@article{sohail_pirzada_guenther_barbieri_sit_menegatti_crook_opperman_khan_2023, title={Cellulose Acetate-Stabilized Pickering Emulsions: Preparation, Rheology, and Incorporation of Agricultural Active Ingredients}, volume={11}, ISSN={["2168-0485"]}, url={https://doi.org/10.1021/acssuschemeng.3c02428}, DOI={10.1021/acssuschemeng.3c02428}, abstractNote={We report the use of cellulose acetate (CA) nanoparticles (NPs) to produce oil in water Pickering emulsions. The CA NP can emulsify various oils and form stable emulsions at concentrations as low as 0.5 wt %. Rheological and microscopic analyses show evidence of interconnected NP aggregate networks between droplets. Yield stress measurements display evidence of “double” yielding. We postulate that the presence of the NP aggregates provides a secondary network between droplet clusters resulting in such behavior. We demonstrate the suitability of the emulsions as agriculture formulations by incorporating an agrochemical, abamectin (Abm), and a plant-growth-promoting microbe (PGPM) in the emulsions. Release assays exhibit sustained Abm release, promising higher efficacy at lower usage volumes. Incorporation of nonsporulating PGPM Pseudomonas simiae in the emulsions shows significantly higher microbe viability compared to controls after 70 days of storage. By demonstrating the application of CA NPs as a sustainable Pickering emulsifier, this study introduces the use of CA as a platform technology for the delivery of diverse agriculture cargos. A comprehensive evaluation of the system is articulated in a fundamental microstructure analysis and a demonstration of practical on-site attributes, including shelf-life stability and functional performance, verified through bioassays and plant growth studies.}, number={42}, journal={ACS SUSTAINABLE CHEMISTRY & ENGINEERING}, author={Sohail, Mariam and Pirzada, Tahira and Guenther, Richard and Barbieri, Eduardo and Sit, Tim and Menegatti, Stefano and Crook, Nathan and Opperman, Charles H. and Khan, Saad A.}, year={2023}, month={Sep}, pages={15178–15191} } @article{rahmanian_pirzada_barbieri_iftikhar_li_khan_2023, title={Mechanically robust, thermally insulating and photo-responsive aerogels designed from sol-gel electrospun PVP-TiO2 nanofibers}, volume={32}, ISSN={["2352-9407"]}, url={https://doi.org/10.1016/j.apmt.2023.101784}, DOI={10.1016/j.apmt.2023.101784}, abstractNote={We present a robust approach for fabricating polyvinylpyrrolidone (PVP)-titania (TiO2) nanofibrous aerogels (NFA) with multifunctional and triggered performances. These low density (∼ 10 mg cm−3) 3D self-supported aerogels having an intrinsically lamellar porous structure (> 99% porosity) are created via solid templating of sol-gel electrospun PVP-TiO2 hybrid nanofibers. The photocatalytic activity of TiO2 allows for on-demand application wherein the aerogel exhibits antibacterial properties upon UV exposure to bacteria such as Escherichia coli and Salmonella enterica. Significantly, while the aerogel sorbs common volatile organic components (VOCs) or oil due to its innate porosity, exposure of the aerogel to ultraviolet (UV) radiation leads to their decomposition. The PVP-TiO2 NFA exhibits a low thermal conductivity (0.062 W m−1 K−1) together with considerable mechanical flexibility up to strains of 50% with >90% recovery, without the need for post-processing. The photo-responsive attributes combined with mechanical resilience, oleophilicity and thermal insulation properties render these aerogels viable candidates for a diverse range of applications. We discuss such property enhancements in terms of the interaction between PVP and TiO2 and aerogel microstructure.}, journal={APPLIED MATERIALS TODAY}, author={Rahmanian, Vahid and Pirzada, Tahira and Barbieri, Eduardo and Iftikhar, Sherafghan and Li, Fanxing and Khan, Saad A.