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 prote

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
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Example Embodiment

[0020] Example 1

[0021] (1) Activation of Graphene Oxide (GO)

[0022] 1.1mgGO was resuspended in 1mL of deionized water and sonicated at room temperature for 3h to fully disperse;

[0023] 2. Add 500 mM (pH 6.1) MES500μL, then at the same time 30mg / mlNHS500μL and 10mg / mlEDC600uL, mix well, and stir quickly at room temperature for 30min;

[0024] 3. The material is washed repeatedly with 500 mM (pH 6.1) MES to remove residual NHS and EDC, and the final volume is 1 mL.

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

[0026] 1. Add 15mg PEI to the above materials and shake at room temperature for 1h;

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

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

[0029] 1. Take 500ugPA and dissolve it in 500mM (pH6.1) MES1mL;

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

[0031] 3. The solution is activa...

Example Embodiment

[0037] Example 2

[0038] In the activation scheme of graphene oxide (GO), the NHS concentration is changed to 50 mg / ml, and the others are the same as in Example 1, and GO-PA composite materials can be prepared.

Example Embodiment

[0039] Example 3

[0040] In the graphene oxide (GO) activation scheme, the NHS concentration is changed to 40 mg / ml, and the others are the same as in Example 1, and GO-PA composite materials can 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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