Large-scale manufacturing method of diagnostic magnetic beads and biological application thereof

A manufacturing method and large-scale technology, applied in biochemical equipment and methods, microbiological measurement/inspection, DNA preparation, etc., can solve problems such as difficult to achieve large-scale manufacturing and cumbersome steps

Active Publication Date: 2021-04-30
苏州白垩纪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing technology has been able to achieve the laboratory production of tens to hundreds of grams of nucleic acid extraction magnetic beads in the laboratory, but the steps are cumbersome or use a large amount of organic solvents or use specific equipment, making it difficult to achieve large-scale manufacturing

Method used

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  • Large-scale manufacturing method of diagnostic magnetic beads and biological application thereof
  • Large-scale manufacturing method of diagnostic magnetic beads and biological application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] Example 1: Preparation method of diagnostic magnetic beads M1

[0124] 1) preparing magnetically cross-linked povidone particles;

[0125] 2) modifying the porous inorganic silica layer with the magnetic cross-linked povidone described in step 1 to prepare silicon-based magnetic cross-linked povidone particles;

[0126] 3) modifying the silicone layer on the silicon-based magnetic cross-linked povidone particles described in step 2 to prepare double-layer silicon-based magnetic cross-linked povidone particles.

[0127] Wherein, the specific synthesis steps of the cross-linked povidone particles in the step 1) include:

[0128] 1-1) 100 parts by mass of crospovidone particles with a particle size of 5-10 μm, crospovidone particles, 400 parts by mass of glycerin, 600 parts by mass of ethanol and 1000 parts by mass of water were mixed and mechanically stirred for 1 hour to disperse;

[0129] 1-2) After feeding nitrogen for 1 hour, add 100 parts by mass of ferrous ammoni...

Embodiment 2

[0138] Embodiment 2: the preparation method of diagnostic magnetic beads M2

[0139] 1) preparing magnetically cross-linked povidone particles;

[0140] 2) modifying the porous inorganic silica layer with the magnetic cross-linked povidone described in step 1 to prepare silicon-based magnetic cross-linked povidone particles;

[0141] 3) modifying the silicone layer on the silicon-based magnetic cross-linked povidone particles described in step 2 to prepare double-layer silicon-based magnetic cross-linked povidone particles.

[0142] Wherein, the specific synthesis steps of the cross-linked povidone particles in the step 1) include:

[0143] 1-1) 150 parts by mass of crospovidone particles with a particle size of 10-20 μm, 200 parts by mass of glycerin, 500 parts by mass of ethanol and 800 parts by mass of water are mixed and mechanically stirred for 1 hour until dispersed;

[0144] 1-2) After feeding nitrogen for 1 hour, add 150 parts by mass of ferrous ammonium sulfate, 280...

Embodiment 3

[0153] Embodiment 3: Magnetic leakage test of diagnostic magnetic beads M1, M2 and commercialized magnetic beads

[0154] The diagnostic magnetic beads M1, M2 provided in Example 1 and the MagneSil magnetic beads of Promega Company (product number MD1441), the Dynabeads MyOne of Thermo Company TM Silane (Product No. 37002D) was diluted with sterile water to 10mg / mL, and 1mL of the above-mentioned diluted magnetic bead suspension was taken. After repeated washing with deionized water for 3 times, 1mL of 0.5M HCl was added and reacted at room temperature for 1 hour at 1500rpm. The supernatant was absorbed by magnetic separation and the content of iron ions in the supernatant was detected by atomic absorption. Table 1 shows that the diagnostic magnetic beads M1 and M2 have low magnetic leakage, while the commercial magnetic beads MagneSil and MyOne TM The magnetic leakage of Silane is very obvious.

[0155] Table 1: Magnetic leakage test of different magnetic beads

[0156] ...

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Abstract

The invention belongs to the technical field of biological magnetic beads, and discloses a large-scale manufacturing method and the biological application of diagnostic magnetic beads. The preparation method comprises the following steps: 1) preparing magnetic cross-linked povidone particles; 2) modifying the porous inorganic silicon dioxide layer with the magnetic cross-linked povidone to prepare silicon-based magnetic cross-linked povidone particles; and 3) modifying an organic silicon layer with the silicon-based magnetic cross-linked povidone particles to prepare the double-layer silicon-based magnetic cross-linked povidone particles.

Description

technical field [0001] The invention relates to the technical field of biological magnetic beads, in particular to a large-scale manufacturing method of diagnostic magnetic beads and its biological application. Background technique [0002] The description of the background technology of the present invention belongs to the relevant technology related to the present invention, and is only used to illustrate and facilitate the understanding of the content of the present invention. prior art on the filing date. [0003] Due to the characteristics of superparamagnetism, large specific surface area and biocompatibility, biomagnetic beads have a wide range of applications in bioseparation engineering, especially in nucleic acid extraction. At present, the main magnetic beads used in the pretreatment process of nucleic acid extraction in molecular diagnosis are "silica magnetic beads", "magnetic silicon particles" and "silicon-based magnetic beads". As the core, the magnetic par...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/6806C08L39/06C08K3/22
CPCC12N15/1013C12Q1/6806C08K3/22C08K2201/01C08K2003/2275C08L39/06C12Q2523/308C12Q2521/537Y02A50/30
Inventor 张佳斌
Owner 苏州白垩纪生物科技有限公司
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