}, year={2023}, month={Jun} } @article{chu_shastry_barbieri_prodromou_greback-clarke_smith_moore_kilgore_cummings_pancorbo_et al._2023, title={Peptide ligands for the affinity purification of adeno-associated viruses from HEK 293 cell lysates}, volume={7}, ISSN={["1097-0290"]}, DOI={10.1002/bit.28495}, abstractNote={Adeno-associated viruses (AAVs) are the vector of choice for delivering gene therapies that can cure inherited and acquired diseases. Clinical research on various AAV serotypes significantly increased in recent years alongside regulatory approvals of AAV-based therapies. The current AAV purification platform hinges on the capture step, for which several affinity resins are commercially available. These adsorbents rely on protein ligands-typically camelid antibodies-that provide high binding capacity and selectivity, but suffer from low biochemical stability and high cost, and impose harsh elution conditions (pH < 3) that can harm the transduction activity of recovered AAVs. Addressing these challenges, this study introduces peptide ligands that selectively capture AAVs and release them under mild conditions (pH = 6.0). The peptide sequences were identified by screening a focused library and modeled in silico against AAV serotypes 2 and 9 (AAV2 and AAV9) to select candidate ligands that target homologous sites at the interface of the VP1-VP2 and VP2-VP3 virion proteins with mild binding strength (KD ~ 10-5 -10-6 M). Selected peptides were conjugated to Toyopearl resin and evaluated via binding studies against AAV2 and AAV9, demonstrating the ability to target both serotypes with values of dynamic binding capacity (DBC10% > 1013 vp/mL of resin) and product yields (~50%-80%) on par with commercial adsorbents. The peptide-based adsorbents were finally utilized to purify AAV2 from a HEK 293 cell lysate, affording high recovery (50%-80%), 80- to 400-fold reduction of host cell proteins (HCPs), and high transduction activity (up to 80%) of the purified viruses.}, journal={BIOTECHNOLOGY AND BIOENGINEERING}, author={Chu, Wenning and Shastry, Shriarjun and Barbieri, Eduardo and Prodromou, Raphael and Greback-Clarke, Paul and Smith, Will and Moore, Brandyn and Kilgore, Ryan and Cummings, Christopher and Pancorbo, Jennifer and et al.}, year={2023}, month={Jul} } @article{zhang_barbieri_lebarre_rameez_mostafa_menegatti_2023, title={Peptonics: A new family of cell-protecting surfactants for the recombinant expression of therapeutic proteins in mammalian cell cultures}, volume={10}, ISSN={["1860-7314"]}, DOI={10.1002/biot.202300261}, abstractNote={Abstract Polymer surfactants are key components of cell culture media as they prevent mechanical damage during fermentation in stirred bioreactors. Among cell‐protecting surfactants, Pluronics are widely utilized in biomanufacturing to ensure high cell viability and productivity. Monodispersity of monomer sequence and length is critical for the effectiveness of Pluronics—since minor deviations can damage the cells—but is challenging to achieve due to the stochastic nature of polymerization. Responding to this challenge, this study introduces Peptonics, a novel family of peptide and peptoid surfactants whose monomer composition and sequence are designed to achieve high cell viability and productivity at a fraction of chain length and cost of Pluronics. A designed ensemble of Peptonics was initially characterized via light scattering and tensiometry to select sequences whose phase behavior and tensioactivity align with those of Pluronics. Selected sequences were evaluated as cell‐protecting surfactants using Chinese hamster ovary (CHO) cells expressing therapeutic monoclonal antibodies (mAb). Peptonics IH‐T1010, ih‐T1010, and ih‐T1020 afforded high cell density (up to 3 × 10 7 cells mL −1 ) and viability (up to 95% within 10 days of culture), while reducing the accumulation of ammonia (a toxic metabolite) by ≈10% compared to Pluronic F‐68. Improved cell viability afforded high mAb titer (up to 5.5 mg mL −1 ) and extended the production window beyond 14 days; notably, Peptonic IH‐T1020 decreased mAb fragmentation and aggregation ≈5%, and lowered the titer of host cell proteins by 16% compared to Pluronic F‐68. These features can improve significantly the purification of mAbs, thus increasing their availability at a lower cost to patients.