Shengchang Tang

Anomalous Self-Diffusion and Sticky Rouse Dynamics in Associative Protein Hydrogels

Natural and synthetic materials based on associating polymers possess diverse mechanical behavior, transport properties and responsiveness to external stimuli. Although much is known about their dynamics on the molecular and macroscopic level, knowledge of self-diffusive dynamics of the network-forming constituents remains limited. Using forced Rayleigh scattering, anomalous self-diffusion is observed in model associating protein hydrogels originating from the interconversion between species that diffuse in both the molecular and associated state. The diffusion can be quantitatively modeled using a two-state model for polymers in the gel, where diffusivity in the associated state is critical to the super diffusive behavior. The dissociation time from bulk rheology measurements was 2-3 orders of magnitude smaller than the one measured by diffusion, because the former characterizes submolecular dissociation dynamics, whereas the latter depicts single protein molecules completely disengaging from the network. Rheological data also show a sticky Rouse-like relaxation at long times due to collective relaxation of large groups of proteins, suggesting mobility of associated molecules. This study experimentally demonstrates a hierarchy of relaxation processes in associating polymer networks, and it is anticipated that the results can be generalized to other associative systems to better understand the relationship of dynamics among sticky bonds, single molecules, and the entire network.

Site-Specific Conjugation of RAFT Polymers to Proteins via Expressed Protein Ligation

Site-specific protein conjugates with RAFT polymers were synthesized using expressed protein ligation. Stable micelles were formed from both linear block copolymer and Y-shaped conjugates.

Effect of polymer chemistry on globular protein–polymer block copolymer self-assembly

Bioconjugates of the model red fluorescent protein mCherry and synthetic polymer blocks with different hydrogen bonding functionalities show that the chemistry of the polymer block has a large effect on both ordering transitions and the type of nanostructures formed during bioconjugate self-assembly. The phase behaviours of mCherry-b-poly(hydroxypropyl acrylate) (PHPA) and mCherry-b-poly(oligoethylene glycol acrylate) (POEGA) in concentrated aqueous solution show that changes in polymer chemistry result in increase in the order–disorder transition concentrations (CODTs) by approximately 10–15 wt% compared to a previously studied globular protein–polymer block copolymer, mCherry-b-poly(N-isopropylacrylamide) (PNIPAM). The CODTs are always minimized for symmetric bioconjugates, consistent with the importance of protein–polymer interactions in self-assembly. Both mCherry-b-PHPA and mCherry-b-POEGA also form phases that have not previously been observed in other globular protein–polymer conjugates: mCherry-b-PHPA forms a cubic phase that can be indexed to Iad and mCherry-b-POEGA displays coexistence of lamellae and a cubic Iad structure over a narrow range of concentration and temperature. Several common behaviours are also revealed by comparison of different polymer blocks. With increasing concentration and temperature, ordered phases always appear in the order lamellar, cubic/PL, and hexagonal, although not all phases are observed in all materials. High concentration solutions (near 80 wt%) also undergo a re-entrant order–disorder transition to form nematic liquid crystalline phases, regardless of the polymer block chemistry.

Responsive block copolymer photonics triggered by protein-polyelectrolyte coacervation.

Ionic interactions between proteins and polyelectrolytes are demonstrated as a method to trigger responsive transitions in block copolymer (BCP) photonic gels containing one neutral hydrophobic block and one cationic hydrophilic block. Poly(2-vinylpyridine) (P2VP) blocks in lamellar poly(styrene-b-2-vinylpyridine) block copolymer thin films are quaternized with primary bromides to yield swollen gels that show strong reflectivity peaks in the visible range; exposure to aqueous solutions of various proteins alters the swelling ratios of the quaternized P2VP (QP2VP) gel layers in the PS-QP2VP materials due to the ionic interactions between proteins and the polyelectrolyte. Parameters such as charge density, hydrophobicity, and cross-link density of the QP2VP gel layers as well as the charge and size of the proteins play significant roles on the photonic responses of the BCP gels. Differences in the size and pH-dependent charge of proteins provide a basis for fingerprinting proteins based on their temporal and equilibrium photonic response. The results demonstrate that the BCP gels and their photonic effect provide a robust and visually interpretable method to differentiate different proteins.

Controlling topological entanglement in engineered protein hydrogels with a variety of thiol coupling chemistries

Topological entanglements between polymer chains are achieved in associating protein hydrogels through the synthesis of high molecular weight proteins via chain extension using a variety of thiol coupling chemistries, including disulfide formation, thiol-maleimide, thiol-bromomaleimide and thiol-ene. Coupling of cysteines via disulfide formation results in the most pronounced entanglement effect in hydrogels, while other chemistries provide versatile means of changing the extent of entanglement, achieving faster chain extension, and providing a facile method of controlling the network hierarchy and incorporating stimuli responsivities. The addition of trifunctional coupling agents causes incomplete crosslinking and introduces branching architecture to the protein molecules. The high-frequency plateau modulus and the entanglement plateau modulus can be tuned by changing the ratio of difunctional chain extender to the trifunctional branching unit. Therefore, these chain extension reactions show promise in delicately controlling the relaxation and mechanical properties of engineered protein hydrogels in ways that complement their design through genetic engineering.

High-velocity micro-particle impact on gelatin and synthetic hydrogel

The high-velocity impact response of gelatin and synthetic hydrogel samples is investigated using a laser-based microballistic platform for launching and imaging supersonic micro-particles. The micro-particles are monitored during impact and penetration into the gels using a high-speed multi-frame camera that can record up to 16 images with nanosecond time resolution. The trajectories are compared with a Poncelet model for particle penetration, demonstrating good agreement between experiments and the model for impact in gelatin. The model is further validated on a synthetic hydrogel and the applicability of the results is discussed. We find the strength resistance parameter in the Poncelet model to be two orders of magnitude higher than in macroscopic experiments at comparable impact velocities. The results open prospects for testing high-rate behavior of soft materials on the microscale and for guiding the design of drug delivery methods using accelerated microparticles.