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Journal articles on the topic 'Biomanufacturing applications'

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Author: Grafiati
Published: 10 December 2022
Last updated: 28 January 2023

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1

Mann, Madison M., Toriana N. Vigil, Samantha M. Felton, William E. Fahy, Mason A. Kinkeade, Victoria K. Kartseva, Mary-Jean C. Rowson, Abigail J. Frost, and Bryan W. Berger. "Proteins in Synthetic Biology with Agricultural and Environmental Applications." SynBio 1, no. 1 (November 21, 2022): 77–88. http://dx.doi.org/10.3390/synbio1010006.

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Synthetic biology tools have become increasingly prevalent as we look to nature for biological approaches to complex problems. With an ever-growing global population, issues of food safety and security, as well as addressing pollution and striving for sustainability are of the utmost importance. In this review, we first highlight synthetic biology techniques such as directed evolution as a toolset for protein engineering and show direct applications for food safety and security. Moreover, we offer an introduction to creative approaches for biosensor design and development and spotlight a few innovative examples. Finally, we address biomanufacturing with direct applications, as well as biomanufacturing to improve natural processes.
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2

Sugita, Naohiko, and Mamoru Mitsuishi. "Special Issue on Biomanufacturing." International Journal of Automation Technology 8, no. 1 (January 5, 2014): 73. http://dx.doi.org/10.20965/ijat.2014.p0073.

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The development of medical devices and systems is essential for improving quality of life and reducing global healthcare costs. Machine tools are increasingly used in the medical, automotive, airplane, and electronics fields thanks to advances in manufacturing technology. The processing of artificial implants and biomaterials, for example, and parts of medical devices such as endoscopes are manufactured with multiaxis machine tools. This demand is expected to increase as society ages. Equipment used in diagnostics and surgery has also developed rapidly. Despite the use of advanced diagnostics such as computed tomography (CT) and magnetic resonance imaging (MRI), however, surgery still largely depends on the skill and sense of the surgeon. Advanced manufacturing technologies are thus needed to achieve these desired attributes. Biomanufacturing requires expertise in basic manufacturing processes such as cutting, electrophysical and chemical processes, forming, and abrasive processes. These, in turn, must be integrated into machine design, surface modification, precision engineering, and metrology within the overarching frameworks of design, life cycle engineering and assembly, production systems, and organization. Biomanufacturing is thus defined as the application of design and manufacturing technologies for reducing cost while advancing safety, quality, efficiency and speed in healthcare services and biomedical sciences. Biomanufacturing provides an excellent platform for converging innovations in precision engineering, nanotechnology, biotechnology, information technology, and cognitive sciences. This special issue presents the latest in research advances, practical and theoretical applications, and case studies on biomanufacturing. The papers featured in this issue provide aid in the development of next-generation manufacturing technologies. We thank the authors for their invaluable contributions and the reviewers for their ever- useful advice. We know you will find this special issue both fascinating and worthwhile.
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Yuan, Jianhua, Jianglin Cao, Fei Yu, Jie Ma, Dong Zhang, Yijing Tang, and Jie Zheng. "Microbial biomanufacture of metal/metallic nanomaterials and metabolic engineering: design strategies, fundamental mechanisms, and future opportunities." Journal of Materials Chemistry B 9, no. 33 (2021): 6491–506. http://dx.doi.org/10.1039/d1tb01000j.

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Biomanufacturing metal/metallic nanomaterials with ordered micro/nanostructures, controllable functions, and promising properties is of great importance in both fundamental studies and practical applications.
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Vilkhovoy, Michael, Abhinav Adhikari, Sandra Vadhin, and Jeffrey D. Varner. "The Evolution of Cell Free Biomanufacturing." Processes 8, no. 6 (June 8, 2020): 675. http://dx.doi.org/10.3390/pr8060675.

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Cell-free systems are a widely used research tool in systems and synthetic biology and a promising platform for manufacturing of proteins and chemicals. In the past, cell-free biology was primarily used to better understand fundamental biochemical processes. Notably, E. coli cell-free extracts were used in the 1960s to decipher the sequencing of the genetic code. Since then, the transcription and translation capabilities of cell-free systems have been repeatedly optimized to improve energy efficiency and product yield. Today, cell-free systems, in combination with the rise of synthetic biology, have taken on a new role as a promising technology for just-in-time manufacturing of therapeutically important biologics and high-value small molecules. They have also been implemented at an industrial scale for the production of antibodies and cytokines. In this review, we discuss the evolution of cell-free technologies, in particular advancements in extract preparation, cell-free protein synthesis, and cell-free metabolic engineering applications. We then conclude with a discussion of the mathematical modeling of cell-free systems. Mathematical modeling of cell-free processes could be critical to addressing performance bottlenecks and estimating the costs of cell-free manufactured products.
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Afghah, Ferdows, Caner Dikyol, Mine Altunbek, and Bahattin Koc. "Biomimicry in Bio-Manufacturing: Developments in Melt Electrospinning Writing Technology Towards Hybrid Biomanufacturing." Applied Sciences 9, no. 17 (August 28, 2019): 3540. http://dx.doi.org/10.3390/app9173540.

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Melt electrospinning writing has been emerged as a promising technique in the field of tissue engineering, with the capability of fabricating controllable and highly ordered complex three-dimensional geometries from a wide range of polymers. This three-dimensional (3D) printing method can be used to fabricate scaffolds biomimicking extracellular matrix of replaced tissue with the required mechanical properties. However, controlled and homogeneous cell attachment on melt electrospun fibers is a challenge. The combination of melt electrospinning writing with other tissue engineering approaches, called hybrid biomanufacturing, has introduced new perspectives and increased its potential applications in tissue engineering. In this review, principles and key parameters, challenges, and opportunities of melt electrospinning writing, and particularly, recent approaches and materials in this field are introduced. Subsequently, hybrid biomanufacturing strategies are presented for improved biological and mechanical properties of the manufactured porous structures. An overview of the possible hybrid setups and applications, future perspective of hybrid processes, guidelines, and opportunities in different areas of tissue/organ engineering are also highlighted.
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6

Chan, Weng Wan, Fang Yu, Quang Bach Le, Sixun Chen, Marcus Yee, and Deepak Choudhury. "Towards Biomanufacturing of Cell-Derived Matrices." International Journal of Molecular Sciences 22, no. 21 (November 3, 2021): 11929. http://dx.doi.org/10.3390/ijms222111929.

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Cell-derived matrices (CDM) are the decellularised extracellular matrices (ECM) of tissues obtained by the laboratory culture process. CDM is developed to mimic, to a certain extent, the properties of the needed natural tissue and thus to obviate the use of animals. The composition of CDM can be tailored for intended applications by carefully optimising the cell sources, culturing conditions and decellularising methods. This unique advantage has inspired the increasing use of CDM for biomedical research, ranging from stem cell niches to disease modelling and regenerative medicine. However, while much effort is spent on extracting different types of CDM and exploring their utilisation, little is spent on the scale-up aspect of CDM production. The ability to scale up CDM production is essential, as the materials are due for clinical trials and regulatory approval, and in fact, this ability to scale up should be an important factor from the early stages. In this review, we first introduce the current CDM production and characterisation methods. We then describe the existing scale-up technologies for cell culture and highlight the key considerations in scaling-up CDM manufacturing. Finally, we discuss the considerations and challenges faced while converting a laboratory protocol into a full industrial process. Scaling-up CDM manufacturing is a challenging task since it may be hindered by technologies that are not yet available. The early identification of these gaps will not only quicken CDM based product development but also help drive the advancement in scale-up cell culture and ECM extraction.
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7

Neville, Jonathan J., Joe Orlando, Kimberly Mann, Bethany McCloskey, and Michael N. Antoniou. "Ubiquitous Chromatin-opening Elements (UCOEs): Applications in biomanufacturing and gene therapy." Biotechnology Advances 35, no. 5 (September 2017): 557–64. http://dx.doi.org/10.1016/j.biotechadv.2017.05.004.

