Basic protein entrapment vector, preparation method of entrapment vector and entrapment method

A protein and alkaline technology, which is applied in biochemical equipment and methods, peptide/protein components, pharmaceutical formulations, etc., can solve the problems of high-efficiency entrapment of basic proteins, difficulty in achieving a large amount of entrapment, etc., and achieve easy entrapment or modification, low equipment cost, good research and application prospects

Inactive Publication Date: 2016-08-17
WENZHOU INST OF BIOMATERIALS & ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This is because in neutral aqueous solution, both calcium carbonate and basic protein are positively charged, and they have the same electrical property and mutually repel each other during co-precipitation or adsorption loading, making it difficult to achieve a large amount of loading.
[0007] Therefore, through searching, there are no reports on the use of calcium carbonate to efficiently entrap basic proteins under mild conditions at home and abroad.

Method used

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  • Basic protein entrapment vector, preparation method of entrapment vector and entrapment method

Examples

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Embodiment 1

[0032] This embodiment relates to a method for encapsulating chymotrypsin in calcium carbonate microspheres doped with sodium heparin.

[0033] (1) Mix 2ml of calcium chloride solution with a concentration of 0.5M and 1ml of sodium heparin with a concentration of 6mg / ml, stir for 1~5min, then quickly add 3ml of carbonic acid with a concentration of 0.33M at 900 rpm Sodium solution, stirred for 35-40s, let stand for 15min, centrifuged to collect the calcium carbonate precipitate containing heparin sodium, washed 3 times with water. The prepared calcium carbonate particles containing heparin sodium have a diameter of about 2-5 μm.

[0034] (2) Take an appropriate amount of the above-mentioned calcium carbonate particles containing heparin sodium and place them in a 1.5ml centrifuge tube, add 1ml of chymotrypsin solution (pH=7.2) with a concentration of 1mg / ml, incubate at 4°C for 30min, and centrifuge , lyophilized to obtain heparin sodium calcium carbonate particles loaded wit...

Embodiment 2

[0036] This embodiment relates to a method for encapsulating lysozyme in calcium carbonate microspheres doped with sodium heparin.

[0037](1) After mixing 2ml of calcium chloride solution with a concentration of 0.5M and 1ml of heparin sodium with a concentration of 6mg / ml, stir for 1~5min, then quickly add 3ml of sodium carbonate with a concentration of 0.33M at 1000 rpm solution, stirred for 35-40s, and stood still for 25 minutes, and the generated calcium carbonate precipitate containing heparin sodium was collected by centrifugation, and washed with water for 3 times. The prepared calcium carbonate microspheres doped with sodium heparin have a diameter of about 2-5 μm, see figure 1 .

[0038] (2) Take an appropriate amount of calcium carbonate microspheres doped with heparin sodium and put them in a 1.5ml centrifuge tube, add 1ml of 1mg / ml lysozyme solution (pH=7.2), incubate at 10°C for 60min, and centrifuge , lyophilized to obtain lysozyme-doped calcium carbonate micr...

Embodiment 3

[0040] This embodiment relates to a method for entrapping lysozyme in calcium carbonate microspheres doped with dextran sulfate.

[0041] (1) Mix 2ml of calcium chloride solution with a concentration of 0.5M and 1ml of dextran sulfate with a concentration of 6mg / ml, stir for 1~5min, and then quickly add 3ml with a concentration of 0.33M at 1200 rpm Sodium carbonate solution, stirred for 35~40s, let stand for 20min, centrifuged to collect the calcium carbonate precipitate doped with dextran sulfate, washed 3 times with water. The prepared calcium carbonate particles doped with dextran sulfate have a diameter of about 2-5 μm.

[0042] (2) Take an appropriate amount of calcium carbonate doped with dextran sulfate and place it in a 1.5ml centrifuge tube, add 1ml of lysozyme solution (pH=7.2) with a concentration of 1mg / ml, and incubate at 25°C for 90min. After centrifugation and freeze-drying, dextran sulfate-doped calcium carbonate microspheres loaded with lysozyme can be obtain...

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Abstract

The application discloses a basic protein entrapment carrier, a preparation method of the entrapment carrier and a entrapment method. At room temperature, the polyanion electrolyte and calcium salt are stirred and mixed, and carbonic acid is added rapidly during rapid stirring or ultrasonication. The salt co-precipitates to form microspheres, and then the polyanion electrolyte-doped calcium carbonate microspheres are obtained after washing, centrifuging and drying. Then mix a certain concentration of basic protein solution with calcium carbonate microspheres, incubate with low-temperature stirring or shaking for 15-90 minutes, centrifuge, remove the supernatant, freeze-dry to obtain calcium carbonate microspheres loaded with basic protein, and the particle size distribution Narrow, good dispersion, good biocompatibility. The advantage of this application is that it can carry basic protein with high concentration and high activity under mild conditions.

Description

technical field [0001] The invention belongs to the field of biological materials, and specifically refers to a basic protein loading carrier, a preparation method of the loading carrier and a loading method. Background technique [0002] Calcium carbonate is a new type of inorganic material with a large amount of use and a wide range of uses. It is widely used in many fields such as coatings, rubber, plastics, papermaking, ink, food, cosmetics and medicine. The practical application of calcium carbonate is closely related to its properties (such as morphology, particle size, Zeta potential, specific surface area, etc.). [0003] Biodegradable medical polymer materials are used for diagnosis, treatment and organ regeneration. In recent years, they have been widely used in the fields of drug controlled release carriers, and their research occupies a very important position in the fields of biotechnology, life science and medicine. [0004] With the development of layer-by-la...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61K38/48A61K38/47
CPCA61K38/4826A61K38/47C12Y302/01017C12Y304/21001
Inventor 昝兴杰史鹏忠
Owner WENZHOU INST OF BIOMATERIALS & ENG
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