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Biotechnology Progress

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https://www.readbyqxmd.com/read/28271638/comparison-of-spectroscopy-technologies-for-improved-monitoring-of-cell-culture-processes-in-miniature-bioreactors
#1
Ruth C Rowland-Jones, Frans van den Berg, Andrew J Racher, Elaine B Martin, Colin Jaques
Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the through-put of such studies; however, there are limitations with their use. One important constraint is the limited number of offline samples that can be taken compared to those taken for monitoring cultures in larger-scale bioreactors. The small volume of miniature bioreactor cultures (15mL) is incompatible with the large sample volume (600µL) required for bioanalysers routinely used...
March 8, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28268261/large-scale-synthesis-of-tert-butyl-3r-5s-6-chloro-3-5-dihydroxyhexanoate-by-a-stereoselective-carbonyl-reductase-with-high-substrate-concentration-and-product-yield
#2
Zhi-Qiang Liu, Zhong-Liang Hu, Xiao-Jian Zhang, Xiao-Ling Tang, Feng Cheng, Ya-Ping Xue, Ya-Jun Wang, Lin Wu, Dan-Kai Yao, Yi-Teng Zhou, Yu-Guo Zheng
To biosynthesize the (3R,5S)-CDHH in an industrial scale, a newly synthesized stereoselective short chain carbonyl reductase (SCR) was successfully cloned and expressed in Escherichia coli. The fermentation of recombinant E. coli harboring SCR was carried out in 500 L and 5000 L fermenters, with biomass and specific activity of 9.7 g DCW/L, 15749.95 U/g DCW and 10.97 g DCW/L, 19210.12 U/g DCW, respectively. The recombinant SCR was successfully applied for efficient for (3R,5S)-CDHH production. The scale-up synthesis of (3R,5S)-CDHH was performed in 5000 L bioreactor with 400 g/L of (S)-CHOH at 30°C, resulting in a space-time yield of 13...
March 7, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28268250/antioxidant-effect-of-thiazolidine-molecules-in-cell-culture-media-improves-stability-and-performance
#3
Jennifer Kuschelewski, Alisa Schnellbaecher, Sascha Pering, Maria Wehsling, Aline Zimmer
The ability of cell culture media components to generate reactive species as well as their sensitivity to oxidative degradation, affects the overall stability of media and the behavior of cells cultured in vitro. This study investigates the influence of thiazolidine molecules, formed from the condensation between cysteine and alpha-ketoacids, on the stability of these complex mixtures and on the performance of cell culture processes aiming to produce therapeutically relevant monoclonal antibodies. Results presented in this study indicate that 2-methyl-1,3-thiazolidine-2,4-dicarboxylic acid and 2-(2-carboxyethyl)-1,3-thiazolidine-2,4-dicarboxylic acid, obtained by condensation of cysteine with pyruvate or alpha-ketoglutarate respectively, are able to stabilize cell culture media formulations, in particular redox sensitive molecules like folic acid, thiamine, L-methionine (met) and L-tryptophan (trp)...
March 7, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28263434/permeability-enhancement-of-escherichia-coli-by-single-walled-carbon-nanotube-treatment
#4
Abdollah Mosleh, Anna Heintz, Ki-Taek Lim, Jin-Woo Kim, Robert Beitle
This research investigated the use of single-walled carbon nanotubes (SWNTs) as an additive to increase the permeability of a bacterial cell wall. Recombinant Escherichia coli BL21 (DE3) that expressed β-lactamase were exposed to SWNTs under various levels of concentration and agitation. Activity of β-lactamase in the culture fluid and Transmission Electron Microscopy (TEM) were used to determine the amount of released protein, and visually examine the permeability enhancement of the cells. It was found that β-lactamase release in the culture fluid occurred in a dose dependent manner with treatment by SWNTs and was also dependent on agitation rate...
