framework region, the complementarity determining regions being compri