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Biotechnology Progress

Sean Yu, Pranav Joshi, Yi Ju Park, Kyeong-Nam Yu, Moo-Yeal Lee
Layer-by-layer cell printing is useful in mimicking layered tissue structures inside the human body and has great potential for being a promising tool in the field of tissue engineering, regenerative medicine, and drug discovery. However, imaging human cells cultured in multiple hydrogel layers in 3D-printed tissue constructs is challenging as the cells are not in a single focal plane. Although confocal microscopy could be a potential solution for this issue, it compromises the throughput which is a key factor in rapidly screening drug efficacy and toxicity in pharmaceutical industries...
December 14, 2017: Biotechnology Progress
Retno Wahyu Nurhayati, Yoshihiro Ojima, Takeaki Dohda, Masahiro Kino-Oka
The increasing application of regenerative medicine has generated a growing demand for stem cells and their derivatives. Single-use bioreactors offer an attractive platform for stem cell expansion owing to their scalability for large-scale production and feasibility of meeting clinical-grade standards. The current work evaluated the capacity of a single-use bioreactor system (1 L working volume) for expanding Meg01 cells, a megakaryocytic (MK) progenitor cell line. Oxygen supply was provided by surface aeration to minimize foaming and orbital shaking was used to promote oxygen transfer...
December 11, 2017: Biotechnology Progress
Athul Rajesh, Devon Zimmerman, Kyle Spivack, Osheiza Abdulmalik, Jacob Elmer
Since donated red blood cells must be constantly refrigerated, they are not available in remote areas and battlefields. We have previously shown that the hemoglobin of the earthworm Lumbricus terrestris (LtEc) is an effective and safe substitute for donated blood that is stable enough to be stored for long periods at the relatively high temperatures that may be encountered in remote areas. The goal of this study was to increase the thermal stability of LtEc by covalently cross-linking LtEc with glutaraldehyde (gLtEc)...
December 11, 2017: Biotechnology Progress
Peggy Ko, Shahram Misaghi, Zhilan Hu, Dejin Zhan, Joni Tsukuda, Mandy Yim, Mark Sanford, David Shaw, Masaru Shiratori, Brad Snedecor, Michael Laird, Amy Shen
In the past few decades, a large variety of therapeutic antibodies and proteins have been expressed in Chinese hamster ovary (CHO) cells. This mammalian expression system is robust, scalable, relatively inexpensive, and importantly allows for post-translational modifications that are important for some therapeutic proteins. Historically, CHO cell lines were derived from colonies of cells grown in semi-solid or liquid plates using either serum-containing or serum-free media. Current advancements in cell sorting and imaging technologies have allowed for isolating and imaging single cell progenitors at the seeding step, significantly increasing the probability of isolating clonally-derived cell lines...
December 11, 2017: Biotechnology Progress
Karthik Veeravalli, Tony Schindler, Emily Dong, Masaki Yamada, Ryan Hamilton, Michael W Laird
Microaerobic (oxygen limited) conditions are advantageous for several industrial applications since a majority of the carbon atoms can be directed for synthesis of desired products. Oxygen limited conditions, however, can result in high levels of undesirable by-products such as acetate which subsequently can have an impact on biomass and product yields. The molecular mechanisms involved in acetate accumulation under oxygen limited conditions are not well understood. Our results indicate that a majority of the genetic modifications known to decrease acetate under aerobic conditions results in similar or even higher acetate under oxygen limitation...
November 30, 2017: Biotechnology Progress
Glen Giese, Ambrose Williams, Maricel Rodriguez, Josefine Persson
Production of knob and hole dual light chain bispecific antibodies poses several unique challenges for development of a feasible industrial scale manufacturing process. We developed an efficient process for the assembly and purification of knob and hole dual light chain bispecific antibodies. Two distinct half-antibodies targeting two different antigens were expressed separately in E. coli cells and captured independently using Protein A chromatography. When combined, the knob and hole mutations in the CH3 domains promoted heterodimer formation...
November 29, 2017: Biotechnology Progress
Jamie Sharp, Tim Wgm Spitters, Patrick Vermette
Few studies report whole pancreatic tissue culture, as it is a difficult task using traditional culture methods. Here, a factorial design was used to investigate the singular and combinational effects of flow, dissolved oxygen concentration (D.O.) and pulsation on whole mechanically-disrupted rat pancreata in a perfusion bioreactor. Whole rat pancreata were cultured for 72h under defined bioreactor process conditions. Secreted insulin was measured and histological (haematoxylin and eosin (H&E)) as well as immunofluorescent insulin staining were performed and quantified...
