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Biotechnology Journal

Andrea Palermo, Laura K Weber, Simone Rentschler, Awale Isse, Martyna Sedlmayr, Karin Herbster, Volker List, Jürgen Hubbuch, Felix F Löffler, Alexander Nesterov-Müller, Frank Breitling
Vaccinations are among the most potent tools to fight infectious diseases. However, cross-reactions are an ongoing problem and there is an urgent need to fully understand the mechanisms of the immune response. For the development of a methodological workflow, the linear epitopes in the immune response to the tetanus toxin is investigated in sera of 19 vaccinated Europeans applying epitope mapping with peptide arrays. The most prominent epitope, appearing in nine different sera (923 IHLVNNESSEVIVHK937 ), is investigated in a substitution analysis to identify the amino acids that are crucial for the binding of the corresponding antibody species - the antibody fingerprint...
September 18, 2017: Biotechnology Journal
Marta Alves da Silva, Albino Martins, Ana R Costa-Pinto, Nélson Monteiro, Susana Faria, Rui L Reis, Nuno M Neves
Many efforts are being directed worldwide to the treatment of OA-focal lesions. The majority of those efforts comprise either the refinement of surgical techniques or combinations of biomaterials with various autologous cells. Herein, we tested electrospun polycaprolactone (PCL) nanofibrous meshes for cartilage tissue engineering. For that, articular chondrocytes (hACs) isolated from human osteoarthritic joints and Wharton's Jelly Stem Cells (hWJSCs) were cultured on electrospun nanofiber meshes, without adding external growth factors...
September 13, 2017: Biotechnology Journal
Alexander C Schmitz, Christopher J Hartline, Fuzhong Zhang
Monoclonal antibodies currently dominate the biopharmaceutical market with growing sales having reached 80 billion USD in 2016. As most top-selling mAbs are approaching the end of their patent life, biopharmaceutical companies compete fiercely in the biosimilars market. These two factors present a strong motivation for alternative process strategies and process optimization. In this work a novel purification strategy for monoclonal antibodies comprising phenylboronic acid multimodal chromatography for capture followed by polishing by ion exchange monolithic chromatography and packed bed hydrophobic interaction chromatography is presented and compared to the traditional protein-A-based process...
September 13, 2017: Biotechnology Journal
Eva Setter-Lamed, Sarah Moraïs, Johanna Stern, Raphael Lamed, Edward A Bayer
Cellulose deconstruction can be achieved by three distinct enzymatic paradigms: free enzymes, multifunctional enzymes and self-assembled, multi-enzyme complexes (cellulosomes). To study their comparative efficiency, the simple and efficient cellulolytic system of the aerobic bacterium, Thermobifida fusca, was developed as an enzymatic model. In previous studies, most of its cellulases were successfully converted to the cellulosomal mode and exhibited high cellulolytic activities, except for Cel6B, a key exoglucanase of the T...
September 13, 2017: Biotechnology Journal
Karthikeyan Narayanan, Sachin Mishra, Satnam Singh, Ming Pei, Balazs Gulyas, Parasuraman Padmanabhan
The field of regenerative medicine integrates advancements made in stem cells, molecular biology, engineering and clinical methodologies. Stem cells serve as a fundamental ingredient for therapeutic application in regenerative medicine. Apart from stem cells, engineering concepts have equally contributed to the success of stem cell based applications in improving human health. The purpose of various engineering methodologies is to develop regenerative and preventive medicine to combat various diseases and deformities...
September 13, 2017: Biotechnology Journal
António Lima Grilo, Marilia Mateus, M Raquel Aires-Barros, Ana M Azevedo
Monoclonal antibodies currently dominate the biopharmaceutical market with growing sales having reached 80 billion USD in 2016. As most top-selling mAbs are approaching the end of their patent life, biopharmaceutical companies compete fiercely in the biosimilars market. These two factors present a strong motivation for alternative process strategies and process optimization. In this work a novel purification strategy for monoclonal antibodies comprising phenylboronic acid multimodal chromatography for capture followed by polishing by ion exchange monolithic chromatography and packed bed hydrophobic interaction chromatography is presented and compared to the traditional protein-A-based process...
September 13, 2017: Biotechnology Journal
Benjamin C U Tai, Chan Du, Shujun Gao, Andrew C A Wan
Cell-based therapies for cartilage repair are continually being developed to treat osteoarthritis. The cells are either introduced directly by intra-articular injection or via a cell-seeded matrix scaffold. Here, we developed poly(vinylalcohol)-based membranes to be used for mesenchymal stem cell implantation in cartilage repair procedures, having controllable physicochemical properties such as porosity, mechanical strength and permeability, and a unique self-sealing property. The membranes possess a bilayer structure with a less porous layer providing mechanical strength and selective permeability, exhibit an elastic modulus of between 0...