}, journal={BIOTECHNOLOGY JOURNAL}, author={Zhang, Ka and Barbieri, Eduardo and Lebarre, Jacob and Rameez, Shahid and Mostafa, Sigma and Menegatti, Stefano}, year={2023}, month={Oct} } @article{shastry_chu_barbieri_greback-clarke_smith_cummings_minzoni_pancorbo_gilleskie_ritola_et al._2023, title={Rational design and experimental evaluation of peptide ligands for the purification of adeno-associated viruses via affinity chromatography}, volume={9}, ISSN={["1860-7314"]}, DOI={10.1002/biot.202300230}, abstractNote={Adeno-associated viruses (AAVs) have acquired a central role in modern medicine as delivery agents for gene therapies targeting rare diseases. While new AAVs with improved tissue targeting, potency, and safety are being introduced, their biomanufacturing technology is lagging. In particular, the AAV purification pipeline hinges on protein ligands for the affinity-based capture step. While featuring excellent AAV binding capacity and selectivity, these ligands require strong acid (pH <3) elution conditions, which can compromise the product's activity and stability. Additionally, their high cost and limited lifetime has a significant impact on the price tag of AAV-based therapies. Seeking to introduce a more robust and affordable affinity technology, this study introduces a cohort of peptide ligands that (i) mimic the biorecognition activity of the AAV receptor (AAVR) and anti-AAV antibody A20, (ii) enable product elution under near-physiological conditions (pH 6.0), and (iii) grant extended reusability by withstanding multiple regenerations. A20-mimetic CYIHFSGYTNYNPSLKSC and AAVR-mimetic CVIDGSQSTDDDKIC demonstrated excellent capture of serotypes belonging to distinct clones/clades - namely, AAV1, AAV2, AAV5, AAV6, AAV8, and AAV9. This corroborates the in silico models documenting their ability to target regions of the viral capsid that are conserved across all serotypes. CVIDGSQSTDDDKIC-Toyopearl resin features binding capacity (≈1014 vp mL-1 ) and product yields (≈60%-80%) on par with commercial adsorbents, and purifies AAV2 from HEK293 and Sf9 cell lysates with high recovery (up to 78%), reduction of host cell proteins (up to 700-fold), and high transduction activity (up to 65%).}, journal={BIOTECHNOLOGY JOURNAL}, author={Shastry, Shriarjun and Chu, Wenning and Barbieri, Eduardo and Greback-Clarke, Paul and Smith, William K. and Cummings, Christopher and Minzoni, Arianna and Pancorbo, Jennifer and Gilleskie, Gary and Ritola, Kimberly and et al.}, year={2023}, month={Sep} } @article{kilgore_minzoni_shastry_smith_barbieri_wu_lebarre_chu_o'brien_menegatti_2023, title={The downstream bioprocess toolbox for therapeutic viral vectors}, volume={1709}, ISSN={["1873-3778"]}, DOI={10.1016/j.chroma.2023.464337}, abstractNote={Viral vectors are poised to acquire a prominent position in modern medicine and biotechnology owing to their role as delivery agents for gene therapies, oncolytic agents, vaccine platforms, and a gateway to engineer cell therapies as well as plants and animals for sustainable agriculture. The success of viral vectors will critically depend on the availability of flexible and affordable biomanufacturing strategies that can meet the growing demand by clinics and biotech companies worldwide. In this context, a key role will be played by downstream process technology: while initially adapted from protein purification media, the purification toolbox for viral vectors is currently undergoing a rapid expansion to fit the unique biomolecular characteristics of these products. Innovation efforts are articulated on two fronts, namely (i) the discovery of affinity ligands that target adeno-associated virus, lentivirus, adenovirus, etc.; (ii) the development of adsorbents with innovative morphologies, such as membranes and 3D printed monoliths, that fit the size of viral vectors. Complementing these efforts are the design of novel process layouts that capitalize on novel ligands and adsorbents to ensure high yield and purity of the product while safeguarding its therapeutic efficacy and safety; and a growing panel of analytical methods that monitor the complex array of critical quality attributes of viral vectors and correlate them to the purification strategies. To help explore this complex and evolving environment, this study presents a comprehensive overview of the downstream bioprocess toolbox for viral vectors established in the last decade, and discusses present efforts and future directions contributing to the success of this promising class of biological medicines.