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Abstract Almost all commercial proteins are purified using ammonium sulfate precipitation. Protein-polymer conjugates are synthesized from pure starting materials, and the struggle to separate conjugates from polymer, native protein, and from isomers has vexed scientists for decades. We have discovered that covalent polymer attachment has a transformational effect on protein solubility in salt solutions. Here, protein-polymer conjugates with a variety of polymers, grafting densities, and polymer lengths are generated using atom transfer radical polymerization. Charged polymers increase conjugate solubility in ammonium sulfate and completely prevent precipitation even at 100% saturation. Atomistic molecular dynamic simulations show the impact is driven by an anti-polyelectrolyte effect from zwitterionic polymers. Uncharged polymers exhibit polymer length-dependent decreased solubility. The differences in salting-out are then used to simply purify mixtures of conjugates and native proteins into single species. Increasing protein solubility in salt solutions through polymer conjugation could lead to many new applications of protein-polymer conjugates.
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Enlightening advances in polymer bioconjugate chemistry: light-based techniques for grafting to and from biomacromolecules
Photochemistry has revolutionized the field of polymer–biomacromolecule conjugation. Ligation reactions necessitate biologically benign conditions, and photons have a significant energy advantage over what is available thermally at ambient temperature, allowing for rapid and unique reactivity. Photochemical reactions also afford many degrees of control, specifically, spatio-temporal control, light source tunability, and increased oxygen tolerance. Light-initiated polymerizations, in particular photo-atom-transfer radical polymerization (photo-ATRP) and photoinduced electron/energy transfer reversible addition–fragmentation chain transfer polymerization (PET-RAFT), have been used for grafting from proteins, DNA, and cells. Additionally, the spatio-temporal control inherent to light-mediated chemistry has been utilized for grafting biomolecules to hydrogel networks for many applications, such as 3-D cell culture. While photopolymerization has clear advantages, there are factors that require careful consideration in order to obtain optimal control. These factors include the photocatalyst system, light intensity, and wavelength. This Perspective aims to discuss recent advances of photochemistry for polymer biomacromolecule conjugation and potential considerations while tailoring these systems.
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- Award ID(s):
- 1904631
- PAR ID:
- 10233888
- Date Published:
- Journal Name:
- Chemical Science
- Volume:
- 11
- Issue:
- 20
- ISSN:
- 2041-6520
- Page Range / eLocation ID:
- 5142 to 5156
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1039/d0sc01544j
