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Biotechnology Journal

Tomás Aparicio, Víctor de Lorenzo, Esteban Martínez-García
The higher intracellular ATP levels of genome-edited strains of P. putida that result from deleting various energy-consuming functions has been exploited for expanding the window of thermal tolerance of this bacterium. Unlike instant growth halt and eventual death of the naturally-occurring strain P. putida KT2440 at 42 °C, the EM42 variant maintained growth and viability of most of the population at the higher temperature for at least 6 h. We took advantage of this quality for implementing a robust thermo-inducible heterologous expression device in this species...
November 12, 2018: Biotechnology Journal
Thomas Fischöder, Claudia Wahl, Christian Zerhusen, Lothar Elling
The availability of nucleotide sugars is considered as bottleneck for Leloir-glycosyltransferases mediated glycan synthesis. A breakthrough for the synthesis of nucleotide sugars was the development of salvage pathway like enzyme cascades with high product yields from affordable monosaccharide substrates. In this regard, we aim at high enzyme productivities of these cascades by a repetitive batch approach. We report here for the first time that the exceptional high enzyme cascade stability facilitates the synthesis of Uridine-5'-diphospho-α-D-galactose (UDP-Gal), Uridine-5'-diphospho-N-acetylglucosamine (UDP-GlcNAc), and Uridine-5'-diphospho-N-acetylgalactosamine (UDP-GalNAc) in a multi-gram scale by repetitive batch mode...
October 26, 2018: Biotechnology Journal
Jean-Pierre Alcaraz, Philippe Cinquin, Donald K Martin
This review takes an approach to implanted medical devices that considers whether the intention of the implanted device is to have any communication of energy or materials with the body. The first part describes some specific examples of 3 different classes of implants, analysed with regards to the type of signal sent to cells. Through several examples, we describe that a one way signalling to the body leads to encapsulation or degradation. In most cases, those phenomena do not lead to major problems. However, encapsulation or degradation are critical for new kinds of medical devices capable of duplex communication, which we define in this review as symbiotic devices...
October 26, 2018: Biotechnology Journal
Claire M Palmer, Hal S Alper
Polyketides are a unique class of molecules with attractive bioactive and chemical properties. As a result, biorenewable production is being explored with these molecules as potential pharmaceutical, fuel and material precursors. In particular, type III polyketide synthases enable access to a diverse class of chemicals using a relatively simple biochemical synthesis pathway. In this review, we highlight recent advances in the engineering of microbial hosts for the production of type III PKS-derived polyketides...
October 25, 2018: Biotechnology Journal
Kalimuthu Kalishwaralal, Woo Young Kwon, Ki Soo Park
Exosomes, membrane-bound phospholipid vesicles having diameters of 50-200 nm, are secreted by all cell types and circulate in human body fluids. These vesicles are known to carry cellular constituents that are specific to the originating cells (e.g., cytoplasmic/membrane proteins, RNA, and DNA). Thus, exosomes, which are both structurally stable and abundant, are robust indicators of cancers and, as a result, they have been utilized to monitor this disease in a manner that is less invasive than gold standard tissue biopsies...
October 25, 2018: Biotechnology Journal
Timothée Baudequin, Cécile Legallais, Fahmi Bedoui
Bone remodelling is strongly linked to external mechanical signals. Such stimuli are widely used in vitro for bone tissue engineering by applying mechanical solicitations to cell cultures so as to trigger specific cell responses. However, the literature highlights considerable variability in devices and protocols. We reviewed here the major biological, mechanical and technical parameters implemented for in vitro tensile loading applications. The objective was to identify which values were used most, and whether there was an optimal protocol to obtain a functional tissue-engineering construct...
October 23, 2018: Biotechnology Journal
Julius Grzeschik, Desislava Yanakieva, Lukas Roth, Simon Krah, Steffen C Hinz, Adrian Elter, Tina Zollmann, Gerhard Schwall, Stefan Zielonka, Harald Kolmar
Yeast surface display emerged as a viable tool for the generation of human and murine monoclonal antibodies. This platform technology enables the careful definition of selection conditions, the potential for high-throughput screening, as well as the isolation of antibodies recognizing predefined epitopes. In this study we demonstrate the applicability of yeast surface display in combination with fluorescence-activated cell sorting (FACS) for the isolation of antigen-specific chicken-derived antibodies. To this end, yeast-displayed recombinant antibody libraries from splenic mRNA of chickens immunized with epidermal growth factor receptor (EGFR) and human chorionic gonadotropin (hCG) were constructed as single chain variable fragments (scFv) by overlap extension polymerase chain reaction...
