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Targeted nanobubble for detecting small cell lung cancer and preparing method and application thereof

A technology of small cell lung cancer and microbubbles, applied in preparations for in vivo experiments, pharmaceutical formulas, echo/ultrasonic imaging agents, etc.

Inactive Publication Date: 2016-07-06
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no research on the construction of nano-microbubbles targeted by

Method used

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  • Targeted nanobubble for detecting small cell lung cancer and preparing method and application thereof
  • Targeted nanobubble for detecting small cell lung cancer and preparing method and application thereof
  • Targeted nanobubble for detecting small cell lung cancer and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0023] Example 1 Preparation of targeted nanobubbles for detection of small cell lung cancer

[0024] 1) Using the membrane hydration method, 36 mg of dipalmitin lecithin (DPPC), 7 mg of distearoylphosphatidylethanolamine (DSPE), and 2 mg of dipalmitic acid (DPPA) were completely dissolved in 8 ml of chloroform. Let the chloroform volatilize naturally in a fume hood to form a phospholipid film. Add 8 mL of hydration solution (PBS:glycerol=9:1 (volume ratio)) to the above-mentioned film-forming petri dish, and hydrate on a shaker for 60 minutes at room temperature. Mix the washed phospholipid membrane with the hydration solution and transfer it to a 50mL centrifuge tube to form liposomes (such as figure 1 Shown).

[0025] 2) Dilute 10μg of anti-progastrin releasing peptide monoclonal antibody with 10mM PBS (Nacl8g / L, Kcl0.2g / L, Na2HPO41.44g / L) to 50μL, add 1μL of 0.5M EDTA solution; use 60mg Mercaptoethylamine was dissolved in 500μL PBS, 10μL of 0.5M EDTA solution was added, the a...

Example Embodiment

[0027] Example 2 Identification of antibody links in targeted nano-microvesicles

[0028] Mix 100ul each of ordinary nano-microbubbles and targeted nano-microbubbles with Dylight488-labeled goat anti-mouse IgG4ul. Under a fluorescence microscope (1000×), it can be seen that the surface of targeted nano-microbubbles emits bright green fluorescence, while the surface of ordinary nano-microbubbles No obvious fluorescence (such as Figure 4 Shown).

Example Embodiment

[0029] Example 3 Combination experiment of targeted nano-microbubbles and small cell lung cancer (H446 cells) cells

[0030] 1.5×10 per hole 4 Cells were inoculated in a 6-well plate covered with a cover glass, cultured overnight in an incubator, fixed with 4% paraformaldehyde at room temperature, and the climbing piece was added to 30ul of targeted nano-microbubbles and ordinary blank nano-microbubbles, 37℃ React in the incubator for 1 hour, observe the binding of the targeted nano-microbubbles and cells under a microscope at 1000 times, and calculate the adhesion rate. Targeting nano-microbubbles adhere tightly around H446 cells and are arranged regularly along the cell membrane. The average cell adhesion rate is (90.2±3.24)%; while ordinary blank nano-microbubbles do not bind to H446 cells (such as Figure 5 Shown).

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Abstract

The invention provides a targeted nanobubble for detecting small cell lung cancer and a preparing method and application thereof.The targeted nanobubble comprises a lipid dimolecular shell and inert gases inside the lipid dimolecular shell, wherein an anti-progastrin releasing peptide monoclonal antibody aiming at the small cell lung cancer is connected to the lipid dimolecular shell.The average grain diameter of the nanobubbles is 300-400 nm.The preparing method comprises the steps that dipalmitoyl phosphatidylcholine, distearoyl phosphatidyl ethanolamine and dipalmitoyl phosphate are completely dissolved in chloroform to generate a lipidosome with the filming-rehydration method, the anti-progastrin releasing peptide monoclonal antibody aiming at the small cell lung cancer is opened with mercaptoethylamine for incubation with the lipidosome, so that a stable thioether bond is formed, then perfluoropropane gas is introduced, and the targeted nanobubble containing the anti-progastrin releasing peptide monoclonal antibody aiming at the small cell lung cancer is obtained.The targeted nanobubble can be applied to color doppler ultrasonic detection of small cell lung cancer.

Description

technical field [0001] The invention relates to a targeted nano-microbubble, in particular to a preparation method and application of a targeted nano-microbubble for detecting small cell lung cancer. Background technique [0002] Small cell lung cancer (small cell lung cancer, SCLC) accounts for about 20% of lung cancer, and the two-year survival rate is less than 5%. It has the highest degree of malignancy in lung cancer, and its cause is unknown. It is a neuroendocrine tumor that can produce a variety of ectopic hormones. Routine examination The diagnosis of SCLC by X-ray, CT and MRI lacks specificity, and SCLC is often at an advanced stage when it is diagnosed (KalemkerianGP, eta1. Smallcelllungcancer. JNatlComprCancNetw2013, 11(1):78-98.). [0003] In 1981, McDonald et al. found that SCLC cell lines can secrete gastrin-releasing peptide (GRP) and contain GRP receptors (McDonaldTJ, eta1. , but its half-life in the blood is only two minutes, progastrin-releasing peptide (...

Claims

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Application Information

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IPC IPC(8): A61K49/22
CPCA61K49/221A61K49/223
Inventor 王伟王金萍许静梁爱华
Owner SHANXI UNIV
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