KR-102963572-B1 - ANTIBODIES SPECIFICALLY BINDING TO ENVELOPE PROTEIN OF SOYBEAN MOSAIC VIRUS AND USES THEREOF

Abstract

The present invention relates to an antibody that specifically binds to a soybean mosaic virus (SMV) envelope protein, and more specifically, to an antibody specific to a soybean mosaic virus envelope protein that can be usefully utilized for the diagnosis of soybean mosaic virus infection, and to the use thereof.

Inventors

• 이미기
• 이은빈
• 권명희

Assignees

• 재단법인 경기도경제과학진흥원

Dates

Publication Date

20260513

Application Date

20231113

Claims (11)

1. An antibody or an antigen-binding fragment thereof that specifically binds to a soybean mosaic virus (SMV) envelope protein, comprising: a light chain complementarity determining region 1 (LCDR1) consisting of the amino acid sequence of SEQ ID NO. 1, a light chain complementarity determining region 2 (LCDR2) consisting of the amino acid sequence of SEQ ID NO. 2, and a light chain complementarity determining region 3 (LCDR3) consisting of the amino acid sequence of SEQ ID NO. 3; a heavy chain complementarity determining region 1 (HCDR1) consisting of the amino acid sequence of SEQ ID NO. 4; a heavy chain complementarity determining region comprising the amino acid sequence of SEQ ID NO. 5; and a heavy chain complementarity determining region comprising the amino acid sequence of SEQ ID NO. 6.
2. In Article 1, An antibody or antigen-binding fragment thereof that specifically binds to a soy mosaic virus envelope protein, comprising a light chain variable region consisting of the amino acid sequence of SEQ ID NO. 7; and a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO. 8.
3. In Article 1, An antibody or an antigen-binding fragment thereof, wherein the antibody specifically binding to the above soy mosaic virus envelope protein is a monoclonal antibody.
4. A composition for detecting soybean mosaic virus comprising an antibody that specifically binds to the soybean mosaic virus envelope protein of claim 1 or an antigen-binding fragment thereof.
5. A kit for detecting soy mosaic virus comprising an antibody that specifically binds to the soy mosaic virus envelope protein of claim 1 or an antigen-binding fragment thereof.
6. In Article 5, The above kit is a kit for detecting soybean mosaic virus, which is a rapid diagnostic kit based on lateral flow immunoassay (LFA).
7. In Article 5, The above kit is a kit for detecting soy mosaic virus, comprising: an antibody or an antigen-binding fragment thereof that specifically binds to the envelope protein of the soy mosaic virus of claim 1; a secondary antibody condensed with a label capable of binding to the antibody or the antigen-binding fragment thereof of claim 1; and a chromogenic substrate.
8. In Article 7, The above label is from the group consisting of horseradish peroxidase (HRP), alkaline phosphatase (AP), glucose oxidase, luciferase, β-D-galactosidase, malate dehydrogenase (MDH), acetylcholinesterase, colloidal gold, metal nanoparticles, silica nanoparticles, organic fluorescent materials, fluorescent proteins, quantum dots, radioactive materials, isotopes, dyes, carbon nanotubes, graphene, and fullerene. A kit for detecting soybean mosaic virus, which is selected.
9. In Article 7, A kit for detecting soybean mosaic virus, wherein the chromogenic substrate is selected from the group consisting of DAB (diaminobenzidine), AEC (3-amino-9-ethylcarbazole), BCIP/NBT (5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium), BCIP/INT (5-bromo-4-chloro-3-indolyl phosphate/iodonitrotetrazolium), NF (New fuchsin), FRT (Fast Red TR Salt), TMB (3,3',5,5'-tetramethyl benzidine), ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) and OPD (o-phenylenediamine).
10. A step of contacting a sample with an antibody that specifically binds to the soybean mosaic virus envelope protein of claim 1 or an antigen-binding fragment thereof; and A method for detecting soy mosaic virus, comprising the step of detecting an antigen-antibody complex formed by binding with the antibody or its antigen-binding fragment.
11. In Article 10, Confirmation of the formation of the above antigen-antibody complex is performed using enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), sandwich ELISA, Western blotting, immunodot blot assay, immunofluorescence assay (IFA), immunoochemiluminescence assay, immunohistochemistry, immunochromatography, lateral flow immunoassay (LFA), colorimetric immunoassay, spectrometric immunoassay, Raman spectroscopic immunoassay, surface plasmon resonance immunoassay, and interferometer. A method for detecting soybean mosaic virus, comprising any one method selected from the group consisting of interferometric immunoassay, visual assessment, protein chip technology, and biosensor.

