Near-infrared light-emitting polypeptide self-assembled gold nano-material, preparation method and application thereof

Abstract

The invention discloses a near-infrared light-emitting polypeptide self-assembly gold nano material, a preparation method and application thereof. The preparation method is simple in synthesis process, low in cost and easy for industrial production. The prepared nano material is an assembly, polypeptide is self-assembled to form a hollow nano fibrous structure, the stability is good, the biotoxicity is low, the gene loading capacity is high, gold nanoparticles are connected to nano fibers in a granular mode, the near-infrared luminescence property is achieved, and visualization research is facilitated. The invention further provides application of the nano material in gene transfection and cell imaging. According to the invention, the nano material is high in gene loading capacity and goodin plasmid transfection applicability, and plasmids with fluorescent labels can be used for transfecting cells to study the plasmid transfection effect; and due to the near-infrared luminescence characteristic, the near-infrared light-emitting polypeptide self-assembly gold nano material can also be used for researching cell imaging, can be used for observing the distribution condition of a to-be-observed substance in cells and the process of entering and exiting the cells, and has important significance in the aspects of bioengineering and gene therapy.

Description

technical field

[0001] The invention belongs to the field of functional optical nanometer materials, and in particular relates to a near-infrared luminescent polypeptide self-assembled gold nanometer material and its preparation method and application. Background technique

[0002] Gene therapy is a new research field in contemporary medicine and biology. It introduces normal genes or therapeutic DNA sequences into target cells in a certain way, regulates the expression of defective genes in cells from the genetic level or corrects and replaces them with normal genes. Defective gene to play a therapeutic role. In order to achieve the desired therapeutic effect, scientists have been studying effective ways to deliver plasmid DNA. At present, the vectors for introducing plasmid DNA into target cells mainly include viral vectors and non-viral vectors. The three mainstream viral vectors include lentiviral vectors, retroviral vectors, and adenoviral vectors, which have high tran...

Examples

Embodiment 1

[0066] The polypeptide c(ARARARAD)-Acp-C shown in formula I was designed and synthesized by Hefei Guopeptide Biotechnology Co., Ltd.

[0067] At room temperature, add 84 μL of chloroauric acid (73 mM), 272 μL of NaOH (0.1M) and 1336.8 μL of water into a 5mLEP tube, then add 51.2 μL of the polypeptide (6 mM), after standing at room temperature for 30 minutes, add 256 μL of pH It is neutral glutathione (24mM), let stand at room temperature for 1 hour, then react with magnetic stirring in an oil bath at 95°C for 12 hours, the magnetic stirring speed is 1500rpm, the solution turns from light yellow to colorless and transparent, and finally to brown yellow , stop the reaction, use a dialysis bag to dialyze in neutral pH water for 2 days to remove excess small molecules, and finally use a 10kDa ultrafiltration tube to ultrafilter and concentrate the dialysate to obtain the target product, which is stored in a 4°C refrigerator for later use.

[0068] Investigate the stability of the ...

Embodiment 2

[0083]At room temperature, add 84 μL of chloroauric acid (73 mM), 272 μL of NaOH (0.1M) and 1336.8 μL of water into a 5mLEP tube, then add 51.2 μL of polypeptide (same as in Example 1), after standing at room temperature for 40 minutes, add 256 μL of pH It is neutral glutathione (24mM), let it stand at room temperature for 1 hour, then react with magnetic stirring in an oil bath at 90°C for 30 hours, the magnetic stirring speed is 1500rpm, the solution is from light yellow to colorless and transparent, and finally to brown yellow , stop the reaction, use a dialysis bag to dialyze in neutral pH water for 2 days to remove excess small molecules, and finally use a 10kDa ultrafiltration tube to ultrafilter and concentrate the dialysate to obtain the target product, which is stored in a 4°C refrigerator for later use.

[0084] Figure 10 It is the fluorescence excitation spectrum and fluorescence emission spectrum of near-infrared luminescent polypeptide self-assembled gold nanomat...

Embodiment 3

[0086] At room temperature, add 21 μL of chloroauric acid (73mM), 68 μL of NaOH (0.1M) and 341.88 μL of water into a 2mL EP tube, then add 5.12 μL of polypeptide (same as Example 1), after standing at room temperature for 30 minutes, add 64 μL Glutathione (24mM) with neutral pH, let it stand at room temperature for 1 hour, then reacted with magnetic stirring in an oil bath at 95°C for 8 hours, the magnetic stirring speed was 1500rpm, the solution turned from light yellow to colorless and transparent, and finally to brown Yellow, stop the reaction, use a dialysis bag to dialyze in neutral pH water for 2 days to remove excess small molecules, and finally use a 10kDa ultrafiltration tube to concentrate the dialysate to obtain the target product, and store it in a 4°C refrigerator for later use.

