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

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Problems solved by technology

At present, most commercial reagents for antibody enrichment use agarose and trehalose as carriers to immobilize recombinant streptococcal protei

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

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Example Embodiment

[0018] Example 1

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

[0020] 1.1mgGO was resuspended in 1mL of deionized water and ultrasonicated for 3h at room temperature to fully disperse;

[0021] 2. Add 500μL of 500mM 2-(N-morpholinyl)ethanesulfonic acid 4-morpholineethanesulfonic acid (pH6.1) (MES), then 16mg / mlNHS500μL and 4mg / mlEDC500uL at the same time, mix well, and stir quickly at room temperature for 30min;

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

[0023] 4. The solution is the activated GO.

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

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

[0026] 2. After centrifugation at 16400×g / 10min, the supernatant is removed, and 500mM (pH6.1) MES is added to wash three times. The resulting material is graphene oxide-recombinant streptococcal protein...

Example Embodiment

[0031] Example 2

[0032] In the activation scheme of graphene oxide (GO), the NHS concentration is changed to 24 mg / ml, and the others are the same as in Example 1, and GOPG non-covalent composite materials can be prepared.

Example Embodiment

[0033] Example 3

[0034] In the activation scheme of graphene oxide (GO), the NHS concentration is changed to 20 mg / ml, and the others are the same as in Example 1, and GOPG non-covalent composite materials can 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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