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Method for separating protein adopting superparamagnetism silicon nanometer granule static adsorption

A superparamagnetic and nanoparticle technology, which is applied in the application field of nanomaterials and nanobiology, can solve the problems of cumbersome pretreatment of ion exchange columns and non-specific adsorption of proteins, so as to improve the action speed and reduce the operation intensity. Effect

Inactive Publication Date: 2006-12-13
HUNAN UNIV
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  • Summary
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  • Description
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AI Technical Summary

Problems solved by technology

These methods have certain deficiencies when they are used for protein separation. For example, the gel used in the gel filtration method generally requires complex pretreatment, requires certain operating techniques, and requires vacuum and low temperature conditions to prevent gelation. Denaturation; the pretreatment of the ion-exchange column of ion-exchange chromatography is cumbersome; the separation of proteins by affinity chromatography still has non-specific adsorption, etc.

Method used

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  • Method for separating protein adopting superparamagnetism silicon nanometer granule static adsorption
  • Method for separating protein adopting superparamagnetism silicon nanometer granule static adsorption
  • Method for separating protein adopting superparamagnetism silicon nanometer granule static adsorption

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Effect test

Embodiment 1

[0035] Embodiment 1: the extraction of cytochrome C in the mixed solution of bovine serum albumin (acidic protein, pI4.6) and cytochrome C (basic protein, pI10.6):

[0036] 1. Adjustment of the pH value of the mixed solution of cytochrome C and bovine serum albumin: at room temperature, use 1 / 15 mol / L disodium hydrogen phosphate solution to adjust the pH value of the mixed protein solution to 8.0.

[0037] 2. Formation of superparamagnetic pure silicon shell nanoparticles-cytochrome C complex: Add 15mg superparamagnetic pure silicon shell nanoparticles to 5.0mL mixed protein solution, mix well and After incubation for 0.50 hours, a complex of superparamagnetic pure silicon shell nanoparticles-cytochrome C is formed.

[0038] 3. Separation and collection of superparamagnetic pure silicon shell nanoparticles-cytochrome C complex: using the superparamagnetism of superparamagnetic pure silicon shell nanoparticles, under the action of an external magnetic field, the superparamagnet...

Embodiment 2

[0042] Example 2: Cytochrome C in a mixed solution of bovine serum albumin (acidic protein, pI4.6), immunoglobulin G (basic protein, pI8.0) and cytochrome C (basic protein, pI10.6) Extraction of:

[0043] 1. Adjustment of the pH value of the mixed solution of bovine serum albumin, immunoglobulin G and cytochrome C: at room temperature, use 1 / 15 mol / L disodium hydrogen phosphate solution to adjust the pH value of the mixed protein solution to 8.0.

[0044] 2. Formation of superparamagnetic pure silicon shell nanoparticles-cytochrome C complex: Add 3.0mg superparamagnetic pure silicon shell nanoparticles to 1.0mL mixed protein solution, mix thoroughly at room temperature of 15-25°C and After incubation for 0.50 hours, a complex of superparamagnetic pure silicon shell nanoparticles-cytochrome C is formed.

[0045] 3. Separation and collection of superparamagnetic pure silicon shell nanoparticles-cytochrome C complex: using the superparamagnetism of superparamagnetic pure silicon...

Embodiment 3

[0049] Embodiment 3: the extraction of bovine serum albumin in the mixed solution of bovine serum albumin (acidic protein, pI4.6) and cytochrome C (basic protein, pI10.6):

[0050] 1. Adjustment of the pH value of the mixed solution of cytochrome C and bovine serum albumin: at room temperature, use 1 / 15 mol / L potassium dihydrogen phosphate solution to adjust the pH value of the mixed protein solution to 5.0.

[0051] 2. Formation of superparamagnetic aminated silicon shell nanoparticles-bovine serum albumin complex: Add 9.0 mg of superparamagnetic aminated silicon shell nanoparticles to 3.0 mL of mixed protein solution, at room temperature of 15-25 °C, Mix well and incubate for 0.50 hour to form a superparamagnetic aminated silicon shell nanoparticle-bovine serum albumin complex.

[0052] 3. Separation and collection of superparamagnetic aminated silicon shell nanoparticles-bovine serum albumin complex: use the superparamagnetism of superparamagnetic aminated silicon shell nan...

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Abstract

The invention discloses the method of superparamagnetism siliceous skin nanometer separating protein. The invention relates the two kinds of superparamagnetism siliceous skin nanometer powders: negative charge superparamagnetism siliceous skin nanometer powder and positive charge superparamagnetism siliceous skin nanometer powder. By controlling pH, the method uses the former one to separate and extract alkaline protein, and uses the latter one to separate and extract acid protein. The invention has the advantages of improving action speed and reducing operation intensity. The invention has the advantages of low cost, and fast speed.

Description

technical field [0001] The invention relates to the application field of nanometer materials and the field of nanobiology, in particular to the application and development of superparamagnetic silicon shell nanoparticles in biomedical research for the electrostatic adsorption and separation of proteins. Background technique [0002] In recent years, with the completion of the Human Genome Project, the analysis of huge human genome information, the development of proteomics research, and the acquisition of data on protein expression levels have attracted much attention and attention from researchers. As the premise and foundation of proteomics research, the separation and purification of proteins has very important biological significance. At present, people have established methods such as gel filtration, colloid electrophoresis, and ultrafiltration by using the different sizes of protein molecules, and developed methods such as ion exchange chromatography, gel electrophores...

Claims

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Application Information

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IPC IPC(8): C07K1/14
Inventor 王柯敏何晓晓陈英杰谭蔚泓吴萍
Owner HUNAN UNIV
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