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Biotechnology Progress

Marc Bieberbach, Peter Kosiol, Alexander Seay, Moritz Bennecke, Björn Hansmann, Stefan Hepbildikler, Volkmar Thom
The downstream process development of novel antibodies (Abs) is often challenged by virus filter fouling making a better understanding of the underlying mechanisms highly desirable. The present study combines the protein characterization of different feedstreams with their virus filtration performance using a novel high throughput filtration screening system. Filtration experiments with Ab concentrations of up to 20 g/L using either low interacting or hydrophobically interacting pre-filters indicate the existence of two different fouling mechanisms, an irreversible and a reversible one...
January 10, 2019: Biotechnology Progress
Binglin Li, Jiao Wang, Jing Li, Xiaoli Zhang, Wenyu Yu, Huan Wang, Dandan Duan, Binxia Zhao
The efficient and environmentally friendly aqueous-solid systems employed Triton-X-100-modified silica as the "artificial interface" to adsorb phosphatidylcholine (PC) in purely aqueous solutions and silica-adsorbed PC was successfully used for phospholipase D (PLD)-mediated transphosphatidylation. Three kinds of silicas with different sizes were employed to investigate advantages and disadvantages of batch and continuous technologies for PLD-catalyzed transphosphatidylation in aqueous-solid systems...
January 10, 2019: Biotechnology Progress
Nils A Brechmann, Per-Olov Eriksson, Kristofer Eriksson, Sven Oscarsson, Jos Buijs, Atefeh Shokri, Göran Hjälm, Véronique Chotteau
High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development of a new process for affinity purification of monoclonal antibodies (mAbs) from non-clarified CHO cell broth using a pilot-scale magnetic separator. The LOABeads had a maximum binding capacity of 65 mg/ml and an adsorption capacity of 25-42 mg IgG/ml bead in suspension for an IgG concentration of 1 to 8 g/L. Pilot-scale separation was initially tested in a mAb capture step from 26 L clarified harvest. Small-scale experiments showed that similar mAb adsorptions were obtained in cell broth containing 40 ×106 cells/ml as in clarified supernatant...
January 10, 2019: Biotechnology Progress
Richa Bafana, Saravanadevi Sivanesan, R A Pandey
Present study used Aspergillus terreus strain C1 isolated from mangrove soil for itaconic acid (IA) production from potato starch waste. Fermentation parameters were optimized by classical one factor approach and statistical experimental designs, such as Plackett-Burman and response surface designs. Anionic deionization of potato waste was found to be a very effective, economic and easy way of improving IA production. The increase in IA production by deionization was found to correlate with removal of phosphate...
January 9, 2019: Biotechnology Progress
Malihatosadat Gholamzadeh, Nahid Hosseinzadeh Gharajeh, Mohammad Amin Hejazi
No abstract text is available yet for this article.
December 28, 2018: Biotechnology Progress
Tomofumi Tadauchi, Cynthia Lam, Laura Liu, Yizhou Zhou, Danming Tang, Salina Louie, Brad Snedecor, Shahram Misaghi
Chinese hamster ovary (CHO) cells are conventionally used to generate therapeutic cell lines via random integration (RI), where desired transgenes are stably integrated into the genome. Targeted integration (TI) approaches, which involve integration of a transgene into a specific locus in the genome, are increasingly utilized for CHO cell line development (CLD) in recent years. None of these CLD approaches, however, are suitable for expression of toxic or difficult-to-express molecules, or for determining the underlying causes for poor expression of some molecules...
December 28, 2018: Biotechnology Progress
Saumya Ahlawat, Mehak Kaushal, Basavaraj Palabhanvi, Muthusivaramapandian Muthuraj, Gargi Goswami, Debasish Das
The present study demonstrates a process engineering strategy to achieve high butanol titer and productivity from wild type Clostridium acetobutylicum MTCC 11274. In the first step, two different media were optimized with the objectives of maximizing the biomass and butanol productivity respectively. In the next step, attributes of these two media compositions were integrated to design a two stage fed-batch process which resulted in maximal butanol productivity of 0.55 g L-1 h-1 with titer of 13.1 g L-1 . Further, two stage fed-batch process along with combinatorial use of magnesium limitation and calcium supplementation resulted in highest butanol titer and productivity of 16...
