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Biotechnology Progress

Yang Cao, Daniel Obeng, Guodong Hui, Luting Xue, Yukun Ren, Xianjie Yu, Fei Wang, Chad Atwell
This paper studies the Generalized Mahalanobis Distance (GMD) approach proposed by Hoffelder(1) which measures the dissimilarity of two multivariate Gaussian distributions with arbitrary covariance matrices and unequal sample sizes. This investigation demonstrated that, with appropriate adjustment, the GMD approach can achieve the targeted nominal Type I error and provide sufficient power for testing equivalence between two profile populations. The adjusted GMD approach was applied to examine the equivalence of harvest profiles between a 12L small scale model and 2000L manufacturing scale in a transfer study performed at Sanofi Speciality Care Framingham Biologics...
October 16, 2017: Biotechnology Progress
V Kouba, R Darmal, D Vejmelkova, P Jenicek, J Bartacek
The adaptation of anammox (ANaerobic AMMonium OXidation) to low temperatures (10-15°C) is crucial for sustaining energy-efficient nitrogen removal from the mainstream of municipal wastewater. But, current adaptation methods take months or even years. To speed up the adaption of anammox to low temperatures, this study describes a new approach: exposing anammox microorganisms to an abrupt temporary reduction of temperature i.e. cold shock. Anammox biomass in a moving bed biofilm reactor was subjected to three consecutive cold shocks (reduction from 24±2 to 5...
October 14, 2017: Biotechnology Progress
Lei Fang, Marc Guell, George M Church, Blaine A Pfeifer
The establishment of erythromycin production within the heterologous host E. coli marked an accomplishment in genetic transfer capacity. Namely, over 20 genes and 50 kb of DNA was introduced to E. coli for successful heterologous biosynthetic reconstitution. However, the prospect for production levels that approach those of the native host requires the application of engineering tools associated with E. coli. In this report, metabolic and genomic engineering were implemented to improve the E. coli cellular background and the plasmid platform supporting heterologous erythromycin formation...
September 28, 2017: Biotechnology Progress
Ghassan M Sulaiman, Amer T Tawfeeq, Marwa D Jaaffer
Copper oxide nanoparticles (CUNPs) were synthesized using Olea europaea leaf extract as reducing and protecting agent. The formation of nanoparticles was observed through a color change from yellowish to brownish black. The CUNPs were confirmed with UV-Vis spectrophotometer, which revealed a peak absorbance at 289 nm. The synthesized CUNPs were characterized by XRD, FTIR, SEM, and TEM. The XRD pattern revealed that CUNPs were crystalline in nature with a diameter around 20 nm. FTIR spectral analysis showed that CUNPs were capped with plant constituents...
September 28, 2017: Biotechnology Progress
Jung-Eun Lee, Praveen V Vadlani, Yadhu N Guragain, Ka-Yiu San, Doo-Hong Min
Switchgrass is a promising feedstock to generate fermentable sugars required for the sustainable operation of biorefineries because of their abundant availability, easy cropping system, and high cellulosic content. The objective of this study was to investigate the potentiality of switchgrass as an alternative sugar supplier for free fatty acid (FFA) production using engineered Escherichia coli strains. Recombinant E. coli strains successfully produced FFAs using switchgrass hydrolysates. A total of about 3 g/L FFAs were attained from switchgrass hydrolysates by engineered E...
September 28, 2017: Biotechnology Progress
Roberto Taboada-Puig, Thelmo A Lú-Chau, María T Moreira, Gumersindo Feijoo, Juan M Lema, Kurt Fagerstedt, Taina Ohra-Aho, Tiina Liitiä, Harri Heikkinen, Jarmo Ropponen, Tarja Tamminen
The objective of this study was to evaluate the ability of one versatile peroxidase and the biocatalytically generated complex Mn(III)-malonate to polymerize coniferyl alcohol (CA) to obtain dehydrogenation polymers (DHPs) and to characterize how closely the structures of the formed DHPs resemble native lignin. Hydrogen peroxide was used as oxidant and Mn(2+) as mediator. Based on the yields of the polymerized product, it was concluded that the enzymatic reaction should be performed in aqueous solution without organic solvents at 4...