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8

Kis, Zoltán, Hugo Sant'Ana Pereira, Takayuki Homma, Ryan M. Pedrigi, and Rob Krams. "Mammalian synthetic biology: emerging medical applications." Journal of The Royal Society Interface 12, no. 106 (May 2015): 20141000. http://dx.doi.org/10.1098/rsif.2014.1000.

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In this review, we discuss new emerging medical applications of the rapidly evolving field of mammalian synthetic biology. We start with simple mammalian synthetic biological components and move towards more complex and therapy-oriented gene circuits. A comprehensive list of ON–OFF switches, categorized into transcriptional, post-transcriptional, translational and post-translational, is presented in the first sections. Subsequently, Boolean logic gates, synthetic mammalian oscillators and toggle switches will be described. Several synthetic gene networks are further reviewed in the medical applications section, including cancer therapy gene circuits, immuno-regulatory networks, among others. The final sections focus on the applicability of synthetic gene networks to drug discovery, drug delivery, receptor-activating gene circuits and mammalian biomanufacturing processes.
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9

Papathanasiou, Maria M., Baris Burnak, Justin Katz, Nilay Shah, and Efstratios N. Pistikopoulos. "Assisting continuous biomanufacturing through advanced control in downstream purification." Computers & Chemical Engineering 125 (June 2019): 232–48. http://dx.doi.org/10.1016/j.compchemeng.2019.03.013.

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10

Skylar-Scott, Mark A., Sebastien G. M. Uzel, Lucy L. Nam, John H. Ahrens, Ryan L. Truby, Sarita Damaraju, and Jennifer A. Lewis. "Biomanufacturing of organ-specific tissues with high cellular density and embedded vascular channels." Science Advances 5, no. 9 (September 2019): eaaw2459. http://dx.doi.org/10.1126/sciadv.aaw2459.

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Engineering organ-specific tissues for therapeutic applications is a grand challenge, requiring the fabrication and maintenance of densely cellular constructs composed of ~108 cells/ml. Organ building blocks (OBBs) composed of patient-specific–induced pluripotent stem cell–derived organoids offer a pathway to achieving tissues with the requisite cellular density, microarchitecture, and function. However, to date, scant attention has been devoted to their assembly into 3D tissue constructs. Here, we report a biomanufacturing method for assembling hundreds of thousands of these OBBs into living matrices with high cellular density into which perfusable vascular channels are introduced via embedded three-dimensional bioprinting. The OBB matrices exhibit the desired self-healing, viscoplastic behavior required for sacrificial writing into functional tissue (SWIFT). As an exemplar, we created a perfusable cardiac tissue that fuses and beats synchronously over a 7-day period. Our SWIFT biomanufacturing method enables the rapid assembly of perfusable patient- and organ-specific tissues at therapeutic scales.
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11

Patrício, Tatiana, Antonio Gloria, and Paulo J. Bártolo. "PCL and PCL/PLA Scaffolds for Bone Tissue Regeneration." Advanced Materials Research 683 (April 2013): 168–71. http://dx.doi.org/10.4028/www.scientific.net/amr.683.168.

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This paper investigates the use of PCL and PCL/PLA scaffolds, produced using a novel additive biomanufacturing system called BioCell Printing, for bone tissue engineering applications. Results show that the BioCell Printing system produces scaffolds with regular and reproducible architecture, presenting no toxicity and enhancing cell attachment and proliferation. It was also possible to observe that the addition of PLA to PCL scaffolds strongly improves the biomechanical performance of the constructs.
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12

Buyel, J. F., E. Stöger, and L. Bortesi. "Targeted genome editing of plants and plant cells for biomanufacturing." Transgenic Research 30, no. 4 (March 1, 2021): 401–26. http://dx.doi.org/10.1007/s11248-021-00236-z.

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AbstractPlants have provided humans with useful products since antiquity, but in the last 30 years they have also been developed as production platforms for small molecules and recombinant proteins. This initially niche area has blossomed with the growth of the global bioeconomy, and now includes chemical building blocks, polymers and renewable energy. All these applications can be described as “plant molecular farming” (PMF). Despite its potential to increase the sustainability of biologics manufacturing, PMF has yet to be embraced broadly by industry. This reflects a combination of regulatory uncertainty, limited information on process cost structures, and the absence of trained staff and suitable manufacturing capacity. However, the limited adaptation of plants and plant cells to the requirements of industry-scale manufacturing is an equally important hurdle. For example, the targeted genetic manipulation of yeast has been common practice since the 1980s, whereas reliable site-directed mutagenesis in most plants has only become available with the advent of CRISPR/Cas9 and similar genome editing technologies since around 2010. Here we summarize the applications of new genetic engineering technologies to improve plants as biomanufacturing platforms. We start by identifying current bottlenecks in manufacturing, then illustrate the progress that has already been made and discuss the potential for improvement at the molecular, cellular and organism levels. We discuss the effects of metabolic optimization, adaptation of the endomembrane system, modified glycosylation profiles, programmable growth and senescence, protease inactivation, and the expression of enzymes that promote biodegradation. We outline strategies to achieve these modifications by targeted gene modification, considering case-by-case examples of individual improvements and the combined modifications needed to generate a new general-purpose “chassis” for PMF.
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13

Juarez, Paloma, Vikram Virdi, Ann Depicker, and Diego Orzaez. "Biomanufacturing of protective antibodies and other therapeutics in edible plant tissues for oral applications." Plant Biotechnology Journal 14, no. 9 (February 13, 2016): 1791–99. http://dx.doi.org/10.1111/pbi.12541.

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14

Guha, Suvajyoti, Leonard F. Pease, Kurt A. Brorson, Michael J. Tarlov, and Michael R. Zachariah. "Evaluation of electrospray differential mobility analysis for virus particle analysis: Potential applications for biomanufacturing." Journal of Virological Methods 178, no. 1-2 (December 2011): 201–8. http://dx.doi.org/10.1016/j.jviromet.2011.09.012.

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15

Nogueira, Diogo E. S., Joaquim M. S. Cabral, and Carlos A. V. Rodrigues. "Single-Use Bioreactors for Human Pluripotent and Adult Stem Cells: Towards Regenerative Medicine Applications." Bioengineering 8, no. 5 (May 17, 2021): 68. http://dx.doi.org/10.3390/bioengineering8050068.

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Research on human stem cells, such as pluripotent stem cells and mesenchymal stromal cells, has shown much promise in their use for regenerative medicine approaches. However, their use in patients requires large-scale expansion systems while maintaining the quality of the cells. Due to their characteristics, bioreactors have been regarded as ideal platforms to harbour stem cell biomanufacturing at a large scale. Specifically, single-use bioreactors have been recommended by regulatory agencies due to reducing the risk of product contamination, and many different systems have already been developed. This review describes single-use bioreactor platforms which have been used for human stem cell expansion and differentiation, along with their comparison with reusable systems in the development of a stem cell bioprocess for clinical applications.
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16

Domingos, Marco, Dinuccio Dinucci, Stefania Cometa, Michele Alderighi, Paulo Jorge Bártolo, and Federica Chiellini. "Polycaprolactone Scaffolds Fabricated via Bioextrusion for Tissue Engineering Applications." International Journal of Biomaterials 2009 (2009): 1–9. http://dx.doi.org/10.1155/2009/239643.

