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Electrochemical aptamer detection method of glycoprotein

A detection method and glycoprotein technology, applied in the field of electrochemistry, can solve the problems of high cost of enzymatic signal amplification, complex labeling process and high cost, and achieve the effects of short detection time, high sensitivity and low cost.

Pending Publication Date: 2022-07-08
GUANGZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, enzymatic signal amplification has disadvantages such as high cost, poor stability, and complicated labeling process; nanomaterial-assisted signal amplification, because it involves the synthesis of nanomaterials and surface biological functionalization, has disadvantages such as complex operation and high cost.

Method used

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  • Electrochemical aptamer detection method of glycoprotein
  • Electrochemical aptamer detection method of glycoprotein
  • Electrochemical aptamer detection method of glycoprotein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Feasibility study of the electrochemical aptamer detection method for glycoproteins.

[0029] 0.1 μM nucleic acid aptamer modified with thiol groups at the end was added dropwise to the surface of the gold electrode and incubated for 15 min; then, the remaining active sites on the electrode surface were blocked with 2 mM mercaptohexanol; then, the electrode surface was mixed with 10 ng / mL glycoprotein. Incubate for 30 min and soak in 0.1 mM 4-(ferroceneacetamido)phenyl)boronic acid (Fc-PBA) solution for 30 min; finally, insert the resulting electrode into 25 mM lithium perchlorate solution, using a square wave volt Anfa (SWV) for testing, from figure 2 It can be seen that for the fully modified gold electrode, a strong oxidation peak can be observed at 0.4V; if no nucleic acid aptamer, glycoprotein or Fc-PBA is added to the electrode surface modification, no obvious oxidation peak can be observed. oxidation peak. Therefore, the resulting detection signal c...

Embodiment 2

[0030] Example 2: Selectivity study of electrochemical aptamer detection method for glycoproteins.

[0031] To evaluate the selectivity of the electrochemical aptamer detection method for glycoproteins, alpha-fetoprotein (AFP) was selected as the target glycoprotein, and 10 ng / mL AFP, 100 ng / mL γ-globulin (Glo), 100 ng / mL human serum albumin were collected (HSA), 100ng / mL carcinoembryonic antigen (CEA), 100ng / mL prostate specific antigen (PSA), 100ng / mL carbohydrate antigen 15-3 (CA15-3) or 1.0μg / mL glucose (glucose) the peak current. from Figure 5 It can be seen that only the target glycoprotein (ie AFP) can generate a strong oxidative current signal, while the oxidative current generated by the other detected interference components is similar to that of the blank control without the target glycoprotein. Obviously, the glycoprotein electrochemical aptamer detection method of the present invention has good selectivity.

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Abstract

The invention relates to the technical field of electrochemistry, in particular to an electrochemical aptamer detection method of glycoprotein, which comprises the following steps: immobilizing a nucleic acid aptamer for specific recognition and capture of glycoprotein, adding a boric acid derivative of an electroactive probe after the nucleic acid aptamer is combined with target glycoprotein, quantitative analysis of the content of the target glycoprotein can be realized by measuring an oxidation-reduction signal of the electroactive probe. According to the electrochemical aptamer detection method of the glycoprotein, the electroactive probe site is labeled on the glycoprotein captured by the aptamer in a targeting manner by virtue of the affinity recognition interaction of the borate, and the electrochemical aptamer detection method can be used for high-sensitivity and high-selectivity electrochemical detection of other glycoproteins or sugar-containing substances only by replacing the sequence of the used aptamer; the method has the excellent characteristics of simplicity and convenience in operation, low cost, short detection time, high sensitivity, good selectivity and the like, and high-sensitivity electrochemical detection of glycoprotein can be realized without adopting an additional signal amplification strategy.

Description

technical field [0001] The invention relates to the technical field of electrochemistry, in particular to an electrochemical aptamer detection method for glycoproteins. Background technique [0002] The basic principles of existing commercial glycoprotein detection methods are antigen-antibody immune recognition reaction. However, antibodies have many shortcomings, such as complex preparation process, large batch-to-batch variation, difficult modification, high price, and poor stability. In contrast, nucleic acid aptamers have broader application prospects in the highly selective detection of glycoproteins due to their excellent properties such as chemical synthesis, easy modification, low cost and good stability. Electrochemical methods have attracted much attention in the detection of glycoproteins due to their advantages of simple equipment, fast response, low cost, high sensitivity, good selectivity, and easy miniaturization. [0003] In order to achieve highly sensiti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
CPCG01N27/26G01N27/3277G01N2458/00
Inventor 牛利胡琼
Owner GUANGZHOU UNIVERSITY
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