Protein bioluminescence imaging sensor based on perylene bisimide star-shaped polymers

A multi-arm polymer, imaging sensor technology, applied in the field of analytical chemistry, can solve the problems of the background matrix not having the possibility to develop into a portable device, unfavorable sensor deviceization, etc., to avoid biological matrix interference, low cost, and stability. Good results

Inactive Publication Date: 2017-06-20
CHONGQING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 perylene bisimide star-shaped polymers
  • Protein bioluminescence imaging sensor based on perylene bisimide star-shaped polymers
  • Protein bioluminescence imaging sensor based on perylene bisimide star-shaped polymers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of perylene imide polymer monomer.

[0043] (1) Preparation of N-acryloyl-L-phenylalanine (NALP): 3.3038g phenylalanine (0.02mol), 0.01g2,6-di-tert-butyl-p-cresol, 0.8g NaOH (0.02 mol) and 20mL of distilled water were placed in a 100ml round bottom flask to obtain a clear solution (pH12), stirred in an ice bath, and 1.63ml of acryloyl chloride (0.02mol) was added dropwise to the solution. After the dropwise addition was complete, the mixture was allowed to warm up 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) all uses 0.02 mol of the corresponding amino acid as a raw material, following the same preparation steps as NAL...

Embodiment 2

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

[0047] 6 kinds of perylene imide multi-arm polymers, i.e. P1, P2, P3, P4, P5 and P6, are prepared as follows:

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

Embodiment 3

[0060] Example 3: Protein bioluminescent imaging sensor based on peryleneimide multi-arm polymer

[0061] Please refer to the appendix for the overlap of the luciferase emission spectrum and the excitation spectrum of the polymer P1-P6 Figure 9 A, the results of detecting BRET signal in the detection wavelength range of 500-700nm are as follows Figure 9 Shown in B.

[0062] Construct a sensing unit, 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 ratio of the BRET acceptor perylene imide multi-arm polymer to the donor luciferase. The stoichiometric ratio of luciferase makes the luciferase concentration constant, gradually increases the polymer concentration, and records the change of BRET efficiency. When the curve becomes stable and no longer rises, the BRET efficiency at this point is the maximum BRET efficiency. The stoichiometric ratio of BRET acceptor and BRET donor lucife...

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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 perylene imide multi-arm polymer. Background technique [0002] Protein detection is an important topic 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 diversity and complexity of target analyte structures, the detection of proteins becomes an extremely challenging problem. Array sensor, also known as "chemical nose / tongue", uses the fingerprints formed by the cross-response between multiple sensing units and analytes to realize the identification of multiple substances and complex mixtures, which is fast, simple and selective High, high sensitivity, good stability and other advantages provide an ideal strategy for protein detection. In the past few years,...

Claims

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Application Information

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Patent Type & Authority Applications(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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