September 11, 2017: Biotechnology Journal
Devi Kasi, Hee Ju Nah, Christy Catherine, Eung-Soo Kim, Kyubeom Han, Jong-Cheon Ha, Dong-Myung Kim
High-yield production of soluble protein is a common concern in diverse fields of biotechnology. In this study, a strategy of using an engineered nucleotide sequence of ubiquitin for enhancing the production of soluble proteins in a cell-free synthesis system is presented. When examined for a series of proteins that otherwise were poorly expressed, N-terminal fusion with ubiquitin significantly increased both the expression levels and solubility of the translational products. The effect of ubiquitin fusion was also markedly augmented by engineering the nucleotide sequence of ubiquitin, leading to several fold enhancements in soluble production of target proteins...
September 11, 2017: Biotechnology Journal
Lars Freier, Eric von Lieres
A novel algorithm for robust multi-objective process optimization under stochastic variability of environmental variables is introduced and applied to a case study in gradient elution chromatography. Process variability is accounted for by simultaneously optimizing several scenarios with random but fixed values of the environmental variables. These iterative optimizations are synchronized by planning the same experiments for all scenarios. Experiments are designed by maximizing the cumulative expected hypervolume improvement as predicted by several Gaussian process regression models...
September 9, 2017: Biotechnology Journal
Bao-Ngoc Pham-Hoang, Pascale Winckler, Yves Waché
Quercetin is a fragile bioactive compound. Several works have tried to preserve it by encapsulation but the form of encapsulation (mono- or supra-molecular structure, tautomeric form), though important for stability and bioavailability, remains unknown. The present work aims at developing a fluorescence lifetime technique to evaluate the structure of quercetin during encapsulation in a vector capsule that has already proven efficiency, yeast cells. Molecular stabilization was observed during a four-month storage period...
September 9, 2017: Biotechnology Journal
Hideto Nakabayashi, Masahiro Kawahara, Teruyuki Nagamune
As receptor tyrosine kinases (RTKs) play important roles in cell-fate control of various cell types, engineered RTKs that could respond to inexpensive ligands might drastically reduce the cost of producing desired cells for various applications in regenerative medicine. We developed several engineered RTKs named "signalobodies" in which the ligand-recognition domain of RTKs is replaced by single-chain Fv for enabling recognition of a specific antigen. However, the remaining concern was the dysregulation of antigen-dependent on/off signaling of the signalobodies...
September 7, 2017: Biotechnology Journal
Danielle J Maddock, Monica L Gerth, Wayne M Patrick
There is substantial interest in engineering microorganisms to produce industrial chemicals that are currently derived from petroleum. One of these petrochemicals is butanone, which could be produced from microbially synthesized 2,3-butanediol through the action of a suitable dehydratase enzyme. Unfortunately, however, there are no known enzymes that natively catalyze this reaction. In this work, we set out to engineer the B12 -dependent glycerol dehydratase from Klebsiella pneumoniae (KpGDHt), in order to increase its activity for the conversion of meso-2,3-butanediol into butanone...
September 7, 2017: Biotechnology Journal
Xinjuan Ning, Xinran Wang, Yuanting Wu, Qianjin Kang, Linquan Bai
The type-I polyketide Ansamitocin P-3 (AP-3) is a potent antitumor agent. Its production is most likely hampered by the required multiple substrate supplies and complicated post-PKS modifications in Actinosynnema pretiosum subsp. pretiosum ATCC 31280. For titer improvement, gene ansa30, encoding for a glycosyltransferase competing for the N-demethyl-AP-3 (PND-3) intermediate for AP-3 biosynthesis, was initially inactivated. In the mutant NXJ-22, the AP-3 titer was increased by 66% along with an obvious accumulation of PND-3, indicating that the N-methylation is a rate-limiting step...
September 7, 2017: Biotechnology Journal
Nikhil Kateja, Devashish Kumar, Avinash Godara, Vijesh Kumar, Anurag S Rathore
Achieving consistent product quality of a biotherapeutic is a major target for any biopharmaceutical manufacturer, even more for a biosimilar producer as comparability with the innovator product is a regulatory expectation. The complexity of biotherapeutic products and their tedious manufacturing processes, however, make this a non-trivial exercise. The primary motivation of this work is to develop an integrated chromatographic platform for purification of monoclonal antibody (mAb) therapeutics that can deliver the desired separation of both charge variants and aggregates, in addition to the process related impurities like host cell proteins (HCP) and host cell DNA...
