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Biotechnology Progress

Michael B Hippach, Ian Schwartz, Jian Pei, Jimmy Huynh, Yasuaki Kawai, Marie M Zhu
During a CHO cell culture production process, important parameters are generally well controlled by a feedback mechanism (PID loop) in order to ensure consistency in both productivity and product quality. These parameters typically include pH, dissolved oxygen (DO), and temperature. While most of these parameters are very well controlled within their specific deadband, stable DO control can be challenging. Oscillations in DO concentration are not uncommon and these fluctuations can be exacerbated with an efficient mass transfer aeration strategy...
September 12, 2018: Biotechnology Progress
David Cetlin, Melanie Pallansch, Coral Fulton, Esha Vyas, Aesha Shah, Taka Sohka, Arun Dhar, Luke Pallansch, Daniel Strauss
Viruses can arise during the manufacture of biopharmaceuticals through contamination or endogenous expression of viral sequences. Regulatory agencies require "viral clearance" validation studies for each biopharmaceutical prior to approval. These studies aim to demonstrate the ability of the manufacturing process at removing or inactivating virus and are conducted by challenging scaled-down manufacturing steps with a "spike" of live virus. Due to the infectious nature of these live viruses, "spiking studies" are typically conducted in specialized biological safety level-2 facilities...
September 8, 2018: Biotechnology Progress
Christopher Thompson, Kelly Wilson, Christopher Larkin, Jeong Lee, William K Wang, Michaela Wendeler
This work presents the optimization and critical evaluation of continuous capture chromatography in the downstream process of a recombinant enzyme. For the upstream manufacturing of this molecule, a perfusion process was implemented due to benefits for product quality and productivity. This process is, however, characterized by low titer and significant changes over the course of the harvest duration in terms of active enzyme concentration and impurity content. We evaluated the feasibility and benefits of a continuous capture operation...
August 9, 2018: Biotechnology Progress
Káren Gercyane O Bezerra, Raquel D Rufino, Juliana M Luna, Leonie A Sarubbo
The cosmetic industry is currently one of the fasting growing sections of the economy in many countries. The recent tendency toward the use of cosmetics of a natural origin has driven the industry to seek alternatives to synthetic components in the formulation of products. Biosurfactants are natural compounds that have considerable potential for application in the formulation of safe, effective cosmetics as a replacement for commonly used chemical tensioactive agents. The present review provides essential information on the physicochemical and biological properties of saponins and microbial biosurfactants employed in cosmetic products, with a focus on the use of these natural compounds in shampoos, addressing the current state of research and patents involving biosurfactants for this purpose...
July 27, 2018: Biotechnology Progress
Abhiram Arunkumar, Junyan Zhang, Nripen Singh, Sanchayita Ghose, Zheng Jian Li
This work examines the ultrafiltration behavior of partially retained proteins like lysozyme and completely retained proteins like monoclonal antibodies using single pass tangential flow filtration (SPTFF) modules with different screen channels and molecular weight cut-offs. When the staging of the SPTFF used the same membrane area in each stage, there was no impact of the module screened channel or the buffer matrix on the final concentration achieved for completely retained monoclonal antibodies. A hybrid configuration containing 30 kDa membranes and 50 kDa membranes increased the maximum achievable concentration for both the monoclonal antibodies used in this work, at the same time, allowing a two-fold to four-fold increase in normalized feed flow-rate through the system compared to only the 30 kDa or only the 50 kDa membranes...
July 18, 2018: Biotechnology Progress
Eva Hackemann, Hans Hasse
Hydrophobic interaction chromatography (HIC) is often used for purifying proteins. The prediction of the adsorption equilibria in such systems is a difficult task as they depend on the pH value and the influence of the salts, which are added for controlling the solvent properties. This is especially tedious when salt mixtures are used, which can have positive or negative cooperative effects on the adsorption. A mathematical model to describe these complex effects of the salts was recently introduced by our group...
July 16, 2018: Biotechnology Progress
M C Tamayo-Ordóñez, B A Ayil-Gutiérrez, Y J Tamayo-Ordóñez, L C Rodríguez-Zapata, M Monforte-González, E A De la Cruz-Arguijo, M J García-Castillo, L F Sánchez-Teyer
Several of the over 200 known species of Agave L. are currently used for production of distilled beverages and biopolymers. The plants live in a wide range of stressful environments due to their resistance to abiotic stress (drought, salinity, extreme temperature) and pathogens, which gives the genus potential for germplasm conservation and biotechnological applications that may minimize economic losses due to global climate change. However, the limited knowledge in the genus of genome structure and organization hampers development of potential improved biotechnological applications by means of genetic manipulation and biocatalysis...