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The most promising approach in Tissue Engineering involves the seeding of porous, biocompatible/biodegradable scaffolds, with donor cells to promote tissue regeneration. Additive biomanufacturing processes are increasingly recognized as ideal techniques to produce 3D structures with optimal pore size and spatial distribution, providing an adequate mechanical support for tissue regeneration while shaping in-growing tissues. This paper presents a novel extrusion-based system to produce 3D scaffolds with controlled internal/external geometry for TE applications.The BioExtruder is a low-cost system that uses a proper fabrication code based on the ISO programming language enabling the fabrication of multimaterial scaffolds. Poly(ε-caprolactone) was the material chosen to produce porous scaffolds, made by layers of directionally aligned microfilaments. Chemical, morphological, and in vitro biological evaluation performed on the polymeric constructs revealed a high potential of the BioExtruder to produce 3D scaffolds with regular and reproducible macropore architecture, without inducing relevant chemical and biocompatibility alterations of the material.
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Scognamiglio, Viviana, Maria Teresa Giardi, Daniele Zappi, Eleftherios Touloupakis, and Amina Antonacci. "Photoautotrophs–Bacteria Co-Cultures: Advances, Challenges and Applications." Materials 14, no. 11 (June 2, 2021): 3027. http://dx.doi.org/10.3390/ma14113027.

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Photosynthetic microorganisms are among the fundamental living organisms exploited for millennia in many industrial applications, including the food chain, thanks to their adaptable behavior and intrinsic proprieties. The great multipotency of these photoautotroph microorganisms has been described through their attitude to become biofarm for the production of value-added compounds to develop functional foods and personalized drugs. Furthermore, such biological systems demonstrated their potential for green energy production (e.g., biofuel and green nanomaterials). In particular, the exploitation of photoautotrophs represents a concrete biorefinery system toward sustainability, currently a highly sought-after concept at the industrial level and for the environmental protection. However, technical and economic issues have been highlighted in the literature, and in particular, challenges and limitations have been identified. In this context, a new perspective has been recently considered to offer solutions and advances for the biomanufacturing of photosynthetic materials: the co-culture of photoautotrophs and bacteria. The rational of this review is to describe the recently released information regarding this microbial consortium, analyzing the critical issues, the strengths and the next challenges to be faced for the intentions attainment.
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18

Li, Guowei, Xinlei Wei, Ranran Wu, Wei Zhou, Yunjie Li, Zhiguang Zhu, and Chun You. "Stoichiometric Conversion of Maltose for Biomanufacturing by In Vitro Synthetic Enzymatic Biosystems." BioDesign Research 2022 (July 1, 2022): 1–11. http://dx.doi.org/10.34133/2022/9806749.

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Maltose is a natural α-(1,4)-linked disaccharide with wide applications in food industries and microbial fermentation. However, maltose has scarcely been used for in vitro biosynthesis, possibly because its phosphorylation by maltose phosphorylase (MP) yields β-glucose 1-phosphate (β-G1P) that cannot be utilized by α-phosphoglucomutase (α-PGM) commonly found in in vitro synthetic enzymatic biosystems previously constructed by our group. Herein, we designed an in vitro synthetic enzymatic reaction module comprised of MP, β-phosphoglucomutase (β-PGM), and polyphosphate glucokinase (PPGK) for the stoichiometric conversion of each maltose molecule to two glucose 6-phosphate (G6P) molecules. Based on this synthetic module, we further constructed two in vitro synthetic biosystems to produce bioelectricity and fructose 1,6-diphosphate (FDP), respectively. The 14-enzyme biobattery achieved a Faraday efficiency of 96.4% and a maximal power density of 0.6 mW/cm2, whereas the 5-enzyme in vitro FDP-producing biosystem yielded 187.0 mM FDP from 50 g/L (139 mM) maltose by adopting a fed-batch substrate feeding strategy. Our study not only suggests new application scenarios for maltose but also provides novel strategies for the high-efficient production of bioelectricity and value-added biochemicals.
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Jeon, Young Chan, Anh Duc Nguyen, and Eun Yeol Lee. "Bioproduction of Isoprenoids and Other Secondary Metabolites Using Methanotrophic Bacteria as an Alternative Microbial Cell Factory Option: Current Stage and Future Aspects." Catalysts 9, no. 11 (October 24, 2019): 883. http://dx.doi.org/10.3390/catal9110883.

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Methane is a promising carbon feedstock for industrial biomanufacturing because of its low price and high abundance. Recent advances in metabolic engineering and systems biology in methanotrophs have made it possible to produce a variety of value-added compounds from methane, including secondary metabolites. Isoprenoids are one of the largest family of secondary metabolites and have many useful industrial applications. In this review, we highlight the current efforts invested to methanotrophs for the production of isoprenoids and other secondary metabolites, including riboflavin and ectoine. The future outlook for improving secondary metabolites production (especially of isoprenoids) using metabolic engineering of methanotrophs is also discussed.
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Duchi, Serena, Stephanie Doyle, Timon Eekel, Cathal D. O’Connell, Cheryl Augustine, Peter Choong, Carmine Onofrillo, and Claudia Di Bella. "Protocols for Culturing and Imaging a Human Ex Vivo Osteochondral Model for Cartilage Biomanufacturing Applications." Materials 12, no. 4 (February 20, 2019): 640. http://dx.doi.org/10.3390/ma12040640.

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Cartilage defects and diseases remain major clinical issues in orthopaedics. Biomanufacturing is now a tangible option for the delivery of bioscaffolds capable of regenerating the deficient cartilage tissue. However, several limitations of in vitro and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. Ex vivo models are of great value for translating in vitro tissue engineered approaches into clinically relevant conditions. Our aim is to obtain a viable human osteochondral (OC) model to test hydrogel-based materials for cartilage repair. Here we describe a detailed step-by-step framework for the generation of human OC plugs, their culture in a perfusion device and the processing procedures for histological and advanced microscopy imaging. Our ex vivo OC model fulfils the following requirements: the model is metabolically stable for a relevant culture period of 4 weeks in a perfusion bioreactor, the processing procedures allowed for the analysis of 3 different tissues or materials (cartilage, bone and hydrogel) without compromising their integrity. We determined a protocol and the settings for a non-linear microscopy technique on label free sections. Furthermore, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical ex vivo tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for in vivo testing.
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Salmi, Mika. "Additive Manufacturing Processes in Medical Applications." Materials 14, no. 1 (January 3, 2021): 191. http://dx.doi.org/10.3390/ma14010191.

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Additive manufacturing (AM, 3D printing) is used in many fields and different industries. In the medical and dental field, every patient is unique and, therefore, AM has significant potential in personalized and customized solutions. This review explores what additive manufacturing processes and materials are utilized in medical and dental applications, especially focusing on processes that are less commonly used. The processes are categorized in ISO/ASTM process classes: powder bed fusion, material extrusion, VAT photopolymerization, material jetting, binder jetting, sheet lamination and directed energy deposition combined with classification of medical applications of AM. Based on the findings, it seems that directed energy deposition is utilized rarely only in implants and sheet lamination rarely for medical models or phantoms. Powder bed fusion, material extrusion and VAT photopolymerization are utilized in all categories. Material jetting is not used for implants and biomanufacturing, and binder jetting is not utilized for tools, instruments and parts for medical devices. The most common materials are thermoplastics, photopolymers and metals such as titanium alloys. If standard terminology of AM would be followed, this would allow a more systematic review of the utilization of different AM processes. Current development in binder jetting would allow more possibilities in the future.
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Brookwell, August, Javin P. Oza, and Filippo Caschera. "Biotechnology Applications of Cell-Free Expression Systems." Life 11, no. 12 (December 8, 2021): 1367. http://dx.doi.org/10.3390/life11121367.

