Fibroneclastin composition freeze-dried micro-core with skin barrier repairing effect and preparation method thereof
A skin barrier repair and elastin technology, which is applied in skin care preparations, medical preparations containing active ingredients, pharmaceutical formulas, etc., to achieve the effect of easy absorption and reduced activity loss
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[0022] The preparation method of the fibronectin involved in the present invention is as follows:
[0023] (1) Obtain the fibronectin (FN) gene through GenBank query, insert the FN gene sequence into pMD19-TSimple Vector, its 5' end is connected with the NotⅠ restriction site (GCGGCCGC), and the 3' end is connected with the XbaⅠ restriction site (TCTAGA), thus obtaining the plasmid pMD19-T-FN;
[0024] (2) Use endonuclease Not I and endonuclease Xba I to perform double enzyme digestion on plasmid pYES2 / CT-MFα and plasmid pMD19-T-FN, respectively. The enzyme digestion reaction system is:
[0025] Double Enzyme Digestion System Volume (μL) Quick Cut Not I 2.0 QuickCut Xba I 2.0 10xQuickCut Green Buffer 5 pYES2 / CT-MFa or FN fragment 15 wxya 2 o
26 Total Volume 50
[0026] Enzyme digestion reaction was carried out at 37°C for 3 hours in a metal bath, and the digestion products were detected by 2% agarose gel electrophoresis, ...
Embodiment 1
[0042] A fibronectin elastin composition freeze-dried microcore with skin barrier repairing effect, which is made of the following raw materials in weight percentage: 15ppm fibronectin, 15ppm platelet-derived growth factor, 15ppm metallothionein type IV, elastin Protein 15ppm, filaggrin 20ppm, mannitol 6%, glycine 6%, trehalose 1%, and the balance is water.
[0043] The preparation method of described freeze-dried microcore comprises the following steps:
[0044] (1) mixing each raw material of formula quantity to obtain mixed solution;
[0045] (2) Using a high-precision fluid dispensing system, the mixed solution is dropped into liquid nitrogen stored in a Dewar bottle drop by drop at a rate of 10 μl to freeze into microcores;
[0046] (3) Take out the microchips and put them into vials, put 10 frozen microchips into each vial, and freeze-dry them. The program control of the freeze-drying is shown in Table 1.
[0047] Table 1 freeze-drying program parameter table
[0048]...
Embodiment 2
[0051] A fibronectin elastin composition freeze-dried microcore with skin barrier repairing effect, which is prepared from the following raw materials in weight percentage: 10 ppm of fibronectin, 10 ppm of platelet-derived growth factor, 10 ppm of metallothionein type IV, elastin Protein 10ppm, filaggrin 40ppm, mannitol 6%, glycine 6%, trehalose 1%, and the balance is water.
[0052] The preparation method of the freeze-dried microcore is the same as in Example 1.
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