Bioengineering protein plaster as well as preparation method and application thereof
A bioengineering and protein technology, applied in pharmaceutical formulations, drug combinations, antitumor drugs, etc., can solve the problems of increasing the incidence of complications, inconvenient daily operations, etc., achieving strong adhesion performance, simple and fast treatment methods, and reducing The effect of tumor recurrence
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[0040] In the present invention, the preparation method of the smearable bioengineered protein plaster provided by the present invention includes:
[0041] A) adding positively charged elastin into deionized water and mixing evenly to obtain a homogeneous protein solution;
[0042] B) adding the negatively charged anionic surfactant to deionized water, and mixing uniformly to obtain a homogeneous surfactant solution;
[0043] C) adding the gold nanorod solution to the protein solution, shaking and mixing evenly, then adding the surfactant solution, shaking and centrifuging to obtain a bioengineered protein plaster.
[0044] In the above preparation method provided by the present invention, the types of elastin-like and alkyl-based surfactants are the same as those mentioned above, and will not be repeated here.
[0045] Further, the concentration of the elastin-like solution is preferably 100-300 mg / mL, specifically 100 mg / mL, 200 mg / mL or 300 mg / mL.
[0046] The concentrati...
Embodiment 1
[0064] Example 1: Expression and purification of K144cys
[0065] Step 1: Obtain a strain stably expressing K144cys. The pET25b prokaryotic expression vector was constructed with the gene sequence of K144cys protein, and then transformed into E.coli BLR (DE3) E. coli, induced expression and screened to obtain a stable expression strain.
[0066] Step 2: Expression of K144cys. The strains were cultured in LB medium and then inoculated into TB medium. After reaching the exponential growth phase, IPTG was added to induce protein expression at 28.5 °C for 12 h, and the cells were washed and collected by centrifugation, and stored at -80 °C.
[0067] Step 3: Purification of K144cys. After the bacteria were crushed by high pressure, the supernatant was collected by centrifugation, filtered and sterilized, and finally purified by a nickel affinity chromatography column, a cation exchange chromatography column, and a desalting column with a protein purifier to finally obtain the K14...
Embodiment 2
[0068] Example 2: Synthesis of Gold Nanorods
[0069] The first step: prepare a gold nanorod growth solution, as follows: Weigh 7g CTAB and 1.234g sodium oleate, add them to a 1L glass flask, add 250mL deionized water, and heat to 50°C in an oil bath to stir and dissolve . After it was completely dissolved, the temperature of the oil bath was adjusted to 30°C for stabilization. Prepare 24mL AgNO 3 An aqueous solution (concentration of 4 mM) was added to the above glass flask and allowed to stand for 15 min. Prepare 250mL HAuCl 4 The aqueous solution (concentration of 1 mM) in a volumetric flask was added to the above glass flask and stirred for 1 h 30 min.
[0070] The second step: prepare the gold seed solution, as follows: prepare 5 mL of CTAB aqueous solution (concentration of 0.2 mM) in a 100 mL glass flask, and place it in a 30° C. oil bath to keep warm. Prepare 5mL of HAuCl 4 An aqueous solution (0.5 mM concentration) was added to the above 100 mL glass flask and s...
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