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Quantum dot-based protein imprinted polymer microsphere as well as preparation and application thereof

A Western blotting and polymer technology, applied in the fields of materials science and engineering and bioseparation engineering, can solve the problems of difficult to ensure quantum dots embedding, cumbersome modification process, difficult to control, etc., and achieve rich molecular imprinting/protein related research, high Identify selective, highly selective effects

Active Publication Date: 2017-11-24
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the fluorescent sensors based on quantum dots are embedded in silica particles or modified on the surface of silica particles, and then synthesized on silica by free radical polymerization or sol-gel process. Preparation of molecularly imprinted layers on the surface. On the one hand, these methods are difficult to ensure that quantum dots are uniformly embedded in highly cross-linked imprinted polymers. In addition, most of them require a cumbersome modification process, which is not only time-consuming but also difficult to control.

Method used

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  • Quantum dot-based protein imprinted polymer microsphere as well as preparation and application thereof
  • Quantum dot-based protein imprinted polymer microsphere as well as preparation and application thereof
  • Quantum dot-based protein imprinted polymer microsphere as well as preparation and application thereof

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Experimental program
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Effect test

Embodiment 1

[0034] a. Preparation of CdTe quantum dots co-stabilized by thioglycolic acid and glutathione: Weigh 38.3mg of tellurium powder and 40mg of sodium borohydride, add 1.5mL of ethanol and 0.5mL of water to isolate the system from oxygen. Heat the reaction in a water bath until the black tellurium powder disappears completely and the supernatant turns lavender. Weigh 92.4 mg of cadmium nitrate and add it to 75 mL of water, then add 25 μL of thioglycolic acid and 110 mg of glutathione, adjust the pH to 9 with 1M sodium hydroxide solution, blow with nitrogen for 30 minutes, and then take the lavender supernatant prepared above Add 1mL of the liquid into the cadmium nitrate solution, under the protection of nitrogen, reflux for 2 hours, and obtain the CdTe quantum dot liquid co-stabilized by green thioglycolic acid and glutathione (TGA-GSH).

[0035] b. Preparation of phycocyanin imprinted polymer: Take 5 mL of TGA-GSH co-stabilized quantum dot solution prepared above, add it to 10 m...

Embodiment 2

[0038] Take 10 μL of the CdTe quantum dots, phycocyanin imprinted polymers (PC MIPs), non-imprinted polymers (PC NIPs) and bovine hemoglobin imprinted polymers (BHb MIPs) obtained by diluting 1000 times with ultrapure water respectively. and non-imprinted polymer (BHb NIPs) solutions, each dilution was dispersed on a copper grid washed with ethanol, after drying, the copper grid loaded with the above-mentioned diluted substances was observed with a transmission electron microscope (see figure 2 A-E); if figure 2 As shown in A, the particle size of quantum dots is about 2nm, such as figure 2 As shown in B, the particle size of PC MIPs is about 5nm, and the dispersion is very good. The thickness of the imprinted layer of phycocyanin is about 3nm. Such a thin imprinted layer is very beneficial for obtaining higher detection sensitivity and faster response of. Such as figure 2 As shown in C, the particle size and shape of PC NIPs are similar to those of quantum dots, indica...

Embodiment 3

[0040] After ultrasonically dispersing the prepared phycocyanin imprinted polymer (PC MIPs) solution, 3 parallel copies were taken, and 1 mL of PC MIPs solution was added to a weighed 1.5 mL centrifuge tube, dried and then weighed. By calculating the ratio of the mass difference before and after the three parallel samples to the volume, the average value is the concentration of the PC MIPs solution, and the concentration is 4.6 mg / mL.

[0041] During detection, 0.2 mL of the PC MIPs solution with a concentration of 4.6 mg / mL obtained above was taken, and the total volume of the measurement solution was 0.5 mL, that is, the concentration of PC MIPs was 1.84 mg / mL. In the same way, the concentration of bovine blood red fluorescent nanoimprinted polymers (BHb MIPs) in the detection process was 1 mg / mL.

[0042] Prepare a series of PBS buffer solutions with pH values ​​of 6.0, 6.5, 7.0, 7.5, 8.0, 8.5 and 9.0. In the first group, take 0.2 mL of phycocyanin imprinted polymer (PC MI...

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Abstract

The invention belongs to the fields of material science and engineering and biological separation engineering, and particularly relates to a quantum dot-based protein imprinted polymer microsphere as well as preparation and application thereof. The CdTe quantum dot is directly used as a functional monomer through taking mercaptoacetic acid (TGA) and glutathione (GSH) together as a stabilizer, protein is a template molecule, dopamine is a cross-linking agent, and the imprinted polymer microsphere is obtained through the auto polymerization of the dopamine. The quantum dot-based protein imprinted polymer microsphere as well as the preparation and the application thereof provided by the invention have the advantages that fluorescence signals of the quantum dots are utilized for detection, and the functionalized quantum dots are directly taken as functional monomers, so that the quantum dots are contained in imprinted holes. Therefore, the detection sensitivity can be improved. Meanwhile, an imprinted polymer layer is obtained through the auto polymerization of the dopamine, and the imprinting process is further simplified. The quantum dot-based protein imprinted polymer microsphere is used for analysis and detection on phycocyanin in a sea water and lake water sample and bovine hemoglobin in a bovine urine sample, the adding standard recovery rate can reach 90 percent or higher, and the reuse performance is good.

Description

technical field [0001] The invention belongs to the fields of material science and engineering and biological separation engineering, and specifically relates to a quantum dot-based protein imprinted polymer microsphere and its preparation and application. Background technique [0002] Molecularly imprinted polymers have been rapidly developed in the fields of environment and life due to their high selectivity, simple preparation, strong resistance to harsh environments, good stability, and low cost. Imprinting objects also gradually develop from small molecular compounds and ions to biological macromolecules such as proteins, viruses, and cells. At present, small molecular compounds and ion imprinting have been well developed, but biomacromolecular imprinting, especially protein imprinting, is still a very challenging research field in the field of molecular imprinting. The main reasons are: 1) protein molecules are large in size and very It is difficult to enter and exit ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G73/06C08J9/26C08K9/04C08K3/30G01N21/64C09K11/88C09K11/02
Inventor 李金花王晓艳陈令新于佳洛
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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