Description

Antibodies specifically binding to the envelope protein of soybean mosaic virus (SMV) and uses thereof The present invention relates to an antibody that specifically binds to the envelope protein of the soybean mosaic virus (SMV) and to the use thereof. Plant viruses cause diseases in numerous plants and inflict severe economic damage across the entire agricultural production process, including reduced yields and lower quality of various crops. In particular, with the recent increase in international trade of food and various plant products, foreign pathogenic plant viruses previously absent domestically are being introduced, leading to a trend of increasingly severe damage. Consequently, the prevention, treatment, and quarantine of viruses affecting major crops are critical tasks in agriculture. Furthermore, unlike diseases caused by fungi or bacteria, diseases caused by plant viruses cannot be prevented or treated by spraying pesticides. Therefore, precise and rapid diagnosis of viral infection in plants or seeds is a prerequisite for managing viral diseases and preventing damage to cultivated crops in advance. Meanwhile, plant viruses have a very wide host range and can cause secondary infections through various routes such as contact, vectors, or the use of infected seeds. Furthermore, diagnosis and control are difficult because there are almost no effective control methods or chemicals available. Therefore, an efficient control method involves establishing a system for early diagnosis and source removal of plant viruses, and the importance of this is steadily increasing. Soybeans (*Glycine max*) are an important source of edible oil and plant protein. Soybean viral diseases generally cause yield losses of 10 to 30%, and in severe cases, 50 to 100%. Soybean mosaic virus (SMV) is a leguminous virus first discovered in Korea and belongs to the genus Carmovirus (family Tombusviridae). Soybean plants infected with SMV exhibit bright yellow mosaic symptoms on young leaves, causing a reduction in crop yield. According to a recent survey on viral distribution in soybean-growing regions, SMV (73.1%), Soybean yellow mosaic virus (7.3%), Soybean yellow mosaic virus (2.6%), and Peanut stunt virus (4.1%) are distributed in Korea. Another study showed that SMV exhibited the highest incidence rate (84.9%) when infection types such as triple or double infections were not taken into account. One method to prevent the spread of viruses is to isolate infected plants from healthy ones through rapid diagnosis. Among the various methods developed to diagnose plant viruses, Reverse Transcription Polymerase Chain Reaction (RT-PCR) is primarily used in Korea for diagnosing soybean mosaic virus infections. However, the RT-PCR method has disadvantages, such as increased diagnostic costs and time, as well as the need for skilled technicians and expensive equipment. To overcome these drawbacks, a diagnostic method for soybean mosaic virus using Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) has been developed and is evaluated as efficient in terms of speed, accuracy, and cost. Despite its many advantages, RT-LAMP technology has several limitations. Since RT-LAMP requires 4 to 6 primers of different lengths, experimental design is difficult, and it is not suitable for studying novel genes. Additionally, an understanding of the structural characteristics of the target gene is required. Furthermore, RT-LAMP has the disadvantage of making it difficult to effectively control virus transmission because equipment such as temperature controllers or heating blocks must be brought to the site. Furthermore, to effectively respond to diseases caused by infections such as viruses and to prevent their progression and spread, it is necessary to diagnose the presence of infection quickly and accurately. If the infection can be diagnosed during the incubation period before symptoms appear, the spread of the infectious disease can be effectively prevented, thereby averting significant damage. Meanwhile, Korean Registered Patent No. 1250388 discloses a ‘specific primer for multiple diagnosis of soybean viruses and its use’ capable of diagnosing alfalfa mosaic virus (AMV), soybean yellow mottle mosaic virus (SYMMV), unreported novel virus (USV3, tentatively named: soybean yellow common mosaic virus, SYCMV), peanut stunt virus (PSV), soybean dwarf virus (SbDV), and soybean mosaic virus (SMV), and Korean Registered Patent No. 0857043 discloses a ‘specific primer for diagnosing soybean mosaic virus,’ but there is no description of the ‘antibody that specifically binds to a soybean mosaic virus envelope protein and its use’ of the present invention. Accordingly, the inventors of the present invention completed the present invention by securing an antibody corresponding to the envelope protein of the soybean mosaic virus during the process of developing a field diagnostic kit for the soybean mosaic virus, and by confirming that the