Paper base dual-imprinting material capable of selectively recognizing protein as well as preparation method and application of paper base dual-imprinting material

A technology for imprinting materials and proteins, which is applied in the field of environmental analysis, can solve the problems of lack of popularization and safety of protein recognition technology, and achieve the effect of portable rapid analysis, increased adsorption capacity, and good practical application value

Active Publication Date: 2018-05-29
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention solves the problem of lack of popularization and safety of protein recognition technology, and provides a paper-based double-imprinted material and its preparation method and application

Method used

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  • Paper base dual-imprinting material capable of selectively recognizing protein as well as preparation method and application of paper base dual-imprinting material
  • Paper base dual-imprinting material capable of selectively recognizing protein as well as preparation method and application of paper base dual-imprinting material
  • Paper base dual-imprinting material capable of selectively recognizing protein as well as preparation method and application of paper base dual-imprinting material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation of graphene oxide-hemin complex: graphene oxide (0.83mmol, sheet diameter 50-200nm) and hemin (0.015mmol) are dissolved in 20ml 50% ethanol solution, with 1mol / L hydroxide Sodium solution adjusts the pH to 10.0, after ultrasonication for 30 minutes, reflux at 100°C for 1 hour and then cool to form a graphene oxide-hemin complex, in which graphene oxide and hemin form a ππ connection through physical adsorption ;

[0040] Disperse graphene oxide-hemin (graphene oxide 0.85mmol, hemin 0.015mmol) and thyroglobulin (0.001mmol) in 1ml of phosphate-buffered saline solution, mix thoroughly, and incubate at room temperature for 2-12 hours in the dark. Hour;

[0041] figure 1 An atomic force microscopy image showing the adsorption of thyroglobulin by the graphene oxide-hemin complex.

[0042] The atomic force microscope image shows that due to the adsorption of thyroglobulin on the surface of the graphene oxide-hemin complex, the height increases from 2nm to 12...

Embodiment 2

[0044] (1) Immobilization of graphene oxide-hemin complex on the surface of filter paper fibers

[0045] Graphene oxide-hemin complex (graphene oxide 0.83mmol, hemin 0.015mmol) was immobilized on the surface of filter paper (40mm×40mm) by negative pressure, and the filter paper was cut into several pieces with a punch (8mm×8mm). small discs.

[0046] figure 2 It shows the electron microscope image of the graphene oxide-hemin complex immobilized on the surface of the filter paper material. figure 2 (a) is the SEM picture under the conditions of 20KV, X300 and 50μm; figure 2 (b) is the SEM picture under the conditions of 20KV, X3000 and 5μm.

[0047] (2) Preparation of paper-based double imprinted materials

[0048] (2-1) Dissolve functional monomer acrylamide (0.056mmol), template thyroglobulin (1.515nmol) and 3,3',5,5'-tetramethylbenzidine (0.004mmol) in 1ml of phosphate buffer solution (10 mmol / L, pH=6.8), vortexed for 10 seconds, and incubated at room temperature for...

Embodiment 3

[0056] The paper-based double-imprinted material (8mm×8mm) and the paper-based non-imprinted material (8mm×8mm) prepared in Example 2 were added to 5 mL of thyroglobulin-containing phosphate buffer solution (pH=6.8), and thyroglobulin The concentration of protein is 50-500mg / L respectively. After shaking for 10 hours, use a fluorescence spectrophotometer to measure the content of thyroglobulin in the supernatant, subtract the content of thyroglobulin in the supernatant from the total amount of thyroglobulin, and calculate the paper-based double-blotting material and paper-based Adsorption capacity of non-blotted material for thyroglobulin. The result is as Figure 5 As shown, the adsorption capacity of the paper-based double-imprinted material to thyroglobulin reached 87.78 mg / g, while the adsorption capacity of the non-imprinted polymer as a control was only 45.37 mg / g, indicating that the specific recognition of thyroglobulin The imprinted sites have been formed.

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Abstract

The invention discloses a paper base dual-imprinting material capable of selectively recognizing a protein as well as a preparation method and an application of the paper base dual-imprinting material. The preparation method comprises the steps of immobilizing a graphene oxide-hemin compound on the surface of a filter paper material and preparing the dual-imprinting material on the surface of filter paper. The paper base dual-imprinting material is used for selectively recognizing and quantitatively detecting the protein. The paper base dual-imprinting material has the characteristics of low cost, environmental protection, simplicity in operation, high recyclability, and specific recognition of the protein, and particularly has the excellent characteristics of color development, water retention and hydrophily, and can rapidly detect the protein macromolecule at high throughput.

Description

technical field [0001] The invention relates to the field of environmental analysis, in particular to a paper-based double-imprint material for selectively recognizing proteins, a preparation method and application thereof. Background technique [0002] Thyroglobulin is an iodized glycoprotein in the thyroid gland, which is secreted by thyroid follicular cells, and can be used as a precursor of thyroxine, which is iodized and coupled to generate thyroxine under the action of thyroid peroxidase; It is closely related to the normal physiological function of the thyroid gland, and the level of thyroglobulin is often used as one of the indicators for evaluating thyroid function. [0003] Thyroxine disruptors are a class of foreign substances that interfere with the synthesis, secretion, transport, binding, effect, or clearance of thyroxine by affecting both thyroxine levels and the responsiveness of target cells to hormones. In recent years, there have been many studies on thyr...

Claims

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Application Information

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IPC IPC(8): C08F220/56C08F222/38C08J9/26B01J20/26B01J20/291G01N21/78
CPCB01J20/268B01J20/291C08F220/56C08J9/26C08J2201/0424C08J2333/26G01N21/78C08F222/385
Inventor 荆涛汪秀黄凯梅素容周宜开
Owner HUAZHONG UNIV OF SCI & TECH
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