Graphene oxide-recombined streptococcal protein G non-covalent composite material as well as preparation method and application thereof

A technology of recombinant streptococcus and composite material, which is applied in the field of chemical synthesis, can solve the problem of low amount of immobilized protein and the like, and achieve the effects of simple preparation steps, maintaining activity, and easy mass production

Inactive Publication Date: 2015-12-16
李云峰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most commercial reagents for antibody enrichment use agarose and trehalose as carriers to immobilize recombinant streptococcal protein G that absorbs 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-recombined streptococcal protein G non-covalent composite material as well as preparation method and application thereof
  • Graphene oxide-recombined streptococcal protein G non-covalent composite material as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Activation of graphene oxide (GO)

[0020] 1. 1 mg GO was resuspended in 1 mL of deionized water, ultrasonicated at room temperature for 3 hours to fully disperse;

[0021] 2. Add 500 μL of 500 mM 2-(N-morpholino)ethanesulfonic acid 4-morpholineethanesulfonic acid (pH6.1) (MES), then simultaneously 16 mg / ml NHS 500 μL and 4 mg / ml EDC 500 uL, mix well, and stir rapidly at room temperature for 30 minutes;

[0022] 3. The material was washed repeatedly with 500mM (pH6.1) MES to remove residual NHS and EDC, and the final volume was 1mL;

[0023] 4. The solution is activated GO.

[0024] (2) Graphene oxide immobilized recombinant streptococcal protein G

[0025] 1. According to the ratio of 1:2.5, mix the material GO and PG prepared in the above (1) and incubate overnight at 4°C with shaking;

[0026] 2. Remove the supernatant after centrifugation at 16400×g / 10min, add 500mM (pH6.1) MES to wash three times, and the obtained material is graphene oxide-recombinant strep...

Embodiment 2

[0032] In the activation scheme of graphene oxide (GO), the concentration of NHS was changed to 24 mg / ml, and the others were the same as in Example 1, so that GOPG non-covalent composite material could be prepared.

Embodiment 3

[0034] In the activation scheme of graphene oxide (GO), the concentration of NHS was changed to 20 mg / ml, and the others were the same as in Example 1, so that GOPG non-covalent composite material could be prepared.

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Abstract

The invention relates to a graphene oxide-recombined streptococcal protein G non-covalent composite material as well as a preparation method and application thereof. A recombined streptococcal protein G is immobilized on a graphene oxide material by making use of the characteristic that graphene oxide is high in specific surface area by a non-covalent combination method so as to prepare a high-capacity antibody-enriched material with antibody adsorption bioactivity. The prepared graphene oxide-recombined streptococcal protein G non-covalent composite material can be used in the fields of antibody purification, antibody enrichment, pathogene 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, specifically the preparation and function of a non-covalent composite material based on graphene oxide-recombinant streptococcal protein G application. 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 im...

Claims

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Application Information

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IPC IPC(8): B01J20/24B01J20/30
Inventor 李云峰
Owner 李云峰
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