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

Immunomagnetic bead for aflatoxin B1 enrichment purification and preparation method and application thereof

A technology of aflatoxin and immunomagnetic beads, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of hidden safety hazards, complicated purification and separation operations of aflatoxin B1 samples, low separation efficiency, etc. Improve the accuracy and reliability of detection and the effect of improving the lower limit of detection

Inactive Publication Date: 2016-02-10
BEIJING KWINBON BIOTECH
View PDF9 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to make up for the deficiencies in the prior art, to provide a kind of immunomagnetic beads for the enrichment and purification of aflatoxin B1 and its preparation method and application, so as to solve the complicated operation of purification and separation of aflatoxin B1 samples, the separation Technical problems with low efficiency and large potential safety hazards

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
  • Immunomagnetic bead for aflatoxin B1 enrichment purification and preparation method and application thereof
  • Immunomagnetic bead for aflatoxin B1 enrichment purification and preparation method and application thereof
  • Immunomagnetic bead for aflatoxin B1 enrichment purification and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Preparation of Immunomagnetic Beads for Aflatoxin B1 Enrichment and Purification

[0028] This example provides a method for preparing a conjugate obtained by coupling aflatoxin B1 monoclonal antibody with carboxyl-containing immunomagnetic beads as an immunomagnetic bead for enrichment and purification of aflatoxin B1. The method includes:

[0029] 1. Preparation of aflatoxin B1 monoclonal antibody

[0030] 1. Synthesis of aflatoxin B1 hapten (synthetic route see attached figure 1 ) and identification

[0031] Dissolve 62mg of aflatoxin B1 in 1mL of dimethyl sulfoxide (DMSO), slowly add 25mg of p-phenylenediamine and 0.1mL of pyridine in a mixture of 1mL of DMSO dropwise at 60°C. The solvent is removed, and the p-phenylenediamine monocondensate of aflatoxin B1 is obtained after column chromatography purification, and the hapten of aflatoxin B1 is obtained with a yield of 90%.

[0032] The above-mentioned hapten was identified by H-NMR spectroscopy, and tw...

Embodiment 2

[0060] Example 2 Characteristic Detection of Immunomagnetic Beads

[0061] Take 0.1 mL of immunomagnetic beads enriched in aflatoxin B1 prepared according to Example 1 (concentration: 10 mg / mL) in a 10 mL centrifuge tube, wash the magnetic beads twice with 5 mL of deionized water, and remove the upper beads after magnetic separation. Clear; then add 1mL of the sample to be tested (the aflatoxin B1 standard was prepared with PBS buffer solution to a concentration of 5ng / mL, 10ng / mL, 15ng / mL, 20ng / mL, 25ng / mL, 30ng / mL, 35ng / mL, 40ng / mL aflatoxin B1 solution as the sample to be tested, and PBS buffer as the blank sample to be tested), mix well, capture at 25°C for 20min, mix the magnetic beads during 5min; remove after magnetic separation For the supernatant, wash the magnetic beads twice with 5 mL deionized water to remove interfering impurities. Finally add 1mL methanol for elution, collect the eluate, and use HPLC method to detect according to "GB / T30955-2014 Determination o...

Embodiment 3

[0065] The usage method of embodiment 3 immunomagnetic beads

[0066] 1. Sample pretreatment

[0067] Homogenize the sample with a homogenizer; weigh 5.0±0.05g of the sample into a sample bottle, add 1.0±0.05g of sodium chloride, 25ml of 60% methanol solution, vortex for 5min with a vortexer, or shake on a shaker for 20min, 3000g Above, centrifuge at room temperature (20-25°C / 68-77°F) for 5 minutes; (if the centrifugation conditions are not available, this step can also be replaced by the following operation: take 10ml of supernatant after standing and filter) absorb 5ml (equivalent to 1g sample) Centrifuge the supernatant / filtrate, add 5ml deionized water, mix well, and set aside. (for magnetic bead capture).

[0068] 2. Immunomagnetic bead capture

[0069] Take 0.2ml of aflatoxin B1 immunomagnetic beads in a 10ml centrifuge tube, wash the beads twice with 5ml of deionized water, and separate the washing solution with a magnetic separation rack each time (stand still on th...

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

PropertyMeasurementUnit
Particle sizeaaaaaaaaaa
Login to View More

Abstract

The invention relates to an immunomagnetic bead for aflatoxin B1 enrichment purification and a preparation method and application thereof. A carboxyl carboxyl serves as a carrier, an aflatoxin B1 monoclonal antibody serves as a recognition midbody, the immunomagnetic bead coupled with the aflatoxin B1 monoclonal antibody is prepared through the process of activating, coupling, washing and closing, and aflatoxin B1 in a detecting sample can be efficiently captured and enriched through incubation of the immunomagnetic bead in proper buffer liquid under certain conditions. The immunomagnetic bead for aflatoxin B1 enrichment purification has the advantages of increasing the concentration of aflatoxin B1 in the sample to be detected, improving the lower limit of detection, eliminating interference of impurities, improving detecting accuracy and reliability, shortening sample treatment time and achieving fast detection.

Description

technical field [0001] The invention relates to a process for enriching and purifying aflatoxin B1 samples, in particular to an immune magnetic bead used for enriching and purifying aflatoxin B1 and its preparation method and application. Background technique [0002] Aflatoxin B1 (AflatoxinB1 is abbreviated as AFB1) is a derivative of dihydrofuran oxinone, which contains a difuran ring and an oxinone (coumarin). Aflatoxin B1 is one of the most carcinogenic chemicals known. Aflatoxin B1 is highly toxic to humans and some animals, and its toxic effect is mainly damage to the liver. In natural food, aflatoxin B1 is the most common and most harmful. AQSIQ stipulates that aflatoxin B1 is one of the mandatory inspection items for most foods. China's food hygiene standards stipulate the allowable amount of aflatoxin B1 in several major contaminated foods. The allowable amount of aflatoxin B1 in corn, peanuts, and peanut oil is ≤20 μg / kg; other edible oils are ≤10 μg / kg. kg; for...

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): G01N33/543
CPCG01N33/54326
Inventor 朱亮亮贾芳芳徐念琴罗晓琴何方洋冯静杨春艳吴小胜
Owner BEIJING KWINBON BIOTECH
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