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Magnetic signal probe of magnetic relaxation time immunosensor and application of magnetic signal probe

An immune sensor and magnetic signal technology, applied in the field of magnetic relaxation time immune sensor detection, can solve the problems that cannot meet the detection of trace harmful substances in complex samples, lack of magnetic signal probes, poor stability, etc., and achieve improved detection Efficiency and stability, broad application prospects, and multiple amplification effects

Active Publication Date: 2020-05-26
富德赛科技(武汉)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the biggest disadvantage of the current magnetic relaxation time immunosensor is low sensitivity and poor stability, which cannot meet the detection of trace harmful substances in complex samples.
[0007] The main reason for the poor sensitivity of traditional magnetic relaxation time immunosensors is the lack of magnetic signal probes with excellent performance

Method used

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  • Magnetic signal probe of magnetic relaxation time immunosensor and application of magnetic signal probe
  • Magnetic signal probe of magnetic relaxation time immunosensor and application of magnetic signal probe
  • Magnetic signal probe of magnetic relaxation time immunosensor and application of magnetic signal probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1. Preparation of multifunctional nano-magnetic probes in this magnetic relaxation immunosensor

[0054] (1) "Ab-MNP 150 - Preparation of polylysine" conjugates

[0055] Add 20 μL EDC (10 mg / ml) and 10 μL NHS (10 mg / ml) to 100 μL ractopamine monoclonal antibody (Ab) solution (2.5 mg / mL), and vortex slowly at room temperature for 5-15 minutes. Add 500 μL MNP to the above mixed solution 150 (2mg / mL), and slowly vortex the reaction for 50-60min. Remove unreacted antibody by magnetic separation, and resuspend with 500 μL PBS (0.01M, pH=7.4) buffer to obtain Ab-MNP 150 Conjugates. Add 20μL EDC (10mg / ml) and 10μL NHS (10mg / ml) to 100μL polylysine (20mg / mL) solution, shake slowly for 10-20min, then add to the Ab-MNP 150 In the conjugate resuspension solution, slowly vortex the reaction for 1-2h. Remove unreacted polylysine by magnetic separation, and finally wash and resuspend with PBS (0.01M, pH=7.4) buffer to obtain Ab-MNP 150 - Polylysine conjugates, stored a...

Embodiment 2

[0069] Example 2. The construction of this magnetic relaxation immunosensor and the comparison with the sensitivity of traditional methods

[0070] 100 μL ractopamine complete antigen (50 μg / mL) was coated on an ELISA plate, incubated at 37° C. for 1-2 h, washed 2-4 times with PBST (0.05% Tween-20) buffer, and set aside. 100 μL Ab-MNP 150 -Polylysine-DOTA-Gd 3+ The magnetic probe was added into a gradient concentration of ractopamine (Rac) standard solution, and reacted at 37°C for 15-20min. Rac-Ab-MNP obtained by magnetic separation 150 -Polylysine-DOTA-Gd 3+ Conjugate and unreacted Ab-MNP 150 -Polylysine-DOTA-Gd 3+ Magnetic probes, and add them to the above ELISA plate, and react at room temperature for 20-60min. Wash 2-4 times with PBST buffer to remove unreacted Ab-MNP 150 -Polylysine-DOTA-Gd 3+ magnetic probe. Then add 200 μ L of PBST (5% Tween-20) buffer, keep 3min, collect the eluate, and carry out T 2 Determination of the signal.

[0071] In this embodiment,...

Embodiment 3

[0073] Embodiment 3. The determination of specificity and recovery rate of this magnetic relaxation immunosensor

[0074] (1) In the specificity test, clenbuterol, albuterol, chloramphenicol, and neomycin were used as interfering substances to test the sensitivity of the sensor. The concentrations of these analogs were 10 ng / mL and the concentration of ractopamine was 0.5 ng / mL. Such as Figure 12 As shown, only ractopamine can induce T 2 Significant changes in the value, other analogues have a negligible effect on the magnetic signal.

[0075] 2) The recovery rate was studied by the standard addition method, that is, different concentrations of ractopamine (0, 0.05, 0.1, 0.5, 1, 5, 10, 50, 100 ng / mL) were added to the blank sample. As shown in Table 1, the detection recovery rate (90%-110%) of ractopamine also shows the accuracy of the method.

[0076] Table 1. The sensor detects the recovery rate of ractopamine in the blank sample

[0077] Added concentration o...

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Abstract

The invention discloses a magnetic signal probe of a magnetic relaxation time immunosensor. The probe comprises a Gd < 3 + > chelate, polylysine and a superparamagnetic nano-particle of an antibody which are coupled. By coupling polylysine and an antibody to the surfaces of superparamagnetic nano-particles,the antibody-superparamagnetic nanoparticle-polylysine conjugate with the multi-dimensionalspace reticular dendritic structure is obtained; then, succinimide-tetraazacyclododecane tetraacetic acid with chelating performance is coupled to the surface of polylysine, and paramagnetic Gd < 3 +> ions are added; gd < 3 + > is captured through DOTA, and finally, a large number of Gd < 3 + > ions are chelated on the surfaces of the superparamagnetic nano-particles, so that the magnetic relaxation time immunosensing magnetic signal probe with multiple signal amplification is obtained. According to the invention, magnetic separation and magnetic sensing integrated nano magnetic particles andparamagnetic Gd < 3 + > ions are controllably assembled into the multifunctional nano magnetic probe for the first time, so that multiple amplification of sensing signals is realized, and the sensitivity and detection speed of the sensor are greatly improved.

Description

technical field [0001] The invention belongs to the field of food safety detection and clinical diagnosis analysis. It specifically relates to a nano-magnetic particle as a carrier, and layer-by-layer coupling of Gd 3+ The magnetic relaxation time immunosensing magnetic signal probe for multiple signal amplification of ions and antibodies, and the invention also relates to a magnetic relaxation time immunosensing detection method based on the magnetic probe. Background technique [0002] Food safety is a hot topic of concern. In recent years, the frequent occurrence of food safety incidents has brought huge troubles and losses to the country and the people. Food safety issues mainly include microbial contamination, food additives, excessive pesticide and veterinary drug residues, and heavy metal pollution. Although my country has formulated national standards and corresponding detection methods for different pollutants, food safety problems caused by food-borne pathogenic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/569G01N33/74
CPCG01N33/5434G01N33/54346G01N33/56911G01N33/74G01N33/9413G01N2333/585G01N2446/20G01N2446/80G01N2446/84G01N2469/10
Inventor 陈翊平董永贞
Owner 富德赛科技(武汉)有限公司
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