Compositions and Methods for Improved Encapsulation of Functional Proteins in Polymeric Vesicles

A composition, comprising:

• a suspension of a polymersome-encapsulated functional protein, wherein the suspension is prepared by:
• thermally blending a quantity of a block copolymer with a quantity of a low molecular weight polyethylene glycol (PEG) for at least 30 minutes, wherein the thermal blending is performed at 90-100° C.;
• mixing and cooling a resulting PEG/polymer formulation to room temperature;
• adding an aliquot of a solution of the functional protein to a sample containing the PEG/polymer formulation, wherein a ratio of the added aliquot to the PEG/polymer formulation is between around 0.5:1 and around 1.5:1 by volume;
• and
• performing at least three dilution steps such that polymersomes that are generated are progressively saturated with the functional protein, wherein each dilution step comprises:
• adding to the sample an additional amount of the solution of the functional protein;
• mixing a resulting dispersion of the functional protein in the PEG/polymer formulation;
• and
• sonicating the resulting dispersion for at least 30 minutes.

A method of treating a subject, the method comprising:

• preparing a suspension of a polymersome-encapsulated functional protein, wherein preparing the suspension comprises:
• thermally blending a quantity of a block copolymer with a quantity of a low molecular weight polyethylene glycol (PEG) for at least 30 minutes, wherein the thermal blending is performed at 90-100° C.;
• mixing and cooling a resulting PEG/polymer formulation to room temperature;
• adding an aliquot of a solution of the functional protein to a sample containing the PEG/polymer formulation, wherein a ratio of the added aliquot to the PEG/polymer formulation is between around 0.5:1 and around 1.5:1 by volume;
• and
• performing at least three dilution steps such that polymersomes that are generated are progressively saturated with the functional protein, wherein each dilution step comprises:
• adding to the sample an additional amount of the solution of the functional protein;
• mixing a resulting dispersion of the functional protein in the PEG/polymer formulation;
• and
• sonicating the resulting dispersion for at least 30 minutes;
• and
• administering to the subject a suspension of a polymersome-encapsulated functional protein, wherein the suspension is prepared by:

A method of preparing a suspension of a polymersome-encapsulated functional protein, comprising:

• thermally blending a quantity of a block copolymer with a quantity of a low molecular weight polyethylene glycol (PEG);
• adding an aliquot of a solution of the functional protein to a sample containing the PEG/polymer formulation;
• and
• performing at least one dilution step such that polymersomes that are generated are progressively saturated with the functional protein.