Construction method and application of polydopamine-mediated modification-free portable conductivity immunosensor with adjustable linear range

An immune sensor and polydopamine technology, applied in the field of analysis and detection, can solve problems such as poor stability and reproducibility, and achieve the effect of improving field operability, simple operation, and good stability

Pending Publication Date: 2022-07-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of method is limited by factors such as the amount of modification and modification conditions, resulting in poor stability and reproducibility. Therefore, developing a conductivity method that does not require electrode modification will greatly improve the stability of the method.

Method used

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  • Construction method and application of polydopamine-mediated modification-free portable conductivity immunosensor with adjustable linear range
  • Construction method and application of polydopamine-mediated modification-free portable conductivity immunosensor with adjustable linear range
  • Construction method and application of polydopamine-mediated modification-free portable conductivity immunosensor with adjustable linear range

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Modification of magnetic beads with biological recognition molecules

[0063] 1. Activation of magnetic beads

[0064] 1) Take 2 mg of magnetic beads (average diameter 1 μm) in a centrifuge tube, wash twice with 500 μL MEST (10 mM MES, 0.05% Tween20, pH 6.0), and remove the supernatant by magnetic separation;

[0065] 2) Prepare 5mg / mL EDC solution and 5mg / mL NHS solution with 10mM MES (pH 6.0);

[0066] 3) Add 100 μL EDC (5 mg / mL) and 50 μL NHS (5 mg / mL) to the centrifuge tube containing magnetic beads, mix well with a vortexer to fully suspend the magnetic beads, dilute to 500 μL with MES, and place in a On a rotary mixer, activate at 37°C for 30min;

[0067] 4) Magnetic separation, remove the supernatant, add 500 μL of MEST, and transfer the magnetic beads to a new centrifuge tube;

[0068] 5) Magnetic separation, remove the supernatant, wash twice with 500 μL of MEST, magnetic separation, remove the supernatant.

[0069] After the above steps, the carb...

Embodiment 2

[0077] Example 2 Synthesis and Biomodification of Gold Core Platinum Shell Nanometers

[0078] 1. Synthesis of gold core platinum shell nanozyme

[0079] In order to avoid the impurity of the prepared nanoparticles, the utensils required for the experiment should be soaked in freshly prepared aqua regia (concentrated nitric acid:concentrated hydrochloric acid: 1:3) for 3h before the experiment, rinsed with ultrapure water, and dried for use. use. The preparation of AuNPs introduced the microwave synthesis method, and the preparation of AuNPs could be completed in 5 minutes. Add 200 μL HAuCl4 (48 μM) and 9.8 mL ultrapure water to the reaction kettle, and keep boiling for 1 min under microwave heating at 100 °C. Then, the magnetic stirring was turned on, and 1 mL of 1 wt% sodium citrate was rapidly added at one time, kept at 100 °C for 4 min, and then cooled to room temperature to obtain a red AuNPs suspension. Au@PtNPs were synthesized by platinum precipitation on the surface ...

Embodiment 3

[0085] Example 3 Optimization of reaction conditions for conductivity immunosensors without signal amplification strategy

[0086] Different mass (20 μg, 50 μg, 100 μg, 150 μg) of DON complete antigen was coupled to the magnetic beads, and other experimental steps were as described above. The result is as figure 2 As shown, the conductivity immunosensor has the best response effect when the complete antigen concentration is 20 μg.

[0087] Different concentrations (1 μg / mL, 2 μg / mL, 3 μg / mL, 4 μg / mL) of DON monoclonal antibody were used for competitive immune reaction, and other experimental steps were as described above. The result is as Figure 4 As shown, the conductivity immunosensor has the best response effect when the concentration of spittoxin monoclonal antibody is 2 μg / mL.

[0088] Dopamine hydrochloride solutions with different concentrations (1mg / mL, 3mg / mL, 5mg / mL, 10mg / mL) were set up, and other experimental steps were as described above. The result is as ...

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Abstract

The invention relates to a construction method and application of a linear range-adjustable polydopamine-mediated modification-free portable conductivity immunosensor, in the process of catalyzing hydrogen peroxide by nano-enzyme, dopamine hydrochloride can be rapidly polymerized to generate polydopamine, rich functional groups on the surface of the polydopamine have a good complexing effect on metal ions (such as Fe < 3 + >, Cu < 2 + > and the like), and the polydopamine-mediated modification-free portable conductivity immunosensor can be used for detecting metal ions (such as Fe < 3 + >, Cu < 2 + > and the like). The metal ions have strong conductivity signals, so that the correlation between the nano-enzyme and the change value of the conductivity signals is established. And in combination with immunoreaction, the content of the target object can control the content of the nano-enzyme combined on the immunomagnetic beads, so that the conductivity signal of the metal ions is regulated and controlled, and the electric signal is read out through a portable conductivity meter to realize quantitative analysis of the target object. An electrode does not need to be modified, and the problems that a traditional electrochemical immunosensor needs to be modified with the electrode, so that stability is poor, and operation is complex are solved.

Description

technical field [0001] The invention belongs to the field of analysis and detection, and in particular relates to a construction method and application of a modified-free portable conductivity immune sensor mediated by polydopamine with adjustable linear range. Background technique [0002] Physical and chemical analysis and biochemical methods are the main quantitative methods in the field of analysis technology. The sample pretreatment steps of physical and chemical analysis methods are relatively complicated, professional technicians are required, and the detection cost is high, which restricts the application of rapid analysis and detection in the field. Biochemical methods can be further divided into microbial analysis and immunoassay. Microbiological analysis is affected by many factors, cannot be accurately quantified, and has poor repeatability. Immunoassays mainly include enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay and colloidal gold i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/06G01N27/327G01N33/543G01N33/558
CPCG01N27/06G01N27/3275G01N33/54326G01N33/558
Inventor 陈翊平冯牛董永贞鲁鹏魏巧玲
Owner HUAZHONG AGRI UNIV
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