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Cell-free systems are a rapidly expanding platform technology with an important role in the engineering of biological systems. The key advantages that drive their broad adoption are increased efficiency, versatility, and low cost compared to in vivo systems. Traditionally, in vivo platforms have been used to synthesize novel and industrially relevant proteins and serve as a testbed for prototyping numerous biotechnologies such as genetic circuits and biosensors. Although in vivo platforms currently have many applications within biotechnology, they are hindered by time-constraining growth cycles, homeostatic considerations, and limited adaptability in production. Conversely, cell-free platforms are not hindered by constraints for supporting life and are therefore highly adaptable to a broad range of production and testing schemes. The advantages of cell-free platforms are being leveraged more commonly by the biotechnology community, and cell-free applications are expected to grow exponentially in the next decade. In this study, new and emerging applications of cell-free platforms, with a specific focus on cell-free protein synthesis (CFPS), will be examined. The current and near-future role of CFPS within metabolic engineering, prototyping, and biomanufacturing will be investigated as well as how the integration of machine learning is beneficial to these applications.
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Lizana-Vasquez, Gaby D., Luis F. Arrieta-Viana, Janet Mendez-Vega, Aldo Acevedo, and Madeline Torres-Lugo. "Synthetic Thermo-Responsive Terpolymers as Tunable Scaffolds for Cell Culture Applications." Polymers 14, no. 20 (October 17, 2022): 4379. http://dx.doi.org/10.3390/polym14204379.

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The use of tailored synthetic hydrogels for in vitro tissue culture and biomanufacturing provides the advantage of mimicking the cell microenvironment without issues of batch-to-batch variability. To that end, this work focused on the design, characterization, and preliminary evaluation of thermo-responsive, transparent synthetic terpolymers based on N-isopropylacrylamide, vinylphenylboronic acid, and polyethylene glycol for cell manufacturing and in vitro culture applications. Polymer physical properties were characterized by FT-IR, 1H-NMR, DLS, rheology, and thermal-gravimetric analysis. Tested combinations provided polymers with a lower critical solution temperature (LCST) between 30 and 45 °C. Terpolymer elastic/shear modulus varied between 0.3 and 19.1 kPa at 37 °C. Cellular characterization indicated low cell cytotoxicity on NIH-3T3. Experiments with the ovarian cancer model SKOV-3 and Jurkat T cells showed the terpolymers’ capacity for cell encapsulation without interfering with staining or imaging protocols. In addition, cell growth and high levels of pluripotency demonstrated the capability of terpolymer to culture iPSCs. Characterization results confirmed a promising use of terpolymers as a tunable scaffold for cell culture applications.
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Guo, Yuanke, Yang Liao, Jing Wang, Chen Ma, Jialun Qin, Jiao Feng, Yan Li, Xin Wang, and Kequan Chen. "Methylotrophy of Pichia pastoris: Current Advances, Applications, and Future Perspectives for Methanol-Based Biomanufacturing." ACS Sustainable Chemistry & Engineering 10, no. 5 (January 27, 2022): 1741–52. http://dx.doi.org/10.1021/acssuschemeng.1c07755.

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Kao, Chen-Yuan, and Eleftherios T. Papoutsakis. "Extracellular vesicles: exosomes, microparticles, their parts, and their targets to enable their biomanufacturing and clinical applications." Current Opinion in Biotechnology 60 (December 2019): 89–98. http://dx.doi.org/10.1016/j.copbio.2019.01.005.

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Kunjapur, Aditya M., Michael G. Napolitano, Eriona Hysolli, Karen Noguera, Evan M. Appleton, Max G. Schubert, Michaela A. Jones, Siddharth Iyer, Daniel J. Mandell, and George M. Church. "Synthetic auxotrophy remains stable after continuous evolution and in coculture with mammalian cells." Science Advances 7, no. 27 (July 2021): eabf5851. http://dx.doi.org/10.1126/sciadv.abf5851.

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Understanding the evolutionary stability and possible context dependence of biological containment techniques is critical as engineered microbes are increasingly under consideration for applications beyond biomanufacturing. While synthetic auxotrophy previously prevented Escherichia coli from exhibiting detectable escape from batch cultures, its long-term effectiveness is unknown. Here, we report automated continuous evolution of a synthetic auxotroph while supplying a decreasing concentration of essential biphenylalanine (BipA). After 100 days of evolution, triplicate populations exhibit no observable escape and exhibit normal growth rates at 10-fold lower BipA concentration than the ancestral synthetic auxotroph. Allelic reconstruction reveals the contribution of three genes to increased fitness at low BipA concentrations. Based on its evolutionary stability, we introduce the progenitor strain directly to mammalian cell culture and observe containment of bacteria without detrimental effects on HEK293T cells. Overall, our findings reveal that synthetic auxotrophy is effective on time scales and in contexts that enable diverse applications.
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Xu, Junjie, Shijiao Ma, Haolan Zheng, Bo Pang, Shuli Li, Feng Li, Lin Feng, and Jiesheng Tian. "Biomanufacturing Biotinylated Magnetic Nanomaterial via Construction and Fermentation of Genetically Engineered Magnetotactic Bacteria." Bioengineering 9, no. 8 (July 30, 2022): 356. http://dx.doi.org/10.3390/bioengineering9080356.

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Biosynthesis provides a critical way to deal with global sustainability issues and has recently drawn increased attention. However, modifying biosynthesized magnetic nanoparticles by extraction is challenging, limiting its applications. Magnetotactic bacteria (MTB) synthesize single-domain magnetite nanocrystals in their organelles, magnetosomes (BMPs), which are excellent biomaterials that can be biologically modified by genetic engineering. Therefore, this study successfully constructed in vivo biotinylated BMPs in the MTB Magnetospirillum gryphiswaldense by fusing biotin carboxyl carrier protein (BCCP) with membrane protein MamF of BMPs. The engineered strain (MSR−∆F−BF) grew well and synthesized small-sized (20 ± 4.5 nm) BMPs and were cultured in a 42 L fermenter; the yield (dry weight) of cells and BMPs reached 8.14 g/L and 134.44 mg/L, respectively, approximately three-fold more than previously reported engineered strains and BMPs. The genetically engineered BMPs (BMP−∆F−BF) were successfully linked with streptavidin or streptavidin-labelled horseradish peroxidase and displayed better storage stability compared with chemically constructed biotinylated BMPs. This study systematically demonstrated the biosynthesis of engineered magnetic nanoparticles, including its construction, characterization, and production and detection based on MTB. Our findings provide insights into biomanufacturing multiple functional magnetic nanomaterials.
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Viana, Tânia, Sara Biscaia, Emilia Dabrowska, Margarida C. Franco, Pedro Carreira, Pedro Morouço, and Nuno Alves. "A Novel Biomanufacturing System to Produce Multi-Material Scaffolds for Tissue Engineering: Concept and Preliminary Results." Applied Mechanics and Materials 890 (April 2019): 283–89. http://dx.doi.org/10.4028/www.scientific.net/amm.890.283.

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To produce multi-material scaffolds for Tissue Engineering accurate techniques are needed in order to obtain three-dimensional constructs with clinically appropriate size and structural integrity. This paper presents a novel biomanufacturing system that can fabricate 3D scaffolds with precise shape and porosity which is achieved through the control of all fabrication modules by an integrated computational platform. The incorporation of a clean flow unit and a camera allows to obtain scaffolds in a clean environment and provides a monitoring tool to analyse constructs during the production, respectively. In this research work is demonstrated that the new system enables the fabrication of multi-material 3D structures using poly (e-caprolactone) and sodium alginate for potential use in Tissue Engineering applications.
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Meyer, Conary, Yusuke Nakamura, Blake J. Rasor, Ashty S. Karim, Michael C. Jewett, and Cheemeng Tan. "Analysis of the Innovation Trend in Cell-Free Synthetic Biology." Life 11, no. 6 (June 11, 2021): 551. http://dx.doi.org/10.3390/life11060551.

