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Preparation method and application of PDL1 aptamer functionalized SERS (Surface Enhanced Raman Scattering) probe

An aptamer and functionalized technology, which can be used in instruments, analytical materials, measuring devices, etc., can solve the problem of inability to measure blood samples, and achieve the effect of good application prospects, good selectivity and high sensitivity

Pending Publication Date: 2022-05-27
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method needs to centrifuge the cells during detection, and it cannot be used for direct measurement of blood samples.

Method used

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  • Preparation method and application of PDL1 aptamer functionalized SERS (Surface Enhanced Raman Scattering) probe
  • Preparation method and application of PDL1 aptamer functionalized SERS (Surface Enhanced Raman Scattering) probe
  • Preparation method and application of PDL1 aptamer functionalized SERS (Surface Enhanced Raman Scattering) probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] like figure 1 As shown, the embodiment of the present application discloses a preparation method of an aptamer-functionalized SERS probe of PDL1, comprising the following steps:

[0072] S1. Preparation of CD63 probe

[0073] S1-1. Magnetic composite material Au / TiO 2 / Fe 3 o 4 preparation of

[0074] a. Preparation of Fe 3 o 4 Nanospheres: under magnetic stirring, 1.0g FeCl 3 ·6H 2 O, 0.43 g of trisodium citrate and 2 g of NaAc (sodium acetate) were completely dissolved in 33.3 mL of ethylene glycol, and then the above mixture was transferred to an autoclave lined with polytetrafluoroethylene and heated at 200 °C for 10 hours, The obtained black product was washed 3 times with deionized water and ethanol, and dried in a sealed glass bottle for 12 hours to obtain Fe 3 o 4 nanospheres.

[0075] b. Modified Fe 3 o 4 Nanospheres: 0.075g dry Fe 3 o 4 Dissolve nanospheres and 0.3mL ammonia water with a mass concentration of 28% in 90% ethanol, heat the above m...

Embodiment 2

[0090] like figure 1 As shown, the embodiment of the present application discloses a preparation method of an aptamer-functionalized SERS probe of PDL1, comprising the following steps:

[0091] S1. Preparation of CD63 probe

[0092] S1-1. Magnetic composite material Au / TiO 2 / Fe 3 o 4 preparation of

[0093] a. Preparation of Fe 3 o 4 Nanospheres: under magnetic stirring, 1.0g FeCl 3 ·6H 2 O, 0.4 g of trisodium citrate and 1.8 g of NaAc (sodium acetate) were completely dissolved in 33.3 mL of ethylene glycol, and then the above mixture was transferred to a Teflon-lined autoclave and heated at 180 °C for 9 h , the obtained black product was washed 3 times with deionized water and ethanol, and dried in a sealed glass bottle for 10 h to obtain Fe 3 o 4 nanospheres.

[0094] b. Modified Fe 3 o 4 Nanospheres: 0.075g dry Fe 3 o 4 Dissolve nanospheres and 0.3mL of 30% ammonia water in mass concentration in 90% ethanol, heat the above mixture to 40°C, and continuously m...

Embodiment 3

[0109] like figure 1 As shown, the embodiment of the present application discloses a preparation method of an aptamer-functionalized SERS probe of PDL1, comprising the following steps:

[0110] S1. Preparation of CD63 probe

[0111] S1-1. Magnetic composite material Au / TiO 2 / Fe 3 o 4 preparation of

[0112] a. Preparation of Fe 3 o 4 Nanospheres: under magnetic stirring, 1.0g FeCl 3 ·6H 2 O, 0.45 g of trisodium citrate and 2.2 g of NaAc (sodium acetate) were completely dissolved in 33.3 mL of ethylene glycol, and then the above mixture was transferred to a Teflon-lined autoclave and heated at 190 °C for 8 hours, the obtained black product was washed 3 times with deionized water and ethanol, and dried in a sealed glass bottle for 11h to obtain Fe 3 o 4 nanospheres.

[0113]b. Modified Fe 3 o 4 Nanospheres: 0.075g dry Fe 3 o 4 Dissolve nanospheres and 0.3mL ammonia water with a mass concentration of 25% in 90% ethanol, heat the above mixture to 50°C, and continuo...

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Abstract

The invention relates to the technical field of biosensing and material preparation thereof, in particular to a preparation method and application of an aptamer functionalized SERS (Surface Enhanced Raman Scattering) probe of PDL1, comprising preparation of an aptamer modified CD63 probe by using Au / TiO2 / Fe3O4 nanospheres, preparation of an aptamer modified PDL1 probe by using Ag / 4-ATP / Au SERS tags, and application of the probe in SERS specific detection of PDL1 by using a'sandwich 'method. According to the application, the exosome PDL1 is tested and detected by simultaneously using SERS (Surface Enhanced Raman Scattering) and ELISA (Enzyme-Linked Immunosorbent Assay), so that the method is proved to measure the expression time The analysis result of an original serum sample shows that the diagnostic sensitivity of the SERS method for detecting the circulating exosome PDL1 can reach 98%, which indicates that the SERS method has excellent sensitivity and accuracy. The aptamer probe is combined with an SERS sandwich method to quantify the PDL1 biomarker in serum, has the advantages of high specificity, high sensitivity, good selectivity and the like, is simple, convenient and rapid to operate, and has a good application prospect in monitoring of PDL1 / PD1-based immunotherapy.

Description

technical field [0001] The invention relates to the technical field of biosensing and material preparation thereof, in particular to a preparation method and application of a PDL1 aptamer functionalized SERS probe. Background technique [0002] Malignant cells bypass immunity through multiple pathways including the release of cytokines, such as tumor growth factor-β or interleukin 10 (IL-10), or altering tumor-associated macrophages and differential expression of cell-surface markers Systemically observed that this disrupts the balance with malignant cells by stimulating a state of immunosuppression that favors cancer metastasis. Cancer cells further recruit regulatory T cells (commonly known as Tregs), which help downregulate tumor antigen expression and promote apoptosis. This leads to the activation of immunosuppressive checkpoints (i.e., PDL1, CTLA-4, VISTA, and B7-H4), which bind to co-inhibitory receptors in T cells to limit the immune response system. The combined r...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/68G01N21/65
CPCG01N33/543G01N33/68G01N21/658
Inventor 黄青穆罕默德刘超
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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