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CN-122011182-A - Anti-Der p 2 antibody and application thereof

CN122011182ACN 122011182 ACN122011182 ACN 122011182ACN-122011182-A

Abstract

The invention relates to the field of allergen detection, in particular to an antibody for resisting Der p2 and application thereof. The invention provides an anti-Der p2 antibody, the heavy chain variable region of which comprises the amino acid sequence of a complementarity determining region, which is shown as SEQ ID No.1, SEQ ID No.2, SEQ ID No.3 and SEQ ID No.3, or the amino acid sequence of the heavy chain variable region is shown as SEQ ID No. 8. The protein of the invention utilizes the Der p2 protein which is the main allergen of dust mites of users to immunize New Zealand rabbits to obtain a group of monoclonal antibodies with high affinity and high sensitivity to Der p 2. The antibody is further subjected to humanized IgE transformation and is applied to magnetic particle chemiluminescence quantitative detection reagents of Der p2 specific IgE. The reagent can establish a standard curve for independent calibration of Der p2, so that accurate and repeatable absolute quantitative analysis is realized, and the detection sensitivity can reach 0.1 IU/mL. The detection method based on the anti-Der p2 antibody provided by the invention has important value for clinical diagnosis of anaphylactic reaction.

Inventors

  • HAN TONGTONG
  • WANG MU
  • LIU SHASHA

Assignees

  • 上海领检科技有限公司
  • 科尼格沃斯(无锡)医学科技有限公司

Dates

Publication Date
20260512
Application Date
20260206

Claims (10)

  1. 1. An anti-Der p 2 antibody is characterized in that the anti-Der p 2 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of a complementarity determining region, namely CDR1 shown in SEQ ID No.1, CDR2 shown in SEQ ID No.2 and CDR3 shown in SEQ ID No.3, or the amino acid sequence of the heavy chain variable region is shown in SEQ ID No. 8.
  2. 2. The anti-Der p2 antibody according to claim 1, wherein the light chain variable region comprises the complementarity determining region having the amino acid sequence of CDR1 shown in SEQ ID No.4, the CDR2 having the amino acid sequence of KAS shown in SEQ ID No.5, wherein KAS in the CDR2 sequence represents lysine, alanine and serine, respectively, or the light chain variable region having the amino acid sequence of SEQ ID No.9, preferably wherein the amino acid sequence of CDR1, CDR2 and CDR3 in the heavy chain variable region is located at positions 25-32, 50-56 and 93-108, respectively, and/or wherein the amino acid sequence of CDR1, CDR2 and CDR3 in the light chain variable region is located at positions 28-33, 51-53 and 90-101, respectively.
  3. 3. The anti-Der p 2 antibody of claim 1, wherein the anti-Der p 2 antibody comprises a heavy chain variable region comprising the amino acid sequence of complementarity determining regions CDR1 shown in SEQ ID No.1, CDR2 shown in SEQ ID No.2, CDR3 shown in SEQ ID No.3, and a light chain variable region comprising the amino acid sequence of complementarity determining regions CDR1 shown in SEQ ID No.4, CDR2 shown in KAS, CDR3 shown in SEQ ID No.5, or the amino acid sequence of the heavy chain variable region shown in SEQ ID No.8, and the amino acid sequence of the light chain variable region shown in SEQ ID No. 9.
  4. 4. The anti-Der p2 antibody of claim 1, wherein the amino acid sequence of the heavy chain variable region of the anti-Der p2 antibody is shown in SEQ ID No.6 and/or the amino acid sequence of the light chain variable region is shown in SEQ ID No. 7.
  5. 5. The anti-Der p 2 antibody of claim 1, wherein the anti-Der p 2 antibody further comprises a human IgE heavy chain constant region or a human Kappa chain constant region, preferably wherein the amino acid sequence of the human IgE heavy chain constant region is shown in SEQ ID No.10 and/or wherein the amino acid sequence of the human Kappa chain constant region is shown in SEQ ID No. 11.
  6. 6. The anti-Der p2 antibody according to claim 1, wherein the amino acid sequence of the heavy chain region of the anti-Der p2 antibody is shown in SEQ ID No.12 and/or the amino acid sequence of the light chain region of the anti-Der p2 antibody is shown in SEQ ID No. 13.
  7. 7. An isolated biological material, wherein the biological material is selected from one or more of the following: an isolated nucleic acid molecule encoding the anti-Der p 2 antibody of any one of claims 1-6; a nucleic acid construct comprising the nucleic acid molecule of feature 1) and an expression vector; an isolated engineered cell comprising the nucleic acid construct of feature 2) or the nucleic acid molecule of feature 1).
  8. 8. A test kit comprising an anti-Der p 2 antibody according to any one of claims 1-6.
  9. 9. Use of an anti-Der p 2 antibody according to any one of claims 1-6 or a biomaterial according to claim 7 for the preparation of a detection product, preferably a hypersensitivity disease detection product.
  10. 10. The use according to claim 9, wherein the hypersensitivity disease detection product is a dust mite allergen detection product, preferably the dust mite allergen detection product is a dust mite allergen component Der p2 specific IgE antibody detection product.

