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Method for covalent coupling protein on amino magnetic bead surface

An amino magnetic bead and surface covalent technology, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve the problems of magnetic bead aggregation and low coupling efficiency, and achieve good repeatability and repeatability , Improve the effect of protein coupling efficiency

Inactive Publication Date: 2009-01-21
北京倍爱康生物技术有限公司
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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for covalently coupling proteins with high efficiency on the surface of amino magnetic beads, so as to solve the problems of low coupling efficiency and magnetic bead aggregation of glutaraldehyde coupling method
Suspension reaction at room temperature for 15 minutes

Method used

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  • Method for covalent coupling protein on amino magnetic bead surface
  • Method for covalent coupling protein on amino magnetic bead surface
  • Method for covalent coupling protein on amino magnetic bead surface

Examples

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Embodiment

[0025] Example: Amino Magnetic Beads Coupling Sheep Anti-Mouse Polyclonal Antibody

[0026] (1) Activation of magnetic beads: Maleimide-containing chemical groups are generated on the surface of magnetic beads. Activation methods include:

[0027] (1.1) Magnetic bead pretreatment: Take 100 mg of amino magnetic bead colloidal solution and settle (magnetic separation) under the action of a magnetic field for 10 minutes, remove the supernatant, and wash the settled magnetic beads twice with magnetic bead activation buffer, 5 ml each time.

[0028] (1.2) Suspend the washed magnetic beads with 1 ml of activation buffer, add the activator Sulfo-SMCC, and suspend at room temperature for 15 minutes. The reaction concentration of Sulfo-SMCC is 1 mM.

[0029] (1.3) Add 10 microliters of 1M glycine buffer solution with pH 7.3, suspend and react at room temperature for 10 minutes, and terminate the activation reaction.

[0030] (1.4) Magnetically separate for 10 minutes, remove the supe...

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Abstract

The invention provides a method for covalence coupling protein molecules onto surfaces of amino ceramic beads. The method is to use various activators to carry out activation for the amino ceramic beads and the protein molecules respectively, both of which are subsequently mixed to immediately accomplish the high-efficient covalence coupling of protein molecules onto the surfaces of the ceramic beads. Because of the different activators held by the activated protein molecules and the activated ceramic beads, no reaction occurs between the activated protein molecule and the activated ceramic beads, thereby efficiently avoiding the accumulation among protein molecules or the accumulation among the ceramic beads. In addition, the rapid and high-efficient reaction between mercapto group produced from activation and the activated maleimide group ensures the high coupling rate and good repeatability. The coupled ceramic bead can be widely applied to the fields of immune test, and cell selection.

Description

technical field [0001] The invention belongs to the technical field of magnetic materials, and particularly provides a method for covalently coupling proteins with high efficiency on the surface of amino magnetic beads. Magnetic beads coupled to proteins can be used for immunoassays, cell sorting, etc. Background technique [0002] Magnetic beads, also known as magnetic particles or magnetic particles, are a kind of spherical composite materials with a diameter of nanometer or micrometer. The core of the magnetic bead is composed of superparamagnetic materials such as ferroferric oxide or ferric oxide, and the periphery is coated with polymer materials such as polystyrene or dextran. The core substance endows the magnetic beads with superparamagnetism, so that the magnetic beads can move to the direction of the magnetic field under the action of an external magnetic field to achieve the purpose of separation; the peripheral polymer materials can be activated by physical or ...

Claims

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Application Information

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IPC IPC(8): C07K1/10
Inventor 仝文斌陈立杰雷焱
Owner 北京倍爱康生物技术有限公司
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