Solid-phase synthesis of protein-polymers on reversible immobilization supports

Abstract

Facile automated biomacromolecule synthesis is at the heart of blending synthetic and biologic worlds. Full access to abiotic/biotic synthetic diversity first occurred when chemistry was developed to grow nucleic acids and peptides from reversibly immobilized precursors. Protein–polymer conjugates, however, have always been synthesized in solution in multi-step, multi-day processes that couple innovative chemistry with challenging purification. Here we report the generation of protein–polymer hybrids synthesized by protein-ATRP on reversible immobilization supports (PARIS). We utilized modified agarose beads to covalently and reversibly couple to proteins in amino-specific reactions. We then modified reversibly immobilized proteins with protein-reactive ATRP initiators and, after ATRP, we released and analyzed the protein polymers. The activity and stability of PARIS-synthesized and solution-synthesized conjugates demonstrated that PARIS was an effective, rapid, and simple method to generate protein–polymer conjugates. Automation of PARIS significantly reduced synthesis/purification timelines, thereby opening a path to changing how to generate protein–polymer conjugates.

Food-Derived Opioid Peptides in Human Health: A Review

“…Meanwhile, the SPPS approach requires peptide synthesis using resin as a support for a growing peptide chain. An amino acid's reactive side chain and α-amino group are first covered (mostly using the fluorenylmethoxycarbonyl protecting group (Fmoc) or tert-Butoxycarbonyl (Boc)) and the amino acids C-terminus is bound to the resin [152]. The N-terminal protecting group is normally removed (or cleaved) by using trifluoroacetic acid (Boc) or by 20% piperidine in N-N-dimethylformamide (Fmoc); then, the resin is washed before the introduction of subsequent amino acids.…”

Protein–polymer interaction: Transfer loading at interfacial region of PES‐based membrane and BSA

“…The phenomenon of protein adsorption on a polymer membrane is seen in a wide variety of applications . The applications include tissue engineering applications both in vitro and in vivo , biophysics, pharmaceutical sciences, and sensors .…”

Next generation protein‐polymer conjugates

“…In a recent advance, an understanding of how to control the reactivity of surface amino groups with activated ATRP initiators has emerged using blocking agents . Another technique is to specifically target the amino group on the N ‐terminus by reducing the reaction pH . Carbodiimide coupling is also frequently used to conjugate an acid‐containing CTA or ATRP initiator to amine‐containing protein residues.…”

“…This method could be applied to solid phase synthesis for simplified purification and processing. Solid phase protein‐polymer synthesis, in which polymer was grown from protein immobilized on agarose beads, was recently reported . Reversible covalent immobilization of proteins on agarose beads via pH‐responsive chemistry was the basis for the p rotein‐ A TRP on r eversible i mmobilization s upports (named PARIS ).…”