}, journal={JOURNAL OF CHROMATOGRAPHY A}, author={Kilgore, Ryan and Minzoni, Arianna and Shastry, Shriarjun and Smith, Will and Barbieri, Eduardo and Wu, Yuxuan and Lebarre, Jacob P. and Chu, Wenning and O'Brien, Juliana and Menegatti, Stefano}, year={2023}, month={Oct} } @article{wang_hosseini_shastry_barbieri_chu_menegatti_daniele_2023, title={Toward the quantification of adeno-associated virus titer by electrochemical impedance spectroscopy}, DOI={10.1109/BioSensors58001.2023.10281105}, journal={2023 IEEE BIOSENSORS CONFERENCE, BIOSENSORS}, author={Wang, Junhyeong and Hosseini, Mahshid and Shastry, Shriarjun and Barbieri, Eduardo and Chu, Wenning and Menegatti, Stefano and Daniele, Michael A.}, year={2023} } @article{rahmanian_ebrahim_razavi_abdelmigeed_barbieri_menegatti_parsons_li_pirzada_khan_2023, title={Vapor phase synthesis of metal-organic frameworks on a nanofibrous aerogel creates enhanced functionality}, volume={11}, ISSN={["2050-7496"]}, url={https://doi.org/10.1039/D3TA05299K}, DOI={10.1039/d3ta05299k}, abstractNote={Vapor-phase synthesis of metal–organic frameworks (MOFs) on nanofibrous aerogels provides a hierarchically porous and mechanically robust material platform for use in a multitude of applications, from carbon dioxide capture to heavy metal removal.}, journal={JOURNAL OF MATERIALS CHEMISTRY A}, author={Rahmanian, Vahid and Ebrahim, Muhammed Ziauddin Ahmad and Razavi, Seyedamin and Abdelmigeed, Mai and Barbieri, Eduardo and Menegatti, Stefano and Parsons, Gregory N. and Li, Fanxing and Pirzada, Tahira and Khan, Saad A.}, year={2023}, month={Nov} } @article{wang_pirzada_xie_barbieri_hossain_opperman_pal_wei_parsons_khan_2022, title={Creating hierarchically porous banana paper-metal organic framework (MOF) composites with multifunctionality}, volume={28}, ISSN={["2352-9407"]}, url={https://doi.org/10.1016/j.apmt.2022.101517}, DOI={10.1016/j.apmt.2022.101517}, abstractNote={We report a robust approach to integrate metal-organic frameworks (MOF) via vapor phase synthesis on a cost-effective and mechanically durable fibrous banana paper (BP) substrate developed from lignocellulosic biomass. The unique hollow fibrous structure of BP combined with the methodology used produces MOF-fiber composites with uniform MOF distribution and enhanced functionalities, with minimal use of organic solvents. The BP-MOF composites demonstrate a high surface area of 552 m2/g and uniform surface growth of MOF on them. Mechanical strength and bending flexibility of the substrate is well retained after the MOF growth, while the hollow tubular nature and hierarchical porosity of the BP facilitate gas diffusion. The BP-MOF composites demonstrate strong antibacterial activity with 99.2% of E.coli destroyed within the first hour of incubation. Preliminary studies with smartphone-based volatile organic compound (VOC) sensor show enhanced 1-octen-3-ol vapor absorption on BP-MOF, indicating its potential for VOC capture and sensing. We believe that the sustainable nature and flexibility of the lignocellulosic BP substrate taken together with uniform growth of MOF on the hierarchically porous BP impart impressive attributes to these composites, which can be explored in diverse applications.}, journal={APPLIED MATERIALS TODAY}, publisher={Elsevier BV}, author={Wang, Siyao and Pirzada, Tahira and Xie, Wenyi and Barbieri, Eduardo and Hossain, Oindrila and Opperman, Charles H. and Pal, Lokendra and Wei, Qingshan and Parsons, Gregory N. and Khan, Saad A.