March 6, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28249362/effects-of-lysosomal-biotherapeutic-recombinant-protein-expression-on-cell-stress-and-protease-and-general-host-cell-protein-hcp-release-in-chinese-hamster-ovary-cells
#5
Damiano Migani, C Mark Smales, Daniel G Bracewell
Recombinant human Acid Alpha Glucosidase (GAA) is the therapeutic enzyme used for the treatment of Pompe disease, a rare genetic disorder characterised by GAA deficiency in the cell lysosomes(1) . The manufacturing process for GAA can be challenging, in part due to protease degradation. The overall goal of this study was to understand the effects of GAA overexpression on cell lysosomal phenotype and host cell protein (HCP) release, and any resultant consequences for protease levels and ease of manufacture. To do this we first generated a human recombinant GAA producing stable CHO cell line and designed the capture chromatographic step anion exchange (IEX)...
March 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28248011/a-novel-extracellular-low-temperature-active-phytase-from-bacillus-aryabhattai-rs1-with-potential-application-in-plant-growth
#6
Authors Moushree Pal Roy, Subhabrata Datta, Shilpi Ghosh
Bacillus aryabhattai RS1 isolated from rhizosphere produced an extracellular, low temperature active phytase. The cultural conditions for enzyme production were optimized to obtain 35 U ml(-1) of activity. Purified phytase had specific activity and molecular weight of 72.97 U mg(-1) and ∼40 kDa, respectively. The enzyme was optimally active at pH 6.5 and 40°C and was highly specific to phytate. It exhibited higher catalytic activity at low temperature, retaining over 40% activity at 10°C. Phytase was more thermostable in presence of Ca(2+) ion and retained 100% residual activity on pre-incubation at 20-50°C for 30 min...
March 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28248010/effects-of-ferrous-iron-on-the-performance-and-microbial-community-in-aerobic-granular-sludge-in-relation-to-nutrient-removal
#7
Gulsum Yilmaz, Ender Cetin, Umit Bozkurt, Karin Aleksanyan Magden
Lab-scale experiments were conducted to investigate the effects of ferrous iron on nutrient removal performance and variations in the microbial community inside aerobic granular sludge for 408 days. Two reactors were simultaneously operated, one without added ferrous iron (SBR1), and one with 10 mg Fe(2+) L(-1) of added ferrous iron (SBR2). 1 mg Fe(2+) L(-1) of added ferrous iron was applied to SBR1 starting from the 191(st) day to observe the resulting variations in the nutrient removal performance and the microbial community...
March 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28248007/antibody-purification-from-cho-cell-supernatant-using-new-multimodal-membranes
#8
Juan Wang, Jinxiang Zhou, Yogender K Gowtham, Sarah W Harcum, Scott M Husson
This contribution describes strategies to purify monoclonal antibodies from Chinese hamster ovary (CHO) cell culture supernatant using newly designed multimodal membranes (MMMs). The MMMs were used for the capture step purification of human IgG1 following a size-exclusion desalting column to remove chaotropic salts that interfere with IgG binding. The MMM column attained higher dynamic binding capacity than a Protein A resin column at an equivalent residence time of 1 min. The two step MMM chromatography process achieved high selectivity for capturing hIgG1 from the CHO cell culture supernatant, though the desalting step resulted in product dilution...
March 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28247994/a-novel-soft-sensor-approach-for-estimating-individual-biomass-in-mixed-cultures
#9
Kyle A Stone, Devarshi Shah, Min Hea Kim, Nathan R M Roberts, Q Peter He, Jin Wang
Due to many advantages associated with mixed cultures, the application of mixed cultures in biotechnology has expanded rapidly in recent years. At the same time, many challenges remain for effective mixed culture applications. One obstacle is how to efficiently and accurately monitor the individual cell populations. The current approaches on individual cell mass quantification are suitable for off-line, infrequent characterization. In this work, we propose a fast and accurate 'soft sensor' approach for estimating individual cell concentrations in mixed cultures...
March 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28247962/hydrogel-based-three-dimensional-cell-culture-for-organ-on-a-chip-applications
#10
REVIEW
Seung Hwan Lee, Kyu Young Shim, Bumsang Kim, Jong Hwan Sung
Recent studies have reported that three-dimensionally cultured cells have more physiologically relevant functions than two-dimensionally cultured cells. Cells are three-dimensionally surrounded by the extracellular matrix (ECM) in complex in vivo microenvironments and interact with the ECM and neighboring cells. Therefore, replicating the ECM environment is key to the successful cell culture models. Various natural and synthetic hydrogels have been used to mimic ECM environments based on their physical, chemical, and biological characteristics, such as biocompatibility, biodegradability, and biochemical functional groups...
