Method for cell surface functional coating modifying

A cell surface and functional technology, applied in the field of microorganisms, can solve the problems of disrupting cell membrane function, low experimental safety factor, and few favorable results, and achieve the effect of improving hydrophilicity, avoiding the possibility of pollution, and reducing pollution

Active Publication Date: 2018-04-13
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

With the development of advanced molecular biology techniques, the method of changing yeast metabolism through genetic engineering has been explored to express receptors or glycoproteins on the cell surface, but they have certain limitations and can only be obtained in natural biochemical pathways. May also disrupt cell membrane function
At the same time, some physical and chemical methods have also been used to improve the stability of cells, such as cell immobilization, medium optimization, ultraviolet mutagenesis, adding organic solvents, etc., but their experimental cycle is too long, there are few favorable results, and the experiment is safe. low coefficient

Method used

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  • Method for cell surface functional coating modifying
  • Method for cell surface functional coating modifying

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Effect test

Embodiment 1

[0027] In this embodiment, the method of modifying the prepared cell surface functional coating is as follows:

[0028] Select yeast as an example, collect the cultured Yarrowia lipolytica cells by centrifugation, put them into different concentrations of dopamine solutions with different pH, shake the reaction on a shaker, and make dopamine self-polymerize on the cell surface to form A tight protective shell.

[0029] Specific steps include:

[0030] 1. Prepare Tris-hydrochloric acid buffer solution with pH=6.0~8.04. Liquid A: 0.05mol / LNa 2 HPO 4 12H 2 O solution, weigh 17.907g of disodium hydrogen phosphate, add distilled water to 1000ml. Liquid B: 0.05mol / LKH 2 PO 4 Solution, weigh potassium dihydrogen phosphate, 6.8045g, add distilled water to 1000ml. Mix A and B in different proportions to obtain the required pH buffer, see Table 1.

[0031] Table 1:

[0032] pH

A solution (ml)

Liquid B (ml)

8.04

47.5

2.5

7.73

45

5

...

Embodiment 2

[0037] In this embodiment, the method of modifying the prepared cell surface functional coating is as follows:

[0038] Collect the cultured Yarrowia lipolytica cells by centrifugation, put them into the dopamine solution prepared with Tris-hydrochloride buffer solution, and shake the reaction on a shaker to make dopamine self-polymerize on the cell surface to form a tight protective shell .

[0039] Specifically include the following steps:

[0040] 1. Put Yarrowia lipolytica ATCC201249 from the preservation tube into the YPD test tube to activate for 6-24 hours, then transfer to the YPD shake flask, culture at 30°C, 225rpm to OD=0.8-1.0, and collect the bacteria by centrifugation at 3000rpm .

[0041] 2. Configure Thris-hydrochloric acid buffer solution with pH=8.04, 95% of 0.05mol / L Na 2 HPO 4 12H 2 O and 5% of 0.05mol / L KH 2 PO 4 mix. To configure a 3.0g / L dopamine solution, weigh 0.15g of dopamine hydrochloride and put it in the ultra-clean bench for ultraviolet r...

Embodiment 3

[0046]In this embodiment, the method of modifying the prepared cell surface functional coating is as follows:

[0047] Collect the cultured Bacillus subtilis by centrifugation, put them into the dopamine solution prepared with Tris-hydrochloric acid buffer solution, shake the reaction on a shaker, and make the dopamine self-polymerize on the cell surface to form a tight protective shell .

[0048] Specifically include the following steps:

[0049] 1. Put Bacillus subtilis CGMCC NO: 9269 into the LB test tube from the seed preservation tube to activate for 6-24 hours, then transfer to the LB shaker flask, cultivate at 37°C and 225rpm until OD=0.8-1.0, and collect by centrifugation at 4000rpm bacteria.

[0050] 2. Configure Thris-hydrochloric acid buffer solution with pH=7.5, 95% of 0.05mol / L Na 2 HPO 4 12H 2 O and 5% of 0.05mol / L KH 2 PO 4 mix. To configure a 2.0g / L dopamine solution, weigh 0.1g of dopamine hydrochloride and put it in the ultra-clean bench for ultraviol...

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Abstract

The invention discloses a method for cell surface functional coating modifying. The method comprises the following steps: putting cells into a dopamine solution prepared by a Tris-hydrochloric acid buffer solution; performing oscillating reaction for 20 minutes to 8 hours; enabling dopamine to generate oxidative polymerization on a cell membrane surface; enabling a dopamine quinone compound and dopamine to generate disproportionated cross-linking reaction; forming a composite layer on the cell surface; and modifying the cell surface functional coating for enabling the cell to keep cell activity in an unfriendly environment. The method is gentle in condition and is simple to operate; and moreover, the whole process is completed in an aqueous solution, so that possibility of generating pollution on environment by the solvent is avoided; and functional coating modification, on the whole cell surface, by dopamine is achieved through one step.

Description

technical field [0001] The invention relates to a method for modifying a cell surface functional coating, which belongs to the technical field of microorganisms. Background technique [0002] With bioelectronic devices, biosensors, cell-based sensors, and theory all about the underlying cell, there is a need for cells that are capable of long-term viability and functionality in this environment. With the development of advanced molecular biology techniques, the method of changing yeast metabolism through genetic engineering has been explored to express receptors or glycoproteins on the cell surface, but they have certain limitations and can only be obtained in natural biochemical pathways. May also disrupt cell membrane function. At the same time, some physical and chemical methods have also been used to improve the stability of cells, such as cell immobilization, medium optimization, ultraviolet mutagenesis, adding organic solvents, etc., but their experimental cycle is to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/16C12R1/125C12R1/19
CPCC12N1/16C12N1/20
Inventor 姜岷王玉珍方艳信丰学章文明
Owner NANJING UNIV OF TECH
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