Rapid Oxygen-Tolerant Synthesis of Protein-Polymer Bioconjugates via Aqueous Copper-Mediated Polymerization

Abstract

The synthesis of protein−polymer conjugates usually requires extensive and costly deoxygenation procedures, thus limiting their availability and potential applications. In this work, we report the ultrafast synthesis of polymer−protein bioconjugates in the absence of any external deoxygenation via an aqueous copper-mediated methodology. Within 10 min and in the absence of any external stimulus such as light (which may limit the monomer scope and/or disrupt the secondary structure of the protein), a range of hydrophobic and hydrophilic monomers could be successfully grafted from a BSA macroinitiator, yielding well-defined polymer−protein bioconjugates at quantitative yields. Our approach is compatible with a wide range of monomer classes such as (meth) acrylates, styrene, and acrylamides as well as multiple macroinitiators including BSA, BSA nanoparticles, and betagalactosidase from Aspergillus oryzae. Notably, the synthesis of challenging protein−polymer−polymer triblock copolymers was also demonstrated, thus significantly expanding the scope of our strategy. Importantly, both lower and higher scale polymerizations (from 0.2 to 35 mL) were possible without compromising the overall efficiency and the final yields. This simple methodology paves the way for a plethora of applications in aqueous solutions without the need of external stimuli or tedious deoxygenation.

Fully Oxygen-Tolerant Visible-Light-Induced ATRP of Acrylates in Water: Toward Synthesis of Protein-Polymer Hybrids

“…These problems can be overcome by attaching polymers to the surface of proteins using the grafting-from or grafting-to approach. − Protein-polymer hybrids (PPHs) can exhibit greater stability and reduced immunogenicity . Examples of polyacrylamide- and polymethacrylate-based PPHs are abundant in the literature. , In contrast, the synthesis of acrylate-based PPHs remains relatively underexplored, and the few available examples in the literature are limited to the use of RAFT polymerization or ATRP methods. ,− …”

Synthesis of Multifunctional Protein‐Polymer Conjugates via Oxygen‐tolerant, Aqueous Copper‐Mediated Polymerization, and Bioorthogonal Click Chemistry

“…Interestingly, clicking was shown to be quantitative in the oxygen‐tolerant, one‐pot approach. The results disclosed herein provide easy and scalable [ 21 ] methodologies to synthesize tailored polymer‐protein bioconjugates and pave the way for future opportunities and applications [ 38 ] such as the synthesis of prodrugs (through clicking drugs to be hydrolysed and liberated within the body) or multicatalysts (through clicking organic catalysts).…”

“…To avoid the use of light, [20] a rapid, oxygen-tolerant synthesis of protein-polymer bioconjugates via aqueous copper-mediated polymerization was also established. [21] One of the major advantages of these approaches is their proven tolerance for a very wide range of monomers (both hydrophilic and hydrophobic in nature) including styrenes, acrylates, methacrylates, and acrylamides, as well as their facile implementation to a variety of proteins or even protein nanoparticles. [18,19,21] One underlying goal in the evolution of protein-polymer conjugates aimed at applications for emerging technologies, is the facile synthesis of multifunctional protein-polymer biohybrids and biohybrid nanocarriers.…”

“…[21] One of the major advantages of these approaches is their proven tolerance for a very wide range of monomers (both hydrophilic and hydrophobic in nature) including styrenes, acrylates, methacrylates, and acrylamides, as well as their facile implementation to a variety of proteins or even protein nanoparticles. [18,19,21] One underlying goal in the evolution of protein-polymer conjugates aimed at applications for emerging technologies, is the facile synthesis of multifunctional protein-polymer biohybrids and biohybrid nanocarriers. A key approach to this direction focuses on the development of simple and efficient protocols to synthesize, modify, and control the polymer moiety.…”

Green Hydrogel Synthesis: Emphasis on Proteomics and Polymer Particle-Protein Interaction

“…The protein-coupled CTA then allowed further polymerization in water to form protein-synthetic polymer conjugates from monomers, including NIPAM. The copper-mediated CRP provides a route to polymer–protein bioconjugate synthesis without organic cosolvents [ 161 ]. Additionally, using a CRP technique, in this case photoinduced electron/energy transfer reversible addition–fragmentation chain-transfer (PET-RAFT), Liu et al developed bilayer hydrogel wound dressings by polymerizing two different monomer formulations in water using visible light, with erythrosin B as a photosensitizer and 2-(dodecylthiocarbonothioylthio) propionic acid (DOPAT) as the chain-transfer agent.…”