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Preparation method and application of functional erythrocyte membrane

An erythrocyte membrane and cell membrane technology, which is applied in the field of preparation of functional erythrocyte membranes, can solve the problem of no RGD peptides and technical reports on erythrocyte biocompatibility treatment, etc. Effect

Inactive Publication Date: 2019-02-12
武汉市卉研生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no technical report on the biocompatibility treatment of RGD peptide and red blood cells (RBC)

Method used

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  • Preparation method and application of functional erythrocyte membrane
  • Preparation method and application of functional erythrocyte membrane
  • Preparation method and application of functional erythrocyte membrane

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preparation example Construction

[0029] A kind of preparation method of functional erythrocyte membrane of the present invention: dissolve 0.6mg of erythrocyte membrane in deionized water 1ml or phosphate buffer, described phosphate buffer is NaH 2 PO 4 &Na 2 HPO 4 , pH 7.4, 100 mM, ultrasonically or stir the membrane solution and DSPE-PEG-RGD (phospholipid PEG polypeptide, active targeting polypeptide) to obtain a functionalized red blood cell membrane.

[0030] The erythrocyte membrane solution and DSPE-PEG-RGD are ultrasonically or stirred at a ratio of 1:10-10:1.

[0031] The temperature of ultrasonic and stirring is 15-45℃, the time is 10s-2h; the ultrasonic power is 10-500W; the stirring speed is 50r / min-1000r / min;

[0032] Application of functional erythrocyte membrane, the functional erythrocyte membrane is applied in nanomaterials; used as a carrier material to transport drugs or biomolecules for medical treatment and cosmetic applications; as a modified camouflage material wrapped on the surface ...

Embodiment 1

[0034] Add 1.5mg of EDTA (ethylenediaminetetraacetic acid) to every mL of fresh whole blood of male ICR mice, and centrifuge at 800g for 5 minutes at 4°C to remove plasma and buffy coat. The resulting erythrocyte membrane was washed with cold PBS (NaH 2 PO 4 and Na 2 HPO 4 , pH7.4, 100mM) centrifuged and washed 3 times. The resulting erythrocyte membrane was dissolved in (NaH 2 PO 4 and Na 2 HPO 4 , pH 7.4, 25mM), ice bath for 30 minutes. Centrifuge at 15000rpm for 5 minutes to remove hemoglobin, and further use PBS (NaH 2 PO 4 and Na 2 HPO 4 , pH 7.4, 100mM) and washed twice by centrifugation. The obtained erythrocyte membrane (0.6 mg) was dissolved in deionized water (1 ml) or phosphate buffer (NaH 2 PO 4 and Na 2 HPO 4 , pH 7.4, 100mM), the membrane solution and DSPE-PEG-RGD (0.06 mg, 0.6mg or 6mg) were sonicated at 37°C for 30 minutes at an ultrasonic power of 200W to obtain a functional erythrocyte membrane.

Embodiment 2

[0036] Functional erythrocyte membranes obtained from 1 ml of whole blood were mixed with gold nanoparticles (50 μg mL -1 ), at 200W with ultrasonic power, at 37°C, ultrasonic for 10 minutes, centrifuged at 10000rpm for 5 minutes to remove excess erythrocyte membrane, and obtain functional erythrocyte membrane-wrapped gold nanoparticles.

[0037] B16-F10 melanoma cells were subcutaneously grafted into 6-month-old BALB / c nude mice, and functionally encapsulated gold nanoparticles were injected into the tail vein at 5 mg / kg for 4 hours, and then irradiated with infrared laser for 10 minutes (1064nm, 1W / cm 2 ), photothermal therapy for melanoma.

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Abstract

The invention discloses a preparation method of a functional erythrocyte membrane. The preparation method is characterized in that 0.6 mg of erythrocyte membrane is dissolved into 1 ml of deionized water or phosphate buffer liquid, wherein the phosphate buffer liquid is NaH2PO4 and Na2HPO4, the pH (potential of Hydrogen) value is 7.4, and the concentration is 100 mM; and a membrane solution and DSPE-PEG-RGD are subjected to ultrasonic wave or stirring, and the functional erythrocyte membrane can be obtained. The preparation method is easy to operate, the final functional erythrocyte membrane can serve as a carrier material to transport drugs or biological molecules to conduct medical and cosmetology and serves as a decoration camouflage material to coat the surface of other materials to realize decoration and camouflage, accordingly, accumulation of nano materials or active biological molecules at specific parts is improved, and the treatment and application effects are enhanced.

Description

technical field [0001] The invention belongs to the field of biomedicine and cosmetics, more specifically it relates to a preparation method and application of a functional erythrocyte membrane. Background technique [0002] Red blood cells (RBC) are the most numerous blood cells in the blood, and they are naturally biocompatible, biodegradable, and have no biological immunity. Erythrocyte membranes are better able to protect the activity of the carried substances and have a longer and more controllable lifetime. These properties make "carrier erythrocytes" a valuable transport vehicle. [0003] In recent years, the surface modification technology of erythrocyte membrane has been greatly developed. These new erythrocyte membrane-camouflaged nanoparticles can not only be used as drug delivery carriers, but also can design a variety of nanomedicines, such as vaccines. [0004] RGD peptide (arginine-glycine-aspartic acid) is the preferred sequence for applications involving b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/46A61K47/18A61K41/00A61P35/00A61K8/99A61K8/64A61Q19/00
CPCA61K8/64A61K8/99A61K41/0052A61K47/183A61K47/46A61K2800/10A61Q19/00A61P35/00
Inventor 吴陈勇曾丽红张保东林国民
Owner 武汉市卉研生物科技有限公司
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