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Cell-free synthetic biology is a maturing field that aims to assemble biomolecular reactions outside cells for compelling applications in drug discovery, metabolic engineering, biomanufacturing, diagnostics, and education. Cell-free systems have several key features. They circumvent mechanisms that have evolved to facilitate species survival, bypass limitations on molecular transport across the cell wall, enable high-yielding and rapid synthesis of proteins without creating recombinant cells, and provide high tolerance towards toxic substrates or products. Here, we analyze ~750 published patents and ~2000 peer-reviewed manuscripts in the field of cell-free systems. Three hallmarks emerged. First, we found that both patent filings and manuscript publications per year are significantly increasing (five-fold and 1.5-fold over the last decade, respectively). Second, we observed that the innovation landscape has changed. Patent applications were dominated by Japan in the early 2000s before shifting to China and the USA in recent years. Finally, we discovered an increasing prevalence of biotechnology companies using cell-free systems. Our analysis has broad implications on the future development of cell-free synthetic biology for commercial and industrial applications.
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Mizutani, Masayoshi, and Tsunemoto Kuriyagawa. "Special Issue on Biomedical Applications." International Journal of Automation Technology 11, no. 6 (October 31, 2017): 861. http://dx.doi.org/10.20965/ijat.2017.p0861.

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Interdisciplinary research that integrates medical science, biotechnology, materials science, mechanical engineering, and manufacturing has seen rapid progress in recent years. Not only fundamental research into biological functions but also the development of clinical approaches to treating patients are being actively carried out by experts in different fields. For example, artificial materials, such as those used in orthopedic surgery and dental implants, are being used more widely in medical treatments. In the area of minimally invasive surgery using X-rays, CT, and MRI, medical devices possessing radiolucent and nonmagnetic properties are playing a major role. Medical auxiliary equipment, such as wheelchairs, prosthetic feet, and other objects used to supplement medical treatment, is also critical. To assure that such advances continue into the future, material development and manufacturing processes should eventually satisfy the requirements of medical and biological applications, which are being debated by experts in different fields. The applicable materials should have excellent specific strength and rigidity, high biocompatibility, and good formability. The various needs for material characteristics and functions make interdisciplinary research essential. Mechanical engineering and manufacturing technologies should be further developed to solve problems involved in the establishment of basic principles by integrating the knowledge of materials science, medical science, biology, chemistry, and other fields. This special issue addresses the latest research advances into the biomedical applications of different manufacturing technologies. This covers a wide area, including biotechnologies, biomanufacturing, biodevices, and biomedical technologies. We hope that learning more about these advances will enable the readers to share in the authors’ experience and knowledge of technologies and development. All papers were refereed through careful peer reviews. We would like express our sincere appreciation to the authors for their submissions and to the reviewers for their invaluable efforts, which have ensured the success of this special issue.
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Lim, Ai Chye, Yuhong Zhou, John Washbrook, Nigel John Titchener-Hooker, and Suzanne Farid. "A decisional-support tool to model the impact of regulatory compliance activities in the biomanufacturing industry." Computers & Chemical Engineering 28, no. 5 (May 2004): 727–35. http://dx.doi.org/10.1016/j.compchemeng.2004.02.013.

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McCracken, Reilly, Noor Al-Nazal, Travis Whitmer, Sijia Yi, James M. Wagner, Colin G. Hebert, Matthew J. Lowry, et al. "Rapid In-Process Measurement of Live Virus Vaccine Potency Using Laser Force Cytology: Paving the Way for Rapid Vaccine Development." Vaccines 10, no. 10 (September 22, 2022): 1589. http://dx.doi.org/10.3390/vaccines10101589.

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Vaccinations to prevent infectious diseases are given to target the body’s innate and adaptive immune systems. In most cases, the potency of a live virus vaccine (LVV) is the most critical measurement of efficacy, though in some cases the quantity of surface antigen on the virus is an equally critical quality attribute. Existing methods to measure the potency of viruses include plaque and TCID50 assays, both of which have very long lead times and cannot provide real time information on the quality of the vaccine during large-scale manufacturing. Here, we report the evaluation of LumaCyte’s Radiance Laser Force Cytology platform as a new way to measure the potency of LVVs in upstream biomanufacturing process in real time and compare this to traditional TCID50 potency. We also assess this new platform as a way to detect adventitious agents, which is a regulatory expectation for the release of commercial vaccines. In both applications, we report the ability to obtain expedited and relevant potency information with strong correlation to release potency methods. Together, our data propose the application of Laser Force Cytology as a valuable process analytical technology (PAT) for the timely measurement of critical quality attributes of LVVs.
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Reuel, Nigel F., Brittany Grassbaugh, Sebastian Kruss, J. Zachary Mundy, Cary Opel, Adebola O. Ogunniyi, Kamal Egodage, et al. "Emergent Properties of Nanosensor Arrays: Applications for Monitoring IgG Affinity Distributions, Weakly Affined Hypermannosylation, and Colony Selection for Biomanufacturing." ACS Nano 7, no. 9 (August 15, 2013): 7472–82. http://dx.doi.org/10.1021/nn403215e.

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Gil, Carmen J., Martin L. Tomov, Andrea S. Theus, Alexander Cetnar, Morteza Mahmoudi, and Vahid Serpooshan. "In Vivo Tracking of Tissue Engineered Constructs." Micromachines 10, no. 7 (July 16, 2019): 474. http://dx.doi.org/10.3390/mi10070474.

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To date, the fields of biomaterials science and tissue engineering have shown great promise in creating bioartificial tissues and organs for use in a variety of regenerative medicine applications. With the emergence of new technologies such as additive biomanufacturing and 3D bioprinting, increasingly complex tissue constructs are being fabricated to fulfill the desired patient-specific requirements. Fundamental to the further advancement of this field is the design and development of imaging modalities that can enable visualization of the bioengineered constructs following implantation, at adequate spatial and temporal resolution and high penetration depths. These in vivo tracking techniques should introduce minimum toxicity, disruption, and destruction to treated tissues, while generating clinically relevant signal-to-noise ratios. This article reviews the imaging techniques that are currently being adopted in both research and clinical studies to track tissue engineering scaffolds in vivo, with special attention to 3D bioprinted tissue constructs.
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Mathew, Robins, Robert McGee, Kevin Roche, Shada Warreth, and Nikolaos Papakostas. "Introducing Mobile Collaborative Robots into Bioprocessing Environments: Personalised Drug Manufacturing and Environmental Monitoring." Applied Sciences 12, no. 21 (October 27, 2022): 10895. http://dx.doi.org/10.3390/app122110895.

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Personalised therapeutic drugs are the future of the medical drug sector. For manufacturers, this will require the flexibility to produce many different unique batches within a given facility. This research paper aims to demonstrate the potential of mobile collaborative robots for improving current manufacturing practices in personalised therapeutics. The benefits and challenges of introducing robots in biologics are explored, including current practices, limitations, likely future practices, and the market outlook. Experiments demonstrating the application of a mobile collaborative robot to perform three different routine tasks is presented. These experiments include the transport of centrifugal tubes, manipulation of infusion bags, and scanning of Petri dishes for environmental monitoring. The investigations highlight the potential of collaborative mobile robotic platforms for automating the routine tasks carried out within the biomanufacturing sector.
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Podolsky, Igor A., Susanna Seppälä, Thomas S. Lankiewicz, Jennifer L. Brown, Candice L. Swift, and Michelle A. O'Malley. "Harnessing Nature's Anaerobes for Biotechnology and Bioprocessing." Annual Review of Chemical and Biomolecular Engineering 10, no. 1 (June 7, 2019): 105–28. http://dx.doi.org/10.1146/annurev-chembioeng-060718-030340.

