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Graphene oxide-recombinant streptococcal protein A composite material as well as preparation method and application thereof

A technology of recombinant streptococcus and composite materials, which is applied in the field of chemical synthesis, can solve the problem of low protein immobilization, achieve the effects of simple preparation steps, increase immobilization capacity, and maintain activity

Inactive Publication Date: 2015-12-30
李云峰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, commercial reagents for antibody enrichment mostly use agarose and trehalose as carriers to immobilize recombinant streptococcal protein A that adsorbs antibodies. For example, agarose-immobilized ProteinG is about 80ug / mL, and the amount of immobilized protein is relatively low.

Method used

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  • Graphene oxide-recombinant streptococcal protein A composite material as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] (1) Activation of graphene oxide (GO)

[0022] 1. 1 mg GO was resuspended in 1 mL of deionized water, ultrasonicated at room temperature for 3 hours to fully disperse;

[0023] 2. Add 500mM (pH6.1) MES 500μL, then simultaneously 30mg / ml NHS 500μL and 10mg / ml EDC 600uL, mix well, and stir rapidly at room temperature for 30min;

[0024] 3. The material was washed repeatedly with 500mM (pH6.1) MES to remove residual NHS and EDC, and the final volume was 1mL.

[0025] (2) PEI modification of the activated graphene oxide material

[0026] 1. Add 15mgPEI to the above materials, shake at room temperature for 1h;

[0027] 2. After centrifugation at 16400×g / 10min, remove the supernatant, and the precipitate is GOPEI.

[0028] (3) Activation of recombinant streptococcal protein A

[0029] 1. Dissolve 500ugPA in 500mM (pH6.1) MES1mL;

[0030] 2. Then add 1.5mg NHS and 3.5mg EDC at the same time, mix well, and stir rapidly for 30min;

[0031] 3. The solution is activated PA. ...

Embodiment 2

[0038] In the activation scheme of graphene oxide (GO), the concentration of NHS was changed to 50 mg / ml, and the others were the same as in Example 1, so that GO-PA composite material could be prepared.

Embodiment 3

[0040] In the activation scheme of graphene oxide (GO), the concentration of NHS was changed to 40mg / ml, and the others were the same as in Example 1, and the GO-PA composite material could be prepared.

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Abstract

The invention relates to a graphene oxide-recombinant streptococcal protein composite material as well as a preparation method and an application thereof. By utilizing the characteristic of high specific surface area of the graphene oxide, recombinant streptococcal protein A is immobilized onto the oxide graphene material to prepare a high-capacity antibody enriched material having antibody adsorption bioactivity. The prepared graphene oxide-recombinant streptococcal protein A composite material can be used in the fields such as antibody purification, antibody enrichment, antigen detection, biological sample pretreatment and the like.

Description

technical field [0001] The invention belongs to the technical field of chemical synthesis, and relates to the preparation technology of graphene composite material and its application in protein enrichment, in particular to the preparation and functional application of a composite material based on graphene oxide-recombinant streptococcal protein A. Background technique [0002] Infectious disease pathogens themselves as antigens can produce corresponding specific antibodies in biological organisms. Most of the existing detection methods detect pathogens through the specific recognition between antigens and antibodies, such as enzyme-linked immunosorbent assay, immunofluorescence and immunocolloidal gold technology. wait. These immunological techniques have the characteristics of simple operation and easy analysis of results. However, the sensitivity of immunological detection methods is relatively low, which may easily cause false negative results. In order to improve the ...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/30C07K1/22
Inventor 李云峰
Owner 李云峰
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