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Graphene oxide-recombinant streptococcal protein g composite material and its preparation and application

A technology of recombinant streptococcus and composite materials, which is applied in the field of chemical synthesis, can solve the problems of low antibody adsorption rate, achieve good reproducibility, facilitate large-scale production, and improve the effect of immobilization capacity

Active Publication Date: 2017-09-26
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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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 for absorbing antibodies. Due to the limitation of its own immobilization capacity, its antibody adsorption rate is low, and the adsorption capacity is 27mg / mL Vector left and right (GE Healthcare Life Sciences, 28-9768-01AA)

Method used

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  • Graphene oxide-recombinant streptococcal protein g composite material and its preparation and application
  • Graphene oxide-recombinant streptococcal protein g composite material and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Activation of graphene oxide (GO)

[0030] 1.1 mg GO was resuspended in 1 mL of deionized water, ultrasonicated at room temperature for 3 hours to fully disperse;

[0031] 2. Add 500mM (pH6.1) MES500μL, then simultaneously 30mg / ml NHS500μL and 10mg / mlEDC600uL, mix well, and stir rapidly at room temperature for 30min;

[0032] 3. The material was washed repeatedly with 500 mM (pH 6.1) MES to remove residual NHS and EDC, with a final volume of 1 mL.

[0033] (2) PEI modification of the activated graphene oxide material

[0034] 1. Add 15mg PEI to the above materials and shake at room temperature for 1 hour;

[0035] 2. After centrifugation at 16400×g / 10min, remove the supernatant, and the precipitate is GO@PEI.

[0036] (3) Activation of recombinant streptococcal protein G

[0037] 1. Dissolve 500ug PG in 500mM (pH6.1) MES1mL;

[0038] 2. Then add 1.5mg NHS and 3.5mg EDC at the same time, mix well, and stir rapidly for 30min;

[0039] 3. The solution is activate...

Embodiment 2

[0048] 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-PG composite material could be prepared.

Embodiment 3

[0050] In the activation scheme of graphene oxide (GO), the concentration of EDC was changed to 20mg / ml, and the others were the same as in Example 1, so that GO-PG composite material could be prepared.

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Abstract

The invention relates to a graphene oxide-recombinant streptococcal protein G composite material and its preparation method and application. In this technology, the high specific surface area of ​​graphene oxide is used to immobilize recombinant streptococcal protein G on graphene oxide material through covalent bonding method to prepare high-capacity antibody enrichment material with antibody adsorption biological activity. . The prepared graphene oxide-recombinant streptococcal protein G composite material can be used in fields such as antibody purification, antibody enrichment, pathogen detection, and biological sample pretreatment.

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 G. 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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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/24B01J20/30C07K1/22G01N33/48G01N1/28
Inventor 张丽华李云峰江波杨开广翁叶靖李森武张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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