December 28, 2018: Biotechnology Progress
K M Eskridge, S G Gilmour, L G Posadas
Fields such as, diagnostic testing, biotherapeutics, drug development and toxicology among others, center on the premise of searching through many specimens for a rare event. Scientists in the business of "searching for a needle in a haystack" may greatly benefit from the use of group screening design strategies. Group screening, where specimens are composited into pools with each pool being tested for the presence of the event, can be much more cost efficient than testing each individual specimen...
December 28, 2018: Biotechnology Progress
Sabrina Ait Braham, Fouzia Hussain, Roberto Morellon-Sterling, Shagufta Kamal, Jakub F Kornecki, Oveimar Barbosa, DjamelEdine Kati, Roberto Fernandez-Lafuente
Alcalase was scarcely immobilized on monoaminoethyl-N-aminoethyl (MANAE)- agarose beads at different pH values (less than 20% at pH 7). The enzyme did not immobilize on MANAE-agarose activated with glutaraldehyde at high ionic strength, suggesting a low reactivity of the enzyme with the support functionalized in this manner. However, the immobilization is relatively rapid when using low ionic strength and glutaraldehyde activated support. Using these conditions, the enzyme was immobilized at pH 5, 7 and 9, and in all cases the activity versus Boc-Ala-ONp decreased to around 50%...
December 21, 2018: Biotechnology Progress
Adriana Amalfitano, Cecilia Martini, Giuseppina Nocca, Massimiliano Papi, Marco De Spirito, Maurizio Sanguinetti, Alberto Vitali, Francesca Bugli, Alessandro Arcovito
In the modern view of selective drug delivery of bioactive molecules, the attention is moving onto the setup of the perfect carrier more than in the optimization of the active compound. In this respect, virus-like particles constitute bioinspired nanodevices with the intrinsic ability to transport a large class of molecules, ranging from smart drugs to small interfering RNAs. In this work, we demonstrate the efficacy of a novel construct obtained fusing a self-assembling protein from the human Rotavirus A, VP6, with the Small Ubiquitin Modifier domain (SUMO), that maintains the ability to form nanoparticles and nanotubes and is able to be used as a drug carrier, even without specific targeting epitopes...
December 21, 2018: Biotechnology Progress
Nasib Qureshi, Rogers Harry-O'kuru, Siqing Liu, Badal Saha
Yellow Top (Physaria fendleri) is a plant that belongs to the mustard family. This plant is used to produce seeds that are rich in hydroxy oil. After extraction of oil, the presscake is land-filled. The seedcake is rich in polymeric sugars and can be used for various bioconversions. For the present case, the seedcake or presscake was hydrolyzed with dilute [0.50% (v/v)] H2 SO4 and enzymes to release sugars including glucose, xylose, galactose, arabinose and mannose. Then the hydrolyzate was used to produce acetone butanol ethanol (ABE)...
December 19, 2018: Biotechnology Progress
Alice Zoso, Alessandro Zambon, Onelia Gagliano, Stefano Giulitti, Luca Prevedello, Gian Paolo Fadini, Camilla Luni, Nicola Elvassore
Systemic diseases affect multiple tissues that interact with each other within a network difficult to explore at the body level. However, understanding the inter-dependences between tissues could be of high relevance for drug target identification, especially at the first stages of disease development. In vitro systems have the advantages of accessibility to measurements and precise controllability of culture conditions, but currently have limitations in mimicking human in vivo systemic tissue response. In this work, we present an in vitro model of cross-talk between an ex vivo culture of adipose tissue from an obese donor and a skeletal muscle in vitro model from a healthy donor...
December 13, 2018: Biotechnology Progress
Ying Li, Ivan Manzano, Andrew L Zydney
It is well established that the structure of plasmid DNA is a strong function of solution ionic conditions due to changes in intramolecular electrostatic interactions between the charged phosphate groups along the DNA backbone. Multivalent cations like spermine and spermidine play a critical role in compacting and controlling the structure of supercoiled DNA in living cells. The objective of this work was to investigate the effects of these polyamines on the ultrafiltration of plasmid DNA, including possible opportunities to use these polycations to enhance the purification of specific plasmid isoforms...
December 10, 2018: Biotechnology Progress
Virna M Giménez, Norma Sperandeo, Sonia Faudone, Sandra Noriega, Walter Manucha, Diego Kassuha
The electrospraying technique provides nano and microparticles that can be used as drug delivery systems. The aims of this study were, firstly, to optimize the influent parameters of electrospraying for the manufacture of a Bosentan (BOS) nanoparticulate platform, and secondly, to evaluate its physicochemical properties and in vitro biopharmaceutical behavior. Particles were characterized by scanning electron microscopy (SEM), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), thermogravimetry (TG) and Fourier transformed Infrared spectroscopy (FTIR)...
