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Blood stability nanoparticle-containing solution, preparation method and miRNA marker detection method thereof

A nanoparticle and marker technology, applied in the field of nanomaterials and medical nanoprobes, can solve the problems of inability to meet the early screening of clinical cancer, unstable test results, and high processing technology requirements, and achieve batch and automatic detection, The system has excellent blood stability and simple clinical operation.

Pending Publication Date: 2020-07-03
深圳市鲲鹏未来科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Existing cancer early diagnosis technologies have shortcomings such as poor detection specificity, unstable test results, and high detection costs, making it difficult to popularize in early cancer screening
On the other hand, the traditional miRNA detection methods are mainly Northern hybridization, microarray analysis and fluorescent quantitative PCR. These detection methods are time-consuming, labor-intensive, complex in process, high in processing technology requirements, and high in cost, and cannot meet the requirements of early clinical cancer screening. Require

Method used

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  • Blood stability nanoparticle-containing solution, preparation method and miRNA marker detection method thereof
  • Blood stability nanoparticle-containing solution, preparation method and miRNA marker detection method thereof
  • Blood stability nanoparticle-containing solution, preparation method and miRNA marker detection method thereof

Examples

Experimental program
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Effect test

preparation Embodiment 1

[0042] Dissolve 10 mg of PLGA-PEG-COOH (U.S. Creative PEGworks Company, product number DLG-5k20k21) and 1 mg of DSPE-PEG-COOH (U.S. Avanti Polar lipids company, product number 880135) in 2 mL of chloroform and mix well; Add dropwise to 4% ethanol aqueous solution, and use a cell ultrasonic breaker to continue ultrasonication at a frequency of 20kHz and a power of 90W for 5 minutes while adding, to promote the self-assembly of the two block copolymers and promote the volatilization of the chloroform solution to obtain polymer micelles solution.

[0043] Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) (Merck sigma, Germany, product number E6383) and N-hydroxysuccinimide (NHS) (Germany) successively. Merck sigma company, Cat. No. 130672) (please add the amount of EDC and NHS), aminated complementary DNA at the 5' end of the corresponding cDNA (sequence 5'-GGGAGCTCATAGGCCGGCCCCTGGCTGCAGATG-3') of the custom-synthesized colorectal cancer marker miR-96-5p The ...

preparation Embodiment 2

[0045] Prepare the nanoparticle solution with reference to the preparation example 1 of the present invention, the difference is: the mass ratio of PLGA-PEG-COOH and DSPE-PEG-COOH is 5:1; ultrasonic treatment is performed with 60W power during the self-assembly process; coupled use Dialysis was performed using a dialysis bag with a molecular weight cut-off of 20 kD.

preparation Embodiment 3

[0047] Prepare the nanoparticle solution with reference to the preparation example 1 of the present invention, the difference is: the mass ratio of PLGA-PEG-COOH and DSPE-PEG-COOH is 20:1; ultrasonic treatment is performed with 130W power during the self-assembly process; coupled use Dialysis was performed using a dialysis bag with a molecular weight cut-off of 50 kD.

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Abstract

The invention provides a nanoparticle solution. The nanoparticle solution comprises polymer micelle nanoparticles with blood stability, in particular nanoparticles modified with functional molecules such as nucleotide sequences. The invention also provides a method of preparing the nanoparticle solution, and a method of detecting a micro RNA (miRNA) marker using the nanoparticle solution.

Description

technical field [0001] The present invention relates to the fields of nanomaterials and medical nanoprobes, in particular to a nanoparticle solution comprising polymer micellar nanoparticles with blood stability, especially nanoparticles modified with functional molecules such as nucleotide sequences. The present invention also relates to a method for preparing the nanoparticle solution, and a method for detecting microRNA (miRNA) markers using the nanoparticle solution. Background technique [0002] Due to their unique physical and chemical properties, nanomaterials have good application prospects in the medical field, especially in drug delivery, medical detection, biosensing, and medical material modification. [0003] In the biomedical diagnostic application of nanomaterials, nanoparticles are used to detect disease markers, and diseases are diagnosed by comparing the changes in fluorescence, color, and aggregation state of nanoparticles before and after detection. Amon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6883C12Q1/6834C08J3/02C08L87/00C08L67/04C08L71/02
CPCC12Q1/6886C12Q1/6883C12Q1/6834C08J3/02C12Q2600/178C08J2387/00C08J2367/04C08J2371/02C08J2471/02C12Q2563/155C12Q2563/107
Inventor 赵鹏飞林泉
Owner 深圳市鲲鹏未来科技有限公司
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