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Preparation of a plasmid delivery vector

A plasmid and vector technology used in the field of biomedicine

Active Publication Date: 2019-02-26
HUAIYIN INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no reports of three kinds of nanoparticles combined with their respective advantages and disadvantages and used as plasmid DNA molecular delivery carriers.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Embodiment 1: Preparation of sepiolite nanomaterials

[0012] Soak 10g of sepiolite in 100mL of distilled water for 24 hours. After soaking loosely, add 0.1g of sodium pyrophosphate, stir with a mixer for 2 hours, and let it settle. After a stable precipitate layer appears, add 0.5mol / L HCl to adjust the pH of the system to 6.0, stir thoroughly for 20 minutes, pour out the above suspension, filter it with a vacuum pump, wash it with distilled water several times, and then put the filter cake into Dry it in an oven, grind it, and keep it sealed for later use after autoclaving.

Embodiment 2

[0013] Embodiment 2: the preparation of montmorillonite nanomaterial

[0014] Add 6.0 g of montmorillonite to 100 mL of distilled water, and stir until completely dissolved using a magnetic stirrer. The rotating speed is 5000r / min, centrifuged for 2h. Transfer the precipitate to an evaporating dish, dry it in vacuum at 65°C for 6-10 hours, grind it to 200 mesh, dry it in vacuum at 65°C for 2-4 hours, sterilize it under high pressure and keep it sealed for later use.

Embodiment 3

[0015] Embodiment 3: the preparation of diatomaceous earth

[0016] Weigh 10g of diatomite, suspend it with 50mL of distilled water and gently pour it into a Falcone tube filled with 450mL of distilled water, oscillate ultrasonically for 10min, and recover the suspension after standing for 5min, then centrifuge at 12000rpm for 30s, discard Serum. The particles obtained by centrifugation are suspended with an equal volume of distilled water, filtered with a vacuum pump, washed with distilled water several times, and then the filter cake is dried in an oven and ground. Store at 4°C after autoclaving.

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PUM

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Abstract

The invention discloses a plasmid DNA molecule transmission carrier. The transmission carrier contains an active component which comprises meerschaum-montmorillonite-diatomite mixed nanometer particles. The plasmid DNA molecule transmission carrier has a small particle size, good adsorption capability, good plasmid compounding capability, low cytotoxicity and an effectively improved conversion rate, is a safe and efficient plasmid transmission carrier and has a good application prospect in a plasmid transmission conversion kit.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for preparing a transfer carrier capable of adsorbing plasmids on the surface. Background technique [0002] A plasmid is a small double-stranded DNA circular molecule naturally occurring in bacteria and yeast that replicates as an independent unit. Although it only occupies a small part of the total DNA of the host cell, it carries important genes. Because of the small size of plasmid DNA and the resistance gene, it is widely used in recombinant DNA technology. [0003] The study of plasmid DNA transport vectors has been puzzling scientists from all over the world and has become a key issue for the success of gene therapy. People have been working hard to find suitable carrier materials that can effectively deliver plasmid DNA to target cells and maintain its efficacy for a certain period of time. With the rapid development of nanobiotechnology, as a non-viral gene carrier...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K47/02A61K47/46A61P35/00
Inventor 孔晶柳晓平
Owner HUAIYIN INSTITUTE OF TECHNOLOGY