CN-121824743-B - Rabbit monoclonal antibody for resisting pseudorabies virus glycoprotein gE and application of rabbit monoclonal antibody in immunodetection

Abstract

The invention belongs to the technical field of immunological detection, and particularly relates to a rabbit monoclonal antibody for resisting pseudorabies virus glycoprotein gE and application thereof in immunological detection. The amino acid sequences of CDR1, CDR2 and CDR3 on the light chain variable region of the rabbit monoclonal antibody are respectively shown in SEQ ID NO.3-5, and the amino acid sequences of CDR1, CDR2 and CDR3 on the heavy chain variable region are respectively shown in SEQ ID NO. 8-10. The rabbit monoclonal antibody aiming at PRV glycoprotein gE and an immunological detection system constructed by the antibody provide an important technical means for rapid differential diagnosis and large-scale screening of pigs infected by pseudorabies virus wild virus and vaccine immunized pigs, have the advantages of strong specificity, high accuracy, good sensitivity, simple detection method operation and the like, have wide application prospect in the PRV pathogen purification field, and have important application and popularization values.

Inventors

• CHEN LIPING
• JI FANGXUAN
• WANG YAQUN
• CHEN LIANG
• WANG XIAOGE
• WU HAI

Assignees

• 唯泰科生物科技(武汉)有限公司

Dates

Publication Date

20260512

Application Date

20260311

Claims (10)

1. 1. The rabbit monoclonal antibody for resisting the pseudorabies virus glycoprotein gE is characterized by comprising a light chain variable region and a heavy chain variable region, wherein the amino acid sequences of CDR1, CDR2 and CDR3 on the light chain variable region are respectively shown as SEQ ID NO.3, SEQ ID NO.4 and SEQ ID NO.5, and the amino acid sequences of CDR1, CDR2 and CDR3 on the heavy chain variable region are respectively shown as SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO. 10.
2. 2. The rabbit monoclonal antibody against pseudorabies virus glycoprotein gE according to claim 1, wherein the amino acid sequence of the light chain variable region of the rabbit monoclonal antibody is shown in SEQ ID No.2 and the amino acid sequence of the heavy chain variable region is shown in SEQ ID No. 7.
3. 3. The rabbit monoclonal antibody against pseudorabies virus glycoprotein gE according to claim 2, wherein the amino acid sequence of the light chain of the rabbit monoclonal antibody is shown in SEQ ID NO.1, and the amino acid sequence of the heavy chain is shown in SEQ ID NO. 6.
4. 4. The rabbit monoclonal antibody against pseudorabies virus glycoprotein gE according to claim 1, wherein the rabbit monoclonal antibody is a full-length antibody or an antigen-binding region thereof, wherein the antigen-binding region is selected from the group consisting of a Fab fragment, a F (ab) 2 fragment, an Fv fragment, (Fv) 2 fragment, an scFv fragment and an sc (Fv) 2 fragment.
5. 5. A nucleic acid molecule encoding the rabbit monoclonal antibody against pseudorabies virus glycoprotein gE according to any of claims 1-4.
6. 6. A recombinant vector comprising the nucleic acid molecule of claim 5.
7. 7. An antibody conjugate, characterized in that it consists of a rabbit monoclonal antibody against the pseudorabies virus glycoprotein gE according to any of claims 1-4 and a detection label linked to said rabbit monoclonal antibody.
8. 8. The antibody conjugate of claim 7, wherein the detection label is horseradish peroxidase or acridinium ester.
9. 9. Use of a rabbit monoclonal antibody against pseudorabies virus glycoprotein gE according to any of claims 1-4 or an antibody conjugate according to any of claims 7-8 for the preparation of a detection kit for detecting pseudorabies virus glycoprotein gE or for the preparation of a detection kit for detecting anti-pseudorabies virus glycoprotein gE antibody.
10. 10. A test kit comprising a rabbit monoclonal antibody against pseudorabies virus glycoprotein gE according to any of claims 1-4 or an antibody conjugate according to any of claims 7-8.

