Application of cell penetrating peptide Tat-mediated growth factor in transdermal transfer
A growth factor-mediated technology, applied in the field of transdermal drug delivery systems and cosmetics, can solve the problems of difficult-to-explain macromolecular substances, and achieve the effect of increasing the transdermal rate and quantity
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Embodiment 1
[0021] Expression and purification of HIV-1-containing transactivator protein Tat protein transduction domain (Tat-protein transductiondomain, Tat-PTD 49-57 ) (Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg) and human epidermal growth factor fusion protein Tat-EGF. The gene fragment encoding Tat-EGF fusion protein was obtained by PCR method, and a 6×His tag was added at the 3' end. After the amplified product was treated with restriction endonuclease BamH I and Nde I, it was connected to the Escherichia coli expression plasmid pET-3c (Novagen, co. , co.Ltd., USA), ampicillin (final concentration 100ug / ml) screened positive clones. The correct recombinant plasmid identified by sequencing was extracted and transformed into Escherichia coli BL21(DE3) (Novagen, co. Ltd., USA), and the transformants were selected by ampicillin resistance (final concentration 100ug / ml). At 37°C, after being induced by IPTG (final concentration: 1mmol / L) for 4 hours, the fusion protein was harvested and separ...
Embodiment 2
[0023] Dissolve 0.1 g of hyaluronic acid in about 80 ml of double-distilled water, then add sodium heparin (1 / 4 of the amount of Tat-EGF fusion protein), 4000.2 g of PEG, stir well and then sterilize under high pressure (0.1Mpa, 121°C for 15 minutes ), cooled to room temperature. The Tat-EGF fusion protein obtained in Example 1 with a purity of more than 98% was sterilized by filtration through a 0.22 μm microporous membrane, and the solution was added to the above mixture, the pH was adjusted to 6.8, and sterilized double-distilled water was added to 100 ml.
Embodiment 3
[0025] Take 0.1g of water-soluble chitosan and dissolve it in an appropriate amount of glacial acetic acid (about 1ml), add about 50ml of double-distilled water, adjust the pH to 6, then add 0.1g of EDTA-2Na, 0.2g of PEG 400, dissolve and then add paraben 0.01g each of the ester and ethylparaben (dissolved by heating with an appropriate amount of water first), stir well, then autoclave (0.1Mpa, 121°C for 15 minutes), and cool to room temperature. Add the Tat-EGF fusion protein with a purity of more than 98% obtained in Example 1, and sterilize it through a 0.22 μm microporous membrane filter into the above mixture, adjust the pH to 6.8, and add sterilized double-distilled water to 100 ml. Dispensed in a spray bottle.
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