Straightforward and Effective Protein Encapsulation in Polypeptide-based Artificial Cells

Abstract

A simple and straightforward approach to encapsulating an enzyme and preserving its function in polypeptide-based artificial cells is demonstrated. A model enzyme, glucose oxidase (GOx), was encapsulated by repeated stepwise adsorption of poly(L-lysine) and poly(L-glutamic acid) onto GOx-coated CaCO₃ templates. These polypeptides are known from previous research to exhibit nanometer-scale organization in multilayer films. Templates were dissolved by ethylenediaminetetraacetic acid (EDTA) at neutral pH. Addition of polyethylene glycol (PEG) to the polypeptide assembly solutions greatly increased enzyme retention on the templates, resulting in high-capacity, high-activity loading of the enzyme into artificial cells. Assay of enzyme activity showed that over 80 mg-mL⁻¹ GOx was retained in artificial cells after polypeptide multilayer film formation and template dissolution in the presence of PEG, but only one-fifth as much was retained in the absence of PEG. Encapsulation is a means of improving the availability of therapeutic macromolecules in biomedicine. This work therefore represents a means of developing polypeptide-based artificial cells for use as therapeutic biomacromolecule delivery vehicles.

Keywords:

• Layer-by-layer assembly
• Microencapsulation
• Nano-assembly
• Polyethylene glycol
• Polypeptide