September 7, 2017: Biotechnology Journal
Dmitry Gil, Anastasia Frank-Kamenetskii, John Barry, Vladimir Reukov, Yun Xiang, Arabinda Das, Abhay K Varma, Mark S Kindy, Naren L Banik, Alexey Vertegel
Polymeric nanoparticles have been extensively studied as drug delivery vehicles both in vitro and in vivo for the last two decades. In vitro methods to assess drug release profiles usually utilize degradation of nanoparticles in aqueous medium, followed by the measurement of the concentration of the released drug. This method, however, is difficult to use for drugs that are poorly water soluble. In this study, we describe a protocol for measuring drug release kinetic using albumin solution as the medium. Albumin is a major blood transport protein, which mediates transport of many lipid soluble compounds including fatty acids, hormones, and bilirubin...
September 7, 2017: Biotechnology Journal
Allison I Hoch, Ralph Duhr, Nunzia Di Maggio, Arne Mehrkens, Marcel Jakob, David Wendt
Bone marrow-derived mesenchymal stromal cells (BMSC), when expanded directly within 3D ceramic scaffolds in perfusion bioreactors, more reproducibly form bone when implanted in vivo as compared to conventional expansion on 2D polystyrene dishes/flasks. Since the bioreactor-based expansion on 3D ceramic scaffolds encompasses multiple aspects that are inherently different from expansion on 2D polystyrene, we aimed to decouple the effects of specific parameters among these two model systems. We assessed the effects of the: i) 3D scaffold vs 2D surface, ii) ceramic vs polystyrene materials, and iii) BMSC niche established within the ceramic pores during in vitro culture, on subsequent in vivo bone formation...
September 7, 2017: Biotechnology Journal
Richard Schmid, Ioana-Sandra Tarau, Angela Rossi, Stefan Leonhardt, Thomas Schwarz, Sebastian Schuerlein, Christian Lotz, Jan Hansmann
The cornea is the most-transplanted tissue worldwide. However, the availability and quality of grafts are limited due to the current methods of corneal storage. In this study, a dynamic bioreactor system is employed to enable the control of intraocular pressure and the culture at the air-liquid interface. Thereby, in vivo-like storage conditions are achieved. Different media combinations for endothelium and epithelium are tested in standard and dynamic conditions to enhance the viability of the tissue. In contrast to culture conditions used in eye banks, the combination of the bioreactor and biochrom medium 1 allows to preserve the corneal endothelium and the epithelium...
September 5, 2017: Biotechnology Journal
Fengwu Bai, Hal Alper
Biocatalysts, either microbial cells or enzymes, render biotechnology breakthroughs in fundamentals and applications. Although rational designs with advanced tools and methodologies such as metabolic engineering and synthetic biology are becoming more powerful, conventional strategies including directed/adaptive evolution and process optimization are still indispensable.
September 4, 2017: Biotechnology Journal
Sean Miletic, Martin Hünerberg, Angelo Kaldis, Jacqueline MacDonald, Antoine Leuthreau, Tim McAllister, Rima Menassa
Enterohemorrhagic Escherichia coli (EHEC) are commonly present in the gastrointestinal tract of cattle and cause serious infectious disease in humans. Immunizing cattle against EHEC is a promising strategy to decrease the risk of food contamination; however, veterinary vaccines against EHEC such as Econiche have not been widely adopted by the agricultural industry, and have been discontinued, prompting the need for more cost-effective EHEC vaccines. The objective of this project was to develop a platform to produce plant-made antigens for oral vaccination of ruminants against EHEC...
September 4, 2017: Biotechnology Journal
Fangyu Cheng, Sijin Luozhong, Zhigang Guo, Huimin Yu, Gregory Stephanopoulos
Hyaluronic acid (HA) is a polysaccharide used in many industries such as medicine, surgery, cosmetics and food. To avoid potential pathogenicity caused by its native producer, Streptococcus, efforts have been made to create a recombinant host for HA production. In this work, a GRAS (generally recognized as safe) strain, Corynebacterium glutamicum, was engineered for enhanced biosynthesis of HA via metabolic pathway regulation. Five enzymes (HasA-HasE) involved in the HA biosynthetic pathway were highlighted, and 8 diverse operon combinations, including HasA, HasAB, HasAC, HasAD, HasAE, HasABC, HasABD and HasABE, were compared...
September 4, 2017: Biotechnology Journal
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