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Microfluid preparation method for tumor therapeutic vaccine nanocarriers

A technology of nanoparticles and carrier materials, which is applied in the direction of antineoplastic drugs, chemical instruments and methods, and medical preparations of non-active ingredients. It can solve the problems of insufficient uniformity of nanoparticles, poor controllability and repeatability of experiments, etc. Simple, reproducible, and biologically effective results

Inactive Publication Date: 2016-10-26
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The existing method to prepare PEI and its derivatives and negatively charged protein nanoparticles is a large-volume mixing method by vortex, but this method will be affected by many conditions, different experimenters, different operating methods, etc., so that The controllability and repeatability of the experiment are poor, and the prepared nanoparticles are not uniform enough

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  • Microfluid preparation method for tumor therapeutic vaccine nanocarriers
  • Microfluid preparation method for tumor therapeutic vaccine nanocarriers
  • Microfluid preparation method for tumor therapeutic vaccine nanocarriers

Examples

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preparation example Construction

[0060] Preparation method of nanoparticles loaded with negatively charged proteins

[0061] The preparation method of the nanoparticle loaded with negatively charged protein of the present invention comprises the steps of:

[0062] (1) dissolving the negatively charged protein to obtain a protein solution;

[0063] (2) dissolving the carrier material to obtain a carrier material solution;

[0064] (3) The protein solution and the carrier material solution are pumped into the Tesla structure chip from different channels, and mixed slightly to obtain nanoparticles loaded with negatively charged proteins.

[0065] The negatively charged protein may be an antigenic protein.

[0066] In one embodiment, the negatively charged protein is OVA. In another embodiment, the negatively charged protein is BSA. However, the negatively charged protein is not limited to specific OVA or BSA, as long as the protein is negatively charged. OVA, the full name Ovalbumin, refers to chicken ovalb...

Embodiment 1

[0099] Embodiment 1 adopts Tesla structure chip to prepare PEI-OVA nanoparticle

[0100] (1) First, dissolve the negatively charged protein OVA in a buffer solution of 1 mM HEPES, pH=7.4, and prepare a 10 mg / ml OVA solution;

[0101] (2) dissolving branched PEI with a molecular weight of 25KDa in deionized water to obtain a PEI solution of 0.125 mg / ml to 3.0 mg / ml;

[0102] (3) The OVA solution obtained in step (1) is divided into two 1ml syringes, the PEI solution obtained in step (2) is also added to the 1ml syringe, the air bubbles in the syringe are removed, and the pathway is connected. The OVA solution and the PEI solution were simultaneously pumped into the Tesla structure chip for micro-mixing (perfusion method such as figure 1 As shown), the composite solution of PEI and OVA is obtained, the mass ratio range is 0.01~0.24, wherein the width of the path of the Tesla structure chip is 200 μm, the height is 110 μm, the perfusion volume of the PEI solution channel is 100 ...

Embodiment 2

[0109] Example 2 Study on the Biological Effects of PEI-OVA Nanoparticles Prepared by Tesla Structure Chip

[0110] PEI-OVA nanoparticles were prepared by referring to the method of Example 1, and the mass ratio of PEI to OVA ranged from 0.01 to 0.24. Dilute the prepared PEI-OVA nanoparticle solutions with different mass ratios to an OVA concentration of 0.5 mg / ml, take 30 μl of the PEI-OVA nanoparticle dilution and add it to a 96-well plate with 120 μl of dendritic cell solution, and the cell density is 8×10 5 / ml, co-incubated overnight in a 37°C cell culture incubator, then added 120 μl of B3Z cell solution into the well, and incubated together for 24 hours, and finally detected the concentration of IL-2 secreted by B3Z cells with an ELISA kit to investigate PEI-OVA nano Antigen presentation effect of particles.

[0111] Experimental group: 1640+FBS group, refers to the addition of 1640 and FBS to the medium of dendritic cells.

[0112] Conclusion: Results Figure 4 As ...

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Abstract

The invention belongs to the field of researches of microfluid and microreactor technologies in pharmaceutical preparations and particularly relates to a microfluid preparation method for tumor therapeutic vaccine nanocarriers. According to the microfluid preparation method, Tesla structural chips are adopted for preparation of electronegative protein carrying nanoparticles for the first time. The microfluid preparation method is simple in process, prepared composite particles are small in particle size, uniform in dispersion and high in repeatability, a mass ratio range of nanoparticles formed by combination of polyethyleneimine and derivatives thereof and antigen proteins can be widened, and accordingly application prospect of the nanoparticles to tumor vaccines is broadened.

Description

technical field [0001] The invention belongs to the research field of microfluid and microreactor technology in pharmaceutical preparations, and in particular relates to a microfluidic preparation method of a tumor therapeutic vaccine nanocarrier. Background technique [0002] At present, our common cationic polymers are chitosan, protamine, polylysine, polyhistidine, polyamine dendrites, polypropyleneimine dendrites, polyethyleneimine (PEI) and so on. Among them, polyethyleneimine (PEI) has become a research hotspot in the carrier field since the 20th century. [0003] Polyethyleneimine (PEI) is widely used in nucleic acid carriers because of its positive charge, which can bind nucleic acid to form nanoparticles, protect nucleic acid from degradation, and its "proton sponge effect" can realize endosome escape of nucleic acid. PEI and negatively charged proteins are combined to form nanoparticles through electrostatic interaction. Our previous research has confirmed that t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/34A61K47/42A61K39/00A61P35/00B01L3/00B01J19/00
CPCA61K39/0011B01J19/0093A61K9/5146A61K9/5169B01L3/5027B01L3/502715B01L2200/10B01L2200/0647
Inventor 邱瑶陈剑李真珍奚春明陈龙陆海燕李方
Owner SHANGHAI JIAO TONG UNIV
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