Microalgae-derived biofuels have potential advantages over other renewable, crop-based resources; however, large-scale production is not currently economical due, in part, to challenges in the harvesting step. In this article, we present a novel approach for the dewatering and harvesting of microalgae using flocculants that can be recovered and recycled. Polyampholytes with molecular charges dependent upon pH (ranging from net positively- to net negatively-charged) are used as a model flocculant system and provide reversible electrostatic interactions with the negatively-charged algal cells. These pH-dependent properties allow the polyampholytic flocculants to efficiently desorb from concentrated biomass and, unlike most commercial flocculants that have permanently charged functionalities, be recovered and recycled for further dewatering processes. The behavior of the model polyampholytic flocculants is characterized for the dewatering of Chlorella vulgaris (UTEX 395). The reversible and recyclable flocculants achieve >99% flocculation efficiencies, are recovered at more than 98 wt% yields after biomass dewatering, and can be recycled over five times for flocculation.

Multi-cellular aggregates are found in healthy and diseased tissues, and while cell-cell contact is important for regulating many cell functions, cells also interact, to varying degrees, with extra-cellular matrix (ECM) proteins. Islets of Langerhans are one such example of cell aggregates in contact with ECM, both at the periphery of the cluster and dispersed throughout. While several studies have investigated the effect of reintroducing contact with ECM proteins on islet cell survival and function, the majority of these experiments only allow contact with the exterior cells. Thus, cell-culture platforms that enable the study of ECM-cell interactions throughout multi-cellular aggregates are of interest. Here, local presentation of ECM proteins was achieved using hydrogel microwell arrays to incorporate protein-laden microparticles during formation of MIN6 β-cell aggregates. Varying the microparticle seeding density reproducibly controlled the number of microparticles incorporated within three-dimensional aggregates (i.e., total amount of protein). Further, a relatively uniform spatial distribution of laminin- and fibronectin-coated microparticles was achieved throughout the x-, y-, and z-directions. Multiple ECM proteins were presented to β-cells in concert by incorporating two distinct populations of microparticles throughout the aggregates. Finally, scaling the microwell device dimensions allowed for the formation of two different sized cell-particle aggregates, ~80 and 160 μm in diameter. While the total number of microparticles incorporated per aggregate varied with size, the fraction of the aggregate occupied by microparticles was affected only by the microparticle seeding density, indicating that uniform local concentrations of proteins can be preserved while changing the overall aggregate dimensions.

This article presents the design and characterization of polyamphoteric flocculants for the separations of biocolloidal suspensions of importance in the production of biopharmaceuticals, microalgal cultures for nutraceuticals and biofuels, and wastewater treatment. The polyamphoteric flocculants consist of tunable, mixed charges dependent upon system pH, thereby providing strong electrostatic attraction to the diversely-charged surfaces of cellular suspensions. Enhanced flocculation efficiencies are achieved, as compared to cationic polyelectrolyte flocculants, and result from the ability of polyampholytes to adsorb to a diverse range of charge character and operate over an extended range of pH conditions.