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Industrial biotechnology has the potential to decrease our reliance on petroleum for fuel and bio-based chemical production and also enable valorization of waste streams. Anaerobic microorganisms thrive in resource-limited environments and offer an array of novel bioactivities in this regard that could revolutionize biomanufacturing. However, they have not been adopted for widespread industrial use owing to their strict growth requirements, limited number of available strains, difficulty in scale-up, and genetic intractability. This review provides an overview of current and future uses for anaerobes in biotechnology and bioprocessing in the postgenomic era. We focus on the recently characterized anaerobic fungi (Neocallimastigomycota) native to the digestive tract of large herbivores, which possess a trove of enzymes, pathways, transporters, and other biomolecules that can be harnessed for numerous biotechnological applications. Resolving current genetic intractability, scale-up, and cultivation challenges will unlock the potential of these lignocellulolytic fungi and other nonmodel micro-organisms to accelerate bio-based production.
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Schnellbaecher, Alisa, Anton Lindig, Maxime Le Mignon, Tim Hofmann, Brit Pardon, Stephanie Bellmaine, and Aline Zimmer. "Degradation Products of Tryptophan in Cell Culture Media: Contribution to Color and Toxicity." International Journal of Molecular Sciences 22, no. 12 (June 9, 2021): 6221. http://dx.doi.org/10.3390/ijms22126221.

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Biomanufacturing processes may be optimized by storing cell culture media at room temperature, but this is currently limited by their instability and change in color upon long-term storage. This study demonstrates that one of the critical contributing factors toward media browning is tryptophan. LC-MS technology was utilized to identify tryptophan degradation products, which are likely formed primarily from oxidation reactions. Several of the identified compounds were shown to contribute significantly to color in solutions but also to exhibit toxicity against CHO cells. A cell-culture-compatible antioxidant, a-ketoglutaric acid, was found to be an efficient cell culture media additive for stabilizing components against degradation, inhibiting the browning of media formulations, and decreasing ammonia production, thus providing a viable method for developing room-temperature stable cell culture media.
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Arab, Bahareh, Adam Westbrook, Murray Moo-Young, and Chih-Hsiung Perry Chou. "A Toolkit for Effective and Successive Genome Engineering of Escherichia coli." Fermentation 9, no. 1 (December 23, 2022): 14. http://dx.doi.org/10.3390/fermentation9010014.

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The bacterium Escherichia coli has been well-justified as an effective workhorse for industrial applications. In this study, we developed a toolkit for flexible genome engineering of this microorganism, including site-specific insertion of heterologous genes and inactivation of endogenous genes, such that bacterial hosts can be effectively engineered for biomanufacturing. We first constructed a base strain by genomic implementation of the cas9 and λRed recombineering genes. Then, we constructed plasmids for expressing gRNA, DNA cargo, and the Vibrio cholerae Tn6677 transposon and type I-F CRISPR-Cas machinery. Genomic insertion of a DNA cargo up to 5.5 kb was conducted using a transposon-associated CRISPR-Cas system, whereas gene inactivation was mediated by a classic CRISPR-Cas9 system coupled with λRed recombineering. With this toolkit, we can exploit the synergistic functions of CRISPR-Cas, λRed recombineering, and Tn6677 transposon for successive genomic manipulations. As a demonstration, we used the developed toolkit to derive a plasmid-free strain for heterologous production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by genomic knock-in and knockout of several key genes with high editing efficiencies.
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Fernandes, Rita P., José M. Escandell, Ana C. L. Guerreiro, Filipa Moura, Tiago Q. Faria, Sofia B. Carvalho, Ricardo J. S. Silva, Patrícia Gomes-Alves, and Cristina Peixoto. "Assessing Multi-Attribute Characterization of Enveloped and Non-Enveloped Viral Particles by Capillary Electrophoresis." Viruses 14, no. 11 (November 17, 2022): 2539. http://dx.doi.org/10.3390/v14112539.

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Virus-based biopharmaceutical products are used in clinical applications such as vaccines, gene therapy, and immunotherapy. However, their manufacturing remains a challenge, hampered by the lack of appropriate analytical tools for purification monitoring or characterization of the final product. This paper describes the implementation of a highly sensitive method, capillary electrophoresis (CE)-sodium dodecyl sulfate (SDS) combined with a laser-induced fluorescence (LIF) detector to monitor the impact of various bioprocess steps on the quality of different viral vectors. The fluorescence labelling procedure uses the (3-(2-furoyl) quinoline-2-carboxaldehyde dye, and the CE-SDS LIF method enables the evaluation of in-process besides final product samples. This method outperforms other analytical methods, such as SDS–polyacrylamide gel electrophoresis with Sypro Ruby staining, in terms of sensitivity, resolution, and high-throughput capability. Notably, this CE-SDS LIF method was also successfully implemented to characterize enveloped viruses such as Maraba virus and lentivirus, whose development as biopharmaceuticals is now restricted by the lack of suitable analytical tools. This method was also qualified for quantification of rAAV2 according to the International Council for Harmonisation guidelines. Overall, our work shows that CE-SDS LIF is a precise and sensitive analytical platform for in-process sample analysis and quantification of different virus-based targets, with a great potential for application in biomanufacturing.
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Song, Guotian, Fengli Wu, Yanfeng Peng, Xiaolong Jiang, and Qinhong Wang. "High-Level Production of Catechol from Glucose by Engineered Escherichia coli." Fermentation 8, no. 7 (July 21, 2022): 344. http://dx.doi.org/10.3390/fermentation8070344.

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Catechol (CA) is an aromatic compound with important applications in the fine chemical and pharmaceutical fields. As an alternative strategy to petroleum-based chemical synthesis, the production of catechol by using microbial cell factories has attracted great interest. However, the toxicity of catechol to microbial cells significantly limits the efficient production of bio-based catechol via one-step fermentation. Therefore, in this study, a two-step strategy for the efficient synthesis of CA was designed. Protocatechuic acid (PCA) was first efficiently produced by the engineered Escherichia coli strain AAA01 via fermentation, and then PCA in the fermentative broth was converted into CA by the whole-cell biocatalyst AAA12 with PCA decarboxylase. By optimizing the expression of flavin isoprenyl transferases and protocatechuic acid decarboxylases, the titer of CA increased from 3.4 g/L to 15.8 g/L in 12 h through whole-cell biocatalysis, with a 365% improvement; after further optimizing the reaction conditions for whole-cell biocatalysis, the titer of CA achieved 17.7 g/L within 3 h, which is the highest titer reported so far. This work provides an effective strategy for the green biomanufacturing of toxic compounds by Escherichia coli cell factories.
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Tabuchi, Takeshi, and Yohei Yokobayashi. "High-throughput screening of cell-free riboswitches by fluorescence-activated droplet sorting." Nucleic Acids Research 50, no. 6 (March 7, 2022): 3535–50. http://dx.doi.org/10.1093/nar/gkac152.