July 16, 2018: Biotechnology Progress
T Abbate, L Dewasme, L Dewasme
In this study, a dynamic model of a Vero cell culture-based dengue vaccine production process is developed. The approach consists in describing the process dynamics as functions of the whole living (uninfected and infected) biomass whereas previous works are based on population balance approaches. Based on the assumption that infected biomass evolves faster than other variables, the model can be simplified using a slow-fast approximation. The structural identifiability of the model is analysed using differential algebra as implemented in the software DAISY...
July 16, 2018: Biotechnology Progress
Lei Wang, R Kenneth Marcus
In preparative chromatography, overloading effects are a fact of life. However, peak distortion is problematic in analytical scale chromatography, especially in the case of bio-macromolecule separations. Capillary-channeled polymer fibers have been employed for fast protein separations in reversed phase, ion exchange, and hydrophobic interaction liquid chromatography. Although the primary advantage of the phase is operation at high linear velocities (~100 mm s-1 ) without van Deemter C-term limitations, the limited specific surface area (<5 m2 g-1 ) suggests that a thorough understanding of overloading effects is needed...
July 16, 2018: Biotechnology Progress
Kim Le, Christopher Tan, Shivani Gupta, Trupti Guhan, Hedieh Barkhordarian, Jonathan Lull, Jennitte Stevens, Trent Munro
Generating a highly productive cell line is resource intensive and typically involves long timelines, due to the need to screen large numbers of candidates in protein production studies. This has led to miniaturization and automation strategies to allow for reductions in resources and higher throughput. Current approaches rely on the use of standard cell culture vessels and bulky liquid handling equipment. New nanofludic technologies offer novel solutions to surpass these limits, further miniaturizing cell culture volumes (105 times smaller) by growing cells on custom nanofluidic chips...
July 16, 2018: Biotechnology Progress
Yuji Haraguchi, Katsuhisa Matsuura, Yuki Kagawa, Akiyuki Hasegawa, Hirotsugu Kubo, Tatsuya Shimizu
This study reports a rapid fabrication system of a morphologically and functionally communicative 3-D cell-dense tissue without scaffolds by centrifugation. The tight adhesion between C2C12 myoblasts and culture surface was accelerated without significant cell damage by centrifugation (80 x g, 37°C, 30 min). A thicker tissue created on a temperature-responsive culture surface was harvested by decreasing temperature. The 3D myoblast tissues having approximately 200 µm-thickness were created at 1.5 h [centrifugation (80 x g, 37°C) for 30 min and tissue harvest for 1 h]...
July 16, 2018: Biotechnology Progress
J P Little, G J Pettet, D W Hutmacher, D Loessner
BACKGROUND: There is a relative paucity of research that integrates materials science and bioengineering with computational simulations to decipher the intricate processes promoting cancer progression. Therefore, a first-generation computational model, SpheroidSim, was developed that includes a biological data set derived from a bioengineered spheroid model to obtain a quantitative description of cell kinetics. RESULTS: SpheroidSim is a 3D agent-based model simulating the growth of multicellular cancer spheroids...
July 16, 2018: Biotechnology Progress
Jonathan J Chen, Lyndsey N Schmucker, Donald P Visco
Blood Clotting Factor XI is an important actor in the clotting mechanism: it activates downstream zymogen involved in the clotting process. It can be targeted for activation or inhibition depending on treatment goals to enhance or inhibit clotting. In terms of anti-thrombosis treatment, Factor XI has emerged as a promising target to focus on. In this work, an iterative virtual high-throughput screening pipeline was proposed that can supplement current efforts to find inhibitors. The first iteration identified 11 compounds to test with 3 active for a hit-rate of 27...
July 16, 2018: Biotechnology Progress
Bruna Regina Zaccarim, Fernanda de Oliveira, Michel Rodrigo Zambrano Passarini, Alysson Wagner Fernandes Duarte, Lara Durães Sette, Angela Faustino Jozala, Maria Francisca Simas Teixeira, Valéria de Carvalho Santos Ebinuma
The population's interest in health products is increasing day by day. Thus, the demand for natural products as additives for food and pharmaceutical commodities is also growing. Colorants are additives that provide color to products and can be produced by microorganisms via various bioprocesses. In the search for new sources of natural colorants, fungi have been employed to produce novel and safer natural colorants. Therefore, the main goal of this study is to describe a Talaromyces species able to produce natural colorants and investigate the nutritional parameters for colorant production using statistical tools...
