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

Time-resolved fluorescent immunochromatographic card for detecting vomitoxin and acetylated derivatives thereof

A technology of time-resolved fluorescence and acetylated derivatives, applied in the fields of immunochromatography and food safety detection, can solve the problem of no patent disclosure, and achieve the effects of high accuracy, many surface functional groups, and bright fluorescence intensity

Inactive Publication Date: 2018-10-02
江苏省苏微微生物研究有限公司
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there are very few domestic patents related to the time-resolved fluorescence immunochromatography card for deoxynivalenol; especially in the rapid quantitative determination of the total amount of deoxynivalenol and its acetylated derivatives, there has been no patent disclosure so far

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
  • Time-resolved fluorescent immunochromatographic card for detecting vomitoxin and acetylated derivatives thereof
  • Time-resolved fluorescent immunochromatographic card for detecting vomitoxin and acetylated derivatives thereof
  • Time-resolved fluorescent immunochromatographic card for detecting vomitoxin and acetylated derivatives thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: Preparation of carboxypolystyrene fluorescent microspheres

[0049] ①Polymerizable chelate monomer (BHHCT-NHCH 2 CH=CH 2 ) preparation

[0050] Dissolve 1.57 g of allylamine and 9.85 g of BHHCT in 200 mL of absolute ethanol, add 1.02 g of triethylamine ((CH 3 CH 2 ) 3 N), stirred and reacted at 0 °C for 8 hours; then the solvent was evaporated to dryness, the solid was washed several times with ice water, and dried overnight with anhydrous magnesium sulfate; recrystallized twice in 95% ethanol to obtain 6.06 g of bright yellow solid crystals, Yield 61.5%.

[0051] ②Polymerizable fluorescent chelate monomer (Eu 3+ -BHHCT-NHCH 2 CH=CH 2 ) preparation

[0052] Dissolve 5.0 g of chelate BHHCT-NHCH in 0.05 M pH7.8 carbonate buffer 2 CH=CH 2 , formulated as 2.0×10 -5 mol / L standard solution. Then add 2.0×10 -7 mol / L EuCL 3 solution, making BHHCT-NHCH in the reaction system 2 CH=CH 2 The concentration is 2.0×10 -7 mol / L, react in a constant t...

Embodiment 2

[0057] Example 2: Preparation of DON Monoclonal Antibody 1 and Antibody 2

[0058] ① Preparation of ascites 1 and 2

[0059] Two weeks before cell culture, 10-week-old female BALB / C mice were injected with 0.5 mL of paraffin for pre-stimulation, and separated into cages for marking. The anti-vomitoxin hybridoma cell lines SW-3 F4 and SW-3 G12 were taken out from the cryopreservation state, respectively used in DMEM medium containing 20% ​​fetal bovine serum, 25 cm 2 The culture flask was revived for 3 days, and then expanded to the logarithmic growth phase and the number of cells to be injected with DMEM medium containing 15% calf serum.

[0060] Select SW-3 F4 and SW-3 G12 cells in the logarithmic growth phase, remove the culture medium and replace with serum-free DMEM medium, blow off the adherent cells in the culture bottle, and put them into a 50 mL sterilized centrifuge tube , centrifuge at 800 rpm for 5 minutes, remove the supernatant and re-suspend the cells in DMEM, ...

Embodiment 3

[0092] Example 3: Labeling DON monoclonal antibody 1 and antibody 2 with nano-europium fluorescent microspheres

[0093] ①Microsphere cleaning

[0094] Take 100 μL of carboxylated polystyrene fluorescent microspheres (particle size 85 nm) in a 2.0 mL centrifuge tube, add 200 μL of MES buffer (50 mM, pH6.0), mix well, and centrifuge at 17,500 rpm for 15 minutes to remove the supernatant. Sonicate the latex microspheres at the bottom to resuspend in 500 µL of the same MES buffer.

[0095] ②Activation of microspheres

[0096] Add 20 μL of NHS (50 mg / ml) and EDC (50 mg / ml) reaction solutions freshly prepared with MES buffer (50 mM, pH 6.0) dropwise into the centrifuge tube, vortex to mix, and place in the mixing tube Incubate for 15 minutes at room temperature with rotation.

[0097] ③Coupling of microspheres and antibodies

[0098] After incubation, wash twice with 500 μL MES buffer, centrifuge at 17,500 rpm for 15 minutes to remove the supernatant, then add 0.5 mg antibody t...

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
Diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a time-resolved fluorescent immunochromatographic card for detecting vomitoxin and acetylated derivatives thereof, and belongs to the fields of immunochromatography and food safety detection. The chromatographic card is composed of a sample pad, a labeling pad, a nitrocellulose membrane, water absorbing filter paper and a PVC bottom plate. The labeling pad is fixed with a nano europium fluorescent microsphere-labeled vomitoxin monoclonal antibody 1 and antibody 2. The antibody 1 is highly specific to vomitoxin and 15-acetyl-deoxynivalenol; the antibody 2 is highly specific to vomitoxin and 3-acetyl-deoxynivalenol. The chromatographic card is used for detection based on an antigen antibody immunology principle. The chromatographic card is combined with a portable time-resolved fluorescence immunoassay instrument, is used for rapid quantitative determination of vomitin and the acetylated derivatives thereof in cereals and products thereof, and has high sensitivityand good stability; the detection limit is 0.168 mg / kg, the detection range is 0.17-5 mg / kg, the time is 10-15 min, the operation is easy, professional training is not needed, and the chromatographiccard is widely applicable to needs for various occasions and different levels of personnel .

Description

technical field [0001] The invention relates to a time-resolved fluorescence immunochromatography card for detecting the total amount of vomitoxin and its acetylated derivatives, in particular to a preparation method and a method for rapid detection thereof, and belongs to the technical field of immunochromatography and the field of food safety detection. Background technique [0002] Deoxynivalenol, also known as deoxynivalenol (DON, hereinafter abbreviated in English), belongs to the trichothecene family compound, and its chemical name is 3α, 7α, 15-trihydroxyfusarium-9-ene -8-one, its structure is a tetracyclic sesquiterpene, the molecular formula is C 15 h 20 o 6 , molecular weight 296.3. DON mainly has two acetylated derivatives, namely 3-acetyl deoxynivalenol (3-Ac-DON, abbreviated in English hereinafter) and 15-acetyl deoxynivalenol (15- Ac-DON, hereinafter abbreviated in English); existing measurement data show that these two acetylated derivatives account for ab...

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): G01N33/533G01N33/577
CPCG01N33/533G01N33/577
Inventor 张海涛张一平龚燕董曼佳杨婷婷王红连陆丽婷华洵璐秦海萍叶进匡群
Owner 江苏省苏微微生物研究有限公司
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