Timed-resolved fluoroimmunoassay detection method of allergen peanut protein

A detection method, a technology for allergens, applied in the field of time-resolved immunofluorescence analysis and detection of allergen peanut protein

Inactive Publication Date: 2013-02-06
FOOD INSPECTION CENT OF CIQ SHENZHEN
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protein detection methods of ELISA and LFIA are mainly to detect the total peanut protein or the main peanut allergen (Ara h 1 or Ara h 2) in food. LFIA can only provide yes / no test results, and ELISA can be semi-quantitative or full quantitative. A little more sensitive but takes more time

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
  • Timed-resolved fluoroimmunoassay detection method of allergen peanut protein
  • Timed-resolved fluoroimmunoassay detection method of allergen peanut protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0009] The example is a further description of the time-resolved immunofluorescence analysis and detection method of the allergen peanut protein provided by the present invention, but the embodiments of the present invention are not limited thereto.

[0010] (1) Antibody biotin labeling:

[0011] Replace the original buffer of the purified allergen Ara h1 protein monoclonal antibody 2G9 with phosphate buffer solution (PBS), and then mix the monoclonal antibody 5G4 with NHS-activated biotin (NSH-Biotin) in an appropriate ratio. After light reaction for 30 minutes, use a 50kDa ultrafiltration membrane column to remove NSH-Biotin that may not be involved in the labeling.

[0012] (2) Preparation of coated board:

[0013] The purified anti-peanut allergen Ara h1 protein monoclonal antibody 5G4 was used 50mmol / L Na 2 CO 3 -NaHCO 3 Dilute the pH 9.6 buffer to an appropriate concentration, add 100 μl to each well of a 96-well microtiter plate, and place at 4°C overnight. Discard the coatin...

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 a detection method for detecting an allergen peanut protein ingredient in food and belongs to the technical field of a timed-resolved fluoroimmunoassay (TRFIA) for detecting the content of the allergen peanut protein in the food. The foundation for measurement disclosed by the invention is double antibody sandwich immune responses and a time-resolved immunofluorescence technique. The method comprises the steps of: using a microwell plate of coating anti-peanut allergen Arah1 protein monoclonal antibody 5G4, taking anti-peanut allergen Arah1 protein monoclonal antibody 2G9 marked by biotin as a capture antibody; using EU3(+)- streptavidin to identify double sandwich antigen-antibody compound, measuring the fluorescence intensity cps by a time-resolved fluorescence spectrometer after adding enhancement solution to oscillate, and determining the peanut allergen Arah1 protein content in the sample by comparing with a standard curve, so as to calculate the allergen peanut protein content in the sample because the Arah1 protein in the peanut is 12-16% of total protein of the peanut. The detection method of the allergen peanut protein ingredient in the food provided by the invention is simple and cheap, and high in sensitivity, and has good application prospect in the field of allergen detection in food safety.

Description

Technical field [0001] A detection method for detecting peanut protein components of food allergens belongs to the technical field of time-resolved fluorescence immunoassay (TRFIA), and is used for detecting the content of allergen peanut protein in food. Background technique [0002] Among food allergies, the symptoms of peanut allergic reaction are the most serious. Even a small amount of ingestion can induce obvious peanut allergic reactions, which can lead to anaphylactic shock and allergic to death. Currently, there is no effective treatment for this peanut allergy symptom. Avoiding the ingestion of flowering allergens can effectively prevent the occurrence of severe allergic reactions and has become one of the effective ways to prevent peanut allergy. The food labeling laws of the United States, the European Union and other countries stipulate that peanuts must be labeled as the main allergen ingredient, and some imported foods have been tested for peanut allergens, and ev...

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
Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N21/64G01N33/531
Inventor汤慕瑾刘志刚吕敬章张恒万志刚陈昊翰蔡伟增
OwnerFOOD INSPECTION CENT OF CIQ SHENZHEN