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tangential flow filtration

Sulena Polez, Domenico Origi, Sotir Zahariev, Corrado Guarnaccia, Sergio G Tisminetzky, Nataša Skoko, Marco Baralle
A significant barrier to insulin is affordability. In this manuscript we describe improvements to key steps in the insulin production process in Pichia pastoris that reduce cost and time. The strategy for recovery and processing of human insulin precursor has been streamlined to two steps from bioreactor to the transpeptidation reaction. In the first step the insulin precursor secreted during the methanol induction phase is recovered directly from the culture broth using Tangential Flow Filtration with a Prostak™ module eliminating the laborious and time-consuming multi-step clarification, including centrifugation...
2016: PloS One
Antonia Bruno, Anna Sandionigi, Andrea Galimberti, Eleonora Siani, Massimo Labra, Clementina Cocuzza, Emanuele Ferri, Maurizio Casiraghi
We propose an innovative, repeatable, and reliable experimental workflow to concentrate and detect environmental bacteria in drinking water using molecular techniques. We first concentrated bacteria in water samples using tangential flow filtration and then we evaluated two methods of environmental DNA extraction. We performed tests on both artificially contaminated water samples and real drinking water samples. The efficiency of the experimental workflow was measured through qPCR. The successful applicability of the high-throughput DNA sequencing (HTS) approach was demonstrated on drinking water samples...
October 27, 2016: MicrobiologyOpen
Md Nasimuzzaman, Danielle Lynn, Rebecca Ernst, Michele Beuerlein, Richard H Smith, Archana Shrestha, Scott Cross, Kevin Link, Carolyn Lutzko, Diana Nordling, David W Russell, Andre Larochelle, Punam Malik, Johannes C M Van der Loo
Compared to other integrating viral vectors, foamy virus (FV) vectors have distinct advantages as a gene transfer tool, including their nonpathogenicity, the ability to carry larger transgene cassettes, and increased stability of virus particles due to DNA genome formation within the virions. Proof of principle of its therapeutic utility was provided with the correction of canine leukocyte adhesion deficiency using autologous CD34(+) cells transduced with FV vector carrying the canine CD18 gene, demonstrating its long-term safety and efficacy...
2016: Molecular Therapy. Methods & Clinical Development
N F Santos-Sánchez, R Valadez-Blanco, B Hernández-Carlos, A Torres-Ariño, P C Guadarrama-Mendoza, R Salas-Coronado
Despite microalgae recently receiving enormous attention as a potential source of biodiesel, their use is still not feasible as an alternative to fossil fuels. Recently, interest in microalgae has focused on the production of bioactive compounds such as polyunsaturated fatty acids (PUFA), which provide microalgae a high added value. Several considerations need to be assessed for optimizing PUFA production from microalgae. Firstly, a microalgae species that produces high PUFA concentrations should be selected, such as Nannochloropsis gaditana, Isochrysis galbana, Phaeodactylum tricornutum, and Crypthecodinium cohnii, with marine species gaining more attention than do freshwater species...
October 2016: Applied Microbiology and Biotechnology
Kirsten Schneider-Ohrum, Corinne Cayatte, Yi Liu, Zhaoti Wang, Alivelu Irrinki, Floro Cataniag, Nga Nguyen, Stacie Lambert, Hui Liu, Shahin Aslam, Greg Duke, Michael P McCarthy, Louise McCormick
: With the goal of developing a virus-like particle-based vaccine based on dense bodies (DB) produced by human cytomegalovirus (HCMV) infections, we evaluated scalable culture, isolation, and inactivation methods and applied technically advanced assays to determine the relative purity, composition, and immunogenicity of DB particles. Our results increase our understanding of the benefits and disadvantages of methods to recover immunogenic DB and inactivate contaminating viral particles...
November 15, 2016: Journal of Virology
Wesley Zmolek, Stefanie Bañas, Robyn M Barfield, David Rabuka, Penelope M Drake
Antibody-drug conjugates represent a growing class of biologic drugs that use the targeted specificity of an antibody to direct the localization of a small molecule drug, often a cytotoxic payload. After conjugation, antibody-drug conjugate preparations typically retain a residual amount of free (unconjugated) linker-payload. Monitoring this free small molecule drug component is important due to the potential for free payload to mediate unintended (off-target) toxicity. We developed a simple RP-HPLC/MRM-MS-based assay that can be rapidly employed to quantify free linker-payload...