September 28, 2017: Biotechnology Progress
Andres F Martinez, Richard D McMahon, Marc Horner, William M Miller
Platelet transfusions, with profound clinical importance in blood clotting and wound healing, are entirely derived from human volunteer donors. Hospitals rely on a steady supply of donations, but these methods are limited by a 5-day shelf life, the potential risk of contamination, and differences in donor/recipient histocompatibility. These challenges invite the opportunity to generate platelets ex vivo. Although much progress has been made in generating large numbers of culture-derived megakaryocytes (Mks, the precursor cells to platelets), stimulating a high percentage of Mks to undergo platelet release remains a major challenge...
September 27, 2017: Biotechnology Progress
Joachim Koepff, Holger Morschett, Tobias Busche, Anika Winkler, Jörn Kalinowski, Wolfgang Wiechert, Marco Oldiges
Recently, a comprehensive screening workflow for the filamentous bacterium Streptomyces lividans, a highly performant source for pharmaceutically active agents was introduced. This framework used parallelized cultivation in microtiter plates to efficiently accelerate early upstream process development. Focusing on growth performance, cultivation was successfully scaled-up to 1 liter stirred tank reactors. However, metabolic adaptation was observed on the transcriptomic level as among others, several genes incorporated in light response were up-regulated during bioreactor cultivation...
September 27, 2017: Biotechnology Progress
Ya-Qing Li, Yuan-Shun Liu, Xi-Wang Ying, Hong-Bin Zhou, Zhehua Wang, Sheng-Chang Wu, Jian-Ping Yan, Yu-Ting Jing, Yang Yang
OBJECTIVE: The aim of the study is to evaluate the effects of silencing a disintegrin and metalloproteinase 17 (ADAM17) gene expression by lentivirus-mediated RNA interference (RNAi) in the gefitinib-resistant lung adenocarcinoma cells, and then to explore whether the recombinant lentivirus mediated ADAM17 RNAi reversed the acquired resistance of lung adenocarcinoma to gefitinib in vitro. METHODS: The gefitinib-resistant RPC-9 cells were established and the mutations of EGFR were detected by gene sequencing...
September 27, 2017: Biotechnology Progress
David Shaw, Mandy Yim, Joni Tsukuda, John C Joly, Andy Lin, Brad Snedecor, Michael W Laird, Steven E Lang
In the development of biopharmaceutical products, the expectation of regulatory agencies is that the recombinant proteins are produced from a cell line derived from a single progenitor cell. A single limiting dilution step followed by direct imaging, as supplemental information, provides direct evidence that a cell line originated from a single progenitor cell. To obtain this evidence, a high-throughput automated imaging system was developed and characterized to consistently ensure that cell lines used for therapeutic protein production are clonally-derived...
September 27, 2017: Biotechnology Progress
Zulfiqar Ali Raza, Shahina Riaz, Ibrahim M Banat
Polyhydroxyalkanoates (PHAs) have become an attractive biomaterial in research in the past few years due to their extensive potential industrial applications. Being long chain hydroxyl fatty acid molecules, the PHAs are hydrophobic in nature, and have less functional groups. These features limit their applications in various areas. To enhance their usage, these polymers may need to be modified including surface and chemical modifications. Such modifications may alter their mechanical properties, surface structure, amphiphilic character and rate of degradation to fulfil the requirements for their future applications...
September 27, 2017: Biotechnology Progress
Yizhou Zhou, David Shaw, Cynthia Lam, Joni Tsukuda, Mandy Yim, Danming Tang, Salina Louie, Michael W Laird, Brad Snedecor, Shahram Misaghi
Establishing that a cell line was derived from a single cell progenitor and defined as clonally-derived for the production of clinical and commercial therapeutic protein drugs has been the subject of increased emphasis in cell line development (CLD). Several regulatory agencies have expressed that the prospective probability of clonality for CHO cell lines is assumed to follow the Poisson distribution based on the input cell count. The probability of obtaining monoclonal progenitors based on the Poisson distribution of all cells suggests that one round of limiting dilution may not be sufficient to assure the resulting cell lines are clonally-derived...
September 23, 2017: Biotechnology Progress
Benjamin A F Bláha, Stephen A Morris, Olotu W Ogonah, Vincenzo Crescente, William Rosenberg, Sophie Maucourant, Tarit K Mukhopadhyay
The time and cost benefits of miniaturized fermentation platforms can only be gained by employing complementary techniques facilitating high-throughput at small sample volumes. Microbial cell disruption is a major bottleneck in experimental throughput and is often restricted to large processing volumes. Moreover, for rigid yeast species such as Pichia pastoris, no effective high-throughput disruption methods exist. This study describes the development of an automated, miniaturized, high-throughput, non-contact, scalable platform based on Adaptive Focused Acoustics (AFA) to disrupt P...