October 23, 2018: Biotechnology Journal
Jukka Saarinen, Friederike Gütter, Mervi Lindman, Mikael Agopov, Sara J Fraser-Miller, Regina Scherließ, Eija Jokitalo, Hélder A Santos, Leena Peltonen, Antti Isomäki, Clare J Strachan
A wide variety of nanoparticles are playing an increasingly important role in drug delivery. Label-free imaging techniques are especially desirable to follow the cellular uptake and intracellular fate of nanoparticles. The combined correlative use of different techniques, each with unique advantages, facilitates more detailed and accurate information about such interactions. The synergistic use of correlative coherent anti-Stokes Raman scattering and electron microscopy (C-CARS-EM) imaging offers label-free, chemically-specific and (sub)nanometer spatial resolution for studying nanoparticle uptake into cells as demonstrated in the current study...
October 23, 2018: Biotechnology Journal
Viktor Sandner, Leon P Pybus, Graham McCreath, Jarka Glassey
High-Throughput (HT) technologies such as miniature bioreactors (MBRs) are increasingly employed within the biopharmaceutical manufacturing industry. Traditionally, these technologies have been utilized for discrete screening approaches during pre-clinical development (e.g. cell line selection and process optimization). However, increasing interest is focused towards their use during late clinical phase process characterization studies as a scale-down model (SDM) of the cGMP manufacturing process. In this review, we describe a systematic approach towards SDM development in one of the most widely adopted MBRs, the ambr™ 15 mL and 250 mL (Sartorius Stedim Biotech) systems...
October 23, 2018: Biotechnology Journal
Mark Pagkaliwangan, Jonathan Hummel, Xhorxhi Gjoka, Marc Bisschops, Mark Schofield
The advantages of continuous chromatography with respect to increased capacity are well established. However, the impact of different loading scenarios and total number of columns on the process economics has not been addressed. Here four different continuous multicolumn chromatography (MCC) loading scenarios were evaluated for process performance and economics in the context of a Protein A mAb capture step. To do so, a computational chromatography model was validated experimentally. The model was then used to predict process performance for each of the loading methods...
October 23, 2018: Biotechnology Journal
André L Rodrigues, Carlos A V Rodrigues, Ana Rita Gomes, Sara Filipa Vieira, Sara M Badenes, Maria Margarida Diogo, Joaquim M S Cabral
The development of bioprocesses capable of producing large numbers of human induced pluripotent stem cells (hiPSC) in a robust and safe manner is critical for the application of these cells in biotechnological and medical applications. Scalable expansion of hiPSC is often performed using polystyrene microcarriers, which have to be removed from the cell suspension using a separation step that causes loss of viable cells. In this study, we demonstrate the application of novel xeno-free dissolvable microcarriers (DM) for an efficient and integrated expansion and harvesting of hiPSC...
October 15, 2018: Biotechnology Journal
Johannes Hemmerich, Niklas Tenhaef, Carmen Steffens, Jannick Kappelmann, Marc Weiske, Sebastian Johannes Reich, Wolfgang Wiechert, Marco Oldiges, Stephan Noack
With modern genetic engineering tools, high number of potentially improved production strains can be created in a short time. This results in a bottleneck in the succeeding step of bioprocess development, which can be handled by accelerating quantitative microbial phenotyping. Miniaturization and automation are key technologies to achieve this goal. In this study, a novel workflow for repeated low-volume sampling of BioLector-based cultivation setup is presented. Six samples of 20 µL each can be taken automatically from shaken 48-well microtiter plates without disturbing cell population growth...
October 15, 2018: Biotechnology Journal
Beate Beyer, Nicole Walch, Alois Jungbauer, Nico Lingg
Biosimilars are increasing in economic importance. Just how similar a biosimilar needs to be to gain market approval is currently still decided on a per case basis. The authors try to shed light on one often cited critical quality attribute of monoclonal antibodies, namely charge heterogeneity. Using high resolution electrophoretic and chromatographic methods, the authors are able to separate and quantify the charge variant content of infliximab originator and three biosimilars. Additionally the authors quantified and compared the antigen binding affinity in an SPR based binding assay and analyzed the glycosylation pattern of all four of these infliximab biosimilar products...
October 13, 2018: Biotechnology Journal
Xiaofeng Dai, Yi Mei, Jianqi Nie, Zhonghu Bai
Adoptive T cell immunotherapy, involving the reprogramming of immune cells to target specific cancer or virus-infected cells, has been recognized as a promising novel approach for the treatment of complex diseases. The impressive global momentum of this therapeutic approach has highlighted the urgent need for establishing it as an effective and standardized onco-therapeutic approach in a large manufacturing scale. However, given its heterogeneity and uncertainty in nature, adoptive T cell immunotherapy is associated with a high failure rate that restricts its manufacturing to a limited number of institutions worldwide...
