Protein initiator as well as preparation method and application thereof
A technology of initiator and protein, applied in the direction of coating, etc.
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[0137] The preparation method of the protein initiator of one embodiment, comprises the following steps:
[0138] S1. Dissolve a protein containing a disulfide bond in the molecule and a reducing agent in a buffer solution to obtain a first solution.
[0139] After mixing the protein containing disulfide bonds in the molecule with the reducing agent, the reducing agent can open the disulfide bonds in the protein structure, form free thiol groups, and transform the protein from an α-helix structure to a β-sheet structure. "The performance of self-assembly is beneficial to the subsequent modification of the material surface.
[0140] In one embodiment, the protein containing disulfide bonds in the molecule is selected from at least one of lysozyme, albumin, soybean protein and whey protein.
[0141] In one embodiment, in the first solution, the concentration of the protein containing disulfide bonds in the molecule is 0.1 mg / mL to 100 mg / mL, preferably 0.2 mg / mL to 30 mg / mL, mo...
Embodiment 1
[0251] The preparation method of dibromomaleimide derivative is as follows:
[0252]
[0253] Weigh 2.98g (10mmol) of hydroxyethyldibromomaleimide and dissolve it in 20mL of dichloromethane, add 2.51g (11mmol) of bromoisobutyryl bromide and 1.21g (11mmol) of triethylamine dropwise to it, at room temperature The reaction was carried out for 20 hours. The reaction solution was successively washed with saturated NaHCO 3 washed with deionized water, and the organic phase was dried with anhydrous magnesium sulfate; ethyl acetate and n-hexane were used as eluents, and 3.8 g of a pale yellow solid product was obtained after purification by column chromatography with a yield of 84%. The product obtained in Example 1 is subjected to the measurement of nuclear magnetic resonance spectrum to obtain figure 1 H NMR spectrum: 4.31ppm (2H,-CH 2), 3.92ppm (2H,-CH 2 ),1.87ppm(6H,-CH 3 ).
[0254] The preparation method of the protein initiator (bovine serum albumin (BSA) macromolecula...
Embodiment 2
[0257] The present embodiment provides a BSA-POEGA conjugate and a preparation method thereof:
[0258] (1) Weigh 20mg BSA macromolecular initiator and 480mg monomer OEGA (polyethylene glycol methyl ether acrylate) into the reaction bottle, add 5mL ultrapure water to dissolve it completely, and replace the reaction bottle with nitrogen for 15min of oxygen to obtain a mixed solution.
[0259] (2) Tris(2-dimethylaminoethyl)amine (5 μL, 0.019 mmol) and 1 mL of water were added to the round-bottomed flask, and nitrogen gas was introduced for 5 min to remove the oxygen in the flask; CuBr (2.7 mg, 0.019 mmol) and the flask was placed in an ice-water bath, and the reaction was stirred for 30 min to obtain a solution containing the complexed product.
[0260] (3) The mixed solution obtained in step (1) was transferred to the flask in step (2) through a syringe, and the reaction was continued for 3 hours under ice bath conditions; after the reaction, the reaction solution was dialyzed...
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