March 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28220676/cho-cells-adapted-to-inorganic-phosphate-limitation-show-higher-growth-and-higher-pyruvate-carboxylase-flux-in-phosphate-replete-conditions
#11
Vishwanathgouda Maralingannavar, Dharmeshkumar Parmar, Tejal Pant, Chetan Gadgil, Venkateswarlu Panchagnula, Mugdha Gadgil
Inorganic phosphate (Pi ) is an essential ion involved in diverse cellular processes including metabolism. Changes in cellular metabolism upon long term adaptation to Pi limitation have been reported in E. coli. Given the essential role of Pi , adaptation to Pi limitation may also result in metabolic changes in animal cells. In this study, we have adapted CHO cells producing recombinant IgG to limiting Pi conditions for 75 days. Not surprisingly, adapted cells showed better survival under Pi limitation. Here, we report the finding that such cells also showed better growth characteristics compared to control in batch culture replete with Pi (higher peak density and integral viable cell density), accompanied by a lower specific oxygen uptake rate and cytochrome oxidase activity towards the end of exponential phase...
February 21, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28218499/fine-tuning-sortase-mediated-immobilization-of-protein-layers-on-surfaces-using-sequential-deprotection-and-coupling
#12
Maryam Raeeszadeh-Sarmazdeh, Ranganath Parthasarathy, Eric T Boder
Increasing interest in protein immobilization on surfaces has heightened the need for techniques enabling layer-by-layer protein attachment. Here, we report a technique for controlling enzyme-mediated immobilization of layers of protein on the surface using a genetically-encoded protecting group. An enterokinase-cleavable peptide sequence was inserted at the N-terminus of bifunctional fluorescent proteins containing Sortase A substrate recognition tags at both ends to control Sortase A-mediated protein immobilization on the surface layer-by-layer...
February 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28218470/maximizing-the-functional-lifetime-of-protein-a-resins
#13
Jennifer Zhang, Sethu Siva, Ryan Caple, Rob Gronke, Sanchayita Ghose
Protein A chromatography is currently the industry gold-standard for monoclonal antibody and Fc-fusion protein purification. The high cost of Protein A however makes resin lifetime and resin reuse an important factor for process economics. Typical resin lifetime studies performed in the industry usually examine the effect of resin re-use on binding capacity, yield, and product quality without answering the fundamental question of what is causing the decrease in performance. A two part mechanistic study was conducted in an attempt to decouple the effect of the two possible factors (resin hydrolysis and/or degradation vs...
February 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28188705/design-construction-and-optimization-of-a-novel-modular-and-scalable-incubation-chamber-for-continuous-viral-inactivation
#14
Raquel Orozco, Linus Amarikwa, Stephanie Parker, Scott Godfrey, Lindsay Hernandez, Chinenye Wachuku, Ben Mai, Brian Song, Shashidhar Hoskatti, Jinkeng Asong, Marcus Fiadeiro, Parviz Shamlou, Cameron Bardliving, Jon Coffman
We designed, built or 3D printed, and screened tubular reactors that minimize axial dispersion to serve as incubation chambers for continuous virus inactivation of biological products. Empirical residence time distribution data were used to derive each tubular design's volume equivalent to a theoretical plate (VETP) values at a various process flow rates. One design, the Jig In a Box (JIB), yielded the lowest VETP, indicating optimal radial mixing and minimal axial dispersion. A minimum residence time (MRT) approach was employed, where the MRT is the minimum time the product spends in the tubular reactor...
