CN-121991209-A - Antibody pair for resisting Hantavirus NP protein and application thereof

Abstract

The invention relates to the technical field of biology, in particular to an antibody pair for resisting Hantavirus NP protein and application thereof. The protein detection limit of the established double-antibody sandwich ELISA detection system based on the antibody is 1ng/mL, the linear range, the specificity, the accuracy and the like are all good, and an effective tool is provided for early clinical diagnosis, epidemiological monitoring, viral load assessment and related scientific research of hantavirus.

Inventors

• ZHENG ZHENHUA
• DING QINGWEN
• CHEN DAN
• ZHANG NAILOU
• XU WEN
• ZHENG SHAOLONG
• GAN MIN
• WEI JINBO

Assignees

• 中国科学院武汉病毒研究所
• 广州国家实验室

Dates

Publication Date

20260508

Application Date

20260202

Claims (10)

1. 1. An antibody pair for resisting Hantavirus NP protein, which is characterized in that, The antibody pair comprises a first monoclonal antibody and a second monoclonal antibody, wherein the monoclonal antibodies comprise a light chain variable region and a heavy chain variable region, the light chain variable region comprises a light chain CDR1, a light chain CDR2 and a light chain CDR3, and the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3; The sequence information of the first monoclonal antibody is as follows: The amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 3; the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 4; the amino acid sequence of the light chain CDR3 is shown as SEQ ID NO. 5; The amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 8; the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 9; the amino acid sequence of the heavy chain CDR3 is shown as SEQ ID NO. 10; the sequence information of the second monoclonal antibody is as follows: the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 17; The amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 18; the amino acid sequence of the light chain CDR3 is shown as SEQ ID NO. 19; the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 22; the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 23; the amino acid sequence of the heavy chain CDR3 is shown in SEQ ID NO. 24.
2. 2. The antibody pair of claim 1, wherein, The amino acid sequence of the light chain variable region of the first monoclonal antibody is shown as SEQ ID NO. 2, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 7; the amino acid sequence of the light chain variable region of the second monoclonal antibody is shown as SEQ ID NO. 16, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 21.
3. 3. The antibody pair of claim 2, wherein, The amino acid sequence of the first monoclonal antibody light chain is shown as SEQ ID NO.1, and the amino acid sequence of the heavy chain is shown as SEQ ID NO. 6; The amino acid sequence of the light chain of the second monoclonal antibody is shown as SEQ ID NO. 15, and the amino acid sequence of the heavy chain is shown as SEQ ID NO. 20.
4. 4. Use of an antibody according to any one of claims 1-3 for the preparation of a kit for detecting hantavirus.
5. 5. A nucleic acid molecule encoding the amino acid sequence of the first monoclonal antibody or the second monoclonal antibody of any one of claims 1-3.
6. 6. The nucleic acid molecule of claim 5, wherein, The nucleic acid molecule encoding the first monoclonal antibody light chain variable region sequence is shown in SEQ ID NO. 12; The nucleic acid molecule encoding the heavy chain variable region sequence of the first monoclonal antibody is shown as SEQ ID NO. 14; The nucleic acid molecule encoding the second monoclonal antibody light chain variable region sequence is shown in SEQ ID NO. 26; The nucleic acid molecule encoding the heavy chain variable region sequence of the second monoclonal antibody is shown in SEQ ID NO. 28.
7. 7. The nucleic acid molecule of claim 6, wherein, The nucleic acid molecule for encoding the first monoclonal antibody light chain sequence is shown as SEQ ID NO. 11; The nucleic acid molecule for encoding the heavy chain sequence of the first monoclonal antibody is shown as SEQ ID NO. 13; the nucleic acid molecule encoding the light chain sequence of the second monoclonal antibody is shown as SEQ ID NO. 25; The nucleic acid molecule encoding the heavy chain sequence of the second monoclonal antibody is shown in SEQ ID NO. 27.
8. 8. A kit for detecting hantavirus comprising the antibody pair of any one of claims 1-3.
9. 9. The kit of claim 8, wherein the detection kit is an enzyme-linked immunosorbent kit, an immunohistochemical kit, an immunofluorescent kit, an immunoblotting kit, or a flow cytometry kit.
10. 10. The kit of claim 9, wherein the kit is a double-antibody sandwich enzyme-linked immunosorbent kit, the first monoclonal antibody is a capture antibody, the second monoclonal antibody is a detection antibody, and the second monoclonal antibody is linked to a detection label.

