Aflatoxin B2 aptamer affinity column and preparation method and application thereof

A kind of technology of aflatoxin and aptamer, which is applied in the direction of chemical instruments and methods, biochemical equipment and methods, separation methods, etc., can solve the problem of inability to meet the requirements of rapid screening of large batches of samples, high detection cost, and relatively expensive, etc. problem, achieve the effect of reducing variance, reducing cross-reaction, and low price

Active Publication Date: 2018-07-06
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
View PDF4 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high requirements for sample purity, some pretreatment processes are required, resulting in high detection costs and long cycles, which cannot meet the requirements of rapid screening of

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
  • Aflatoxin B2 aptamer affinity column and preparation method and application thereof
  • Aflatoxin B2 aptamer affinity column and preparation method and application thereof
  • Aflatoxin B2 aptamer affinity column and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0048] Example 1 Aflatoxin B modified with amino group 2 Aptamer preparation aptamer affinity column

[0049] 1. Preparation of cyanogen bromide modified agarose. Use Sepharose 4B as a carrier and activate with cyanogen bromide.

[0050] Mix the agarose with an equal volume of water in a fume hood, and add it to a reactor equipped with a pH electrode and a magnetic stirrer. Add cyanogen bromide to the above agarose solution at an amount of 100 mg cyanogen bromide per mL of agarose solution , Adjust the pH value to 11 with NaOH, control the pH value of the entire reaction at 11, control the temperature at about 20°C, and complete the reaction in 10 minutes; after the reaction, add an equal volume of ice chips to the above reaction solution and pour it quickly Buchner funnel, use 10 times the volume of the agarose solution in cold buffer (200mM Na 2 HPO 4 ,5mM MgCl 2 , PH 8.0) Suction filtration and washing, the hydroxyl on the surface reacts with cyanogen bromide to obtain cyanogen...

Example Embodiment

[0065] Example 2 Utilization of Aflatoxin B 2 Purification of aflatoxin B from peanut samples by aptamer affinity column 2 And its detection

[0066] In this example, aflatoxin B was added quantitatively to a normal peanut sample 2 Standard, then use the aflatoxin B prepared in Example 1 2 The aptamer affinity column was purified, and after purification, it was detected with a high performance liquid chromatography-fluorescence detector to determine the recovery rate. details as follows:

[0067] 1. Peanut sample processing

[0068] 1) Crush the peanut sample.

[0069] 2) Add aflatoxin B to the crushed peanut samples according to the standards of 0.5, 5, 50μg per gram of sample. 2 Standard.

[0070] 3) Weigh 5 g of the sample, add 25 mL of methanol-water (70:30, v / v), and place it on a homogenizer at a high speed of 11000 rpm for 3 minutes.

[0071] 4) Filter with 0.45μm syringe filter membrane.

[0072] 5) Take 5 mL of the filtrate, blow nitrogen to near dryness at 40°C, add 0.5 mL of m...

Example Embodiment

[0084] Example 3 Aflatoxin B 2 Investigation on the reusability and column capacity of aptamer affinity column

[0085] This example uses aflatoxin B 2 , And use the aflatoxin B prepared in Example 1 2 The aptamer affinity column is used for purification. Aflatoxin B retained on the affinity column 2 After eluting with methanol, the concentration was detected by high performance liquid chromatography-fluorescence detector. After the affinity column is rebalanced, the loading and elution are repeated, and the eluted aflatoxin B is determined 2 concentration. The main purpose is to test aflatoxin B 2 Reusability and column capacity of the aptamer affinity column.

[0086] 1. Prepare aflatoxin B with different solubility 2 Standard.

[0087] 2. Take out aflatoxin B 2 The aptamer affinity column is opened, the injection port plug is opened, the injection port is connected with the needle hole of the syringe, and the syringe is inserted into the air control operation frame.

[0088] 3. O...

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 provides an aflatoxin B2 aptamer affinity column and a preparation method thereof, wherein the affinity column uses cyanogens-bromide-modified agarose as a vector, a nucleic acid aptamercapable of recognizing aflatoxin B2 with high affinity and high specificity is covalently coupled to the vector, and the coupled aflatoxin B2 aptamer complex vector is loaded into the affinity column. The affinity column is mainly used for purification and cleaning of the aflatoxin B2 in food, feeds, milk, blood samples, traditional Chinese medicines and other various samples so as to facilitatehigh performance liquid chromatography and fluorescence detection of the aflatoxin B2 in the samples.

Description

technical field [0001] The invention belongs to the technical field of food safety detection, in particular, relates to a kind of aflatoxin B 2 Aptamer affinity column and its preparation method and application. Background technique [0002] Aflatoxin B 2 (Aflatoxin B 2 ,AFB 2 ) is a kind of structurally similar toxic secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus, etc. of the fungal genus. It has strong carcinogenic, teratogenic and mutagenic effects, and is the most stable mycotoxin found so far. , It is not easy to destroy under general food processing conditions, so it has planted a huge hidden danger to the dietary safety of consumers. About 25% of the food in the world may be contaminated by aflatoxin in production, processing, transportation, storage and other links every year. Due to the seriousness of aflatoxin to human health, many countries and international organizations have Toxin residues in food or traditional Chinese med...

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): C12N15/115B01J20/281B01J20/30B01D15/38C07D493/14
CPCB01D15/3804B01J20/281C07D493/14C12N15/115C12N2310/16
Inventor 刘洪美栾云霞陆安祥付海龙王纪华
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap