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Magnetic nano-grade material with cell-targeting property, and biomedical application thereof

A technology of magnetic nanoparticles and derivatives, applied in the direction of medical preparations containing active ingredients, analytical materials, radioactive physical shapes, etc., can solve the problems of not giving full play to the advantages of magnetotactic bacteria

Active Publication Date: 2013-07-24
INST OF GEOLOGY & GEOPHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, biomimetic synthesis in the past still relied too much on severe chemical conditions unfavorable to protein denaturation (temperature up to 90°C or adding polymeric gel to control particle size), and did not exert the high efficiency of magnetotactic bacteria at room temperature and pressure. The advantages of controlling particle size, shape and crystallinity, so how to improve the biomimetic synthesis process and realize the biomimetic synthesis of magnetotactic bacterial membrane proteins is an urgent scientific problem [Arakaki et al., 2003; Arakaki et al., 2007; Prozorov et al .,2007;Arakaki et al.,2010]

Method used

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  • Magnetic nano-grade material with cell-targeting property, and biomedical application thereof
  • Magnetic nano-grade material with cell-targeting property, and biomedical application thereof
  • Magnetic nano-grade material with cell-targeting property, and biomedical application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0144] Biomimetic synthesis of human H subunit magnetic ferritin with intact protein shell conformation

[0145] Using recombinant human ferritin as a template, the full-length cDNA of the H subunit of human ferritin was cloned and constructed on the pET11b plasmid (purchased from Novagen); Plasmids were transformed or co-transformed into bacteria BL21(DE3)pLysS (purchased from Novagen), and IPTG (isopropyl-β-D-thiogalactopyranoside) was added to activate the T7 promoter to induce expression; ultrasound was performed after expression Break up and release the protein; separate and purify the protein; use the purified human H subunit ferritin as a template to combine ferrous salt and oxidant H 2 o 2 Add the solution of recombinant human ferritin to react, control the pH value to 8.5, control the temperature to 65°C, and form strong magnetic nanoparticles inside the recombinant human ferritin; the concentration of ferrous salt is the number of ferrous atoms added each time The ...

Embodiment 2

[0149] Synthesis of magnetic nanomaterials with ferritin-shell-wrapped ferromanganese oxide cores

[0150] Using purified human H subunit ferritin as a template, ferrous salt, manganese salt (the ratio of iron to manganese is 11.5, equivalent to the incorporation of 8% manganese) and oxidant H 2 o 2 Add the solution of recombinant human ferritin for reaction, control the pH value to 8.5, control the temperature at 65°C, and form strong magnetic nanoparticles inside the recombinant human ferritin; finally, the number of iron atoms added to each protein molecule is 4600, manganese The number of atoms is 400; after the reaction is completed, the magnetic particles with complete protein structure are obtained after separation by exclusion chromatography, centrifugation and molecular sieve purification. figure 2 a is an electron micrograph of the ferromanganese oxide nucleus, it can be seen that the formed nucleus is approximately spherical and exhibits good monodispersity; fig...

Embodiment 3

[0152] Biomimetic synthesis of nanomagnetic materials using prokaryotically expressed magnetotactic bacterial membrane protein Mms6

[0153] The whole genome of the magnetotactic bacterium AMB-1 was extracted, the mms6 gene was amplified by PCR, and cloned into the pET15b plasmid (purchased from Novagen). The EcoR I and BamH I restriction sites were used to transform the bacterium BL21(DE3)pLysS ( purchased from Novagen) for prokaryotic expression. The Mms6 protein with His tag was purified by nickel column affinity chromatography. Mix His-Mms6 protein with iron salt solution, adjust the pH to 7-9 with NaOH, react at room temperature for 24 hours to generate hydrated iron oxide compound (ferrihydrite), and then add ferrous salt solution (so that ferrous ions and iron The ratio of ions is 1:2), and continue to react for 24 hours until the color of the solution turns black completely. The obtained magnetic particles are collected by a magnet, washed three times with deoxygenat...

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Abstract

The invention discloses a magnetic nano-grade material with a cell-targeting property, and a biomedical application thereof. The magnetic nano-grade material can be specifically bound with highly expressed transferrin receptors on tissue cell surfaces, and can enter the cells. The material can be bound with various high-transferring-receptor-expression tissue cells with broad spectrum and specificity, such that high-efficiency cell targeting property can be realized on animal models. The material can be used as a magnetic resonance contrast agent and a fluorescent molecular probe in disease diagnosis, and can be used as a medicine carrier in disease treatment.

Description

technical field [0001] The invention belongs to the cross fields of bionic synthesis, nanotechnology, molecular imaging and biomedicine. Specifically, the present invention relates to a magnetic nanomaterial with cell targeting and its biomedical application. Background technique [0002] Magnetic resonance imaging (MRI) has the unique advantages of non-invasiveness and high resolution, so it is a very good imaging tool for early diagnosis of diseases. However, its sensitivity is low, it cannot distinguish between diseased tissue and normal tissue well, and the detection has no specificity. These are some inherent shortcomings of MRI [Terreno et al., 2010]. In order to overcome this shortcoming of low sensitivity, researchers try to synthesize various MRI contrast agents (contrast agents, also known as contrast agents) to shorten the longitudinal relaxation time (longitudinal relaxation time, T 1 ) or transverse relaxation time (transverse relaxation time, T 2 ) to enhanc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/18A61K49/14A61K47/42A61K51/12A61K51/08A61K45/00A61K38/19A61P35/00A61P29/00G01N33/574
CPCA61K49/0041A61K49/0056C07K14/47A61K49/1866A61K49/06A61K49/14A61K47/6923A61P29/00A61P35/00A61K49/1824
Inventor 潘永信曹长乾田兰香蔡垚朱日祥
Owner INST OF GEOLOGY & GEOPHYSICS CHINESE ACAD OF SCI
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