October 1, 2016: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Ralf P Friedrich, Jan Zaloga, Eveline Schreiber, Ildikó Y Tóth, Etelka Tombácz, Stefan Lyer, Christoph Alexiou
Functionalized superparamagnetic iron oxide nanoparticles are frequently used to develop vehicles for drug delivery, hyperthermia, and photodynamic therapy and as tools used for magnetic separation and purification of proteins or for biomolecular imaging. Depending on the application, there are various possible covalent and non-covalent approaches for the functionalization of particles, each of them shows different advantages and disadvantages for drug release and activity at the desired location.Particularly important for the production of adsorptive and covalent bound drugs to nanoparticles is the pureness of the involved formulation...
December 2016: Nanoscale Research Letters
Sophia T Mundle, Michael Kishko, Rachel Groppo, Joshua DiNapoli, John Hamberger, Bryan McNeil, Harry Kleanthous, Mark Parrington, Linong Zhang, Stephen F Anderson
Respiratory syncytial virus (RSV) is an important human pathogen, and is the most frequent viral cause of severe respiratory disease in infants. In addition, it is increasingly being recognized as an important cause of respiratory disease in the elderly and immunocompromised. Although a passive prophylactic treatment does exist for high-risk neonates and children, the overall disease burden warrants the development of a safe and effective prophylactic vaccine for use in otherwise healthy newborns and children...
July 12, 2016: Vaccine
Pierre-Olivier Buclez, Gabriella Dias Florencio, Karima Relizani, Cyriaque Beley, Luis Garcia, Rachid Benchaouir
Recombinant adeno-associated viruses (rAAV) are largely used for gene transfer in research, preclinical developments, and clinical trials. Their broad in vivo biodistribution and long-term efficacy in postmitotic tissues make them good candidates for numerous gene transfer applications. Upstream processes able to produce large amounts of rAAV were developed, particularly those using baculovirus expression vector system. In parallel, downstream processes present a large panel of purification methods, often including multiple and time consuming steps...
2016: Molecular Therapy. Methods & Clinical Development
Alina Venereo-Sanchez, Renald Gilbert, Melanie Simoneau, Antoine Caron, Parminder Chahal, Wangxue Chen, Sven Ansorge, Xuguang Li, Olivier Henry, Amine Kamen
Virus-like particles (VLPs) constitute a promising alternative as influenza vaccine. They are non-replicative particles that mimic the morphology of native viruses which make them more immunogenic than classical subunit vaccines. In this study, we propose HEK-293 cells in suspension culture in serum-free medium as an efficient platform to produce large quantities of VLPs. For this purpose, a stable cell line expressing the main influenza viral antigens hemagglutinin (HA) and neuraminidase (NA) (subtype H1N1) under the regulation of a cumate inducible promoter was developed (293HA-NA cells)...
June 17, 2016: Vaccine
Vyacheslav Furtak, Merja Roivainen, Olga Mirochnichenko, Tatiana Zagorodnyaya, Majid Laassri, Sohail Z Zaidic, Lubna Rehman, Muhammad M Alam, Vladimir Chizhikov, Konstantin Chumakov
An approach is proposed for environmental surveillance of poliovirus by concentrating sewage samples with tangential flow filtration (TFF) followed by deep sequencing of viral RNA. Subsequent to testing the method with samples from Finland, samples from Pakistan, a country endemic for poliovirus, were investigated. Genomic sequencing was either performed directly, for unbiased identification of viruses regardless of their ability to grow in cell cultures, or after virus enrichment by cell culture or immunoprecipitation...
April 14, 2016: Euro Surveillance: Bulletin Européen sur les Maladies Transmissibles, European Communicable Disease Bulletin
Daria Popova, Adam Stonier, David Pain, Nigel J Titchener-Hooker, Suzanne S Farid
Increases in mammalian cell culture titres and densities have placed significant demands on primary recovery operation performance. This article presents a methodology which aims to screen rapidly and evaluate primary recovery technologies for their scope for technically feasible and cost-effective operation in the context of high cell density mammalian cell cultures. It was applied to assess the performance of current (centrifugation and depth filtration options) and alternative (tangential flow filtration (TFF)) primary recovery strategies...
July 2016: Biotechnology Journal
Rizwan R Soomro, Theoneste Ndikubwimana, Xianhai Zeng, Yinghua Lu, Lu Lin, Michael K Danquah
Even though microalgal biomass is leading the third generation biofuel research, significant effort is required to establish an economically viable commercial-scale microalgal biofuel production system. Whilst a significant amount of work has been reported on large-scale cultivation of microalgae using photo-bioreactors and pond systems, research focus on establishing high performance downstream dewatering operations for large-scale processing under optimal economy is limited. The enormous amount of energy and associated cost required for dewatering large-volume microalgal cultures has been the primary hindrance to the development of the needed biomass quantity for industrial-scale microalgal biofuels production...
