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Method for Improving the Response Performance of Graphene Oxide-Based Aptamer Sensors

An aptamer sensor, graphene-based technology, used in instruments, scientific instruments, material excitation analysis, etc., can solve problems such as complex interaction relationships and redundant base recognition

Inactive Publication Date: 2021-06-04
BEIJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the target, aptamer and GO exist in the system of the GO-aptamer complex platform, the interaction relationship between the three is complex, and the dissociation constant K d or K a The redundant bases in the complex system cannot be effectively identified, and the optimal truncated sequence obtained is not necessarily the most suitable for the GO platform, and the response performance of the GO-based aptamer sensor constructed based on this is usually limited.
Therefore, it is of great significance to establish a comprehensive judgment method for aptamer sequence optimization suitable for the GO-aptamer complex platform, but there are also great challenges

Method used

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  • Method for Improving the Response Performance of Graphene Oxide-Based Aptamer Sensors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Aptamer / GO detection platform detects AFB1

[0045] 1.1 Aptamer truncation

[0046] Base removal design was performed on P-AFB1-50 to obtain a series of truncated aptamers, namely P-AFB1-50, P-AFB1-46, P-AFB1-40, P-AFB1-35, P-AFB1- 31, P-AFB1-37, P-AFB1-34 and P-AFB1-30, the truncated aptamer sequences are shown in Table 1:

[0047] Table 1P-AFB1-50 truncated aptamer sequence

[0048]

[0049]

[0050] 1.2 CD method to monitor the degree of configuration change of truncated aptamers before and after the appearance of the target

[0051] In 10mM Tris salt buffer solution (pH 8.0, 120mM NaCl, 20mM CaCl 2 , 5mM KCl), test AFB1 (1.0μg mL -1 ) CD spectra of P-AFB1-50 chains at a concentration of 1.0 μM before and after appearance. Such as figure 1 As shown, after the appearance of AFB1 in the P-AFB1-50 aptamer chain, the positive peak of the CD spectrum at 283 nm was significantly enhanced. Therefore, CD technology can be used to characterize the inter...

Embodiment 2

[0076] Example 2 Aptamer / GO detection platform detects OTA

[0077] 2.1 Sequence truncation

[0078] The P-OTA-61 aptamer was designed by base deletion, and a series of truncated aptamers were obtained, respectively P-OTA-61, P-OTA-48 and P-OTA-36, the sequences of the truncated aptamers See Table 4:

[0079] Table 4 Base sequences of P-OTA-61 series truncated aptamers

[0080]

[0081] 2.2 Using the CD method to test the degree of configuration change of the above-mentioned truncated aptamers before and after the appearance of OTA

[0082] In 10mM Tris salt buffer solution (pH 8.0, 120mM NaCl, 20mM CaCl 2 , 5mM KCl), the test concentration is 1.0μg mL -1 CD spectra of aptamer chains (P-OTA-61, P-OTA-48, P-OTA-36) at a concentration of 1.0 μM before and after OTA appearance.

[0083] Such as Figure 7 As shown, the addition of OTA to the original aptamer P-OTA-61 solution did not cause a change in the CD signal, indicating that the P-OTA-61 chain has a small degree of...

Embodiment 3

[0096] Example 3 Constructing a dual-channel aptamer / GO fluorescence response platform to realize the simultaneous detection of AFB1 and OTA

[0097] In order to balance the interaction between aptamer chains with different properties and GO, 60nM FAM-labeled P-AFB1-40 and 180nM Texas red-labeled P-OTA-36 were mixed, and then different concentrations of GO were added to the solution to adjust the GO concentration. For optimization, after mixing for 3 minutes, use it to test its fluorescence spectrum.

[0098] Such as Figure 11 As shown, with the increase of GO concentration, the fluorescence intensities at the emission wavelengths of the two channels decreased gradually. When the GO concentration increased to 16 μg mL -1 , the quenching efficiency of P-OTA-36-Texas red reached 95%, while that of P-AFB1-40-FAM reached 80%. Therefore, this experiment chooses 16 μg mL -1 GO was used to construct a dual-channel aptamer / GO composite platform for qualitative and quantitative det...

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Abstract

The invention belongs to the technical field of post-screening of aptamers and the technical field of rapid biosensing detection, and in particular relates to a method for improving the response performance of a graphene oxide (GO)-based aptamer sensor. The method for improving the response performance of the graphene oxide-based aptamer sensor of the present invention comprises the following steps: designing a truncated aptamer according to the secondary structure of the original aptamer; monitoring the interaction between the truncated aptamer and the target and Configuration change; determination of the quenching constant between the truncated aptamer and graphene oxide before and after the appearance of the target; comprehensive analysis and judgment from the aspects of probe configuration, binding strength, and adsorption and desorption difficulty, etc., to obtain a suitable graphene oxide platform The aptamer with the best response performance is used to construct a graphene oxide-based aptamer sensing platform. The invention constructs a high-performance GO-based aptamer sensor, and realizes rapid and sensitive detection of a single mycotoxin and multiple mycotoxins.

Description

technical field [0001] The invention belongs to the technical field of post-screening of aptamers and the technical field of rapid biosensing detection, and specifically relates to a method for improving the response performance of a graphene oxide-based aptamer sensor. Background technique [0002] Graphene oxide (GO)-based aptamer sensors are a simple and easy-to-use detection platform, usually using fluorescently labeled aptamers (aptamers) as probes, water-soluble GO as probes for adsorption and fluorescence quenching. carrier. Under normal circumstances, the aptamer probe is tightly adsorbed on the GO surface and the fluorescence is quenched. When the target appears, the specific interaction between the target and the probe makes the aptamer desorb from the GO surface, resulting in significant the fluorescence output signal. The graphene oxide (GO)-based aptamer sensing platform has the characteristics of both aptamers and GO, and has outstanding advantages such as si...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N21/19
CPCG01N21/19G01N21/6428G01N2021/6432G01N2021/6443
Inventor 李运超王杏林何佳乐
Owner BEIJING NORMAL UNIVERSITY
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