Description

Anti-Der p 2 antibody and application thereof Technical Field The invention relates to the field of allergen detection, in particular to an antibody for resisting Der p 2 and application thereof. Background House dust mites (Dermatophagoides pteronyssinus) are global indoor allergens whose class 2 allergen components Der p2 and Der p 1 are the main allergens and play a central role in the pathogenesis of dust mite allergy. Der p2 has unique structural and immunological functions, and its sensitization rate is highly correlated with clinical severity. The precursor of the house dust mite 2 allergen (Der p 2) consists of 146 amino acids, is 129 amino acids after signal peptide removal processing, has a molecular weight of 14KD and has no glycosylation site. The sensitization rate is equivalent to Der p1, and can reach 80-95% in dust mite allergic patients. It is a very stable protein, heat resistant and not easily degradable, which makes it long-lasting in the environment, continuously exposed to the human body. Furthermore, der p2 has very high sequence homology (about 88%) with Der f 2 of another major dust mite, and thus there is a strong cross reaction. Patients sensitized to Der p2 almost always respond positively to Der f 2 as well, which makes Der p2 an excellent marker for diagnosing dust mite allergy. Although Der p1 and Der p2 are both major components, about 10-20% of patients may only be sensitized to one of them. Therefore, the differential detection of Der p1 and Der p2 specific IgE can maximally improve the diagnosis sensitivity of dust mite allergy, and complete depiction of the sensitization spectrum of patients is realized. Similar to Der p1, sensitization to Der p2 is also associated with more severe clinical symptoms (e.g., asthma). Studies have shown that patients who are simultaneously sensitized to Der p1 and Der p2 are generally at higher risk and severity of asthma than patients who are sensitized to only one of them. Therefore, detection results of Der p 2-specific IgE are important basis for risk stratification and clinical phenotyping. Furthermore, der p2 is a key antigen that must be included in dust mite allergen-specific immunotherapy (Allergen Immunotherapy, AIT) formulations. Detection of Der p 2-specific IgE helps predict the efficacy of AIT. Patients with strong positives for Der p2 are more required to receive an effective formulation comprising Der p2 for treatment. Monitoring of changes in Der p 2-specific IgE levels during AIT (as may occur later) may be one of the indicators of immune response monitoring. Der p 2-specific IgE detection is significant, but the reliability of the results is severely limited by standardization issues. All commercial specific IgE detection systems (such as ImmunoCAP, immulite, etc.) currently face a fundamental technical bottleneck, namely the lack of international standards and specific standard curves for single allergen components such as Der p 2, etc., i.e. the quantitative detection of all allergen specific IgE relies on a universal standard curve calibrated with total IgE antibodies. Disclosure of Invention In view of the above-mentioned drawbacks of the prior art, an object of the present invention is to provide an antibody against Der p 2 and use thereof for solving the problems of the prior art. To achieve the above and other related objects, the present invention provides an anti-Der p 2 antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of complementarity determining regions, which is shown as SEQ ID No.1 for CDR1, SEQ ID No.2 for CDR2, and SEQ ID No.3 for CDR3, or the amino acid sequence of the heavy chain variable region is shown as SEQ ID No. 8. Preferably, the light chain variable region comprises the amino acid sequence of the complementarity determining region shown in SEQ ID No.4 as CDR1, the CDR2 sequence of the light chain variable region is shown in KAS, and the CDR2 sequence of the light chain variable region is shown in SEQ ID No.5 as CDR3, wherein KAS in the CDR2 sequence respectively represents lysine, alanine and serine, or the amino acid sequence of the light chain variable region is shown in SEQ ID No. 9. The present invention also provides an isolated biological material selected from one or more of the following: 1) An isolated nucleic acid molecule encoding the aforementioned anti-Der p2 antibody; 2) A nucleic acid construct comprising the aforementioned nucleic acid molecule and an expression vector; 3) An isolated engineered cell comprising the aforementioned nucleic acid construct or the aforementioned nucleic acid molecule. The invention also provides a detection kit comprising the anti-Der p 2 antibody. Preferably, the detection kit further comprises a plurality of anti-human IgE antibodies, magnetic beads, buffers, stabilizers, protectants, preservatives, chelating agents, or surfactants. The inven