Establishment and application of zebra fish allergy detection model

A technology for detecting models and zebrafish, applied in applications, fish farming, climate change adaptation, etc., can solve problems such as subjective judgment indicators and complicated operation process

Active Publication Date: 2013-10-16
HANGZHOU HUANTE BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] 5) Simple—the experimental process is simple, and the sensitization of the drug can be quantitatively analyzed after the zebrafish is treated with the drug, while the traditional animal experiment process is complicated, the judgment index is subjective, and false positive results are prone to occur

Method used

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  • Establishment and application of zebra fish allergy detection model
  • Establishment and application of zebra fish allergy detection model
  • Establishment and application of zebra fish allergy detection model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1. Establishment of Zebrafish Allergy Detection Model

[0063] 1 Zebrafish selection

[0064] Take 4 to 5 pairs of zebrafish parents to mate, according to Westerfield [7] method for hatching embryos. The zebrafish at the optimal treatment stage were observed under a dissecting microscope, and the zebrafish with normal development were picked and moved into 96-well microwell plates.

[0065] 2 Determination of the conditions for establishing the zebrafish allergy detection model.

[0066] (1) Determination of the reaction concentration of the Tryptase detection substrate BAPNA.

[0067] BAPNA is the specific substrate of Tryptase. 10, 20, 30 and 40 mg / ml BAPNA were used to treat 3 dpf zebrafish containing 100 μg / ml C48 / 80 respectively, and the control group BAPNA was used to treat 3 dpf without C48 / 80 zebrafish. The experiment was carried out in a 96-well plate, with 2 zebrafish per well, 100 μl of reaction solution, and 6 wells for each concentration; eac...

Embodiment 2

[0079] Example 2. Zebrafish Allergy Detection Model Evaluation of the allergenicity of different batches of Tween-80.

[0080] 1 Zebrafish Selection

[0081] Take 4 to 5 pairs of zebrafish parents to mate, according to Westerfield [7] method for hatching embryos. The zebrafish at the optimal treatment stage were observed under a dissecting microscope, and the zebrafish with normal development were picked and moved into 96-well microwell plates.

[0082] 2 Tween-80 treatment

[0083] Tween-80 (#20080709 and #20091019) was provided by the China Institute for Food and Drug Control Packaging Materials and Pharmaceutical Excipients. Tween-80 to ddH 2 O was prepared at 100 mg / ml as the mother solution, and three detection concentrations were designed for each batch of Tween-80, which were 1, 10 and 100 μg / ml; the zebrafish treated with BAPNA was used as the solvent control group, and the BAPNA-treated zebrafish containing 100 μg / ml C48 / 80 zebrafish was used as a positive cont...

Embodiment 3

[0089] Example 3. Zebrafish Allergy Detection Model Evaluate the allergenicity of some impurities in Tween-80.

[0090] 1 Zebrafish selection

[0091] Take 4 to 5 pairs of zebrafish parents to mate, according to Westerfield [7] method for hatching embryos. The zebrafish at the optimal treatment stage were observed under a dissecting microscope, and the zebrafish with normal development were picked and moved into 96-well microwell plates.

[0092] 2 Impurity treatment of Tween-80

[0093] The impurities of Tween-80 are chloroethanol, ethylene glycol and 30% H 2 o 2 Provided by the China Institute for Food and Drug Control Packaging Materials and Pharmaceutical Excipients. Three detection concentrations were designed for each impurity, which were 0.01‰, 0.1‰ and 1‰; the zebrafish treated with BAPNA was used as the solvent control group, and the zebrafish treated with BAPNA containing 100 μg / ml C48 / 80 was used as the positive control group; the experiment It was carried o...

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Abstract

The invention relates to an establishment method of a zebra fish allergy detection model, and provides a method for evaluating injection (including adjuvant materials) allergenicity by the model simultaneously. The establishment method of the model mainly comprises the following steps: determining reaction concentration of Tryptase detection substrate BAPNA; determining a developmental stage of zebra fish; determining the quantity of the zebra fish for detection; determining concentration of positive control Compound 48/80, performing microwell plate analysis; and performing statistical analysis. The application of a living zebra fish model for evaluation of the injection (including the adjuvant materials) allergenicity has the advantages of reliability, fastness, efficiency, cheapness, high cost performance and the like, and high-throughput evaluation for the injection (including the adjuvant materials) allergenicity in vivo can be realized; and the establishment method and the application of the zebra fish allergy detection model has important significance on the injection (including the adjuvant materials) safety evaluation.

Description

technical field [0001] The invention relates to an in vivo detection model, in particular to a method for constructing a zebrafish model for simple, rapid and efficient in vivo detection of hypersensitivity reactions and its application. Background technique [0002] Many clinically used drugs are neither antigens nor haptens. These drugs enter the human body without a latent period and without the combination process of antigens and antibodies, and the same clinical manifestations as type I allergy occur rapidly. Since this reaction only occurs in a very small number of individuals who receive therapeutic doses of drugs, there is no sensitization process, and the serum IgE concentration of the patients does not increase, but they show typical symptoms of immediate allergic reactions, so Dukor [1] Called pseudo-drug allergy, or anaphylactoid reactions (anaphylactoid reactions). [0003] So far, most of the knowledge about anaphylactoid reactions has been obtained from clini...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K61/00
CPCY02A40/81
Inventor 李春启劳乔聪俞航萍
Owner HANGZHOU HUANTE BIOLOGICAL TECH CO LTD
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