The global transition to electric vehicles and renewable energy systems has heightened the demand for lithium-ion batteries (LIBs), creating an urgent need for sustainable battery recycling methods to recover critical raw materials, including lithium. Lithium-selective cation-exchange polymeric membranes are one of the emerging options to achieve such lithium recycling. To make this change even greener, instead of using traditional fossil-origin polymers to produce membranes, this research employed bacterial cellulose acetate (BCA), a bio-derived and eco-friendly polymer. By adding 5 wt% N-methyl-N-propylpiperidinium bis(trifluoromethanesulfonyl)imide (PP13-TFSI), an ionic liquid (IL) which is a plasticizer and lithium-ion conductor, and 20 wt% hydrogen manganese oxide (HMO), which is a lithium-selective inorganic filler, four BCA-based membranes (BCA, BCA-IL, BCA-HMO and BCA-IL-HMO) were prepared. The membranes were extensively characterized for their morphology, thermal stability, chemical, and mechanical properties. Subsequently, they were tested in diffusion cells (without applying any external driving force) for ionic conductivity, lithium selectivity, and lithium flux using binary salt mixtures and synthetic LIB leachate. The BCA-IL membrane outperformed other BCA-based membranes in terms of separation factors, achieving values of 10.50 (Li+/Mn2+), 11.75 (Li+/Ni2+), and 10.95 (Li+/Co2+) with a lithium flux of 0.12 mol m-2 h-1 when processing synthetic LIB leachate. Under the same conditions, the BCA-HMO membranes exhibited a higher lithium flux (0.51 mol m-2 h-1) but with lower separation factor values of 3.39 (Li+/Mn2+), 3.62 (Li+/Ni2+), and 3.36 (Li+/Co2+). The use of plant-derived cellulose acetate (CA) as an alternative to BCA was also assessed; however, despite promising ideal lithium selectivity values (for example, 112 for Li+/Ni2+ in the case of CA-HMO membrane), their conductivity was up to two orders of magnitude lower than that of BCA-based membranes. All these findings highlight the promising potential of BCA-based membranes for lithium recovery from lithium-ion battery leachates.
Gene therapies using adeno-associated viruses (AAVs) for central nervous system (CNS) disorders face challenges because host immune responses are not represented in classical preclinical models. Here, we present a human-induced pluripotent stem cell (hiPSC)-derived innate immunocompetent 3D CNS model that recapitulates neuroinflammatory hallmarks, serving as a platform for preclinical gene therapy development. By utilizing various scales of stirred-tank bioreactor systems, we generated neurospheroids (iNSpheroids) composed of neurons, astrocytes, and oligodendrocytes, alongside microglial cells (iMGLs) to mimic the neuroimmune axis. These systems enabled large-scale production of iNSpheroids and subsequent miniaturization for co-culture experiments and screening of inflammatory stimuli, while maintaining a highly controlled environment. The iMGL-iNSpheroids demonstrated active neuron-microglia crosstalk and exhibited distinct inflammatory responses to a series of neuroinflammatory factors. iMGL-iNSpheroids mounted an early response to rAAV9, which is underscored by the activation of inflammatory pathways (e.g., TNF-via NF-κB activation) in glial cell populations. This model offers a valuable tool to dissect neuroinflammatory mechanisms, accelerating gene therapy development.
Rift Valley fever (RVF) is a mosquito-borne zoonosis of major concern for human and animal health, yet no licensed human vaccine exists. Here, we engineered a self-assembling nanoparticle vaccine candidate by genetically fusing the RVF virus glycoprotein Gn to the N-terminus of a hybrid bacterial ferritin, generating nanoparticles that display 24 copies of Gn on their surface. Cryo-electron microscopy at 6 Å resolution confirmed ordered and symmetric presentation of the antigens, consistent with the structural models of ferritin and Gn. When incubated with human monocyte-derived dendritic cells, the Gn-ferritin nanoparticles were efficiently internalized and induced robust expression of maturation markers (e.g., CD54, CD83, CD86) and secretion of pro-inflammatory cytokines (e.g., IL-1β, IL-6, IL-12p40, TNF-α), in contrast to soluble Gn or ferritin controls. These findings demonstrate that ferritin nanoparticles provide a structurally defined and immunologically active platform for RVF virus antigen display, establishing a foundation for the development of safe and effective subunit vaccines against this emerging pathogen.
The clinical efficacy of cell therapy products is intrinsically related to their state of differentiation and maturity. However, current approaches to target cell phenotype and potency lack efficacy, scalability, and cost-effectiveness. Metabolism is a key driver of cell fate, a characteristic that can be explored to design bioprocesses yielding functional cell therapy products. Here, we review recent studies focused on exploring metabolic shifts to improve cell potency and discuss how these strategies can contribute to accelerating bioprocess development and benefit from their translation into scalable, tightly controlled, and affordable manufacturing workflows.
Lentiviral vectors (LV) are the preferred gene therapy tools when sustained, long-term transgene expression is required. However, their clinical application is hindered by high manufacturing costs and complex ex vivo protocols. Unlike adeno-associated viruses (AAV), impurities in LV productions remain poorly characterized. This knowledge gap hampers the development of more efficient purification strategies and safer therapies suitable for in vivo administration. In this study, particles in HEK293T-derived LV feedstocks were characterized using advanced analytics, showcasing a small percentage of intact LV (similar to 6%), and an overwhelming majority (similar to 94%) consisted of nonfunctional impurities such as VSV-G empty particles and extracellular vesicles (EV). To address this, a negative-mode affinity chromatography process was developed to selectively remove impurities while retaining functional LV. Initially, the selectivity of several VHH binders for closely resembling impurities and their lack of interaction with functional vectors was assessed. Spin columns packed with functionalized resins against tetraspanins CD81, CD63 and CD9 were evaluated on phenotyped LV feedstocks (Delta VSV-G, Delta p24, and Delta N-glycans) generated via modified transfections. Standard LV and non-specific binders served as controls. Binder performance was evaluated using ELISAs (p24, VSV-G, CD63, CD81, CD9), digital droplet PCR, and infectivity assays. Among the binders, anti-CD63 emerged as the leading candidate, preserving 87 +/- 6% of infectious, full-genome LV while removing 87 +/- 1% of chimeric, non-infectious particles sharing LV components. This novel negative-mode affinity approach for impurity removal represents a significant advance for LV manufacturing. Enhanced purity and product quality may enable reduced therapeutic doses and improved clinical outcomes, paving the way for next-generation LV purification strategies.