December 8, 2018: Biotechnology Progress
Vincent Leung, Meredith Brooks, Sophia Emerson, M Monsur Ali, Carlos D M Filipe
Rolling circle amplification (RCA), polymerase chain reaction (PCR), and loop-mediated isothermal amplification (LAMP), are powerful tools that can be used for gene manipulation, pathogen detection, and infectious disease diagnostics. However, these techniques require trained personnel, as the pipetting steps involved can lead to contamination and, consequently, erroneous results. Furthermore, many of the reagents used in molecular biology are thermally labile and must be kept within a cold-chain. In this paper, we present a simple and cost-effective method that allows molecular biology reagents to be thermally stabilized into ready-to-use mastermixes via drying in pullulan and trehalose films...
December 8, 2018: Biotechnology Progress
Christopher J Yehl, Mario G Jabra, Andrew L Zydney
Buffer exchange, desalting, and formulation of high value biotherapeutics are currently performed using batch diafiltration; however, this type of tangential flow filtration process may be difficult to implement as part of a fully continuous biomanufacturing process. The objective of this study was to explore the potential of using countercurrent dialysis for continuous protein formulation and buffer exchange. Experiments were performed using concentrated solutions of Immunoglobuin G (IgG) with commercially available hollow fiber dialyzers having 1...
December 6, 2018: Biotechnology Progress
Ricardo F Caroço, Marta Bevilacqua, Ibrahim Armagan, Paloma A Santacoloma, Jens Abildskov, Thomas Skov, Jakob K Huusom
This article explores different opportunities to evaluate quality variation in raw materials from biological origin. Assessment of raw materials attributes is an important step in a bio-based production since fluctuations in quality are a major source of process disturbance. This can be due to a variety of biological, seasonal and supply scarcity reasons. The final properties of a product are invariably linked with the initial properties of the raw material. Thus, the operational conditions of a process can be tuned to drive the product to the required specification based on the quality assessment of the raw material being processed...
December 3, 2018: Biotechnology Progress
T K Karakach, A Dachon, J Choi, C Miguez, L Masson, B Tartakovsky
This study describes the application of the multivariate curve resolution (MCR) analysis technique for real-time analysis of culture fluorescence during recombinant Pichia pastoris cultivation in a bioreactor. Fluorescence spectra were acquired with an on-line dual excitation wavelength fluorometer and then used to develop a real time MCR-based bioprocess monitoring and diagnostics tool. Initial bioreactor experiments using two similar recombinant antibody secreting P. pastoris cell lines showed significant differences in protein production...
December 2, 2018: Biotechnology Progress
Sotirios-Spyridon Vamvakas, John Kapolos, Lambros Farmakis, Fotios Genneos, Maria-Eleni Damianaki, Xenia Chouli, Aggeliki Vardakou, Sofia Liosi, Evgenia Stavropoulou, Eleftheria Leivaditi, Marianthi Fragki, Elina Labrakou, Eleni-Giselda Gashi, Dimitra Demoli
Despite the fact that Saccharomyces cerevisiae has suicide tendencies since its product affects cell function, it is a key player in alcoholic fermentation. The presence of ethanol in the medium affects membrane integrity and fluidity, as well as the rate of ethanol production. Msn2/4p transcription factors are key regulators in stress response and play a critical role in cell response to ethanol challenge. Protein kinase A (tpk1/2/3) is controlling the activation/inactivation of a multitude of proteins through phosphorylation at specific serine residues...
December 2, 2018: Biotechnology Progress
Sven Markert, Carsten Musmann, Peter Hülsmann, Klaus Joeris
Recently, we established an automated microtiter plate (MTP)-based system for suspension cell culture for high throughput (HT) applications in biopharmaceutical process development. In the present report the new system was evaluated regarding its potential to improve clone screening by allowing high-throughput fed-batch cultivation at an early stage. For this purpose, a fully automated procedure was compared to a mainly batch mode-based manual standard process. The new system performed daily measurements of viable cell density and product concentration for a total of 96 clones in biological duplicates that were evaluated for final clone selection within two weeks...
December 2, 2018: Biotechnology Progress
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