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Bacterial nano magnetic corpuscle-glycosphingolipid compound and preparation method and application of compound

A glycosphingolipid and nanomagnetic technology, applied in the field of separation, can solve the problems of inability to obtain protein, incomplete glycosphingolipid, and inability to clarify the mechanism of action and binding site, so as to promote utilization and high drug targeting , high efficiency effect

Inactive Publication Date: 2014-12-17
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, people’s understanding of glycosphingolipids is not complete at present. When exploring the function and mechanism of glycosphingolipids, it is often impossible to elucidate its mechanism of action and binding sites because the proteins that interact with it cannot be obtained.

Method used

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  • Bacterial nano magnetic corpuscle-glycosphingolipid compound and preparation method and application of compound
  • Bacterial nano magnetic corpuscle-glycosphingolipid compound and preparation method and application of compound
  • Bacterial nano magnetic corpuscle-glycosphingolipid compound and preparation method and application of compound

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1 BMPs-GM 3 Preparation of complex

[0030] BMPs treatment: Weigh 1 mg of BMPs and heat in 100 μL of 1% SDS solution at 100°C for 5 minutes to remove BMPs surface proteins; collect BMPs with a magnetic stand, and wash 3 times with 100 mM phosphate (PB, pH=7.5) buffer.

[0031] BMPs-GM 3 Preparation of complexes: Resuspend 1 mg of treated BMPs in 1 ml of 2 mg / mL GM 3 (monosialotrihexosyl ganglioside) solution, 100W ultrasound for 60min (6s ultrasound, 4s interval); after incubation in the dark for 2h, wash BMPs with 100mM phosphate (PB, pH=7.5) buffer three times to obtain BMPs -GM 3 Complex.

[0032] Blocking and storage: Add the obtained complex into blocking solution (10 mM PBS containing 1% BSA, and containing 2% PEG4000, 2% PEG6000), and store at 4°C for later use.

[0033] The preparation method of the SDS solution: weigh 1 g of SDS and dissolve it in 100 mL of water. Described buffer preparation method: get 170mmol Na 2 HPO 4 ,30mmol KH 2 PO 4 Solu...

Embodiment 2

[0034] Example 2 BMPs-GM 1 Preparation of complex

[0035] (1) BMPs treatment: 20 mg of BMPs was weighed and shaken with 1 ml of 20% SDS at room temperature for 2 hours to remove BMPs surface proteins. The BMPs were collected and washed three times with deionized water to obtain the processed BMPs.

[0036] (2) BMPs-GM 1 Preparation of complexes: Resuspend 10mg of BMPs processed in 1ml 20mg / mL GM 1 (Monosialotetrahexosyl ganglioglyceride) solution, 80W ultrasonic for 10min, incubate in the dark for 3h, wash with deionized water to obtain BMPs-GM 1 Glycosphingolipid complexes.

Embodiment 3

[0037] Example 3 BMPs-GM 3 Preparation of glycosphingolipid complexes

[0038] (1) Preparation of raw material BMPs: refer to literature Xiang L, Wei J, Jianbo S et al. Purified and sterilized magnetosomes from Magnetospirillum gryphiswaldense MSR-1 were not toxic to mouse fibroblasts in vitro[J]. Letters in Applied Microbiology, 2007, 45(1):75-81 to prepare.

[0039] (2) BMPs treatment: weigh 10 mg of BMPs, and treat with 1 ml of 0.01% CTAB at 60° C. for 20 minutes to remove BMPs surface proteins. The BMPs were collected and washed three times with deionized water to obtain the processed BMPs.

[0040] (3) BMPs-GM 3 Preparation of complexes: resuspend 10mg of processed BMPs in 10ml 0.1mg / mL GM 3 In the solution, 150W ultrasonic for 15min (ultrasonic 5s, interval 5s), after static reaction for 2h, wash with deionized water, and then get BMP s -GM 3 Glycosphingolipid complexes.

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Abstract

The invention discloses a bacterial nano magnetic corpuscle-glycosphingolipid compound and a preparation method and an application of the compound, and belongs to the fields of separation and medicines. The BMPs-glycosphingolipid compound disclosed by the invention is prepared by the following steps: firstly, removing surface proteins of BMPs by using a surfactant; suspending the BMPs into a glycosphingolipid solution; and carrying out ultrasonic or room-temperature reaction, and incubating to couple the BMPs to the glycosphingolipid, thereby obtaining the bacterial nano magnetic corpuscle-glycosphingolipid compound. Irrelevant protein is effectively prevented from being introduced by removal of the surface proteins, and the protein acting with the glycosphingolipid can be obtained. The compound disclosed by the invention can be used for promoting utilization of the glycosphingolipid by cells, meanwhile, magnetic particles in the compound can play a targeting effect, and the compound is guided to be gathered towards the focus part, so that the drug targeting property is relatively high. After the compound is introduced into the tumor region, magnetic targeting thermal therapy can also be introduced, thereby further treating the focus.

Description

technical field [0001] The invention relates to a preparation method of a bacterial nano magnetosome-glycosphingolipid complex, which belongs to the field of separation and the field of medicine. Background technique [0002] Glycosphingolipids (glycosphingolipids) is a very important class of glycolipids (glycolipids) molecules, it is formed by a ceramide skeleton (composed of sphingosine and fatty chains) connected with one or more sugar chains, in which sphingosine The ceramide formed by the acylation of the amino group by the fatty acid constitutes the hydrophobic part, which is embedded in the lipid bilayer of the cell membrane, while the hydrophilic part formed by the connection of the sugar chain and the primary hydroxyl group of sphingosine extends out of the plasma membrane. Glycosphingolipids are arranged in clusters on the cell membrane, and play an important role in immune response, cell development, cell adhesion and recognition, cell proliferation and different...

Claims

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Application Information

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IPC IPC(8): A61P35/00C07K1/14A61K47/46C07K14/71A61K41/00C12N5/09
Inventor 关锋李想杨刚龙郭佳
Owner JIANGNAN UNIV
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