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Preparation of molecular imprinting sensor for AFP online detection in human serum

A technology of molecular imprinting and human serum, applied in the field of biomedicine, can solve the problems of fluorescence change, and the research has not yet been reported, and achieves the effects of easy modification, good portability and low detection limit.

Active Publication Date: 2019-06-07
LANZHOU INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The literature shows that APBA-PA has both boronic acid groups and fluorescent groups, and can undergo boronic acid affinity with targets containing cis-ortho-dihydroxyl groups under neutral and weakly alkaline conditions to form boronic acid lactones. Its autofluorescence changes (Z.F. Xu, ACS Appl. Mater. Interfaces, 2014, 6 , 1406-1414), has been gradually used in the preparation of fructose, glucose and other sugar small molecule imprinted polymers, but the research on biomacromolecules such as glycoproteins has not been reported

Method used

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  • Preparation of molecular imprinting sensor for AFP online detection in human serum
  • Preparation of molecular imprinting sensor for AFP online detection in human serum
  • Preparation of molecular imprinting sensor for AFP online detection in human serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Preparation of a molecularly imprinted sensor for online detection of AFP in human serum (such as figure 1 shown), including the following steps:

[0041] ⑴ Preparation of (3-2-(4-(acryloyloxy)methyl)-1H-1,2,4-triazol-1-yl)acetamido)phenyl)boronic acid, namely APBA-PA (ACS Appl . Mater. Interfaces., 2014, 6, 1406-1414):

[0042] Weigh 1.488g of 3-aminophenylboronic acid hemisulfate and 2.016g of sodium bicarbonate and dissolve in 80 mL of ultrapure water, then add 3.680g of bromoacetyl bromide dropwise. The reaction solution was reacted in an ice-water bath for 2 h, and then continued to react for 16 h at room temperature. After the reaction, the precipitate was filtered out and redissolved in 20 mL sodium hydroxide solution (0.5 M). The pH of the solution was adjusted to 2.0, and the precipitate was filtered off and lyophilized.

[0043] Weigh 1.37g of the above product and 0.36g of sodium azide and dissolve it in 21 mL of N,N-dimethylformamide. After st...

Embodiment 2

[0062] Example 2 The preparation of a molecular imprinted sensor for online detection of AFP in human serum comprises the following steps:

[0063] ⑴ ~ ⑼ with embodiment 1.

[0064] (10) The cleaned carrier was first immersed in piranha solution, and the surface hydroxyl group activation reaction was carried out at 110 °C for 0.5 h, and then immersed in MPS absolute ethanol solution with a volume concentration of 10% at 75 °C for 20 min, and finally After rinsing with absolute ethanol and drying naturally, the double-bond functionalized carrier was obtained.

[0065] (11) The double-bond functionalized carrier was immersed in APBA-PA absolute ethanol solution with a mass concentration of 40%, and then AIBN equivalent to APBA-PA was added, and free radical polymerization was carried out at 72°C. After 6 h, the reaction was completed. Rinse with absolute ethanol and blow dry with nitrogen to obtain the APBA-PA grafted carrier.

[0066] ⑿ Immerse the APBA-PA grafted carrier int...

Embodiment 3

[0068] Example 3 The preparation of a molecular imprinted sensor for online detection of AFP in human serum comprises the following steps:

[0069] ⑴ ~ ⑼ with embodiment 1.

[0070] (10) The cleaned carrier was first immersed in piranha solution, and the surface hydroxyl group activation reaction was carried out at 90 °C for 2 h, and then immersed in MPS absolute ethanol solution with a volume concentration of 5% at 65 °C for 40 min, and finally After rinsing with absolute ethanol and drying with nitrogen gas, the double bond functionalized carrier was obtained.

[0071] (11) The double-bond functionalized carrier was immersed in APBA-PA absolute ethanol solution with a mass concentration of 20%, and then AIBN equivalent to APBA-PA was added, and free radical polymerization was carried out at 69 °C. After 15 h, the reaction was completed. Rinse with absolute ethanol and blow dry with nitrogen to obtain the APBA-PA grafted carrier.

[0072] ⑿ Immerse the APBA-PA grafted carri...

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Abstract

The invention relates to a preparation method of a molecular imprinting sensor for AFP online detection in human serum. The preparation method comprises the following steps: (1) preparing APBA-PA; (2)preparing a piranha solution; (3) preparing a MPS absolute ethyl alcohol solution; (4) preparing an APBA-PA absolute ethyl alcohol solution; (5) preparing a PB buffer solution; (6) preparing an AFP dilute solution; (7) preparing a TEOS solution; (8) preparing template molecule eluent; (9) cleaning a carrier; (10) orderly immersing the cleaned carrier into the piranha solution and the MPS absoluteethyl alcohol solution; rinsing, and drying to obtain a double-bond functionalized carrier; (11) immersing the double-bond functionalized carrier into the APBA-PA absolute ethyl alcohol solution, adding the AIBN, rinsing and blow-drying to obtain an APBA-PA grafted carrier; (12), immersing the APBA-PA grafted carrier into the AFP dilute solution to react, rinsing and air-drying to obtain the carrier fixed with the AFP on the surface; and (13) immersing the carrier fixed with the AFP on the surface into a TEOS solution to react, eluting, rinsing, and naturally air-drying to obtain the molecular imprinting sensor. The preparation method disclosed by the invention is simple, the cost is low, and the selected sensor is good in selectivity and good in biocompatibility.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the preparation of a molecular imprint sensor for online detection of AFP in human serum. Background technique [0002] Cancer is the second largest killer of human health. According to statistics, nearly ten million people die from cancer every year in the world (R. A. Smith, CA Cancer J. Clin. , 2017, 67 ,100-121). Early diagnosis and early treatment are the key to improving the survival rate of cancer patients. At present, enzyme-linked immunoassay technology is mainly used to detect tumor markers such as AFP (alpha-fetoprotein) in human serum clinically, as an important basis for early screening of cancer. The immunoassay based on natural antibodies is simple and fast, with high sensitivity and good selectivity, but this method has problems such as poor stability of reagents, high cost of obtaining high-quality antibodies, and high requirements for the experience of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/58G01N33/68
Inventor 马润恬赵晓博孙晓宇师彦平
Owner LANZHOU INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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