}, year={2022}, month={Aug} } @article{barbieri_cutright_ramesh_khan_efimenko_genzer_menegatti_2022, title={Potent Antibacterial Composite Nonwovens Functionalized with Bioactive Peptides and Polymers}, volume={8}, ISSN={["2196-7350"]}, url={https://doi.org/10.1002/admi.202201061}, DOI={10.1002/admi.202201061}, abstractNote={Abstract This study presents a set of strategies for producing potent antibacterial fabrics by functionalizing nonwoven fabrics (NWFs) with antimicrobial peptides and polymers (AMPs). The incorporation of AMPs is initially optimized on 2D substrates by evaluating conjugation on a poly(maleic anhydride) copolymer coating versus adsorption on polycationic/anionic films and microgels. The evaluation of the resulting surfaces against S. aureus and E. coli highlights the superior antibacterial activity of poly‐ionic films loaded with daptomycin and polymyxin B as well as microgels featuring controlled release of bacitracin and polymyxin B. These formulations are translated onto spun‐bond polypropylene and poly(ethylene terephthalate) NWFs. The poly‐ionic coatings are either covalently anchored or physically adsorbed onto the surface of the fibers, while the microgels and antibacterial polymers are adsorbed and photo‐crosslinked thereon using a ultraviolet (UV)‐crosslinkable benzophenone‐based polymer. Selected formulations loaded with bacitracin and polymyxin B afford a 10 5 ‐fold reduction of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) in artificial sweat, respectively, on par with commercial antibacterial NWFs. The proposed antibacterial fabric, however, outperforms its commercial counterparts in terms of biocompatibility, showing virtually no adverse effect on human epidermal keratinocytes. Collectively, these results demonstrate affordable and scalable routes for developing antimicrobial NWFs that efficiently eliminate resilient pathogenic bacteria.}, journal={ADVANCED MATERIALS INTERFACES}, author={Barbieri, Eduardo and Cutright, Camden C. and Ramesh, Srivatsan and Khan, Saad A. and Efimenko, Kirill and Genzer, Jan and Menegatti, Stefano}, year={2022}, month={Aug} } @article{fan_barbieri_shastry_menegatti_boi_carbonell_2022, title={Purification of Adeno-Associated Virus (AAV) Serotype 2 from Spodoptera frugiperda (Sf9) Lysate by Chromatographic Nonwoven Membranes}, volume={12}, ISSN={["2077-0375"]}, DOI={10.3390/membranes12100944}, abstractNote={The success of adeno-associated virus (AAV)-based therapeutics in gene therapy poses the need for rapid and efficient processes that can support the growing clinical demand. Nonwoven membranes represent an ideal tool for the future of virus purification: owing to their small fiber diameters and high porosity, they can operate at high flowrates while allowing full access to target viral particles without diffusional limitations. This study describes the development of nonwoven ion-exchange membrane adsorbents for the purification of AAV2 from an Sf9 cell lysate. A strong anion-exchange (AEX) membrane was developed by UV grafting glycidyl methacrylate on a polybutylene terephthalate nonwoven followed by functionalization with triethylamine (TEA), resulting in a quaternary amine ligand (AEX-TEA membrane). When operated in bind-and-elute mode at a pH higher than the pI of the capsids, this membrane exhibited a high AAV2 binding capacity (9.6 × 1013 vp·mL-1) at the residence time of 1 min, and outperformed commercial cast membranes by isolating AAV2 from an Sf9 lysate with high productivity (2.4 × 1013 capsids·mL-1·min-1) and logarithmic reduction value of host cell proteins (HCP LRV ~ 1.8). An iminodiacetic acid cation-exchange nonwoven (CEX-IDA membrane) was also prepared and utilized at a pH lower than the pI of capsids to purify AAV2 in a bind-and-elute mode, affording high capsid recovery and impurity removal by eluting with a salt gradient. To further increase purity, the CEX-IDA and AEX-TEA membranes were utilized in series to purify the AAV2 from the Sf9 cell lysate. This membrane-based chromatography process also achieved excellent DNA clearance and a recovery of infectivity higher that that reported using ion-exchange resin chromatography.}, number={10}, journal={MEMBRANES}, author={Fan, Jinxin and Barbieri, Eduardo and Shastry, Shriarjun and Menegatti, Stefano and Boi, Cristiana and Carbonell, Ruben G.}, year={2022}, month={Oct} } @article{barbieri_sales junior_fonseca murta_silva_2022, title={Study of Methods for the Synthesis of Pyrrole Derivatives and Evaluation of anti-Trypanosoma cruzi Activity}, ISSN={["1984-6835"]}, DOI={10.21577/1984-6835.20220052}, journal={REVISTA VIRTUAL DE QUIMICA}, author={Barbieri, Eduardo and Sales Junior, Policarpo Ademar and Fonseca Murta, Silvane Maria and Silva, Barbara V}, year={2022}, month={Mar} }