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Polymerization Initiated Self-Assembly Method of Protein Conjugates into Nanospheres
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A technology of polymerization initiation and nano-microspheres, which is applied in the direction of nanotechnology, microsphere preparation, nanotechnology, etc., to achieve good application prospects, controllable process and good repeatability
Active Publication Date: 2022-03-22
NANJING UNIV OF SCI & TECH
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Currently, few lipase-polymer conjugates synthesized by PISA have been reported in the literature
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Embodiment 1
[0033] (1) Dissolve lipase with 10mL phosphate buffer solution, make 0.018mmol protein solution, then stir evenly at 25°C on a magnetic stirrer;
[0034] (2) Dissolve N-succinimidyl 2-bromo-2-methylpropionate in 1ml dimethyl sulfoxide solution to form a 0.055mmol organic solution, then stir on a magnetic stirrer at 25°C Uniform;
[0035] (3) Take out the organic solution described in step (2), slowly add it dropwise to the protein solution obtained in step (1), then stir evenly on a magnetic stirrer at 4°C, and react for one day;
[0036] (4) The macroinitiator that step (3) is obtained is dissolved in the aqueous solution, adds water-soluble monomer (HEAA, NIPAM) 50wt% and carries out degassing 15min processing is reaction bottle A, gets another reaction bottle B to pack Deionized water (1ml), catalyst (CuBr) and ligand were uniformly mixed on a stirrer for degassing treatment, and the reaction bottle A was slowly introduced into the reaction bottle B and reacted at room tem...
Embodiment 2
[0043] This example is basically the same as Example 1, the only difference is that a hydrophobic polymer (poly(TBA)) is grafted from the lipase surface to form an amphiphilic protein-polymer conjugate and self-assemble into nanospheres.
[0044] (1) Dissolve lipase with 10mL phosphate buffer solution to make a protein solution of 0.018mmoll, then stir evenly at 25°C on a magnetic stirrer;
[0045] (2) Dissolve N-succinimidyl 2-bromo-2-methylpropionate in 1ml dimethyl sulfoxide solution to form a 0.055mmol organic solution, then stir on a magnetic stirrer at 25°C uniform;
[0046] (3) Take out the organic solution described in step (2), slowly add it dropwise to the protein solution obtained in step (1), then stir evenly on a magnetic stirrer at 4°C, and react for one day;
[0047] (4) Dissolve the macromolecular initiator obtained in step (3) in an aqueous solution, add 40 mg of hydrophobic monomer (TBA) and carry out degassing for 15 minutes to be treated as reaction bottle...
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Abstract
The invention discloses a method for self-assembly of protein conjugates into nano microspheres by polymerization initiation. In this method, the lipase CALB is used as a carrier, modified into a macromolecular initiator by ammonolysis reaction, and the controllable free radical polymerization LRP regulated by Cu(0) is used in aqueous solution and water / methanol mixed solution, respectively, for hydrophilicity and Hydrophobic monomers were polymerized and their activity explored. Under mild action conditions, the polymerization-initiated conjugate self-assembles into nano-microspheres, and improves the catalytic activity of proteins, showing broad application prospects in the field of protein immobilization.
Description
technical field [0001] The invention belongs to the technical field of biomacromolecules, and relates to a method for initiating self-assembly of protein conjugates into nanometer microspheres by polymerization. Background technique [0002] In recent years, polymerization-induced self-assembly (PISA), as a facile method for synthesizing polymeric nanomaterials, has broad application prospects in the fields of catalysis and biotechnology. Compared with the solvent and pH value used in the traditional self-assembly method, PISA has no cumbersome reaction and purification process, but has the characteristics of simplicity, high repetition rate, and high solid content. Studies have shown that in controlled radical polymerization (CRP) , widely used reversible addition-fragmentation chain transfer polymerization (RAFT) to synthesize amphiphilic block polymers with specific morphologies. However, due to the limitations of atom transfer radical polymerization (ATRP) in aqueous ph...
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