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Liquid crystal type aptamer biosensor, preparation method thereof and application of liquid crystal type aptamer biosensor in detection of prostate specific antigens

A biosensor and aptamer technology, applied in the field of chemical detection, can solve the problems of narrow detection linear range, high cost of single- and double-stranded aptamers, and achieve the effects of less harsh storage conditions, lower PSA detection limit, and high sensitivity

Pending Publication Date: 2021-09-17
CHANGCHUN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The literature reports that the PSA liquid crystal aptamer sensor has problems such as narrow detection linear range and high cost of single- and double-chain aptamers.

Method used

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  • Liquid crystal type aptamer biosensor, preparation method thereof and application of liquid crystal type aptamer biosensor in detection of prostate specific antigens
  • Liquid crystal type aptamer biosensor, preparation method thereof and application of liquid crystal type aptamer biosensor in detection of prostate specific antigens
  • Liquid crystal type aptamer biosensor, preparation method thereof and application of liquid crystal type aptamer biosensor in detection of prostate specific antigens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Proportion optimization of the self-assembled APTES / DMOAP hybrid film on the surface of the sensor substrate

[0054] In order to ensure the uniform vertical orientation of the liquid crystal molecules of the liquid crystal sensor 5CB, and to immobilize more aptamer molecules on the glass substrate under the sensor to improve the detection range of PSA, the APTES and APTES / DMOAP hybrid films self-assembled on the surface of the glass substrate under the sensor were carried out. DMOAP ratio optimization.

[0055] (1) The concentration of DMOAP modified on the lower slide was fixed at 0.3% (v / v), and the concentration of APTES was changed to 0.3%, 1.5% and 3.0%, respectively. Soak the cleaned lower glass slide in the mixed solution of absolute ethanol of APTES and DMOAP, soak for 2 hours at 80°C, rinse with a large amount of absolute ethanol and ultrapure water in sequence, and then rinse with N 2 Blow-dry and dry at a constant temperature of 110°C for 1 hour ...

Embodiment 2

[0057] Example 2: Optimization of GA and aptamer concentrations on the sensor substrate surface when APTES / DMOAP=1:1

[0058] In order to immobilize the nucleic acid aptamer with modified amino groups on the surface of the lower glass slide, it is necessary to carry out aldylation treatment on the modified APTES on the surface of the lower glass slide. The bottom slide of the self-assembled APTES / DMOAP hybrid membrane was incubated with 0.1% and 0.5% (v / v) GA, respectively.

[0059] (1) The lower slide of the (APTES-GA) / DMOAP mixed membrane obtained by 0.1% GA treatment was further coupled with an aminated aptamer. Colored and bright areas (see Figure 5 a), Excessive aptamers affect the molecular orientation of 5CB in the copper grid; when the aptamer concentration is 1nmol / L, there are still some light spots in the image (see Figure 5 b) When the concentration of the aptamer drops to 100pmol / L, the polarized light microscopy image appears uniform black (see Figure 5 c)....

Embodiment 3

[0062] In order to increase the number of aptamer molecules immobilized on the surface of the glass slide substrate under the sensor, it is necessary to increase the number of aptamed molecules of APTES. 0.1% and 0.5% (v / v) GA were used to incubate the self-assembled APTES / DMOAP hybrid film on the surface of the slide substrate. 0.1% (v / v) GA formhylated (APTES-GA) / DMOAP composite layer under the slide, the polarized light microscope image appears uniform black (see attached Figure 6 a), further immobilize the aptamer; when the concentration of the aptamer is 100pmol / L, a large area of ​​color and bright area appears in the polarized light microscope image (see Figure 6 b). In order to immobilize more aptamer molecules on the surface of the glass slide under the sensor substrate to improve the sensitivity of the liquid crystal aptamer biosensor to detect PSA, this patent adopts the optimization conditions of Example 2, that is, the self-assembly of the sensor substrate (APT...

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Abstract

The invention relates to a liquid crystal type aptamer biosensor, a preparation method thereof and an application of the liquid crystal type aptamer biosensor in detection of prostate specific antigens. The biosensor comprises an upper slide, a copper grid and a lower slide which are sequentially arranged and treated by certain steps. The invention provides a new method for rapidly detecting the prostate specific antigen (PSA), and the PSA liquid crystal type aptamer sensor has the advantages of good specificity, wide linear range, high sensitivity, no need of marking, convenience and rapidness in detection and the like, and is beneficial to wide application.

Description

technical field [0001] The invention relates to the technical field of chemical detection, in particular to a prostate-specific antigen liquid crystal aptamer biosensor with a wide linear range and low detection limit, a preparation method and quantitative detection application thereof. Background technique [0002] Prostate specific antigen (PSA) is a serine protease with a molecular weight of about 33kDa regulated by androgen. This enzyme is a member of the kallikrein family and is produced in the epithelial cells of the prostate gland and ducts. As the first tumor marker approved by the US FDA, PSA is widely used in the screening and diagnosis of prostate cancer patients, and its sensitivity to prostate cancer is as high as 90%. The PSA concentration secreted by a healthy prostate into the human circulatory system is generally lower than 4.0 ng / mL, and 4.0 ng / mL is the internationally accepted concentration threshold for evaluating prostate cancer. For patients undergoin...

Claims

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Application Information

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IPC IPC(8): G01N21/21G01N21/17G01N33/573
CPCG01N21/21G01N21/17G01N33/573G01N2021/1776G01N2333/96433
Inventor 金丽虹潘兴隆吕九宫牟晓雨张翔宇李晶怡申炳俊夏冰高妍
Owner CHANGCHUN UNIV OF SCI & TECH
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