Description

Antibody pair for resisting Hantavirus NP protein and application thereof Technical Field The invention relates to the technical field of biology, in particular to an antibody pair for resisting Hantavirus NP protein and application thereof. Background Hantavirus (Hantavirus) is a segmented negative-strand RNA virus of the genus hantavirus, the family bunyaviridae, and can cause severe human diseases such as hemorrhagic fever with renal syndrome (HFRS) and Hantavirus Pulmonary Syndrome (HPS). These diseases progress rapidly and have high mortality, while early symptoms of infection (such as fever, headache, myalgia) lack of specificity and are easily confused with common influenza diseases. Therefore, developing rapid and accurate early diagnosis technology is important to realizing timely clinical intervention, reducing the death rate and controlling epidemic spread. At present, serological detection is a common technology for diagnosing hantavirus infection, and the core of the serological detection depends on specific antibodies generated after body infection for recognition. However, antibodies need to accumulate to a detectable threshold for a certain period of time at the beginning of infection, and often cannot be effectively detected within days after infection due to insufficient antibody levels, resulting in "detection dead zones" during the diagnostic window, leading to missed detection or delayed diagnosis for early infected individuals. Although the nucleic acid detection has higher sensitivity, the nucleic acid detection has strict requirements on the experimental environment, instruments and equipment and professional abilities of operators, the detection process consumes longer time, the duration window of the viral RNA in the body fluid of a patient is limited, and the factors limit the popularization and application of the nucleic acid detection in basic medical institutions or large-scale field screening scenes. Therefore, it is needed to develop a detection system capable of directly, sensitively and specifically detecting hantavirus to overcome the limitation of the current hantavirus detection. Disclosure of Invention In view of the above, the invention provides an antibody pair for resisting Hantavirus NP protein and application thereof. The technical scheme of the invention is realized as follows: In a first aspect, the invention provides an antibody pair of anti-hantavirus NP proteins, which comprises a first monoclonal antibody and a second monoclonal antibody, wherein the monoclonal antibodies comprise a light chain variable region (VL) and a heavy chain variable region (VH), the light chain variable region comprises a light chain CDR1, a light chain CDR2 and a light chain CDR3 (LCDR 1-LCDR 3), and the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3 (HCDR 1-HCDR 3); The sequence information of the first monoclonal antibody is as follows: The amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 3; the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 4; the amino acid sequence of the light chain CDR3 is shown as SEQ ID NO. 5; The amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 8; the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 9; the amino acid sequence of the heavy chain CDR3 is shown as SEQ ID NO. 10; the sequence information of the second monoclonal antibody is as follows: the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 17; The amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 18; the amino acid sequence of the light chain CDR3 is shown as SEQ ID NO. 19; the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 22; the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 23; the amino acid sequence of the heavy chain CDR3 is shown in SEQ ID NO. 24. Further, the amino acid sequence of the light chain variable region of the first monoclonal antibody is shown as SEQ ID NO. 2, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 7; the amino acid sequence of the light chain variable region of the second monoclonal antibody is shown as SEQ ID NO. 16, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 21. Further, the amino acid sequence of the first monoclonal antibody light chain is shown as SEQ ID NO.1, and the amino acid sequence of the heavy chain is shown as SEQ ID NO. 6; The amino acid sequence of the light chain of the second monoclonal antibody is shown as SEQ ID NO. 15, and the amino acid sequence of the heavy chain is shown as SEQ ID NO. 20. In a second aspect, the invention provides the use of said antibody for the preparation of a kit for detecting hantavirus. In a third aspect, the invention provides a nucleic acid molecule encoding the amino acid sequence of the first monoclonal antibody or the second monoclonal an