Protein bioluminescence imaging sensor based on peryleneimide multi-arm polymer

A multi-arm polymer and imaging sensor technology, applied in the field of analytical chemistry, can solve the problems that the background matrix does not have the possibility of developing into a portable device, unfavorable sensor deviceization, etc., to avoid biological matrix interference, low cost, and specificity strong effect

Inactive Publication Date: 2019-07-16
CHONGQING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these systems generally require excitation light sources, which inevitably receive interference from the background matrix and do not have the possibility of being developed into portable devices, which is not conducive to the realization of device-based sensors.
Therefore, it is still extremely challenging to develop a novel protein detection array sensor with high accuracy, high sensitivity and simple portability.

Method used

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  • Protein bioluminescence imaging sensor based on peryleneimide multi-arm polymer
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  • Protein bioluminescence imaging sensor based on peryleneimide multi-arm polymer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of peryleneimide polymerized monomers.

[0043] (1) Preparation of N-acryloyl-L-phenylalanine (NALP): 3.3038g of phenylalanine (0.02mol), 0.01g of 2,6-di-tert-butyl-p-cresol, 0.8g of NaOH (0.02 mol) and 20 mL of distilled water were placed in a 100 mL round-bottomed flask to obtain a clear solution (pH 12), which was stirred in an ice bath, and 1.63 mL of acryloyl chloride (0.02 mol) was added dropwise to the solution. After the dropwise addition, the mixture was warmed to room temperature and stirred at room temperature for 1 hour. The clear aqueous solution was then acidified to pH 1-2 with concentrated hydrochloric acid. The precipitate was filtered and recrystallized, yield: 62%.

[0044] (2) N-acryloyl-L-serine (NALS), N-acryloyl-L-aspartic acid (NALA), N-acryloyl-L-leucine (NALL) and N-acryloyl-L -The preparation of histidine (NALH) uses 0.02mol of the corresponding amino acid as raw material, and follows the same preparation steps as NAL...

Embodiment 2

[0046] Example 2: Preparation of peryleneimide multi-arm polymers P1-P6.

[0047] Six peryleneimide multi-arm polymers, namely P1, P2, P3, P4, P5 and P6, are prepared as follows:

[0048] Preparation of P1: The polymerization was carried out in a strictly dry Schlenk tube, to which were added initiator 4Br-PDI (10 mg, 5.4 x 10-3 mmol, 1 equiv), NALP (508 mg, 2.16 mmol, 4 x 100 equiv) and butyl Ketone / methanol / water (2:1:1, 2.0 mL total). The reaction tube was degassed through three freeze-pump-thaw cycles, then pentamethyldivinyltriamine (77.8 mg, 0.45 mmol, 4 x 21 equiv) and CuBr (21.4 mg, 0.15 mmol, 4 x 7 equiv) were added. After stirring at room temperature for 10 minutes to ensure complete formation of the catalyst complex, the polymerization was carried out at 60°C under a nitrogen blanket. After 5 hours of reaction, the reaction was quenched with liquid nitrogen, the reaction mixture was poured into excess ether to precipitate, the precipitate was dissolved in water an...

Embodiment 3

[0060] Example 3: Peryleneimide multi-arm polymer-based protein bioluminescence imaging sensor

[0061] The overlap of the luciferase emission spectrum and the excitation spectrum of polymers P1-P6 is shown in the appendix Figure 9 A, the detection wavelength is 500-700nm band to detect the result of BRET signal as shown in Fig. Figure 9 shown in B.

[0062] A sensing unit is constructed, the sensing unit is a BRET acceptor peryleneimide multi-arm polymer and a BRET donor luciferase, and a titration method is used to determine the relationship between the BRET acceptor peryleneimide multi-arm polymer and the donor luciferase. The stoichiometric ratio of luciferase, the concentration of luciferase is constant, the concentration of polymer is gradually increased, and the change of BRET efficiency is recorded. When the curve becomes stable and no longer rises, the BRET efficiency at this point is the maximum BRET efficiency. Taking the stoichiometric ratio of BRET acceptor an...

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Abstract

The invention relates to a protein bioluminescence imaging sensor based on perylene bisimide star-shaped polymers. By adopting firefly luciferase as a donor and six perylene bisimide star-shaped polymers as a receptor, to-be-detected protein is competitively combined with a receptor polymer by non-covalent interaction, and luciferase of the donor is displaced, so that a bioluminescence resonant energy transfer signal is changed; a signal is acquired by a small animal bioluminescent imager, and response signals of each protein to six sensing units jointly form a unique response mode of the protein, and the unique response mode is similar to a fingerprint and is used for identifying. 10 proteins are identified by using linear discriminant analysis, and the correct rate reaches 100 percent; 76 unknown protein samples are successfully identified by combining with an ultraviolet spectroscopy technology, and the correct rate is as high as 95 percent. According to the method, the signals can be acquired by simple sample adding and shooting without a light source; the method has the advantages of quickness, convenience, good stability, high sensitivity, high specificity, low cost and the like.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and relates to a protein bioluminescence imaging sensor based on a peryleneimide multi-arm polymer. Background technique [0002] Protein detection is an important subject in analytical chemistry, and its application in medicine has also received extensive attention, especially in the diagnosis of disease markers, drug screening, and disease prognosis monitoring. However, due to the structural diversity and complexity of target analytes, the detection of proteins becomes an extremely challenging problem. Array sensors, also known as "chemical nose / tongue", utilize the fingerprints formed by the cross-response between multiple sensing units and analytes to realize the identification of multiple substances and complex mixtures, with fast, simple and selective The advantages of high sensitivity, high stability and good stability provide an ideal strategy for protein detection. In the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 魏为力卢辰玮夏之宁
Owner CHONGQING UNIV
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