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A preparation method and application of carboxyl magnetic beads coated with dextran

A technology of carboxyl magnetic beads and aldehyde dextran, which is applied in the biological field, can solve the problems of low coupling efficiency and high non-specific adsorption, and achieve the effect of increasing coverage and carboxyl content

Active Publication Date: 2017-08-18
BEAVERNANO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The purpose of the present invention is to provide a method for preparing carboxyl magnetic beads based on the surface of amino magnetic beads grafted with dextran and its application, so as to solve the problems of low coupling efficiency and high non-specific adsorption of carboxyl magnetic beads when covalently grafting proteins. Problems and Broadening the Applications of Carboxylated Magnetic Beads

Method used

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  • A preparation method and application of carboxyl magnetic beads coated with dextran
  • A preparation method and application of carboxyl magnetic beads coated with dextran

Examples

Experimental program
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Effect test

Embodiment 1

[0028] The preparation method of carboxyl magnetic beads (dextran and glutaraldehyde co-coupled to amino magnetic beads) in this example is as follows:

[0029] (1) Preparation of aldehyde dextran: Weigh 1 g of dextran (molecular weight: 70KDa), dissolve it in 20 mL of ultrapure water, add 0.1M NaIO4 to the solution to make the molar ratio of dextran to 1:2, Shake at 30°C for 2 hours in the dark to obtain aldehyde dextran; dialyze with ultrapure water for 8 hours or longer to remove unreacted NaIO4, vacuum freeze-dry the dialyzed solution to obtain a white solid that is aldehyde glucan Polysaccharides, keep away from light and refrigerate at 2-8°C;

[0030] (2) Graft the aldehyde dextran and glutaraldehyde obtained in step (1) onto the surface of amino magnetic beads at a mass ratio of 1:5 to prepare aldehyde magnetic beads, and mix the amino magnetic beads well, here The amino magnetic beads are MagpearL NH2, the average particle size is 1μm, the inner core is ferric oxide, ...

Embodiment 2

[0036] The preparation method of the carboxylated magnetic beads (using carboxylated dextran oxidation experiment) in this example is as follows:

[0037] (1) Carboxylation of dextran: Weigh 5 g of dextran (molecular weight: 70 KDa), dissolve it in 10 g of ultrapure water, add 0.08 g of sodium borohydride and 0.1 g of 50% NaOH to the solution, and keep away from light for no more than 25 Stir at ℃ for 4 hours, then add 3.9g 50% NaOH and 1.24g bromoacetic acid, shake at room temperature not exceeding 25℃ for 16 hours, then adjust the pH to neutral with doubled concentrated hydrochloric acid, and add ethanol to make the ratio of the two to 1: 1. Precipitate carboxylated dextran, dissolve the precipitate with 12mL ultrapure water, add 0.04g NaCL, add ethanol to make the ratio of the two 1:1, precipitate carboxylated dextran, repeat the above operation twice, Dissolve the precipitate in 12 mL of ultrapure water, add ethanol to precipitate carboxylated dextran, dry the precipitate ...

Embodiment 3

[0044] The preparation method of the carboxyl magnetic beads (formyl dextran grafted onto the amino magnetic beads alone) in this example, the steps are as follows:

[0045] (1) Oxidized dextran: Weigh 1 g of dextran (molecular weight: 70KDa), dissolve it in 20 mL of ultrapure water, add 0.1M NaIO4 to the solution to make the molar ratio of dextran to 1:2, avoid Light shock for 2 hours to obtain aldehyde dextran; dialyze with ultrapure water for 8 hours or longer to remove unreacted NaIO4, vacuum freeze-dry the dialyzed solution to obtain a white solid that is aldehyde dextran, Keep away from light and refrigerate at 2-8°C;

[0046] (2) Graft the aldehyde-dextran obtained in step (1) onto the surface of amino magnetic beads to prepare aldehyde-based magnetic beads: mix the amino magnetic beads thoroughly, absorb 1 mL (10 mg / mL) of magnetic beads, magnetically separate them, and use ultra- Wash the magnetic beads 2-3 times with pure water, magnetically separate, and discard th...

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Abstract

The invention discloses a method for preparing carboxyl magnetic beads based on glucan coating. According to the method, glucan is dissolved in ultrapure water, sodium periodate is added, reaction is conducted at a dark place for two hours at 30 DEG C, then dialysis purification is conducted with the ultrapure water, vacuum freeze drying is conducted on a solution obtained after dialysis, and then polyaldehyde dextran is obtained; the polyaldehyde dextran is dissolved in PBS and added to amino magnetic beads, glutaraldehyde is added to the amino magnetic beads, oscillation is conducted for four hours away from light at the room temperature, and the polyaldehyde dextran and the glutaraldehyde are grafted to the surfaces of the amino magnetic beads together to generate aldehyde group magnetic beads with the covalent grafting method; glutamic acid is dissolved in PBS and added to the aldehyde group magnetic beads, oscillation is conducted for four hours at the room temperature, then sodium borohydride is added, oscillation reaction is continued for another two hours, and then the carboxyl magnetic beads are obtained. The monodisperse magnetic beads formed by means of the method facilitate downstream grafting of biomolecules without affecting the activity of biomolecules, and the increase of carboxyl density of the surfaces of the magnetic beads enables biomolecule grafting efficiency to be higher.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method and application of carboxyl magnetic beads coated with dextran on the surface of amino magnetic beads. Background technique [0002] Magnetic beads, that is, magnetic microbeads or magnetic microspheres, are a type of spherical composite with superparamagnetic properties and a diameter of nanometer or micrometer scale, which can move to the magnetic field under the action of an external magnetic field, so as to achieve the purpose of separating the target product . The surface of magnetic beads has different functional groups, which can be divided into amino magnetic beads, carboxyl magnetic beads, epoxy magnetic beads and so on according to the different surface functional groups. The functional groups on the surface of magnetic beads can be covalently grafted with various biomolecules or small molecules, which can be used in downstream biological e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/24B01J20/28B01J20/30C07K1/22
CPCB01J20/24B01J20/28009B01J2220/48C07K1/22
Inventor 李培培王辉王知丰黄明贤
Owner BEAVERNANO TECH
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