September 7, 2017: Biotechnology Progress
Melani C Stone, Jon Borman, Gisela Ferreira, P David Robbins
Flowthrough anion exchange chromatography is commonly used as a polishing step in downstream processing of monoclonal antibodies and other therapeutic proteins to remove process-related impurities and contaminants such as host cell DNA, host cell proteins, endotoxin, and viruses. DNA with a wide range of molecular weight distributions derived from Chinese Hamster Ovary cells was used to advance the understanding of DNA binding behavior in selected anion exchange media using the resin (Toyopearl SuperQ-650M) and membranes (Mustang® Q and Sartobind(®) Q) through DNA spiking studies...
September 7, 2017: Biotechnology Progress
A Demirbas, K Groszman, M Pazmiño-Hernandez, D C Vanegas, B Welt, J A Hondred, N T Garland, J C Claussen, E S McLamore
Flavonoids are important value added products for dye sensitized solar cells biosensors, functional foods, medicinal supplements, nanomaterial synthesis, and other applications. Brassica oleracea contains high levels of anthocyanins in leaf sap vacuoles, and there are many viable extraction techniques that vary in terms of simplicity, environmental impact, cost, and extract photochemical/electrochemical properties. The efficiency of value added biotechnologies from flavonoid is a function of anthocyanin activity/concentration and molecule stability (i...
September 7, 2017: Biotechnology Progress
Belén Infanzón, Silvia Cesarini, Josefina Martínez, F I Javier Pastor, Pilar Diaz
Previously isolated and characterized Pseudomonas lipases were immobilized in a low-cost MP-1000 support by a re-loading procedure that allowed a high activity per weight of support. Immobilized LipA, LipC, and LipCmut lipases, and commercial Novozym® 435 were tested for fatty acid methyl ester (FAMEs) synthesis using conventional and alternative feedstocks. Triolein and degummed soybean oils were used as model substrates, whereas waste cooking oil and M. circinelloides oil were assayed as alternative, low cost feedstocks, whose free fatty acid (FFA), and acylglyceride profile was characterized...
September 7, 2017: Biotechnology Progress
Zuzana Antošová, Klára Herkommerová, Iva Pichová, Hana Sychrová
Laccases are enzymes with a broad range of biotechnological applications and have, for example, the ability to oxidize many xenobiotics including synthetic dyes. In order to obtain an efficient laccase for the decolorization of dyes which spoil wastewater from the textile industry, genes encoding three various laccase enzymes were expressed in Saccharomyces cerevisiae. The expression of laccases from ascomycete Myceliophthora thermophila (MtL), and two basidiomycetes Trametes versicolor (TvL) and Trametes trogii (TtL) was optimized via selection of plasmids, promoters, media composition and cultivation conditions...
September 7, 2017: Biotechnology Progress
James Savery, Krista Evans
No abstract text is available yet for this article.
August 26, 2017: Biotechnology Progress
Ya-Jun Wang, Wei Shen, Xi Luo, Zhi-Qiang Liu, Yu-Guo Zheng
t-Butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-2) is a key chiral diol precursor of atorvastatin calcium (Lipitor®). We have constructed a Kluyveromyces lactis aldo-keto reductase mutant KlAKR-Y295W/W296L (KlAKRm) by rational design in previous research, which displayed high activity and excellent diastereoselectivity (dep  > 99.5%) toward t-butyl 6-cyano-(5R)-hydroxy-3-oxohexanoate ((5R)-1). To realize in situ cofactor regeneration, a robust KlAKRm and Exiguobacterium sibiricum glucose dehydrogenase (EsGDH) co-producer E...
August 26, 2017: Biotechnology Progress
Chapman Wright, Christina Alves, Scott Estes, John Pieracci, Rashmi Kshirsagar
The Biogen upstream platform is capable of delivering equivalent quality material throughout the cell line generation process. This allows us to rapidly deliver high quality biopharmaceuticals to patients with unmet medical needs. The drive to reduce time-to-market led the cell engineering group to develop an expression system that can enable this strategy. We have developed a clonal CHO host cell line that can routinely produce consistent antibody material at high titers throughout the cell line generation process...
August 26, 2017: Biotechnology Progress
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