Polymer Structure and Conformation Alter the Antigenicity of Virus-like Particle-Polymer Conjugates.

Covalent conjugation of water-soluble polymers to proteins is critical for evading immune surveillance in the field of biopharmaceuticals. The most common and long-standing polymer modification is the attachment of methoxypoly(ethylene glycol) (mPEG), termed PEGylation, which has led to several clinically approved pharmaceuticals. Recent data indicate that brush-type polymers significantly enhance in vitro and in vivo properties. Herein, the polymer conformation of poly(ethylene glycol) is detailed and compared with those of water-soluble polyacrylate and polynorbornene (PNB) when attached to icosahedral virus-like particles. Small-angle neutron scattering reveals vastly different polymer conformations of the multivalent conjugates. Immune recognition of conjugated particles was evaluated versus PEGylated particles, and PNB conjugation demonstrated the most effective shielding from antibody recognition.

"Graft-to" Protein/Polymer Conjugates Using Polynorbornene Block Copolymers.

A series of water-soluble polynorbornene block copolymers prepared via Ring-Opening Metathesis Polymerization (ROMP) were grafted to proteins to form ROMP-derived bioconjugates. ROMP afforded low-dispersity polymers and allowed for strict control over polymer molecular weight and architecture. The polymers consisted of a large block of PEGylated monoester norbornene and were capped with a short block of norbornene dicarboxylic anhydride. This cap served as a reactive linker that facilitated attachment of the polymer to lysine residues under mildly alkaline conditions. The generality of this approach was shown by synthesizing multivalent polynorbornene-modified viral nanoparticles derived from bacteriophage Qß, a protein nanoparticle used extensively for nanomedicine. The conjugated nanoparticles showed no cytotoxicity to NIH 3T3 murine fibroblast cells. These findings establish protein bioconjugation with functionalized polynorbornenes as an effective alternative to conventional protein/polymer modification strategies and further expand the toolbox for protein bioconjugates.

Protein ROMP: Aqueous Graft-from Ring-Opening Metathesis Polymerization.

Ring-opening metathesis polymerization (ROMP) was carried out from the surface of a protein under aqueous conditions. Grubbs' third generation catalyst was modified with PEGylated pyridyl groups to form a water-soluble species that showed high activity in aqueous buffered solutions at near-neutral pH. The modification of a protein with this catalyst to create a stable macroinitiator for ROMP is described. The protein macroinitiator was then used to polymerize water-soluble norbornenes, resulting in high molecular weight protein/polymer conjugates. Varying polymerization time and monomer concentration demonstrated the kinetics of molecular weight evolution and macroinitiator conversion of graft-from ROMP.