A protein carrier for cell transfection

A protein and carrier technology, applied in the field of protein carrier for cell transfection, can solve the problems of high toxicity and limited transfection efficiency, and achieve the effects of enhanced stability, excellent targeting function, good biocompatibility and degradability

Active Publication Date: 2021-01-05
GUANGZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Several protein carriers that have been reported so far, such as polystyrene microspheres, cationic liposome reagents, polyarginine, polyacetylimide, etc., have the problems of high toxicity or limited transfection efficiency. Therefore, it is necessary to construct A protein carrier with both safety, low toxicity and high-efficiency transfection properties

Method used

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  • A protein carrier for cell transfection
  • A protein carrier for cell transfection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The preparation of embodiment 1 chitosan and poly(2-methacryloyloxyethyl phosphorylcholine) graft (Chi-PMPC-1):

[0025] (1) Preparation of N-phthaloyl-chitosan ((NP-Chi))

[0026] Chitosan (1.0g, 6.20mmol) and phthalic anhydride (2.76g, 18.60mmol) were dispersed in 30ml of DMF containing 5% (v / v), stirred and heated to 120°C under an argon atmosphere . After reacting for 8 hours, it was cooled to room temperature, poured into ice water, and the precipitate was obtained by suction filtration, washed thoroughly with ice methanol, and dried to obtain NP-Chi.

[0027] (2) Preparation of N-phthaloyl-chitosan ATRP macroinitiator (NP-Chi-Br)

[0028] Dissolve N-phthaloyl-chitosan (5.0g) in 30mL DMSO, add 2-bromoisobutyryl bromide (1.0mL) and triethylamine (1.1mL), and stir the reaction at room temperature for 3d After dialysis, filtration, washing and drying, a yellow solid product was obtained, namely NP-Chi-Br.

[0029] (3) Preparation of chitosan ATRP macroinitiator (C...

Embodiment 2

[0033] The preparation of embodiment 2 chitosan and poly(2-methacryloyloxyethyl phosphorylcholine) graft (Chi-PMPC-2)

[0034] (1) Preparation of N-phthaloyl-chitosan (NP-Chi)

[0035] Chitosan (1.0g, 6.20mmol) and phthalic anhydride (2.76g, 18.60mmol) were dispersed in 30ml of DMF containing 5% (v / v), stirred and heated to 120°C under an argon atmosphere . After reacting for 8 hours, it was cooled to room temperature, poured into ice water, and the precipitate was obtained by suction filtration, washed thoroughly with ice methanol, and dried to obtain NP-Chi.

[0036] (2) Preparation of N-phthaloyl-chitosan ATRP macroinitiator (NP-Chi-Br)

[0037] Dissolve N-phthaloyl-chitosan (5.0g) in 30mL DMSO, add 2-bromoisobutyryl bromide (1.0mL) and triethylamine (1.1mL), and stir the reaction at room temperature for 3d After dialysis, filtration, washing and drying, a yellow solid product was obtained, namely NP-Chi-Br.

[0038] (3) Preparation of chitosan ATRP macroinitiator (Chi-...

Embodiment 3

[0042] The preparation of embodiment 3 chitosan and poly(2-methacryloyloxyethyl phosphorylcholine) graft (Chi-PMPC-3):

[0043] (1) Preparation of N-phthaloyl-chitosan (NP-Chi)

[0044] Chitosan (1.0g, 6.20mmol) and phthalic anhydride (2.76g, 18.60mmol) were dispersed in 30ml of DMF containing 5% (v / v), stirred and heated to 120°C under an argon atmosphere . After reacting for 8 hours, it was cooled to room temperature, poured into ice water, and the precipitate was obtained by suction filtration, washed thoroughly with ice methanol, and dried to obtain NP-Chi.

[0045] (2) Preparation of N-phthaloyl-chitosan ATRP macroinitiator (NP-Chi-Br)

[0046] Dissolve N-phthaloyl-chitosan (5.0g) in 30mL DMSO, add 2-bromoisobutyryl bromide (1.0mL) and triethylamine (1.1mL), and stir the reaction at room temperature for 3d After dialysis, filtration, washing and drying, a yellow solid product was obtained, namely NP-Chi-Br.

[0047] (3) Preparation of chitosan ATRP macroinitiator (Chi...

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Abstract

The invention discloses a preparation method of a protein carrier for cell transfection. The preparation method comprises the following steps: reacting phthalic anhydride with chitosan to protect an amino group, then reacting the obtained reaction product with 2-bromoisobutyryl bromide to obtain an ATRP macromolecular initiator, removing a phthalimido group to obtain a Chi-Br macromolecular initiator, and finally carrying out an ATRP controlled polymerization reaction to obtain a chitosan and poly(2-methacryloyloxyethylphosphorylcholine) graft (Chi-PMPC) which can be used as the protein transfection carrier. The host material chitosan used in the present invention is a natural polysaccharide substance, and the grafted poly(2-methacryloyloxyethylphosphorylcholine) also has the imitated characteristic of the lipid layer structure of an erythrocyte outer surface cell membrane, so the prepared chitosan and poly(2-methacryloyloxyethylphosphorylcholine) graft has a good biocompatibility anda good material stability.

Description

technical field [0001] The invention relates to the field of biological materials, in particular to a protein carrier for cell transfection. Background technique [0002] Protein transfection refers to the process of transferring protein into cells, and generally requires the help of a carrier to help the target protein transfect into cells. At present, there are many studies on gene carriers, but little attention has been paid to protein carriers. Several protein carriers that have been reported so far, such as polystyrene microspheres, cationic liposome reagents, polyarginine, polyacetylimide, etc., have the problems of high toxicity or limited transfection efficiency. Therefore, it is necessary to construct A protein carrier with safe, low toxicity and high-efficiency transfection properties. Contents of the invention [0003] In order to overcome the deficiencies and shortcomings of the prior art, the purpose of the present invention is to provide a method for prepar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F251/00C08F230/02C12N5/00
CPCC08F251/00C12N5/00C08F230/02
Inventor 余细勇缪迎羚
Owner GUANGZHOU MEDICAL UNIV
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