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Preparation method and application of tumor-targeted nano-artificial antibody

An artificial antibody and targeting technology, applied in the direction of nanotechnology, medical preparations of non-active ingredients, pharmaceutical formulas, etc., can solve the problems of large batch performance differences, immunogenicity, long screening cycle, etc., and achieve long-term use Lifespan, strong resistance to harsh environments, and the effect of shortening the screening time

Inactive Publication Date: 2019-11-22
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, natural antibodies are mainly obtained by animal immunization, which has the disadvantages of high cost, low preparation efficiency, long screening cycle, variability, difficulty in storage, and immunogenicity. Monoclonal antibodies also have the problem of large batch performance differences. In practical applications has great limitations

Method used

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  • Preparation method and application of tumor-targeted nano-artificial antibody
  • Preparation method and application of tumor-targeted nano-artificial antibody
  • Preparation method and application of tumor-targeted nano-artificial antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Preparation of nano-gold spheres and nano-gold rods

[0038] 1. Preparation of gold nanospheres

[0039] 0.0255g HAuCl 4 Dissolve in 260mL of water, heat to boil, then add 15g of 1% sodium citrate solution to the solution. After the color of the solution remains unchanged, continue to boil for 15min, centrifuge to collect the gold nanospheres, and store them for later use. The particle size and morphology of the synthesized gold nanoparticles were studied by transmission electron microscopy (TEM), such as figure 1 shown.

[0040] 2. Preparation of gold nanorods

[0041] 0.3553 g of CTAB, 0.25 mL of 0.1 M HAuCl 4 and 0.2277 mg of NaBH 4 Mix rapidly for 2 minutes, and the solution turns brown after the mixing is completed. This is the seed solution, which was left to stand at 28°C for 2 hours. Then sequentially add 15mL 0.1M HAuCl 4 , 5.096gAgNO 3, 0.55mL HCl and 42.2712mg ascorbic acid were added to 300mL solution containing 10.9535g CTAB, and then 1....

Embodiment 2

[0042] Example 2: Preparation of Nano-artificial Antibodies Targeting Epidermal Growth Factor Receptor (EGFR)

[0043] 1. Design and synthesis of nanoparticles

[0044] N-isopropylacrylamide (58-X mol%), charged functional monomer (3-acrylamidopropyl) trimethylammonium chloride (X mol%), N-tert-butylacrylamide (35mol %), the crosslinker N,N'-methylenebisacrylamide (7 mol%) and sodium dodecylsulfonate (10 mg) were dissolved in water to give a total monomer concentration of 130 mM. After adding the initiator, polymerization was carried out at 65° C. for 3 hours with a magnetic stirrer under a nitrogen atmosphere. The polymerized solution was purified by dialysis against an excess of pure water, and polymer nanoparticles were obtained after freeze-drying.

[0045] 2. Preliminary screening of artificial antibodies

[0046] The epidermal growth factor receptor (EGFR) was selected as the target, and the molecularly imprinted polymer nanoparticles were used as the biomimetic antib...

Embodiment 3

[0049] Example 3: Preparation of Nano-artificial Antibodies Targeting Epithelial Cell Adhesion Molecule (EpCAM)

[0050] Dissolve 228.6 mg of N-isopropylacrylamide, 23 mg of acrylamide, 82.6 mg of N-tert-butylacrylamide, 50 mg of N,N'-methylenebisacrylamide and sodium dodecylsulfonate SDS (30 mg) In 50mL of water, the total monomer concentration was 65mM, and 2mg of epithelial cell adhesion molecule (EpCAM) polypeptide was added at the same time, the mixed solution was filtered, nitrogen gas was blown for 30min, ammonium persulfate was added, and reacted at 65°C for 10h. The polypeptide template was then eluted with 1 M sodium chloride solution to obtain nanoartificial antibodies against epithelial cell adhesion molecule (EpCAM). Then the reaction mixture was dialyzed for 3 days, and then freeze-dried to obtain the nano-artificial antibody.

[0051] The scanning electron microscope image of the nano-artificial antibody obtained by mixing the above monomer solution and gold na...

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Abstract

The invention discloses a preparation method and an application of a tumor-targeted nano-artificial antibody. A plurality of functional monomers and a cross-linking agent are polymerized according todifferent components to form a gel nano particle artificial antibody, and the affinity and selectivity of the artificial antibody to a tumor marker are regulated and controlled by changing the ratio of the functional monomers. The specificity and selectivity of the nano artificial antibody to the tumor marker are improved by combining a molecular imprinting technology and taking tumor marker holoprotein or a stable polypeptide fragment as a template. After screening, the affinity constant KD value of the nano artificial antibody to the tumor marker reaches 10<-11> M, which is equivalent to that of an antibody. The obtained nano artificial antibody is combined with magnetic nano particles, a monolithic column or a micro-fluidic chip, so that selective recognition and capture of tumor markers, circulating tumor cells and exosomes can be realized; high-sensitivity detection of tumor markers, circulating tumor cells and exosomes is achieved through high-sensitivity Raman spectroscopy, fluorescence quantitative detection, ELISA kits, chemiluminescence kits and other methods.

Description

technical field [0001] The invention belongs to the technical field of biological nanomaterials, and in particular relates to a preparation method and application of a tumor-targeting nanoartificial antibody. Background technique [0002] Cancer is one of the diseases with the highest fatality rate in the world, and its early clinical manifestations are not very specific, so patients are often in the middle and late stages when they see a doctor, and early diagnosis and timely and effective treatment are closely related to the survival rate and quality of life of patients . Early cancer detection can detect precancerous lesions, early cancers and potential invasive cancers, improve the treatment effect of cancer patients, and finally achieve the purpose of prolonging the overall survival of cancer patients. [0003] Tumor markers are produced by tumor tissue or the host's response to tumors, which can reflect and monitor the occurrence and development of tumors, and judge t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F220/54C08F222/38C08F220/34C08F220/56C08F220/06A61K47/32B82Y40/00
CPCA61K47/32B82Y40/00C08F220/54C08F222/385C08F220/34C08F220/56C08F220/06
Inventor 吕永琴李燕李媛
Owner BEIJING UNIV OF CHEM TECH
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