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Preparation method of photoelectrochemical sensor for detecting in-situ ratio of Cry1Ab protein in transgenic crop

A photoelectrochemical and sensor technology, which is applied in the field of biosensors, can solve the problems of small semiconductor band gaps, etc., and achieve the effect of improving selectivity, reducing interference, and increasing electron transfer rate

Active Publication Date: 2021-08-27
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As some common inorganic semiconductor materials, such as CdSe quantum dots (QDs), CdTe QDs, CdS QDs, etc., although the water solubility is good, the semiconductor band gap is small

Method used

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  • Preparation method of photoelectrochemical sensor for detecting in-situ ratio of Cry1Ab protein in transgenic crop
  • Preparation method of photoelectrochemical sensor for detecting in-situ ratio of Cry1Ab protein in transgenic crop
  • Preparation method of photoelectrochemical sensor for detecting in-situ ratio of Cry1Ab protein in transgenic crop

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] according to figure 1 Described preparation process:

[0048] (1) Preparation of Au NRs nanomaterials:

[0049] Au NRs were prepared using a seed-mediated growth method. First, 4.5mL H 2 O, 5 mL 0.2M CTAB solution, 500 μL 5 mM HAuCl 4 solution and 610 μL of 10 mM NaBH on ice 4 The solution is mixed. After stirring for 2 min, the mixture was allowed to react undisturbed for 2 h at 30 °C to obtain a gold seed solution. Then, 24mL 0.2M CTAB, 4.8mL 5mM HAuCl 4 solution, 50 μL 0.1M AgNO 3 solution, 32 μL of 1.2M HCl and 2.8 mL of 10 mM ascorbic acid solution were mixed in a 50 mL vial, and then 48 μL of the seed solution was added. The resulting solution was gently stirred for 15 s and left undisturbed for 20 h. The color of the solution changed from colorless to brick red, indicating the formation of Au NRs.

[0050] (2) Preparation of CdTe QDs nanomaterials:

[0051] CdTe QDs were synthesized using a hydrothermal method. First, synthesize the fresh precursor sod...

Embodiment 2

[0066] (1) Preparation of Au NRs nanomaterials:

[0067] Au NRs were prepared using a seed-mediated growth method. First, 4.5mL H 2 O, 5 mL 0.2M CTAB solution, 500 μL 5 mM HAuCl 4 solution and 610 μL of 10 mM NaBH on ice 4 The solution is mixed. After stirring for 2 min, the mixture was allowed to react undisturbed for 2 h at 30 °C to obtain a gold seed solution. Then, 24mL 0.2M CTAB, 4.8mL 5mM HAuCl 4 solution, 50 μL 0.1M AgNO 3 solution, 32 μL of 1.2M HCl and 2.8 mL of 10 mM ascorbic acid solution were mixed in a 50 mL vial, and then 48 μL of the seed solution was added. The resulting solution was gently stirred for 15 s and left undisturbed for 20 h. The color of the solution changed from colorless to brick red, indicating the formation of Au NRs.

[0068] (2) Preparation of CdTe QDs nanomaterials:

[0069] CdTe QDs were synthesized using a hydrothermal method. First, synthesize the fresh precursor sodium tellurium hydride (NaHTe): add 0.0638g tellurium powder and ...

Embodiment 3

[0084] (1) Preparation of Au NRs nanomaterials:

[0085] Au NRs were prepared using a seed-mediated growth method. First, 4.5mL H 2 O, 5 mL 0.2M CTAB solution, 500 μL 5 mM HAuCl 4 solution and 610 μL of 10 mM NaBH on ice 4 The solution is mixed. After stirring for 2 min, the mixture was allowed to react undisturbed for 2 h at 30 °C to obtain a gold seed solution. Then, 24mL 0.2M CTAB, 4.8mL 5mM HAuCl 4 solution, 50 μL 0.1M AgNO 3 solution, 32 μL of 1.2M HCl and 2.8 mL of 10 mM ascorbic acid solution were mixed in a 50 mL vial, and then 48 μL of the seed solution was added. The resulting solution was gently stirred for 15 s and left undisturbed for 20 h. The color of the solution changed from colorless to brick red, indicating the formation of Au NRs.

[0086] (2) Preparation of CdTe QDs nanomaterials:

[0087] CdTe QDs were synthesized using a hydrothermal method. First, synthesize the fresh precursor sodium tellurium hydride (NaHTe): add 0.0638g tellurium powder and ...

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Abstract

The invention belongs to the technical field of biosensors, and discloses a preparation method of an in-situ ratio photoelectrochemical immunosensor which is applied to detection of Cry1Ab protein in transgenic crops. Au NRs and MB are used as double sensitizers to realize double amplification of PEC signals. On the one hand, Au NRs modified CdTe QDs with an SPR effect is used as a photoelectric layer, so that initial signal amplification is realized; on the other hand, MB serving as a PEC signal enhancer is embedded into dsDNA, and secondary signal amplification is achieved. The concentration of Cry1Ab is quantitatively detected by taking the light current after the target object is added and the light current generated by the Ab2-CdSe QDs-dsDNA sensitized by the MB as a specific value respectively. The two signals are acquired at the same electrode and are mutually referenced, so that the interference of a solution matrix and environmental factors on the electrode is effectively reduced. The detection range of the photoelectrochemical sensor is 0.01-100 ng.mL <-1 >, and the detection limit is as low as 1.4 pg.mL <-1 >. The in-situ ratio PEC sensor constructed by the invention has good stability, selectivity and reproducibility, and realizes sensitive detection of Cry1Ab protein in transgenic crops.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and in particular relates to a preparation method and application of an in-situ ratiometric photoelectrochemical immunosensor based on dual signal amplification, which can be used for sensitive detection of Cry1Ab protein in transgenic crops. Background technique [0002] Photoelectrochemical (PEC) immunoassay is a powerful technique based on the photoelectric effect and antigen-antibody specific recognition. Among them, the PEC sensor uses the electrical signal generated by light excitation as the output signal, which leads to the complete separation of the excitation source and the output signal, ensuring high detection sensitivity and low background. Crystalline proteins produced by Bacillus thuringiensis (such as Cry1Ab protein) are insecticidal toxins used for pest control in transgenic crops, but the abundance of Cry1Ab protein in transgenic crops has aroused great concern about environm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327G01N27/36G01N33/68
CPCG01N27/26G01N27/36G01N27/3278G01N33/68
Inventor 刘东孟淑云由天艳贾帆
Owner JIANGSU UNIV
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