Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Enzyme linked immunosorbent assay kit, preparation method thereof and method for detecting pepsase through same

An enzyme-linked immunosorbent reagent and pepsin technology, which is applied in the field of clinical diagnosis, can solve the problems of single structure of enzyme-linked reaction kit, inconvenient, no use of pepsin chemiluminescent enzyme-linked immunosorbent kit, etc., to improve the convenience of the kit Accuracy, the effect of avoiding complete separation

Active Publication Date: 2015-11-25
中国人民解放军总医院第六医学中心 +1
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no related technology for pepsin detection using chemiluminescent ELISA, and there is no chemiluminescent ELISA kit for pepsin detection.
[0006] In addition, the structure of the traditional enzyme-linked reaction kit is relatively simple and single, which has certain inconvenience during the test operation.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Enzyme linked immunosorbent assay kit, preparation method thereof and method for detecting pepsase through same
  • Enzyme linked immunosorbent assay kit, preparation method thereof and method for detecting pepsase through same
  • Enzyme linked immunosorbent assay kit, preparation method thereof and method for detecting pepsase through same

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0066] A preparation method of an enzyme-linked immunosorbent kit, comprising the following steps:

[0067] Step 1. Preparation of pepsin monoclonal antibody, extracting pepsin in human gastric mucosal secretions, immunizing BALB / c mice with purified pepsin, and preparing the pepsin monoclonal antibody by hybridoma fusion technology;

[0068] Step 2, the preparation of the microplate 6, after diluting the obtained pepsin monoclonal antibody, adding it to the microwell 7 for coating, to obtain the microplate 6;

[0069] Step 3, the preparation of enzyme-labeled pepsin antibody, preparing a horseradish peroxidase solution, mixing the pepsin monoclonal antibody with the horseradish peroxidase solution, and then adding NaBH to the obtained reaction solution 4 The solution is then subjected to ultrafiltration, and the ultrafiltration retentate is retained to obtain the enzyme-labeled pepsin antibody.

[0070] In the preparation method of described ELISA kit,

[0071] In step 1, p...

Embodiment 1

[0080] making process:

[0081] Step 1, preparation of pepsin monoclonal antibody. Use ion exchange chromatography and gel filtration technology to separate and purify pepsin in human gastric juice, immunize BALB / c mice with purified pepsin, prepare monoclonal antibody against human pepsin by hybridoma fusion technology, and use protein affinity Chromatography and gel filtration techniques are used to purify monoclonal antibodies; enzyme-linked immunosorbent techniques are used to screen highly specific diagnostic monoclonal antibodies.

[0082] Step 2, preparation of the enzyme-labeled antibody in the kit. Weigh horseradish peroxidase and dissolve it in ultrapure water to make a solution, let it stand overnight at 3°C ​​to fully dissolve it; take the above solution the next day, and add freshly prepared NaIO to it 4 Aqueous solution, after mixing, place in the refrigerator to avoid light and stir. The reaction solution was taken out and put into a dialysis bag, and the rea...

Embodiment 2

[0088] making process:

[0089] Step 1, preparation of pepsin monoclonal antibody. Use ion exchange chromatography and gel filtration technology to separate and purify pepsin in human gastric juice, immunize BALB / c mice with purified pepsin, prepare monoclonal antibody against human pepsin by hybridoma fusion technology, and use protein affinity Chromatography and gel filtration techniques are used to purify monoclonal antibodies; enzyme-linked immunosorbent techniques are used to screen highly specific diagnostic monoclonal antibodies.

[0090] Step 2, preparation of the enzyme-labeled antibody in the kit. Weigh horseradish peroxidase and dissolve it in ultrapure water to prepare a solution, let it stand overnight at 4°C to fully dissolve it; take the above solution the next day, and add freshly prepared NaIO to it 4 Aqueous solution, after mixing, place in the refrigerator to avoid light and stir. The reaction solution was taken out and put into a dialysis bag, and the re...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an enzyme linked immunosorbent assay kit, a preparation method thereof and a method for detecting pepsase through the same. The enzyme linked immunosorbent assay kit comprises a kit body, an elisa plate and reagent bottles. The elisa plate is provided with a plurality of micro holes which contain pepsase monoclonal antibodies. The elisa plate is contained in the kit body. The reagent bottles are arranged in the kit body and contain a working solution, a standard solution, a buffer solution, a scrubbing solution and a developing solution respectively, wherein the working solution contains an enzyme-labeled pepsase antibody. The label of the enzyme-labeled pepsase antibody is horse radish peroxidase, and labeling is carried out through a sodium periodate method. According to the enzyme linked immunosorbent assay kit, the chemiluminescence enzyme linked immunosorbent technology can be utilized for accurately detecting pepsase, and therefore the purpose of non-invasive diagnosis of the laryngopharyngeal reflux disease is achieved.

Description

technical field [0001] The present invention relates to the technical field of clinical diagnosis, more specifically, the present invention relates to a chemiluminescent enzyme-linked immunosorbent assay kit for detecting pepsin, which can detect the content of pepsin and perform non-invasive diagnosis on laryngopharyngeal reflux disease . Background technique [0002] Laryngopharyngeal reflux disease refers to the general term for a series of symptoms and signs caused by the reflux of gastric contents to the position above the upper esophageal sphincter. Symptoms such as chronic long-term cough, dyspnea, and laryngospasm, as well as mucosal hyperplasia and hypertrophy in the posterior commissure of the vocal cords, diffuse congestion and edema of the vocal cords, and in severe cases, laryngeal signs such as granuloma, disappearance of the laryngeal chamber, and subglottic stenosis. [0003] Laryngopharyngeal reflux disease is the source of some diseases in otolaryngology h...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N21/76G01N33/573G01N33/577
Inventor 李进让吴立峰郭春雨
Owner 中国人民解放军总医院第六医学中心
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com