Search

CN-121991225-A - Monoclonal antibody combination for detecting human IL-33 protein and application thereof

CN121991225ACN 121991225 ACN121991225 ACN 121991225ACN-121991225-A

Abstract

The invention relates to the field of biological detection, in particular to a monoclonal antibody combination for detecting human IL-33 protein and application thereof. The antibody combination comprises 1B3 and 4A10, wherein the amino acid sequence of the complementarity determining region of the heavy chain variable region of the monoclonal antibody 1B3 corresponds to SEQ ID NO.1-SEQ ID NO.3, the CDR amino acid sequence of the light chain variable region corresponds to SEQ ID NO.4-SEQ ID NO.6, the CDR amino acid sequence of the heavy chain variable region of the monoclonal antibody 4A10 corresponds to SEQ ID NO.7-SEQ ID NO.9, and the CDR amino acid sequence of the light chain variable region corresponds to SEQ ID NO.10-SEQ ID NO.12. The antibody combination ensures the production repeatability of the raw materials, provides a reliable basis for preparing diagnosis tools such as ELISA kits with high sensitivity and high stability, and has wide application prospect.

Inventors

  • QIN KUN
  • LU SHUAI
  • LI YUE
  • SUN YUTIAN
  • ZHONG MING
  • SU DU

Assignees

  • 北京溯本源和生物科技有限公司

Dates

Publication Date
20260508
Application Date
20260408

Claims (10)

  1. 1. A monoclonal antibody combination for detecting human IL-33 protein, wherein the monoclonal antibody combination comprises monoclonal antibody 1B3 and monoclonal antibody 4a10; the heavy chain variable region of the monoclonal antibody 1B3 comprises three complementarity determining regions, and the amino acid sequences of the complementarity determining regions are respectively shown as SEQ ID NO.1-SEQ ID NO. 3; the light chain variable region of the monoclonal antibody 1B3 comprises three complementarity determining regions, and the amino acid sequences of the complementarity determining regions are respectively shown in SEQ ID NO.4-SEQ ID NO. 6; The heavy chain variable region of the monoclonal antibody 4A10 comprises three complementarity determining regions, and the amino acid sequences of the complementarity determining regions are respectively shown in SEQ ID NO.7-SEQ ID NO. 9; the light chain variable region of monoclonal antibody 4A10 comprises three complementarity determining regions, and the amino acid sequences of the complementarity determining regions are shown in SEQ ID NO.10-SEQ ID NO.12, respectively.
  2. 2. The monoclonal antibody combination for detecting human IL-33 protein according to claim 1, wherein the amino acid sequence of the heavy chain variable region of the monoclonal antibody 1B3 is shown in SEQ ID NO.13, and the amino acid sequence of the light chain variable region of the monoclonal antibody 1B3 is shown in SEQ ID NO. 14.
  3. 3. The monoclonal antibody combination for detecting human IL-33 protein according to claim 2, wherein the amino acid sequence of the heavy chain variable region of the monoclonal antibody 4A10 is shown in SEQ ID NO.15, and the amino acid sequence of the light chain variable region of the monoclonal antibody 4A10 is shown in SEQ ID NO. 16.
  4. 4. The monoclonal antibody combination for detecting human IL-33 protein according to claim 3, wherein the nucleotide sequence encoding the heavy chain variable region of the monoclonal antibody 1B3 is shown in SEQ ID NO.17 and the nucleotide sequence encoding the light chain variable region of the monoclonal antibody 1B3 is shown in SEQ ID NO. 18.
  5. 5. The monoclonal antibody combination for detecting human IL-33 protein according to claim 4, wherein the nucleotide sequence encoding the heavy chain variable region of monoclonal antibody 4A10 is shown in SEQ ID NO.19 and the nucleotide sequence encoding the light chain variable region of monoclonal antibody 4A10 is shown in SEQ ID NO. 20.
  6. 6. The monoclonal antibody combination for detecting human IL-33 protein according to claim 5, wherein the monoclonal antibody combination specifically recognizes a recombinant protein of human IL-33 protein and a natural protein of human IL-33.
  7. 7. Use of a combination based on the monoclonal antibody of claim 1 for the preparation of a tool for detecting human IL-33 protein.
  8. 8. The use of claim 7, wherein the means comprises reagents, kits, test strips and antibody chips.
  9. 9. The use according to claim 8, wherein the kit comprises a double antibody sandwich ELISA kit.
  10. 10. The use according to claim 9, wherein the ELISA kit employs monoclonal antibody 1B3 as the coating antibody and monoclonal antibody 4a10 as the labeling antibody.