Description

Rabbit monoclonal antibody for resisting pseudorabies virus glycoprotein gE and application of rabbit monoclonal antibody in immunodetection Technical Field The invention relates to the technical field of immunological detection, in particular to a rabbit monoclonal antibody for resisting pseudorabies virus glycoprotein gE and application thereof in immunological detection. Background Pseudorabies (Pseudorabies, PR), also known as ojeszky's disease, is an acute infectious disease caused by pseudorabies virus (Pseudorabies virus, PRV) with fever, itching and central nervous disorder of animals as main symptoms. Pigs are the main infectious source and natural host of PRV, and healthy pigs can be infected by directly contacting with sick pigs and pigs with toxicity. Adult pigs are usually infected implicitly, but live with and expel toxin, which can cause abortion, stillbirth, weak fetus or mummy fetus of pregnant sows, and the infected newborn piglets have fever, inappetence, intractable diarrhea and neurological symptoms, and even die. Once the PRV infection is developed, the PRV infection is difficult to eradicate, and the high-onset and outbreak epidemic characteristics of the PRV infection cause great economic loss to the global pig industry, and the PRV infection becomes one of serious infectious diseases which seriously endanger the healthy development of the pig industry. Preventing and controlling the widespread spread of this disease is a difficult task currently faced. At present, prevention and control of pseudorabies are mainly vaccine immunization prevention, and detection of pseudorabies virus antigen and serum neutralizing antibody is combined to distinguish immunized animals and naturally infected animals, so that wild virus infected pigs are eliminated and purified. Therefore, monitoring and detecting the level of PRV virus or neutralizing antibodies in porcine serum is a very important link in the prevention and treatment of pseudorabies. The surface envelope glycoprotein gE is an important virulence factor on pseudorabies virus, and is an important target point of a plurality of PRV wild strain infection detection kits at present. In addition, the PRV vaccine in the market is a gE gene deletion vaccine, antibodies aiming at gE protein cannot be detected in immunized pig serum after immunization of the gE gene deletion vaccine, and specific antibodies (infectious antibodies) aiming at gE protein can be detected in wild strain infected pig serum, so that the situation that the pig population is infected by PRV wild virus can be intuitively reflected by monitoring the situation of the anti-gE protein antibodies in the pig population, wild virus infected pigs are screened and eliminated in time, and a healthy negative pig population is established. The key of the method is to develop monoclonal antibodies with moderate affinity and specific binding to glycoprotein gE, the suitability of the antibody performance directly determines the sensitivity and specificity of the whole immune detection system, if the affinity of the antibodies is too high, positive serum can not effectively block the binding of labeled antibodies and antigens, false negative results appear, if the affinity of the antibodies is too low, the binding of positive serum and antigens can not be sensitively detected, and false negative results can also appear, if the specificity of the antibodies is poor, cross reaction can occur with other antigens, and non-specific blocking of samples appears, thereby the false positive results appear. Therefore, the method for screening the rabbit monoclonal antibody with high specificity and moderate affinity for resisting the PRV glycoprotein gE has very important practical application value. Disclosure of Invention Aiming at the technical problems existing in the prior art, the invention provides a rabbit monoclonal antibody with good reactivity and recognition specificity to pseudorabies virus glycoprotein gE, and provides a coding gene and an expression vector of the antibody. The invention further provides a kit for preparing and detecting pseudorabies virus glycoprotein gE or an anti-pseudorabies virus glycoprotein gE antibody or application of the antibody in preparing anti-pseudorabies virus medicaments and a corresponding kit. The invention is realized by the following technical scheme: The first aspect of the invention provides a rabbit monoclonal antibody against pseudorabies virus glycoprotein gE, which comprises a light chain variable region and a heavy chain variable region, wherein the amino acid sequences of CDR1, CDR2 and CDR3 on the light chain variable region are respectively shown as SEQ ID NO.3, SEQ ID NO.4 and SEQ ID NO.5, and the amino acid sequences of CDR1, CDR2 and CDR3 on the heavy chain variable region are respectively shown as SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO. 10. Further, the amino acid sequence of the light chain variable region of the rabbit monoclonal a