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Human alpha-defensin peptide enzyme linked immunosorbent assay kit

An immunoadsorption and defensin technology, which can be used in biological testing, material inspection products, measuring devices, etc., and can solve the problems of high price, narrow detection range and low sensitivity.

Inactive Publication Date: 2015-04-22
WUHAN NCE BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commercially available double-antibody sandwich kits for detecting HNP generally have shortcomings such as narrow detection range, low sensitivity, and high price.

Method used

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  • Human alpha-defensin peptide enzyme linked immunosorbent assay kit
  • Human alpha-defensin peptide enzyme linked immunosorbent assay kit
  • Human alpha-defensin peptide enzyme linked immunosorbent assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1 Index detection of a human alpha defensin polypeptide enzyme-linked immunosorbent assay kit

[0080] This kit uses a double-antibody sandwich enzyme-linked immunoassay to determine the level of HNP in serum or plasma samples. The preparation process is as follows: use a 96-well polystyrene reagent plate (PS) as a solid phase carrier, and first place it under ultraviolet light Irradiate for 2 hours to activate the PS plate; pre-coat a certain concentration of anti-human and mouse monoclonal antibody on the microwell strip of the reagent plate, overnight at 4°C, wash the plate and block with bovine serum albumin to make the coated ELISA plate; when testing, add the sample to be tested or different concentrations of HNP protein standard solution, anti-human HNP rabbit polyclonal antibody, and horseradish peroxidase-labeled goat anti-rabbit polyclonal antibody to form the sample to be tested or HNP Standard-rabbit polyclonal antibody-enzyme-labeled goat anti-rabbi...

Embodiment 2

[0106] Embodiment 2 The test kit of the present invention and the test kit of a certain foreign A import company

[0107] The inventor compared the differences in precision, recovery and other aspects of quantitative detection between the human neutrophil α-defensin polypeptide detection kit described in the present invention and the imported kit through the following experiments.

[0108] 1) Precision experiment:

[0109] The human neutrophil α-defensin polypeptide detection kit of the present invention and the kit of an imported company from abroad were respectively used to detect high, medium and low value quality control products. These two kits were used to measure three constant value samples 8564.1pg / mL, 1504.6pg / mL, and 42.4pg / mL respectively, and each sample was repeatedly measured 20 times. The specific steps are the same as in Example 1. The specific data are shown in Table 5. From From the data in Table 5, it can be seen that the precision of the human neutrophil ...

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Abstract

The invention relates to a human alpha-defensin peptide enzyme linked immunosorbent assay kit and a preparation and detection method thereof. The human alpha-defensin peptide enzyme linked immunosorbent assay kit comprises a mouse-anti-human HNP monoclonal antibody coated ELISA plate, an HNP protein freeze-dried standard substance, a rabbit anti-human HNP polyclonal antibody, a horseradish peroxidase labeled goat anti-rabbit immune globulin polyclonal antibody, standard diluent, sample diluent, a 25-fold concentrated washing buffer solution, antibody diluent, a TMB developing solution and an enzyme reaction terminating solution. The preparation method comprises the following steps: coupling alpha-defensin and a carrier protein, adopting a coupling compound with the optimal coupling binding ratio (the molar ratio of HNP to SPDP to mouse serum albumin is 1:5:1, and the molar ratio of HNP to SPDP to rabbit serum albumin is 1:5:1, preparing to obtain a high-sensitivity antibody by using a complete antigen, and assembling the kit by using the obtained antibody. The kit disclosed by the invention has the advantages of high detection sensitivity, accurate result, wide detection range, simplicity and convenience in operation, long preservation time and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a human α-defensin polypeptide enzyme-linked immunosorbent assay kit, preparation and detection methods. Background technique [0002] Defensins belong to the family of antimicrobial peptides and widely exist in various organisms including plants, invertebrates and vertebrates. Defensin protein peptides are cationic small molecular short peptides with a molecular weight of 2-6kDa, containing six conserved cysteine ​​residues, forming three typical intramolecular disulfide bonds, and are divided into α-defensins and 13-defensins Two categories of ingredients. α-defensins mainly exist in the azurophilic granules of human neutrophils, and are also called human neutrophil peptides (Human neutrophol peptides, HNP). There are three main types of human neutrophil defensins, HNP1, HNP2 and HNP3, which can promote neutrophil chemotaxis, enhance macrophage phagocytosis, regulate complement a...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577
CPCG01N33/68G01N33/577
Inventor 韩兵王烁冯玉冰万君花刘俊
Owner WUHAN NCE BIOMEDICAL CO LTD
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