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Abstract Cell-free systems that display complex functions without using living cells are emerging as new platforms to test our understanding of biological systems as well as for practical applications such as biosensors and biomanufacturing. Those that use cell-free protein synthesis (CFPS) systems to enable genetically programmed protein synthesis have relied on genetic regulatory components found or engineered in living cells. However, biological constraints such as cell permeability, metabolic stability, and toxicity of signaling molecules prevent development of cell-free devices using living cells even if cell-free systems are not subject to such constraints. Efforts to engineer regulatory components directly in CFPS systems thus far have been based on low-throughput experimental approaches, limiting the availability of basic components to build cell-free systems with diverse functions. Here, we report a high-throughput screening method to engineer cell-free riboswitches that respond to small molecules. Droplet-sorting of riboswitch variants in a CFPS system rapidly identified cell-free riboswitches that respond to compounds that are not amenable to bacterial screening methods. Finally, we used a histamine riboswitch to demonstrate chemical communication between cell-sized droplets.
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Ye, Jian-Wen, and Guo-Qiang Chen. "Halomonas as a chassis." Essays in Biochemistry 65, no. 2 (July 2021): 393–403. http://dx.doi.org/10.1042/ebc20200159.

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Abstract With the rapid development of systems and synthetic biology, the non-model bacteria, Halomonas spp., have been developed recently to become a cost-competitive platform for producing a variety of products including polyesters, chemicals and proteins owing to their contamination resistance and ability of high cell density growth at alkaline pH and high salt concentration. These salt-loving microbes can partially solve the challenges of current industrial biotechnology (CIB) which requires high energy-consuming sterilization to prevent contamination as CIB is based on traditional chassis, typically, Escherichia coli, Bacillus subtilis, Pseudomonas putida and Corynebacterium glutamicum. The advantages and current status of Halomonas spp. including their molecular biology and metabolic engineering approaches as well as their applications are reviewed here. Moreover, a systematic strain engineering streamline, including product-based host development, genetic parts mining, static and dynamic optimization of modularized pathways and bioprocess-inspired cell engineering are summarized. All of these developments result in the term called next-generation industrial biotechnology (NGIB). Increasing efforts are made to develop their versatile cell factories powered by synthetic biology to demonstrate a new biomanufacturing strategy under open and continuous processes with significant cost-reduction on process complexity, energy, substrates and fresh water consumption.
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Kwee, Edward, Edward E. Herderick, Thomas Adams, James Dunn, Robert Germanowski, Frank Krakosh, Cynthia Boehm, James Monnich, Kimerly Powell, and George Muschler. "Integrated Colony Imaging, Analysis, and Selection Device for Regenerative Medicine." SLAS TECHNOLOGY: Translating Life Sciences Innovation 22, no. 2 (November 11, 2016): 217–23. http://dx.doi.org/10.1177/2211068216676587.

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Stem and progenitor cells derived from human tissues are being developed as cell sources for cell-based assays and therapies. However, tissue-derived stem and progenitor cells are heterogeneous. Differences in observed clones of stem cells likely reflect important aspects of the underlying state of the source cells, as well as future potency for cell therapies. This paper describes a colony analysis and picking device that provides quantitative analysis of heterogeneous cell populations and precise tools for cell picking for research or biomanufacturing applications. We describe an integrated robotic system that enables image acquisition and automated image analysis to be coupled with rapid automated selection of individual colonies in adherent cell cultures. Other automated systems have demonstrated feasibility with picking from semisolid media or off feeder layers. We demonstrate the capability to pick adherent bone-derived stem cells from tissue culture plastic. Cells are efficiently picked from a target site and transferred to a recipient well plate. Cells demonstrate viability and adherence and maintain biologic potential for surface markers CD73 and CD90 based on phase contrast and fluorescence imaging 6 days after transfer. Methods developed here can be applied to the study of other stem cell types and automated culture of cells.
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Chen, Guimin, Anton Stepanenko, Olha Lakhneko, Yuzhen Zhou, Olena Kishchenko, Anton Peterson, Dandan Cui, et al. "Biodiversity of Duckweed (Lemnaceae) in Water Reservoirs of Ukraine and China Assessed by Chloroplast DNA Barcoding." Plants 11, no. 11 (May 30, 2022): 1468. http://dx.doi.org/10.3390/plants11111468.

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Monitoring and characterizing species biodiversity is essential for germplasm preservation, academic studies, and various practical applications. Duckweeds represent a group of tiny aquatic plants that include 36 species divided into 5 genera within the Lemnaceae family. They are an important part of aquatic ecosystems worldwide, often covering large portions of the water reservoirs they inhabit, and have many potential applications, including in bioremediation, biofuels, and biomanufacturing. Here, we evaluated the biodiversity of duckweeds in Ukraine and Eastern China by characterizing specimens using the two-barcode protocol with the chloroplast atpH–atpF and psbK–psbI spacer sequences. In total, 69 Chinese and Ukrainian duckweed specimens were sequenced. The sequences were compared against sequences in the NCBI database using BLAST. We identified six species from China (Spirodela polyrhiza, Landoltia punctata, Lemna aequinoctialis, Lemna minor, Lemna turionifera, and Wolffia globosa) and six from Ukraine (S. polyrhiza, Lemna gibba, Lemna minor, Lemna trisulca, Lemna turionifera, and Wolffia arrhiza). The most common duckweed species in the samples from Ukraine were Le. minor and S. polyrhiza, accounting for 17 and 15 out of 40 specimens, respectively. The most common duckweed species in the samples from China was S. polyrhiza, accounting for 15 out of 29 specimens. La. punctata and Le. aequinoctialis were also common in China, accounting for five and four specimens, respectively. According to both atpH–atpF and psbK–psbI barcode analyses, the species identified as Le. aequinoctialis does not form a uniform taxon similar to other duckweed species, and therefore the phylogenetic status of this species requires further clarification. By monitoring duckweeds using chloroplast DNA sequencing, we not only precisely identified local species and ecotypes, but also provided background for further exploration of native varieties with diverse genetic backgrounds. These data could be useful for future conservation, breeding, and biotechnological applications.
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Traustason, Bergthor, Matthew Cheeks, and Duygu Dikicioglu. "Computer-Aided Strategies for Determining the Amino Acid Composition of Medium for Chinese Hamster Ovary Cell-Based Biomanufacturing Platforms." International Journal of Molecular Sciences 20, no. 21 (November 2, 2019): 5464. http://dx.doi.org/10.3390/ijms20215464.

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Chinese hamster ovary (CHO) cells are used for the production of the majority of biopharmaceutical drugs, and thus have remained the standard industry host for the past three decades. The amino acid composition of the medium plays a key role in commercial scale biologics manufacturing, as amino acids constitute the building blocks of both endogenous and heterologous proteins, are involved in metabolic and non-metabolic pathways, and can act as main sources of nitrogen and carbon under certain conditions. As biomanufactured proteins become increasingly complex, the adoption of model-based approaches become ever more popular in complementing the challenging task of medium development. The extensively studied amino acid metabolism is exceptionally suitable for such model-driven analyses, and although still limited in practice, the development of these strategies is gaining attention, particularly in this domain. This paper provides a review of recent efforts. We first provide an overview of the widely adopted practice, and move on to describe the model-driven approaches employed for the improvement and optimization of the external amino acid supply in light of cellular amino acid demand. We conclude by proposing the likely prevalent direction the field is heading towards, providing a critical evaluation of the current state and the future challenges and considerations.
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Liu, Fengyuan, Srichand Hinduja, and Paulo Bártolo. "User interface tool for a novel plasma-assisted bio-additive extrusion system." Rapid Prototyping Journal 24, no. 2 (March 12, 2018): 368–78. http://dx.doi.org/10.1108/rpj-07-2016-0115.

