When polyelectrolytes and oppositely-charged multivalent ions are mixed in aqueous solutions, they can self-assemble into an array of soft materials and complex fluids, ranging from micro- and nanoparticles, to coacervates, to macroscopic gels. Here, we describe the formation and useful/interesting properties of two such materials: (1) submicron particles formed via ionotropic gelation of the cationic polysaccharide chitosan with tripolyphosphate (TPP); and (2) coacervates prepared from mixtures of the synthetic polycation poly(allylamine hydrochloride) (PAH) with either TPP or pyrophosphate (PPi). For chitosan/TPP particles (which are widely explored as potential drug carriers) we show how, by inhibiting chitosan/TPP binding, monovalent salt (NaCl) can be used to: (1) drastically slow down the rapid ionotropic gelation process to facilitate the experimental analysis of how these particles form; (2) enhance the stability of these particles to aggregation; and (3) achieve improved control over particle size.
Unlike the gel-like chitosan/TPP ionic networks, which are both soft (with 10^3 - 10^4 Pa storage moduli) and water-rich, mixtures of PAH with TPP and PPi form high-modulus, putty-like coacervates with storage moduli above 10^5 Pa and much lower (26 - 40 wt%) water contents. These moduli and water contents evidently reflect the high ionic crosslink densities enabled by the densely-charged and flexible PAH chains, and strong PAH/PPi and PAH/TPP binding (which also imparts these coacervates with long relaxation times). Besides their bulk properties, we show that the coacervates adhere to diverse substrates (both hydrophilic and hydrophobic) and, when used as wet adhesives, deliver short-term tensile adhesion strengths above 10^5 Pa. Further, the dense crosslinking within PAH/PPi and PAH/TPP coacervates makes them strong barriers to solute diffusion and (regardless of the solute-coacervate binding strength) enables them to release small water-soluble molecules over multiple months. These findings suggest that PAH/PPi and PAH/TPP coacervates can provide a simple route to both underwater adhesion and long-term controlled release.
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Effects of Protein Unfolding on Aggregation and Gelation in Lysozyme Solutions
In this work, we investigate the role of folding/unfolding equilibrium in protein aggregation and formation of a gel network. Near the neutral pH and at a low buffer ionic strength, the formation of the gel network around unfolding conditions prevents investigations of protein aggregation. In this study, by deploying the fact that in lysozyme solutions the time of folding/unfolding is much shorter than the characteristic time of gelation, we have prevented gelation by rapidly heating the solution up to the unfolding temperature (~80 °C) for a short time (~30 min.) followed by fast cooling to the room temperature. Dynamic light scattering measurements show that if the gelation is prevented, nanosized irreversible aggregates (about 10–15 nm radius) form over a time scale of 10 days. These small aggregates persist and aggregate further into larger aggregates over several weeks. If gelation is not prevented, the nanosized aggregates become the building blocks for the gel network and define its mesh length scale. These results support our previously published conclusion on the nature of mesoscopic aggregates commonly observed in solutions of lysozyme, namely that aggregates do not form from lysozyme monomers in their native folded state. Only with the emergence of a small fraction of unfolded proteins molecules will the aggregates start to appear and grow.
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- Award ID(s):
- 1856479
- NSF-PAR ID:
- 10276366
- Date Published:
- Journal Name:
- Biomolecules
- Volume:
- 10
- Issue:
- 9
- ISSN:
- 2218-273X
- Page Range / eLocation ID:
- 1262
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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2019 ,57 , 1014–1026
- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.3390/biom10091262
