Protein entrapping method

A protein and solution technology, which is applied in the field of protein loading, can solve the problems of drug waste, low drug release, uninspected particle size, and drug loading, and achieve fast release speed, high drug release, and low drug loading. The effect of high encapsulation rate

Inactive Publication Date: 2016-06-15
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0003] After searching the documents of the prior art, it is found that the Chinese patent publication number is CN1686560A, and the publication date is October 26, 2005. The patent name is chitosan quaternary ammonium salt nanoparticles and its preparation method and use. This patent is a A kind of chitosan quaternary ammonium salt nanoparticle used as drug carrier, its shortcoming is that the release amount of medicine is low, causes the waste of medicine
Second, the particle size, drug loading and other parameters of chitosan quaternary ammonium salt loaded with pharmaceutical protein were not investigated.

Method used

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Examples

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Embodiment 1

[0030] This example involves the preparation and particle size determination of blank nanoparticles. By controlling the concentration, ratio, and drop rate of the reactants, the conditions for forming nanoparticles are determined, and the subsequent exploration steps for preparing drug-loaded nanoparticles are reduced. The specific operation is as follows:

[0031] Step 1, dissolving chitosan quaternary ammonium salt (HACC) in ultrapure water to prepare an aqueous solution of chitosan quaternary ammonium salt with a mass concentration of 1.5-2.3 mg / mL.

[0032] Step 2: Add sodium tripolyphosphate (TPP) aqueous solution with a mass concentration of 0.35-0.6 mg / mL dropwise into the above solution at a rate of 0.2-0.8 mL / min through a constant-flow pump, and stir for 20-40 min after dropping. The volume ratio of sodium tripolyphosphate (TPP) solution to chitosan quaternary ammonium salt solution was controlled in the range of 1:(1~2).

[0033] Step 3, using a high-speed centrifu...

Embodiment 2

[0042] This embodiment relates to the preparation and particle size determination of BSA-loaded nanoparticles, and the specific operations are as follows:

[0043] Step 1, bovine serum albumin (BSA) is dissolved in the chitosan quaternary ammonium salt (HACC) aqueous solution of 2.0mg / mL with different mass concentration 0.5,0.9,1.3mg / mL;

[0044] Step 2: Add 0.45mg / mL sodium tripolyphosphate (TPP) aqueous solution dropwise to the above solution at a rate of 0.2-0.8mL / min through a constant flow pump, and stir for 20-40min after dropping. The volume ratio of sodium tripolyphosphate (TPP) solution to chitosan quaternary ammonium salt solution was controlled in the range of 1:(1~2).

[0045] Step 3, using a high-speed centrifuge to centrifuge at 4500-5000 rpm for 12-18 minutes. Take the supernatant and filter it with a 0.22 μm membrane filter.

[0046] 1 mL of the prepared nanoparticles was directly absorbed, and the particle size was measured by a dynamic light scattering ins...

Embodiment 3

[0050] This embodiment relates to the preparation of loaded rHAm nanoparticles, the specific operations are as follows:

[0051] Step 1, dissolving recombinant amelogenin (rHAm) in 1.5-2.3 mg / mL chitosan quaternary ammonium salt (HACC) aqueous solution at a mass concentration of 0.5 mg / mL;

[0052] Step 2: Add 0.35-0.6 mg / mL sodium tripolyphosphate (TPP) aqueous solution dropwise to the above solution through a constant flow pump at a rate of 0.2-0.8 mL / min, and stir for 20-40 min after the dropping is completed. The volume ratio of sodium tripolyphosphate (TPP) solution to chitosan quaternary ammonium salt solution was controlled in the range of 1:(1~2).

[0053] Step 3, using a high-speed centrifuge to centrifuge at 4500-5000 rpm for 12-18 minutes. The supernatant was taken and filtered with a 0.22 μm filter membrane to obtain rHAm-loaded nanoparticles.

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Abstract

The invention belongs to the technical field of medicine, and discloses a protein loading method, specifically adding a sodium tripolyphosphate solution dropwise into a chitosan quaternary ammonium salt solution containing a protein to be loaded, and then centrifuging and filtering the obtained mixed solution That's it. The method of the invention does not need to use organic solvents, has mild conditions and simple operation, and is very suitable for encapsulating protein drugs. The present invention uses it as a carrier, respectively carrying bovine serum albumin and recombinant amelogenin. The particle size of the bovine serum albumin-loaded nanoparticles is between 205.48-278.68nm, and the final release amount can be as high as close to 100%. The final release of loaded recombinant amelogenin nanoparticles can reach up to 50%.

Description

technical field [0001] The invention relates to a method for preparing nanoparticles in the technical field of medicine, in particular to a protein loading method. Background technique [0002] Chitosan is the only alkaline polysaccharide that occurs naturally. It is low toxic, renewable, and has excellent biodegradability and biocompatibility. Its molecular chain is rich in hydroxyl and amino groups, which are easy to carry out chemical reactions. With the development of polymer materials and pharmacy, chitosan as a new type of preparation auxiliary material is being paid more and more attention by people. However, chitosan is insoluble in water and general organic solvents, but soluble in dilute inorganic acids and some organic acids. The defect that chitosan can only be dissolved in acidic solution limits its application. Because the acidic medium is easy to interfere with the performance of sensitive drugs, and it is easy to cause toxicity to organisms. The use of c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/02A61K47/36A61K38/38A61K38/17A61K9/14
CPCA61K47/02A61K9/143A61K9/146A61K38/1709A61K38/38A61K47/36
Inventor 郭圣荣黄赛燕沈园园
Owner SHANGHAI JIAO TONG UNIV
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