Nanometer immune-magnetic bead, and preparation method thereof and kit thereof

A technology of immunomagnetic beads and kits, which is applied in the field of preparation of nano-immunomagnetic beads, can solve the problems of easy aggregation and precipitation, many operation steps, and low saturation magnetization, and achieve good dispersion stability, simple preparation process, and magnetic response. fast effect

Active Publication Date: 2014-10-15
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, due to its large size, the dispersion stability of magnetic beads in the target solution is poor, and it is easy to agglomerate and precipitate, so the efficiency of separation and enrichment is low
Some studies have shown that by adjusting the preparation method, magnetic beads (200-500nm) with a size close to the nanometer scale can be obtained, which improves the dispersion stability of the magnetic beads and obtains a relatively good separation and enrichment effect, but the preparation method has many steps. High labor cost, not convenient for the promotion of kits
In addition, there are also reports on the preparation method of small-sized magnetic beads (<20nm). Although it solves the problem of dispersion of large-sized magnetic beads, its saturation magnetization is low and the magnetic response is poor, which is not conducive to rapid separation and enrichment operations.

Method used

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  • Nanometer immune-magnetic bead, and preparation method thereof and kit thereof
  • Nanometer immune-magnetic bead, and preparation method thereof and kit thereof
  • Nanometer immune-magnetic bead, and preparation method thereof and kit thereof

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preparation example Construction

[0022] The method for preparing nano-immunomagnetic beads provided by the invention comprises the following steps:

[0023] (1) In an inert atmosphere, inject an alkaline solution into a mixed solution containing an iron source and a stabilizer to obtain magnetic nanoparticles; the volume ratio of the alkaline solution to the mixed solution is 1:(8-10) , the mol ratio of the iron source, the stabilizer and the alkaline substance in the alkaline solution is 1:(5-10):(10-14);

[0024] (2) Modification of antibodies with specific immune activity on the surface of magnetic nanoparticles.

[0025] According to the present invention, the alkaline substance can be any alkali metal hydroxide commonly used in the field, preferably sodium hydroxide and / or potassium hydroxide. In the alkaline solution, there is no particular limitation on the concentration of the alkaline substance, preferably 2-3 mmol / mL.

[0026] According to the present invention, the solvents in the alkaline soluti...

Embodiment 1

[0044] (1) Preparation of magnetic nanoparticles

[0045] Add sodium hydroxide to the diethylene glycol solvent, heat to 110°C under the protection of argon, keep it for 1h, and then cool down to 70°C to obtain a sodium hydroxide solution with a mass concentration of 0.1g / mL;

[0046] Add anhydrous ferric chloride and polyacrylic acid to diethylene glycol solvent (the amount of solvent is 42mL per millimole of anhydrous ferric chloride), and mix under the conditions of mechanical stirring and argon protection. The solution was heated to 220°C and kept for 30 minutes, wherein the molar ratio of anhydrous ferric chloride and polyacrylic acid was 1:10;

[0047] At 220°C, inject 3.5mL of sodium hydroxide solution into 34mL of the mixture of anhydrous ferric chloride and polyacrylic acid within 1s, adjust the temperature to 210°C after injection, and allow the reaction to continue for 1h at this temperature, Naturally cooling the reaction solution to 25°C to obtain a solution cont...

Embodiment 2

[0057] (1) Preparation of magnetic nanoparticles

[0058] Add sodium hydroxide to the diethylene glycol solvent, heat to 110°C under the protection of argon, keep it for 1h, and then cool down to 70°C to obtain a sodium hydroxide solution with a mass concentration of 0.1g / mL;

[0059] Add anhydrous ferric chloride and polyacrylic acid to diethylene glycol solvent (the amount of solvent is 42mL per millimole of anhydrous ferric chloride), and mix under the conditions of mechanical stirring and argon protection. The solution was heated to 220°C and kept for 30 minutes, wherein the molar ratio of anhydrous ferric chloride and polyacrylic acid was 1:5;

[0060] At 220°C, inject 3.5mL of sodium hydroxide solution into 34mL of the mixture of anhydrous ferric chloride and polyacrylic acid within 5s, adjust the temperature to 210°C after injection, and keep the reaction at this temperature for 1h, Naturally cooling the reaction solution to 25°C to obtain a solution containing magneti...

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Abstract

The invention discloses a nanometer immune-magnetic bead, and preparation method and a kit thereof. The method comprises the following steps: 1, injecting an iron source and stabilizer mixed liquid into an alkaline solution in an inert atmosphere to obtain magnetic nano-particles, wherein a volume ratio of the alkaline solution to the mixed liquid is 1:(8-10), and a molar ratio of the iron source to the stabilizer to an alkaline substance in the alkaline solution is 1:(5-10):(10-14); and 2, modifying the surfaces of the magnetic nano-particles surface with an antibody having specific immune activity. The preparation method of the nanometer immune-magnetic bead has a simple preparation process and good repeatability. The kit constructed on the basis of the nanometer immune-magnetic bead has the characteristics of good dispersion stability in a target detection liquid, fast magnetic response, strong specificity, high sensitivity and high target capture rate, and has a certain promotion effect on the improvement of the food safety detection efficiency.

Description

technical field [0001] The invention relates to a preparation method of nanometer immune magnetic beads, nanometer immune magnetic beads prepared by the method and a reagent kit including the nanometer immune magnetic beads. Background technique [0002] Food-borne pathogenic microorganisms have caused harm to human health that cannot be ignored in daily life. Every year, diseases caused by food-borne pathogenic microorganisms break out in various countries and regions in the world. The incidence rate is high, the infectivity is high, and it is not easy to control . How to detect and eliminate hidden dangers in the early stage of food-borne bacterial growth is a problem that has been concerned in the field of food hygiene and safety. Conventional bacterial detection methods have long cycles and low detection limits, which greatly limit the efficiency and effectiveness of food circulation and safety testing. By separating and enriching the low-concentration bacteria in food...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/28B01J20/22B01J20/26B01J20/30B01J13/02B82Y40/00B82Y30/00G01N1/40
Inventor 刘雪峰唐智勇
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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