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Purpose This paper aims to describe the control software of a novel manufacturing system called plasma-assisted bio-extrusion system (PABS), designed to produce complex multi-material and functionally graded scaffolds for tissue engineering applications. This fabrication system combines multiple pressure-assisted and screw-assisted printing heads and plasma jets. Control software allows the users to create single or multi-material constructs with uniform pore size or pore size gradients by changing the operation parameters, such as geometric parameters, lay-down pattern, filament distance, feed rate and layer thickness, and to produce functional graded scaffolds with different layer-by-layer coating/surface modification strategies by using the plasma modification system. Design/methodology/approach MATLAB GUI is used to develop the software, including the design of the user interface and the implementation of all mathematical programing for both multi-extrusion and plasma modification systems. Findings Based on the user definition, G programing codes are generated, enabling full integration and synchronization with the hardware of PABS. Single, multi-material and functionally graded scaffolds can be obtained by manipulating different materials, scaffold designs and processing parameters. The software is easy to use, allowing the efficient control of the PABS even for the fabrication of complex scaffolds. Originality/value This paper introduces a novel additive manufacturing system for tissue engineering applications describing in detail the software developed to control the system. This new fabrication system represents a step forward regarding the current state-of-the-art technology in the field of biomanufacturing, enabling the design and fabrication of more effective scaffolds matching the mechanical and surface characteristics of the surrounding tissue and enabling the incorporation of high number of cells uniformly distributed and the introduction of multiple cell types with positional specificity.
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Patrício, T., M. Domingos, A. Gloria, U. D'Amora, J. F. Coelho, and P. J. Bártolo. "Fabrication and characterisation of PCL and PCL/PLA scaffolds for tissue engineering." Rapid Prototyping Journal 20, no. 2 (March 11, 2014): 145–56. http://dx.doi.org/10.1108/rpj-04-2012-0037.

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Purpose – The main purpose of this research work is to study the effect of poly lactic acid (PLA) addition into poly (e-caprolactone) (PCL) matrices, as well the influence of the mixing process on the morphological, thermal, chemical, mechanical and biological performance of the 3D constructs produced with a novel biomanufacturing device (BioCell Printing). Design/methodology/approach – Two mixing processes are used to prepare PCL/PLA blends, namely melt blending and solvent casting. PCL and PCL/PLA scaffolds are produced via BioCell Printing using a 300-μm nozzle, 0/90° lay down pattern and 350-μm pore size. Several techniques such as scanning electron microscopy (SEM), simultaneous thermal analyzer (STA), nuclear magnetic resonance (NMR), static compression analysis and Alamar BlueTM are used to evaluate scaffold's morphological, thermal, chemical, mechanical and biological properties. Findings – Results show that the addition of PLA to PCL scaffolds strongly improves the biomechanical performance of the constructs. Additionally, polymer blends obtained by solvent casting present better mechanical and biological properties, compared to blends prepared by melt blending. Originality/value – This paper undertakes a detailed study on the effect of the mixing process on the biomechanical properties of PCL/PLA scaffolds. Results will enable to prepare customized PCL/PLA scaffolds for tissue engineering applications with improved biological and mechanical properties, compared to PCL scaffolds alone. Additionally, the accuracy and reproducibility of by the BioCell Printing enables to modulate the micro/macro architecture of the scaffolds enhancing tissue regeneration.
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Aebischer, Megane K., Hugo Gizardin-Fredon, Honorine Lardeux, Dominik Kochardt, Carsten Elger, Markus Haindl, Raphael Ruppert, Davy Guillarme, and Valentina D’Atri. "Anion-Exchange Chromatography at the Service of Gene Therapy: Baseline Separation of Full/Empty Adeno-Associated Virus Capsids by Screening of Conditions and Step Gradient Elution Mode." International Journal of Molecular Sciences 23, no. 20 (October 15, 2022): 12332. http://dx.doi.org/10.3390/ijms232012332.

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Gene therapy is opening unprecedented opportunities for novel therapeutic approaches. Based on the concept of rescuing function mutations by co-expressing the correct gene to allow biological functions to be restored, it requires the use of viral vectors to ensure the proper delivery of therapeutic genes. In this context, recombinant adeno-associated viruses (rAAV) are the most widely used vectors. Their biomanufacturing process requires the insertion of the therapeutic gene into the rAAV (full capsids). However, a percentage of rAAV that do not contain the desired gene (empty capsids), as well as partly filled capsids, might also be produced, potentially impacting the efficiency of the therapy. Therefore, the determination of the rAAV capsids’ full/empty ratio needs to be monitored to ensure consistent product quality and efficacy. Anion-exchange chromatography (AEX) can serve this need. In this contribution, thorough AEX method development, including a mobile phase, a stationary phase and gradient conditions, has highlighted its potential in supporting gene therapy. Taking advantage of the fact that viral capsids follow an “on/off” retention behavior, the application of a step gradient approach to the rAAV serotype 8 (rAAV8) allowed the unprecedented separation of rAAV8 full/empty capsids, with a resolution gain of 3.7 as compared to the resolution obtained with a fully optimized linear gradient. Finally, the developed analytical approach allowed a precise and accurate baseline separation and quantification of full and empty rAAV8 capsids, with the potential to be applied as a high-throughput quality control (QC) method.
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Fricke, Philipp Moritz, Angelika Klemm, Michael Bott, and Tino Polen. "On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases." Applied Microbiology and Biotechnology 105, no. 9 (April 15, 2021): 3423–56. http://dx.doi.org/10.1007/s00253-021-11269-z.

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Abstract Acetic acid bacteria (AAB) are valuable biocatalysts for which there is growing interest in understanding their basics including physiology and biochemistry. This is accompanied by growing demands for metabolic engineering of AAB to take advantage of their properties and to improve their biomanufacturing efficiencies. Controlled expression of target genes is key to fundamental and applied microbiological research. In order to get an overview of expression systems and their applications in AAB, we carried out a comprehensive literature search using the Web of Science Core Collection database. The Acetobacteraceae family currently comprises 49 genera. We found overall 6097 publications related to one or more AAB genera since 1973, when the first successful recombinant DNA experiments in Escherichia coli have been published. The use of plasmids in AAB began in 1985 and till today was reported for only nine out of the 49 AAB genera currently described. We found at least five major expression plasmid lineages and a multitude of further expression plasmids, almost all enabling only constitutive target gene expression. Only recently, two regulatable expression systems became available for AAB, an N-acyl homoserine lactone (AHL)-inducible system for Komagataeibacter rhaeticus and an l-arabinose-inducible system for Gluconobacter oxydans. Thus, after 35 years of constitutive target gene expression in AAB, we now have the first regulatable expression systems for AAB in hand and further regulatable expression systems for AAB can be expected. Key points • Literature search revealed developments and usage of expression systems in AAB. • Only recently 2 regulatable plasmid systems became available for only 2 AAB genera. • Further regulatable expression systems for AAB are in sight.
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Chan, Weng Wan, David Chen Loong Yeo, Vernice Tan, Satnam Singh, Deepak Choudhury, and May Win Naing. "Additive Biomanufacturing with Collagen Inks." Bioengineering 7, no. 3 (July 1, 2020): 66. http://dx.doi.org/10.3390/bioengineering7030066.

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Collagen is a natural polymer found abundantly in the extracellular matrix (ECM). It is easily extracted from a variety of sources and exhibits excellent biological properties such as biocompatibility and weak antigenicity. Additionally, different processes allow control of physical and chemical properties such as mechanical stiffness, viscosity and biodegradability. Moreover, various additive biomanufacturing technology has enabled layer-by-layer construction of complex structures to support biological function. Additive biomanufacturing has expanded the use of collagen biomaterial in various regenerative medicine and disease modelling application (e.g., skin, bone and cornea). Currently, regulatory hurdles in translating collagen biomaterials still remain. Additive biomanufacturing may help to overcome such hurdles commercializing collagen biomaterials and fulfill its potential for biomedicine.
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