February 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28188702/control-of-chitin-nanofiber-production-by-the-lipid-producing-diatom-cyclotella-sp-through-fed-batch-addition-of-dissolved-silicon-and-nitrate-in-a-bubble-column-photobioreactor
#15
Omar Chiriboga, Gregory L Rorrer
Diatoms are single-celled algae that make cell walls of nanopatterned biogenic silica called frustules through metabolic uptake of dissolved silicon and its templated condensation into biosilica. The centric marine diatom Cyclotella sp. also produces intracellular lipids and the valued co-product chitin, an N-acetyl glucosamine biopolymer that is extruded from selected frustule pores as pure nanofibers. The goal of this study was to develop a nutrient feeding strategy to control the production of chitin nanofibers from Cyclotella with the co-production of biofuel lipids...
February 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28188696/applied-in-situ-product-recovery-in-abe-fermentation
#16
REVIEW
Victoria Outram, Carl-Axel Lalander, Jonathan G M Lee, E Timothy Davies, Adam P Harvey
The production of biobutanol is hindered by the product's toxicity to the bacteria, which limits the productivity of the process. In situ product recovery of butanol can improve the productivity by removing the source of inhibition. This paper reviews in situ product recovery techniques applied to the acetone butanol ethanol fermentation in a stirred tank reactor. Methods of in situ recovery include gas stripping, vacuum fermentation, pervaporation, liquid-liquid extraction, perstraction and adsorption, all of which have been investigated for the acetone, butanol and ethanol fermentation...
February 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28188692/bioreactor-scale-up-and-protein-product-quality-characterization-of-piggybac-transposon-derived-cho-pools
#17
Yashas Rajendra, Sowmya Balasubramanian, Robert B Peery, James R Swartling, Neil A McCracken, Dawn L Norris, Christopher C Frye, Gavin C Barnard
Chinese hamster ovary (CHO) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mAbs), bispecific antibodies and Fc-fusion proteins. Creating and characterizing the stable CHO clonally-derived cell lines (CDCLs) needed to manufacture these therapeutic proteins is a lengthy and laborious process. Therefore, CHO pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development. We recently described the generation of CHO pools yielding mAb titers as high as 7...
February 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28188688/evaluation-of-heavy-chain-c-terminal-deletions-on-productivity-and-product-quality-of-monoclonal-antibodies-in-chinese-hamster-ovary-cho-cells
#18
Zhilan Hu, Danming Tang, Shahram Misaghi, Guoying Jiang, Christopher Yu, Mandy Yim, David Shaw, Brad Snedecor, Michael Laird, Amy Shen
Monoclonal antibodies (mAbs) have been well established as potent therapeutic agents and are used to treat many different diseases. During cell culture production, antibody charge variants can be generated by cleavage of heavy chain (HC) C-terminal lysine and proline amidation. Differences in levels of charge variants during manufacturing process changes make it challenging to demonstrate process comparability. In order to reduce heterogeneity and achieve consistent product quality, we generated and expressed antibodies with deletion of either HC C-terminal lysine (-K) or lysine and glycine (-GK)...
February 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28187511/a-novel-cholesterol-lipid-delivery-system-for-murine-myeloma-cell-lines
#19
Gaurav Chauhan, Albert E Schmelzer
Murine myeloma NS0 cells are cholesterol-dependent auxotrophs and require externally provided cholesterol for sustained growth. Traditionally, cholesterol is provided to these cells by supplementing cell culture media with concentrated solution of cholesterol and other water insoluble components dissolved in 200-proof ethanol. However, the solubility of cholesterol in ethanol is limited and for processes requiring large amounts of cholesterol, the consequential increase in added ethanol may negatively impact cell growth...
February 10, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28127952/shock-treatment-of-corn-stover
#20
Austin Bond, Hema Rughoonundun, Eric Petersen, Carol Holtzapple, Mark Holtzapple
Corn stover digestibility was enhanced via shock treatment. A slurry of lime-treated corn stover was placed in a partially filled closed vessel. From the ullage space, either a shotgun shell was fired into the slurry, or a gas mixture was detonated. Various conditions were tested (i.e., pressures, depth, solids concentrations, gas mixtures). A high pressurization rate (108,000 MPa/s shotgun shells; 4,160,000 MPa/s hydrogen/oxygen detonation) was the only parameter that improved enzymatic digestibility. Stoichiometric propane/air deflagration had a low pressurization rate (37...
January 27, 2017: Biotechnology Progress
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