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Nucleic acid protein nanocomposite as well as preparation method and application thereof

A nanocomposite and nucleic acid nanotechnology, which is applied in the direction of peptide/protein components, drug combinations, and pharmaceutical formulations, can solve limitations and other problems, achieve low toxicity and side effects, realize large-scale production, and simple operation process

Active Publication Date: 2018-07-24
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is limited to gold nanoparticles and chemotherapeutic drugs, and only limited to those with chemotherapeutic effects

Method used

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  • Nucleic acid protein nanocomposite as well as preparation method and application thereof
  • Nucleic acid protein nanocomposite as well as preparation method and application thereof
  • Nucleic acid protein nanocomposite as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Preparation of Nucleic Acid-Protein Nanostructured Complexes

[0078] (1) Assembly and purification of nucleic acid nanostructures with modified upper capture strands

[0079] A long scaffold chain is mixed with the designed staple short chain and capture chain, and the pre-designed nucleic acid nanostructure is assembled according to the principle of complementary base pairing through the classical programmed cooling method. The scaffold chain is the M13 phage genome, and the staple chain is a designed short nucleic acid sequence that is complementary to the scaffold chain. The molar ratio is 1:10:10. After the self-assembly is completed, a centrifugal purification column (100kDa molecular weight cut-off) can be used at 5000 rpm for 3 minutes to concentrate, and the liquid retained in the purification column can be collected in a new EP tube.

[0080] (2) Preparation, purification and quantification of nucleic acid chain functional protein coupling products

[0081]...

Embodiment 2

[0111] The difference between this example and Example 1 is that in step (1) when preparing the DNA nanostructure of the modified upper capture strand, the molar ratio of the scaffold strand, the staple strand and the capture strand is set to 1:5:5 , after the self-assembly is completed, use a centrifugal purification column (100kDa molecular weight cut-off) at 8000 rpm for 2 minutes to concentrate, and collect the retained liquid in the purification column into a new EP tube. In step (2), take 50 μL of ribonuclease A (RNase A) with a concentration of 50 μM in a 1.5 mL EP tube, add 5 μL of Sulfo-SMCC solution with a concentration of 25 mM, shake and mix, and then add 180 μL of 1×PBS Buffer solution, vortexed to mix, fast centrifuged for 20 seconds, sealed with a parafilm, and placed on a shaker for 4 hours. Control the molar ratio of RNase A to Sulfo-SMCC to be 1:50. After the reaction was completed, excess Sulfo-SMCC was removed by centrifugation at 2500 rpm for 3 min with a...

Embodiment 3

[0113] The difference between this example and Example 1 is that in step (1) when preparing the DNA nanostructure of the modified upper capture strand, the molar ratio of the scaffold strand, the staple strand and the capture strand is set to 1:7:7 , after the self-assembly is completed, use a centrifugal purification column (100kDa molecular weight cut-off) at 7000 rpm for 2.5min to concentrate, and collect the trapped liquid in the purification column into a new EP tube. In step (2), take 50 μL of 50 μM ribonuclease A (RNase A) in a 1.5 mL EP tube, add 1 μL of 25 mM Sulfo-SMCC solution, shake and mix, and then add 220 μL of 1×PBS Buffer solution, vortexed to mix, fast centrifuged for 15 seconds, sealed with a parafilm, and placed on a shaker for 4.5 hours. Control the molar ratio of RNase A to Sulfo-SMCC to be 1:10. After the reaction was completed, excess Sulfo-SMCC was removed by centrifugation at 2750 rpm for 2.5 min with a G25 adsorption column. The purified Sulfo-SMCC...

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Abstract

The invention provides a nucleic acid protein nanocomposite as well as a preparation method and application thereof. Functional proteins and a coupling reagent are coupled with each other and are thenconnected with a nucleic acid chain, a nucleic acid nanostructure is connected with a capture chain, and the nucleic acid chain is connected with the capture chain by means of complementary base pairing; the nucleic acid protein nanocomposite provided by the invention realizes the precise arrangement of the functional proteins on the nucleic acid nanostructure, and solves the problem that proteindrugs can not easily enter cells as biomacromolecules; the nucleic acid protein nanocomposite can rapidly deliver the loaded protein drugs into tumor cells and achieve the cytotoxic effect so as to achieve the aims of killing cancer cells and inhibiting the vitality of the cancer cells. In addition, the preparation method of the nucleic acid protein nanocomposite is simple and easy to implement,and can precisely control the positions of the functional proteins on the nucleic acid nanostructure; furthermore, the nucleic acid protein nanocomposite has a good anti-tumor effect, and is expectedto be applied to the study of the administration of various biological functional macromolecules and the early diagnosis of diseases.

Description

technical field [0001] The invention belongs to the field of nanomedicine, and relates to a nucleic acid protein nanocomposite and its preparation method and application. Background technique [0002] Proteins play important functions in the living body, such as transport proteins to transport goods, cell surface protein receptors to receive and transmit signals, proteases to catalyze a series of biochemical reactions, etc. When the expression of these proteins with specific functions is abnormal, corresponding diseases will occur, so protein therapy emerges as the times require. Protein therapy refers to the delivery of some functional proteins that exist naturally in the body or protein analogs that have a therapeutic effect on diseases to target sites and perform specific functions. Compared with traditional small molecule chemotherapy drugs, protein therapy provides a new idea for tumor treatment. Studies have shown that ribonuclease A can inhibit the proliferation of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/54A61K38/46A61P35/00
CPCA61K38/465
Inventor 丁宝全段方圆刘建兵蒋乔赵帅
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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