Rapid detection device for Sudan III hapten, coupled antigen, antibody and colloidal gold and its application

A detection device and antigen-coupling technology, which is applied in the direction of measuring devices, food testing, material inspection products, etc., can solve the problems of high technical requirements for operators, failure to display results immediately, complicated operation of instruments and equipment, etc., and achieve convenient and economical use Fast, low-cost results

Active Publication Date: 2020-04-14
四川省食品药品检验检测院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The instruments and equipment used in the above methods are complicated to operate, high in cost, and have high technical requirements for operators, and the results cannot be displayed immediately, so they are not suitable for rapid online detection and monitoring of suspected objects by regulatory authorities and production enterprises

Method used

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  • Rapid detection device for Sudan III hapten, coupled antigen, antibody and colloidal gold and its application
  • Rapid detection device for Sudan III hapten, coupled antigen, antibody and colloidal gold and its application
  • Rapid detection device for Sudan III hapten, coupled antigen, antibody and colloidal gold and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1. Preparation of Sudan Red III Hapten

[0059] Add 3.53g of Sudan Red III, 1.52g of bromoacetic acid, and 0.1g of sodium carbonate to a 100mL three-necked flask in sequence, and react overnight in acetonitrile solution at 90°C. After rotary evaporation was completed, ethyl acetate was extracted and purified through a column to obtain Sudan III hapten.

[0060] figure 2 The mass spectrum of the thus obtained Sudan III hapten is given, and it can be known that the molecular formula of the hapten is as follows:

[0061]

Embodiment 2

[0062] Example 2. Synthesis of Sudan Red III coupled antigen

[0063] The Sudan III hapten obtained in Example 1 was used to prepare the immune antigen. Dissolve 0.1 mmol of Sudan Red III hapten in 2 mL of DMF, and add 27.5 mg of dicyclohexylcarbodiimide (DCC) and 14.4 mg of N-hydroxysuccinimide (NHS) with stirring. Magnetic stirring was carried out overnight at 4°C. After centrifugation, the supernatant was liquid A. Weighed 140 mg of bovine serum albumin (BSA) and hemocyanin (KLH), and dissolved them in 10 mL of 0.1 mol / L PBS (pH 8 .0). Add 1 mL of dimethylformamide (DMF), stir and dissolve to prepare liquid B, gradually drop liquid A into liquid B under magnetic stirring, and react at 4°C for 12 hours. After centrifugation, the supernatant was taken, dialyzed with normal saline at 4°C for 3 days, and the dialysate was changed 3 times a day, so as to obtain Sudan Red III-coupled antigens respectively coupled with bovine serum albumin and hemocyanin. The obtained Sudan Red...

Embodiment 3

[0064] Example 3. Preparation of Sudan Red III Monoclonal Antibody

[0065] The Sudan Red III conjugated antigen prepared in Example 2 was used to prepare the Sudan Red III monoclonal antibody, as follows: 4 6-week-old Kunming mice were immunized with the identified Sudan Red III conjugated antigen, and after three booster immunizations, blood was collected Measure the potency. When the serum titer no longer rises, mice are immunized with twice the dose of antigen without adjuvant. Three days later, the mice are killed by decapitation. The spleen is taken under sterile conditions to prepare spleen cells, which are compared with vigorously growing mouse myeloma cells. Mix in a 50mL centrifuge tube at a ratio of 8:1, add 30mL of serum-free IPMI 1640 medium, centrifuge at 1100r / min for 5 minutes, discard the supernatant, shake the cell mass gently, and place in a 37°C water bath. Slowly add 1mL of 50% PEG-4000 to the cells, drop it within 1 minute, and gently stir the sediment a...

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Abstract

The invention belongs to the field of food safety detection, and more specifically relates to a sudan red III semiantigen with a structure represented by formula I, and also relates to a sudan red IIIcoupling antigen, a sudan red III antibody, and a colloidal gold rapid detection device containing the sudan red III coupling antigen and the sudan red III antibody, and applications thereof. The cross reaction of the colloidal gold rapid detection device with other kinds sudan red is reduced, so that the colloidal gold rapid detection device can be used for detecting sudan red III in food accurately; cost is low; detection is rapid and accurate; and rapid on-line detection and monitoring of suspected objects can be realized.

Description

technical field [0001] The invention belongs to the field of food safety detection. More specifically, the present invention relates to a Sudan Red III hapten, a Sudan Red III conjugated antigen, a Sudan Red III antibody, a colloidal gold rapid detection device comprising the Sudan Red III conjugated antigen and a Sudan Red III antibody, and its application. Background technique [0002] Sudan Red is a class of industrial dyes with an azo structure, including Sudan Red I, II, III and IV. Sudan red is carcinogenic, and it is a coloring agent that is banned from being used in food in my country. The four types of Sudan Red have great similarities, and it is necessary to distinguish the added types in the detection and traceability. At present, when antibodies are used to detect Sudan red compounds, the cross-reactivity is relatively serious, and most of them cannot be distinguished. [0003] In addition, the existing Sudan red detection methods mainly include high performan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/02
CPCG01N33/02G01N33/558
Inventor 余晓琴王炳志严义勇朱海马红圳陈思斯付辉黄瑛李道霞
Owner 四川省食品药品检验检测院
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