2016: Frontiers in Plant Science
Janina Brand, Katharina Voigt, Bianca Zochowski, Ulrich Kulozik
The application of membrane adsorbers instead of classical packed bed columns for protein fractionation is still a growing field. In the case of egg white protein fractionation, the application of classical chromatography is additionally limited due to its high viscosity that impairs filtration. By using tangential flow membrane adsorbers as stationary phase this limiting factor can be left out, as they can be loaded with particle containing substrates. The flow conditions existing in tangential flow membrane adsorbers are not fully understood yet...
March 18, 2016: Journal of Chromatography. A
N A Willoughby, H Bock, M A Hoeve, S Pells, C Williams, G McPhee, P Freile, D Choudhury, P A De Sousa
The broad capacity of pluripotent human embryonic stem cells (hESC) to grow and differentiate demands the development of rapid, scalable, and label-free methods to separate living cell populations for clinical and industrial applications. Here, we identify differences in cell stiffness, expressed as cell elastic modulus (CEM), for hESC versus mesenchymal progenitors, osteoblast-like derivatives, and fibroblasts using atomic force microscopy and data processing algorithms to characterize the stiffness of cell populations...
January 2016: Biomicrofluidics
Felipe Tapia, Daniel Vázquez-Ramírez, Yvonne Genzel, Udo Reichl
With an increasing demand for efficacious, safe, and affordable vaccines for human and animal use, process intensification in cell culture-based viral vaccine production demands advanced process strategies to overcome the limitations of conventional batch cultivations. However, the use of fed-batch, perfusion, or continuous modes to drive processes at high cell density (HCD) and overextended operating times has so far been little explored in large-scale viral vaccine manufacturing. Also, possible reductions in cell-specific virus yields for HCD cultivations have been reported frequently...
March 2016: Applied Microbiology and Biotechnology
Lise Besnard, Virginie Fabre, Michael Fettig, Elina Gousseinov, Yasuhiro Kawakami, Nicolas Laroudie, Claire Scanlan, Priyabrata Pattnaik
Vaccines are derived from a variety of sources including tissue extracts, bacterial cells, virus particles, recombinant mammalian, yeast and insect cell produced proteins and nucleic acids. The most common method of vaccine production is based on an initial fermentation process followed by purification. Production of vaccines is a complex process involving many different steps and processes. Selection of the appropriate purification method is critical to achieving desired purity of the final product. Clarification of vaccines is a critical step that strongly impacts product recovery and subsequent downstream purification...
January 2016: Biotechnology Advances
Douglas S Watson, Kristi R Kerchner, Sean S Gant, Joseph W Pedersen, James B Hamburger, Allison D Ortigosa, Thomas I Potgieter
Tangential flow microfiltration (MF) is a cost-effective and robust bioprocess separation technique, but successful full scale implementation is hindered by the empirical, trial-and-error nature of scale-up. We present an integrated approach leveraging at-line process analytical technology (PAT) and mass balance based modeling to de-risk MF scale-up. Chromatography-based PAT was employed to improve the consistency of an MF step that had been a bottleneck in the process used to manufacture a therapeutic protein...
January 2016: Biotechnology Progress
Devon Zimmerman, Jack Dienes, Osheiza Abdulmalik, Jacob J Elmer
Although donated blood is the preferred material for transfusion, its limited availability and stringent storage requirements have motivated the development of blood substitutes. The giant extracellular hemoglobin (aka erythrocruorin) of the earthworm Lumbricus terrestris (LtEc) has shown promise as a blood substitute, but an efficient purification method for LtEc must be developed to meet the potential large demand for blood substitutes. In this work, an optimized purification process that uses divalent and trivalent metal salts to selectively precipitate human, earthworm, and bloodworm hemoglobin (HbA, LtEc, and GdHb, respectively) from crude solutions was developed...
September 2016: Protein Expression and Purification
Lanlan Cai, Yunlan Yang, Nianzhi Jiao, Rui Zhang
Tangential flow filtration (TFF), which has been widely adopted to concentrate a diverse array of microbes from water, is a promising method of microbial separation or removal. However, it is essential to select an optimal membrane suitable for the specific filtration application. This study evaluated two different scales of TFF systems for concentrating and separating microbes (including bacteria and viruses) from contrasting marine waters. Among bacteria-size membranes, polyvinylidene difluoride (PVDF) membranes showed higher bacterial recovery, but lower viral permeation efficiencies than polyethersulfone (PES) membranes, regardless of environments and scales of TFF...
2015: PloS One
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