[0087] Figure 11 It is the fluorescence emission spectrum diagram of the near-infrared luminescent peptide self-assembled gold nanomaterial, and there is a maximum emission peak at 810n...

technical field

[0001] The invention belongs to the field of functional optical nanometer materials, and in particular relates to a near-infrared luminescent polypeptide self-assembled gold nanometer material and its preparation method and application. Background technique

[0002] Gene therapy is a new research field in contemporary medicine and biology. It introduces normal genes or therapeutic DNA sequences into target cells in a certain way, regulates the expression of defective genes in cells from the genetic level or corrects and replaces them with normal genes. Defective gene to play a therapeutic role. In order to achieve the desired therapeutic effect, scientists have been studying effective ways to deliver plasmid DNA. At present, the vectors for introducing plasmid DNA into target cells mainly include viral vectors and non-viral vectors. The three mainstream viral vectors include lentiviral vectors, retroviral vectors, and adenoviral vectors, which have high tran...

Examples

Embodiment 1

[0066] The polypeptide c(ARARARAD)-Acp-C shown in formula I was designed and synthesized by Hefei Guopeptide Biotechnology Co., Ltd.

[0067] At room temperature, add 84 μL of chloroauric acid (73 mM), 272 μL of NaOH (0.1M) and 1336.8 μL of water into a 5mLEP tube, then add 51.2 μL of the polypeptide (6 mM), after standing at room temperature for 30 minutes, add 256 μL of pH It is neutral glutathione (24mM), let stand at room temperature for 1 hour, then react with magnetic stirring in an oil bath at 95°C for 12 hours, the magnetic stirring speed is 1500rpm, the solution turns from light yellow to colorless and transparent, and finally to brown yellow , stop the reaction, use a dialysis bag to dialyze in neutral pH water for 2 days to remove excess small molecules, and finally use a 10kDa ultrafiltration tube to ultrafilter and concentrate the dialysate to obtain the target product, which is stored in a 4°C refrigerator for later use.

[0068] Investigate the stability of the ...

Embodiment 2

[0083]At room temperature, add 84 μL of chloroauric acid (73 mM), 272 μL of NaOH (0.1M) and 1336.8 μL of water into a 5mLEP tube, then add 51.2 μL of polypeptide (same as in Example 1), after standing at room temperature for 40 minutes, add 256 μL of pH It is neutral glutathione (24mM), let it stand at room temperature for 1 hour, then react with magnetic stirring in an oil bath at 90°C for 30 hours, the magnetic stirring speed is 1500rpm, the solution is from light yellow to colorless and transparent, and finally to brown yellow , stop the reaction, use a dialysis bag to dialyze in neutral pH water for 2 days to remove excess small molecules, and finally use a 10kDa ultrafiltration tube to ultrafilter and concentrate the dialysate to obtain the target product, which is stored in a 4°C refrigerator for later use.

[0084] Figure 10 It is the fluorescence excitation spectrum and fluorescence emission spectrum of near-infrared luminescent polypeptide self-assembled gold nanomat...

Embodiment 3

[0086] At room temperature, add 21 μL of chloroauric acid (73mM), 68 μL of NaOH (0.1M) and 341.88 μL of water into a 2mL EP tube, then add 5.12 μL of polypeptide (same as Example 1), after standing at room temperature for 30 minutes, add 64 μL Glutathione (24mM) with neutral pH, let it stand at room temperature for 1 hour, then reacted with magnetic stirring in an oil bath at 95°C for 8 hours, the magnetic stirring speed was 1500rpm, the solution turned from light yellow to colorless and transparent, and finally to brown Yellow, stop the reaction, use a dialysis bag to dialyze in neutral pH water for 2 days to remove excess small molecules, and finally use a 10kDa ultrafiltration tube to concentrate the dialysate to obtain the target product, and store it in a 4°C refrigerator for later use.

[0087] Figure 11 It is the fluorescence emission spectrum diagram of the near-infrared luminescent peptide self-assembled gold nanomaterial, and there is a maximum emission peak at 810n...

Images