October 11, 2018: Biotechnology Journal
Yunhong Song, Meixia Liu, Leipeng Xie, Chun You, Junsong Sun, Yi-Heng P Job Zhang
Soluble hydrogenase I (SHI) from the hyperthermophilic archaeon Pyrococcus furiosus is a heterotetrameric [NiFe] hydrogenase that catalyzes the reversible reduction of protons by NADPH into hydrogen gas (H2 ). Here, the authors expressed the four αβγδ subunits of SHI encoded by one gene cluster in another hyperthermophilic archaeon, Thermococcus kodakarensis KOD1, which uses its hydrogenase maturation apparatus without the coexpression of native P. furiosus hydrogenase endopeptidases (maturation proteases)...
October 11, 2018: Biotechnology Journal
Huadong Peng, Lizhong He, Victoria S Haritos
The unique strained ring structure in cyclopropane fatty acids (CFA) conveys oxidative stability and lubricity to lipids. These attributes are highly valuable for industrial applications such as cosmetics and specialist lubrication but there is currently no commercial source of the lipid. Here, built on recently engineered strains of Saccharomyces cerevisiae, the authors have developed an efficient strategy for CFA production. Expression of the Escherichia coli cyclopropane fatty acid synthetase (Ec.CFAS) in the engineered yeast resulted in formation of cis-9,10-methylene-hexadecanoic and octadecanoic acids in both the phospholipid (PL) and triacylglycerol (TAG) fractions...
October 8, 2018: Biotechnology Journal
Alexandre Kriznik, Mélissa Yéléhé-Okouma, Jean-Christophe Lec, Guillaume Groshenry, Hélène Le Cordier, Christophe Charron, Marc Quinternet, Hortense Mazon, François Talfournier, Sandrine Boschi-Muller, Jean-Yves Jouzeau, Pascal Reboul
Purification of recombinant proteins remains a bottleneck for downstream processing. The authors engineered a new galectin 3 truncated form (CRDSAT ), functionally and structurally characterized, with preserved solubility and lectinic activity. Taking advantage of these properties, the authors designed an expression vector (pCARGHO), suitable for CRDSAT -tagged protein expression in prokaryotes. CRDSAT binds to lactose-Sepharose with a high specificity and facilitates solubilization of fusion proteins. This tag is structurally stable and can be easily removed from fusion proteins using TEV protease...
October 8, 2018: Biotechnology Journal
Sangwook Chu, Adam D Brown, James N Culver, Reza Ghodssi
Viruses are unique biological agents that infect living host cells through molecular delivery of a genomic cargo. Over the past two decades advancements in genetic engineering and bioconjugation technologies have allowed the unprecedented use of these "unfriendly" biological molecules, as nanoscopic platforms for the advancement of an array of nanotechnology applications. This mini-review focuses on providing a brief summary of key demonstrations leveraging the versatile characteristics of Tobacco mosaic virus (TMV) for molecular assembly and bio-device integration...
October 4, 2018: Biotechnology Journal
Aditya Utturkar, Keith Gillette, Chia-Yun Sun, Mark Pagkaliwangan, Rachel Quesenberry, Mark Schofield
An emphasis on continuous monoclonal antibody (mAb) bioprocessing in the pharmaceutical industry necessitates effective approaches for downstream process development (PD). With a PD strategy, the process parameters are optimized to directly develop streamlined three-step continuous chromatography processes. A design of experiment (DoE) approach with single column (batch mode) is used to simulate a multi-column (continuous mode) purification method and characterize each chromatography step: Protein A capture, anion exchange, and mixed mode cation exchange...
October 4, 2018: Biotechnology Journal
Ana S Nunes, Elisabete C Costa, Andreia S Barros, Duarte de Melo-Diogo, Ilídio J Correia
In vitro 3D cancer spheroids generally exhibit a drug resistance profile similar to that found in solid tumors. Due to this property, these models are an appealing for anticancer compounds screening. Nevertheless, the techniques and methods aimed for drug discovery are mostly standardized for cells cultured in 2D. The development of 2D cell culture models displaying a drug resistant profile is required to mimic the in vivo tumors, while the equipment, techniques and methodologies established for conventional 2D cell cultures can continue to be employed in compound screening...
September 22, 2018: Biotechnology Journal
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