July 14, 2018: Biotechnology Progress
Matheus M Pereira, Mafalda R Almeida, Joana Gomes, Ana F C S Rufino, Marguerita E Rosa, João A P Coutinho, Aminou Mohamadou, Mara G Freire
Given the biotechnology advances observed in recent years in terms of upstream, the development of effective downstream processes becomes mandatory to decrease the associated costs of biotechnological-based products. Although a large interest has been devoted to ionic-liquid-based aqueous biphasic systems (IL-based ABS) as tailored separation platforms, imidazolium-based ILs have been the preferred choice as phase-forming agents. To overcome some toxicity and biodegradability issues associated to imidazolium-based ILs, novel ABS composed of ILs analogues of glycine-betaine (AGB-ILs) are here proposed and investigated...
July 14, 2018: Biotechnology Progress
Thais M de Borba, Taiele B Machado, Adriano Brandelli, Susana J Kalil
The parameters half-life, z value, enthalpy, entropy, and free energy were evaluated in the temperature range of 40 to 55°C for a Bacillus sp. P45 protease present in a medium composed of ionic liquid (IL) and organic solvent. The protease was previously treated in IL [Emim][Tf2 N] and increased activity was observed in four out of five organic solvents tested. The reaction medium containing acetone and IL (1:1 v/v ratio) was more stable than the buffer medium, with half-life of 2.4 h at 55°C. Thermodynamic parameters values showed there was probably less rupture of non-covalent bonds that stabilize the protein structure, which promoted the protease P45 stabilization...
July 10, 2018: Biotechnology Progress
Szymon Skoneczny
The article presents a mathematical model and steady-state characteristics of a continuous-flow two-phase fluidized-bed bioreactor with external aeration. The quantitative description includes biofilm growth on inert carriers, distributions of density, and diffusion coefficients in biofilm, decay of microorganisms, gas-liquid and liquid-biofilm mass transfer, biofilm detachment, biomass thickening and recirculation. The method for the determination of steady-states and the assessment of stability based on the orthogonal collocation method was presented...
June 12, 2018: Biotechnology Progress
Simel Bağder Elmacı, Filiz Özçelik
In this study, it was aimed to enhance the enzymatic hydrolysis of recalcitrant yellow pine wood and subsequently to produce ethanol efficiently through the application of 1-ethyl-3-methylimidazolium acetate (EMIMAc) ionic liquid. The effect of biomass particle size (<2.5 mm, 500-850 μm, 363-500 μm and <363 μm) prior to pretreatment and pretreatment time (15, 30 and 45 min) on the structural properties and enzymatic hydrolysis yield of EMIMAc pretreated pine wood were also investigated. In general, EMIMAc functioned more efficiently at larger biomass particle sizes compared to smaller ones...
June 9, 2018: Biotechnology Progress
Jenna Williamson, Jennifer Miller, Joseph McLaughlin, Rodney Combs, Chia Chu
N-linked glycosylation profiles are routinely characterized on mammalian-derived protein therapeutic products and achieving consistency in the product-associated glycan attributes is an important indicator that the manufacturing process is under control. More importantly, meeting target glycan profile is a common criterion for ensuring product efficacy. During laboratory process development and subsequent scale up for pilot demonstration for a monoclonal antibody program, discrepancies in the molecule's terminal galactosylation level at 2-L, 100-L, and 6,000-L scales were observed...
June 8, 2018: Biotechnology Progress
Rüdiger Ohs, Martin Leipnitz, Marie Schöpping, Antje C Spiess
Thiamine diphosphate (ThDP)-dependent enzymes catalyze a broad range of reactions with excellent enantioselectivity. Among these reactions, carboligations of aldehydes are of particular interest since the products, chiral hydroxy ketones, are valuable building blocks in the pharmaceutical industry. However, the substrates, for example, benzaldehyde, inactivate the biocatalysts, for example the ThDP-dependent benzaldehyde lyase from Pseudomonas fluorescens (PfBAL). Because only few mechanistic kinetic models for carboligation and simultaneous inactivation are available today, we quantitatively determined the reaction kinetics and inactivation of the self-carboligation of benzaldehyde yielding the product (R)-benzoin catalyzed by PfBAL directly from progress curves using model-based experimental analysis...
June 8, 2018: Biotechnology Progress
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