Abstract
New protein delivery systems are needed that efficiently encapsulate proteins and avoid formulation processes that affect protein structure and function. We have developed a protein delivery system termed disulfide-crosslinked polyion micelles (DCPMs), which consist of nanocomplexes formed by electrostatic self-assembly of a protein with a poly(ethylene glycol)-poly(L-lysine) block copolymer (PEG-PLL). The PEG-PLL amines are modified with crosslinkable dithiopyridine groups, using a Michael addition reaction that preserves the positive charges on the PLL chain to optimize polyionic complexation and disulfide crosslinking. DCPMs for vaccine delivery were prepared with ovalbumin and immunostimulatory CpG-DNA and are designed to release the vaccine intracellularly through reduction of disulfide crosslinks. DCPMs were also developed as a long-circulating enzyme carrier that maintains the enzymatic activity of the anti-oxidant enzyme catalase within the micelle core. Ovalbumin and catalase were each modified with SPDP to tether the protein in the micelle core, resulting in a high degree of protein retention under SDS-PAGE. DCPMs efficiently encapsulate and retain functional proteins in a stable polyionic complex and are a versatile delivery system for enzymes, vaccine antigens, and other protein therapeutics.
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