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Method for preparing protein imprinted polymer by removing template molecules under electric field assisted actions

A technology of western blotting and template molecules, which is applied in the field of preparation of western blotting polymers, and achieves the effects of reducing processing costs, high efficiency and lowering production costs.

Inactive Publication Date: 2015-09-16
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there has been no report on the application of the three-electrode system to the preparation of polymers for western blotting

Method used

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  • Method for preparing protein imprinted polymer by removing template molecules under electric field assisted actions

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Experimental program
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Effect test

Embodiment 1

[0013] Firstly, according to the prior art, the hemoglobin polymer is made, and then the following steps are carried out:

[0014] a. The hemoglobin polymer is immobilized on the gold electrode as the working electrode, the saturated calomel electrode is used as the reference electrode, and the platinum electrode is used as the counter electrode, and the working electrode, the reference electrode and the counter electrode constitute a three-electrode system;

[0015] b. Put the three-electrode system into the protein polymer template molecular elution solution composed of 10% (v / v) acetic acid and 10% (v / v) sodium lauryl sulfate solution, because the elution solution The pH value is lower than the isoelectric point of hemoglobin, so apply potentials of 0.1V, 0.4V, 0.7V and 1.0V between the working electrode and the reference electrode, and elute for 30-120min respectively to obtain the hemoglobin imprinted polymer;

[0016] c. Dry the obtained hemoglobin-imprinted polymer unde...

Embodiment 2

[0023] First, according to the prior art, the myoglobin polymer is made, and then the following steps are carried out:

[0024] a. The myoglobin polymer is immobilized on the graphite electrode as the working electrode, the mercurous sulfate electrode is used as the reference electrode, and the carbon electrode is used as the counter electrode, and the working electrode, the reference electrode and the counter electrode constitute a three-electrode system;

[0025] b. Put the three-electrode system into the phosphate buffer solution of pH 9.0 (protein polymer template molecule elution solution), because the pH value of the elution solution is higher than the isoelectric point of myoglobin, so between the working electrode and the A potential of -1.0V was applied between the reference electrodes for 30 minutes to obtain a myoglobin imprinted polymer;

[0026] c. Dry the obtained myoglobin-imprinted polymer under nitrogen gas to obtain completely eluted myoglobin-imprinted polym...

Embodiment 3

[0028] First, according to the prior art, the myoglobin polymer is made, and then the following steps are carried out:

[0029] a. The myoglobin polymer is immobilized on the silver electrode as the working electrode, the calomel electrode is used as the reference electrode, and the platinum electrode is used as the counter electrode, and the working electrode, the reference electrode and the counter electrode constitute a three-electrode system;

[0030] b. Then put the three-electrode system into the phosphate buffer solution (elution solution of protein polymer template molecules) with pH 10.0. A potential of -1.0V was applied between the reference electrodes for 120min to obtain a myoglobin imprinted polymer;

[0031] c. Dry the obtained myoglobin-imprinted polymer under nitrogen gas to obtain completely eluted myoglobin-imprinted polymer.

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Abstract

The invention discloses a method for preparing a protein imprinted polymer by removing template molecules under electric field assisted actions, which has the advantages of time saving, labor saving and high efficiency. The method comprises the following steps: immobilizing a protein polymer onto an electrode as work electrode, wherein the work electrode, a reference electrode and a counter electrode constitute a tri-electrode system; putting the tri-electrode system into a protein polymer template molecule eluting solution, and applying a constant potential for 30-120 minutes between the work electrode and reference electrode to obtain the protein imprinted polymer, wherein if the pH value of the protein polymer template molecule eluting solution is lower than the isoelectric point of the protein, the applied potential is 0.1-1.0V, or else, the applied potential is -1.0 to -0.1V; and drying the obtained protein imprinted polymer in nitrogen.

Description

technical field [0001] The invention relates to a method for preparing a western imprinted polymer, in particular to a method for preparing a western imprinted polymer by saving time, labor and high efficiency by assisted removal of template molecules by an electric field. Background technique [0002] Western imprinted polymer is a new type of polymer functional material that can recognize proteins, which can be used in proteomics to separate and enrich low-abundance proteins from high-abundance proteins; With specific recognition performance, high-sensitivity bionic sensors can be constructed for trace analysis of proteins. The existing method for preparing western imprinted polymers is to form a complex by first interacting with proteins (such as template molecules such as hemoglobin, myoglobin or albumin) and functional monomers (such as hydrogen bonds, van der Waals forces, electrostatic interactions, etc.), and then Under the action of the initiator, the complex and t...

Claims

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Application Information

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IPC IPC(8): C08J9/26
Inventor 孙越兰玉廷杜洪莹冯春梁
Owner LIAONING NORMAL UNIVERSITY
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