Polycarboxyl glycine betaine gene transfection preparation as well as preparation method and application thereof

A polycarboxybetaine and gene transfection technology, which can be applied to other methods of inserting foreign genetic materials, gene therapy, anti-infective drugs, etc. The effect of serum stability

Inactive Publication Date: 2016-01-13
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In this patent, the polycarboxybetaine component is a cationic liposome modified by polycarboxybetaine neutral lipids, which must contain cationic

Method used

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  • Polycarboxyl glycine betaine gene transfection preparation as well as preparation method and application thereof
  • Polycarboxyl glycine betaine gene transfection preparation as well as preparation method and application thereof
  • Polycarboxyl glycine betaine gene transfection preparation as well as preparation method and application thereof

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Embodiment 1

[0045] The preparation of embodiment 1 polycarboxybetaine gene transfection preparation

[0046] 1) Preparation of compound buffer: weigh sodium acetate (16.406g, 0.2mol) and dissolve it in 1L water to obtain 0.2mol / L sodium acetate mother liquor, weigh acetic acid (18.015g, 0.3mol) and dissolve it in 1L water to obtain 0.3mol / L The acetic acid mother liquor of L, gets the sodium acetate mother liquor of 49mL and the acetic acid mother liquor of 51mL and mixes, and obtains the acetic acid-sodium acetate composite buffer solution that pH is 4.6;

[0047] 2) Prepare the blend solution of polycarboxybetaine polymer and complex buffer: weigh 1mgCBD-PCB 20 Dissolve in 0.5mL of acetic acid-sodium acetate complex buffer solution, the dissolution time is 0.5h, and the temperature is 20°C;

[0048] 3) Prepare a compound solution formed by compounding the blend solution and the nucleic acid to be transfected: take 0.04 mL of the blend solution obtained in step (2), dissolve 10.00 μg ...

Embodiment 2

[0050] The preparation of embodiment 2 polycarboxybetaine gene transfection preparation

[0051] 1) Preparation of composite buffer: Weigh disodium hydrogen phosphate (28.392g, 0.2mol) and dissolve it in 1L water to obtain 0.2mol / L disodium hydrogen phosphate mother liquor, weigh acetic acid (35.994g, 0.3mol) and dissolve it in 1L water To obtain 0.3mol / L sodium dihydrogen phosphate mother liquor, get 8mL of disodium hydrogen phosphate mother liquor and 92mL of sodium dihydrogen phosphate mother liquor and mix to obtain a disodium hydrogen phosphate-sodium dihydrogen phosphate composite buffer solution with a pH of 5.8;

[0052] 2) Prepare the blend solution of polycarboxybetaine polymer and complex buffer: weigh 1mgCBD-PCB 20 Dissolve in 1.0mL of acetic acid-sodium acetate complex buffer solution for 1 hour at a temperature of 37°C;

[0053] 3) Prepare the compound solution formed by compounding the blend solution and the nucleic acid to be transfected: take 0.08 mL of the...

Embodiment 3

[0055] The preparation of embodiment 3 polycarboxybetaine gene transfection preparation

[0056] 1) Preparation of compound buffer solution: Weigh potassium hydrogen phthalate (40.844g, 0.2mol) and dissolve it in 1L of water to obtain 0.2mol / L disodium hydrogen phosphate potassium hydrogen phthalate mother liquor, take 5mL of phthalic acid Potassium hydrogen and 2.022mL of 0.2mol / L hydrochloric acid were mixed, and water was added to make the volume to 20mL to obtain a potassium hydrogen phthalate-hydrochloric acid composite buffer solution with a pH of 3.0;

[0057] 2) Prepare a blend solution of polycarboxybetaine polymer and complex buffer: weigh 1 mg of cholesterol-PCB 20 Dissolve in 1.0mL acetic acid-sodium acetate complex buffer solution, the dissolution time is 3h, and the temperature is 25°C;

[0058] 3) Prepare a compound solution formed by compounding the blend solution and the nucleic acid to be transfected: take 0.08 mL of the blend solution obtained in step (2)...

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Abstract

The invention provides a polycarboxyl glycine betaine gene transfection preparation as well as a preparation method and an application thereof. The polycarboxyl glycine betaine gene transfection preparation comprises a gene transfection reagent based on polycarboxyl glycine betaine molecules, and nucleic acid to be subjected to transfection, wherein the gene transfection reagent based on polycarboxyl glycine betaine molecules comprises polycarboxyl glycine betaine polymer, a compound buffer solution and a gene transfection working solution. The gene transfection preparation has the excellent non-specific protein adsorption resisting property, high gene transfection efficiency, as well as pH responsibility, and significant blood stability, the gene transfection efficiency of nucleic acid is obviously improved, and the polycarboxyl glycine betaine gene transfection preparation can be applied to various research fields including gene therapy, gene function, biological pharmacy, drug screening and the like.

Description

technical field [0001] This field belongs to the field of gene transfection reagents, and relates to a polycarboxybetaine gene transfection preparation and its preparation method and application. Background technique [0002] Gene transfection is the transfer of nucleic acid molecules with biological functions to the site of action in cells, and the nucleic acid maintains its biological functions in cells. Since free nucleic acid molecules are easily degraded by ribozymes in tissues or cells, tissue targeting is poor, and nucleic acid molecules themselves have negative charges, resulting in poor cell penetration. The key to the application of nucleic acid molecules is to develop gene transfection reagents with selectivity, safety and high efficiency. At present, vectors commonly used in research and clinical applications are roughly divided into two categories: viral vectors and non-viral vectors. Viral vectors have attracted much attention because of their high efficiency...

Claims

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Application Information

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IPC IPC(8): C12N15/87A61K48/00A61P35/00A61P31/00A61P9/00
Inventor 张欣李燕
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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