Description

Monoclonal antibody combination for detecting human IL-33 protein and application thereof Technical Field The invention relates to the field of biological detection, in particular to a monoclonal antibody combination for detecting human IL-33 protein and application thereof. Background Interleukin 33 (IL-33), which belongs to the Interleukin-1 (IL-1) family, is a pro-inflammatory protein with a structure that is highly similar to IL-1β of the human IL-1 subfamily. IL-33 protein has transcription regulation property, and related research shows that IL-33 is a protein with two functions, and can be used as cell factor and nuclear factor in cell, and is mainly expressed in endothelial cell, epithelial cell and fibroblast-like cell. As an "alerter," IL-33 is released when cells or tissues become damaged, suggesting the onset of inflammation or injury in the body, and activating immune cells expressing the ST2 receptor (IL-1 RL 1), thereby rapidly initiating an immune response. IL-33 signaling is dependent on its complex formation with ST2 and the co-receptor IL-1RAcP, activating NF-. Kappa.B and MAPK signaling pathways, thereby inducing downstream immune responses. IL-33 activates type 2 innate lymphoid cells (ILC 2), helper T cells type 2 (Th 2), mast cells and eosinophils, promotes secretion of Th2 cytokines such as IL-4, IL-5 and IL-13, and plays an important role in Th2 immune response, inflammatory regulation, tissue repair, fibrosis and the like. In addition, IL-33 is involved in innate immune regulation and is closely related to various diseases such as asthma, allergic diseases, inflammatory bowel diseases, autoimmune diseases, cardiovascular diseases and the like, and is an important immune regulation factor and a potential disease biomarker and an intervention target. Therefore, the IL-33 level in serum, plasma, tissue homogenate or cell culture supernatant is accurately and sensitively detected, and the method has important significance for researching disease mechanism, evaluating inflammatory state and monitoring treatment effect. Currently, methods for detecting IL-33 mainly include immunological methods (such as ELISA, immunoblotting, flow cytometry), molecular biological methods (such as qRT-PCR), and biochip technologies. Although the methods have advantages, in practical application, the qRT-PCR has some limitations that the qRT-PCR can only reflect mRNA level and cannot directly reflect protein amount or secretion activity, immunoblotting and flow cytometry are complex in operation, high in standardization difficulty, low-abundance samples are easy to be interfered by background noise, a biochip can realize high-flux detection, but sensitivity and quantitative accuracy are limited, dependence on antibodies or probes is strong, and batch-to-batch differences can influence result consistency. In contrast, enzyme-linked immunosorbent assay (ELISA) is still the main method for detecting IL-33 due to the ability to directly quantify proteins, simple operation and easy standardization. However, the existing part of IL-33 detection products still have the defects in antibody raw materials. Some reagents adopt polyclonal antibodies or monoclonal antibodies with undefined epitope information, so that nonspecific binding or background interference is easy to occur, meanwhile, the pairing effect between different antibodies is not ideal, the detection sensitivity is limited, and the detection is unstable particularly in low-abundance sample detection. In addition, the undefined antibody sequence or inconsistent sources can cause the fluctuation of the performances of kits in different batches, seriously influence the reliability of detection results, and are difficult to meet the requirements of high-standard scientific research and clinical detection. Therefore, the method for obtaining the human IL-33 monoclonal antibody with high affinity, high specificity and definite sequence, and screening out the antibody pairing combination suitable for sandwich ELISA is the key for constructing a stable, reliable and high-performance IL-33 immune detection system. Disclosure of Invention First, the technical problem to be solved In view of the above-mentioned shortcomings and disadvantages of the prior art, the present invention provides a monoclonal antibody combination and application for detecting human IL-33 protein, which solves the technical problems of non-specific binding and limited sensitivity of the antibody raw material in the prior art. (II) technical scheme In order to achieve the above purpose, the main technical scheme adopted by the invention comprises the following steps: In a first aspect, the application provides a monoclonal antibody combination for detecting human IL-33 protein, the monoclonal antibody combination comprising monoclonal antibody 1B3 and monoclonal antibody 4a10; the heavy chain variable region of the monoclonal antibody 1B3 comprises three complementarity determining region