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Fluorescence and/or radionuclide containing nanometer kit for detecting specific activated protease of serum and preparation method and application of kit

A technology of radionuclide and active protease, applied in the field of serum-specific active protease detection nano-kit and its preparation field, can solve the problems of difficulty in evading detection and destruction of the human immune system, sensitivity influence, difficulty in achieving effects, etc., and achieves the operation procedure. Simple and easy to use, extend the validity period, reduce the use of the effect

Active Publication Date: 2014-09-03
朱高红
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the study found that although such nanoparticles increased Fe 3 o 4 Protective layer and siloxane protective layer, but this kind of nanoparticles still cannot withstand the biological corrosion of tissues in the human physiological environment, and it is difficult to escape the detection and destruction of the human immune system.
This will affect the sensitivity of the detection of relevant specific proteases in the application of this nanoparticle; especially when used for introduction and combined with magnetic heat therapy, it is difficult to achieve the desired effect

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] A. Fe / Fe 3 o 4 Preparation of magnetic nanoparticles: Dissolve oleylamine in 1-dodecene with a volume ratio of 50~70 times, heat to 160~230°C, add 10~20ml iron pentacarbonyl under the protection of argon, and keep at 160~230°C Stir at the temperature for 0.5~1.5h, cool naturally to room temperature, the iron-ferric oxide (Fe / Fe 3 o 4 ) After the nanoparticles settle, the supernatant is poured out, and the settled iron-ferric oxide (Fe / Fe 3 o 4 ) Nanoparticles were washed 4-6 times with absolute ethanol, after drying, dispersed in 500ml chloroform solution, added 100g dopamine, stirred for 20 hours, collected nanoparticles by high-speed centrifugation, fully washed with chloroform, and dried spare.

[0138] B. Synthesis of UpA protease-specifically cleaved peptide chain (SGRSA): Dissolve 3 mmol of serine amino acid and 2.9 mmol of HBTU in 18 ml of diisopropylethylamine (DIEA)-dimethyl with a volume ratio of 1:20~25 In the mixed solution of formamide (DMD), add this...

Embodiment 2

[0150] A. Fe / Fe 3 o 4Preparation of magnetic nanorods: Dissolve 1 mmol ferric chloride and 3 mmol cetyltrimethylammonium bromide (CTAB) in a mixed solution of 2 ml deionized water, 6 ml n-octane, and 1 ml butanol. With magnetic stirring, slowly add 5 ml of 28% ammonia solution dropwise. The formed nanorods were collected by high-speed centrifugation, thoroughly washed with water, dried, dispersed in 15 ml of absolute ethanol, added with 0.4 g of sodium borohydride, and reduced to obtain nanorods of zero-valent iron. Disperse the obtained nanorods into 20ml of toluene, add 1ml of (3-aminopropyl) triethoxysilane, reflux for 24 hours, collect the nanorods by high-speed centrifugation, wash them with ethanol and dry them for later use. (see attached Figure 7 )

[0151] B. Synthesis of MMP-1 protease-specifically cleaved peptide chain (VPMS-MRGG): 3 mmol of glycylic acid and 2.9 mmol of HBTU were dissolved in 10 ml of diisopropylethylamine with a volume ratio of 1:20~25 ( DIE...

Embodiment 3

[0156] A. Fe / Fe 3 o 4 Preparation of / Au magnetic nanoparticles: Dissolve oleylamine in 1-dodecene with a volume ratio of 50~70 times, heat to 160~230°C, add 10ml iron pentacarbonyl under the protection of argon, and keep at 160~230°C Stir at the temperature for 0.5~1.5h, cool naturally to room temperature, the iron-ferric oxide (Fe / Fe 3 o 4 ) After the nanoparticles settle, the supernatant is poured out, and the settled iron-ferric oxide (Fe / Fe 3 o 4 ) nanoparticles were washed 4 to 6 times with absolute ethanol, and after drying, the obtained nanoparticles were dispersed into 50ml 2% oleylamine / chloroform solution, then 0.3g chloroauric acid was added, and after stirring for 48 hours, the The gold-coated magnetic nanoparticles were collected, washed thoroughly with ethanol, and dried for later use. (see attached Figure 8 )

[0157] B. Synthesis of MMP-2 protease specifically cleaved peptide chain IPVS-LRSGC: Dissolve 3 mmol of glycine and 2.9 mmol of HBTU in 10 ml of...

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Abstract

The invention discloses a fluorescence and / or radionuclide containing nanometer kit for detecting specific activated protease of serum and a preparation method and application of the kit. The kit is prepared from the following components: magnetic nanoparticles, protease-specific cut peptide chain, and fluorescent molecular and / or radionuclide; the preparation method comprises the steps of preparing the nanoparticles, synthesizing the peptide chain, connecting the fluorescent molecular and / or radionuclide and constructing a nanometer platform. The fluorescence and / or radionuclide containing nanometer kit for detecting specific activated protease of serum can be applied to detection of tumor-specific protease of solid tumors such as breast cancer, non-small cell lung cancer, thyroid cancer, liver cancer, ovarian cancer, endometrial cancer, pancreatic cancer, prostate cancer, glioma and melanoma; all components of the nanometer platform are non-toxic and high in biological applicability, protease-specific cut peptide chain has high-sensitive responsiveness to tumor-specific protease and the detection method is rapid, accurate and specific.

Description

technical field [0001] The invention belongs to the interdisciplinary technical field of materials, biology and analytical chemistry, and specifically relates to a serum-specific active protease detection nano kit containing fluorescence and / or radionuclides, a preparation method and application thereof. Background technique [0002] The reason why magnetic nanoparticles have broad application prospects is that they have many unique effects different from conventional materials, such as quantum size effect, surface effect, small size effect and macroscopic quantum tunneling effect, etc. These effects make magnetic nanoparticles have different Optical, electrical, acoustic, thermal, magnetic, and sensitive properties of conventional materials. [0003] When the particle size of magnetic nanoparticles is smaller than its superparamagnetic critical size, the particles enter a superparamagnetic state without coercivity and remanence. As we all know, for bulk magnetic materials ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/574
CPCG01N33/54326G01N33/54346G01N33/54393G01N33/573G01N33/57407G01N33/60
Inventor 朱高红周朴臣王红旺史蒂芬博斯曼
Owner 朱高红
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