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Application of allergen specific IgE antibody idiotypic heterogeneity intensity in kit for allergen detection

An allergen and heterogeneity technology, applied in the field of clinical medicine/experimental diagnostics, can solve problems such as inappropriateness, expensive testing costs, and high technical requirements for operators, and achieve good correlation results

Pending Publication Date: 2020-02-07
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for cell biology tests, fresh peripheral blood and leukocyte extraction are required, and separate blood collection is required at a predetermined time
At the same time, the entire detection process is relatively complicated, requires flow cytometry, and has high technical requirements for operators, coupled with expensive detection costs, it is not suitable for popularization and use as a routine detection method

Method used

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  • Application of allergen specific IgE antibody idiotypic heterogeneity intensity in kit for allergen detection
  • Application of allergen specific IgE antibody idiotypic heterogeneity intensity in kit for allergen detection
  • Application of allergen specific IgE antibody idiotypic heterogeneity intensity in kit for allergen detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The preparation of embodiment 1 kit

[0042] The kit mainly contains R1 solution, R2 solution and washing buffer, and the specific preparation method is as follows:

[0043] (1) Preparation of R1 solution

[0044] Including the steps of binding different coded fluorescent microspheres (MB, purchased from Merck) coated with SA to different biotin-labeled epitopes (E); the specific method is: 1ml of fluorescent microspheres (concentration: 1mg / mL )+Bio-E (concentration: 1 mg / mL) 100 μL, mix well and incubate with shaking for 1 hour, wash the microspheres, and restore to the original volume with fluorescent microsphere preservation solution. Fluorescent microspheres coated with different epitopes and different codes were mixed in equal proportions, and the concentration of the microspheres was adjusted to 0.01 mg / ml as a working solution to obtain the R1 solution.

[0045] (2) Preparation of R2 solution

[0046] The R2 solution is a phycoerythrin (PE)-labeled mouse anti...

Embodiment 2 Embodiment 1

[0050] The detection method of the use process of kit in embodiment 2 embodiment 1

[0051] The detection steps are as follows:

[0052] (1) Dilute the serum sample with 0.01M pH 7.2 phosphate buffer (containing 5% bovine serum albumin) to dilute the serum sample 1:10, such as 180 μL buffer + 20 μL serum;

[0053] (2) Add 25 μL of 1:10 diluted serum sample, 100 μL of mixed reagent 1 indirectly coated with different antigenic epitopes and different fluorescent codes into a 96-well plate, and incubate on a plate shaker at 37°C for 1 hour to make The sIgE antibodies with different idiotype epitopes in the serum to be tested combine with the corresponding peptides on the surface of the microspheres one by one to form immune complexes;

[0054] (3) Place the 96-well plate on the magnetic field for 3 minutes, discard the unbound serum protein in the supernatant; after washing three times with washing buffer, discard the supernatant and add 100 μL phosphate buffer (same as above) to...

Embodiment 3 application Embodiment 1

[0061] Example 3 Using the kit in Example 1 and the detection method in Example 2 to detect the heterogeneity intensity of milk allergen β-lactoglobulin sIgE idiotype epitope

[0062] The detection process is as follows:

[0063] (1) The information of the selected epitope is as follows:

[0064] Table: Amino acid sequences of selected β-lactoglobulin epitopes

[0065]

[0066] The amino-terminus was labeled with biotin molecules (Bio-E), and Shanghai Jier Biochemical was commissioned to synthesize these 8 peptides.

[0067] (2) Labeling: purchase 8 different coded fluorescent microspheres (MB 01-08) coated with SA, take 1 mL (concentration: 1 mg / mL) of each and add them to EP tubes, then add 100 μL (concentration: 1 mg / mL) of the above Biotin-labeled antigenic epitopes were incubated at 37°C for 1 hour with shaking. Remove unbound polypeptides by a magnetic field, wash with PBST three times, and restore to the original volume with fluorescent microsphere preservation so...

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Abstract

The invention belongs to the field of clinical medicine/experimental diagnostics, relates to experimental diagnosis of allergic diseases, and particularly provides a brand-new experimental diagnosis index, namely allergen-specific IgE (sIgE) antibody idiotypic heterogeneity, for allergic diseases. The allergen sIgE antibody idiotypic heterogeneity intensity is associated with patient clinical manifestations or severity of allergic symptoms. The invention especially relates to an application of allergen specific IgE antibody idiotypic heterogeneity intensity in a kit for allergen detection.

Description

technical field [0001] The invention belongs to the field of clinical medicine / experimental diagnostics, relates to experimental diagnosis of allergic diseases, and specifically provides a brand-new experimental diagnostic index for allergic diseases, that is, idiotype heterogeneity of allergen-specific IgE (sIgE) antibodies. The intensity of allergen sIgE antibody idiotype heterogeneity is related to the clinical manifestations of patients or the severity of allergic symptoms. In particular, it relates to the application of the allergen-specific IgE antibody idiotype heterogeneity intensity in the allergen detection kit. Background technique [0002] (1) Allergic diseases [0003] Hypersensitivity refers to the pathological or abnormal immune response caused by the sensitized body encountering the same antigen stimulation again. According to different clinical manifestations and mechanisms, hypersensitivity reactions are divided into 4 types, among which, type I hypersens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/543G01N33/58G01N15/14
CPCG01N33/533G01N33/54326G01N33/582G01N15/14
Inventor 李会强李军普李柳栩黄伦辉张蓓崔亚琼刘丹丹于洋
Owner TIANJIN MEDICAL UNIV
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