November 29, 2017: Biotechnology Progress
Daniela B Hirsch, María F Baieli, Nicolás Urtasun, Juan M Lázaro-Martínez, Romina J Glisoni, María V Miranda, Osvaldo Cascone, Federico J Wolman
A cation exchange matrix with zwitterionic and multi-modal properties was synthesized by a simple reaction sequence coupling sulfanilic acid to a chitosan based support. The novel chromatographic matrix was physico-chemically characterized by ss-NMR and ζ potential, and its chromatographic performance was evaluated for lysozyme purification from diluted egg white. The maximum adsorption capacity, calculated according to Langmuir adsorption isotherm, was 50.07 ± 1.47 mg g-1 while the dissociation constant was 0...
November 29, 2017: Biotechnology Progress
Marwa I Wahba
The poor mechanical stability of chitosan has long impeded its industrial utilization as an immobilization carrier. In this study, the mechanical properties of chitosan beads were greatly improved through utilizing the slow rate of the sodium bicarbonate-induced chitosan gelation and combining it with the chemical cross-linking action of glutaraldehyde (GA). The GA-treated sodium bicarbonate-gelled chitosan beads exhibited much better mechanical properties and up to 2.45-fold higher observed activity of the immobilized enzyme (β-D-galactosidase (β-gal)) when compared to the GA-treated sodium tripolyphosphate (TPP)-gelled chitosan beads...
November 29, 2017: Biotechnology Progress
Ryo Hamamoto, Hidemi Ito, Makoto Hirohara, Ryongsok Chang, Tomoko Hongo-Hirasaki, Tomohiro Hayashi
Membrane fouling commonly occurs in all filter types during virus filtration in protein-based biopharmaceutical manufacturing. Mechanisms of decline in virus filter performance due to membrane fouling were investigated using a cellulose-based virus filter as a model membrane. Filter performance was critically dependent on solution conditions; specifically, ionic strength. To understand the interaction between immunoglobulin G (IgG) and cellulose, sensors coated with cellulose were fabricated for surface plasmon resonance and quartz crystal microbalance with energy dissipation measurements...
November 29, 2017: Biotechnology Progress
Christine DeMaria, Trent Munro
No abstract text is available yet for this article.
November 28, 2017: Biotechnology Progress
William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Eugene Schaefer
The ambr bioreactors are single-use microbioreactors for cell line development and process optimization. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. While there have been some recent studies evaluating the ambr15™ technology as a scale down model for fed-batch operations, little has been reported for semi-continuous or continuous operation...
November 20, 2017: Biotechnology Progress
Alessandro Mora, Sheng Sam Zhang, Gerald Carson, Bernard Nabiswa, Patrick Hossler, Seongkyu Yoon
Efficient and effective cell line screening is paramount towards a successful biomanufacturing program. Here we describe the implementation of 24-deep well plate screening of CHO lines as part of the cell line development platform at AbbVie. Incorporation of this approach accelerated the identification of the best candidate lines for process development. In an effort to quantify and predict process performance comparability, we compared cell culture performance in 24-deep well plates (24-DWP) and in shake flasks, for a panel of Chinese Hamster Ovary cell lines expressing a monoclonal antibody...
November 18, 2017: Biotechnology Progress
Nipon Sarmah, Revathi D, Sheelu G, Yamuna Rani K, S Sridhar, Mehtab V, Sumana C
Lipases are the industrially important biocatalysts, which are envisioned to have tremendous applications in the manufacture of a wide range of products. Their unique properties such as better stability, selectivity and substrate specificity position them as the most expansively used industrial enzymes. The research on production and applications of lipases is ever growing and there exists a need to have a latest review on the research findings of lipases. The present review aims at giving the latest and broadest overall picture of research and development on lipases by including the current studies and progressions not only in the diverse industrial application fields of lipases, but also with regard to its structure, classification and sources...
October 31, 2017: Biotechnology Progress
Fumiyuki Kobayashi, Sachiko Odake
The effect of ethanol on the inactivation of Saccharomyces pastorianus by a two-stage system with low pressure carbon dioxide microbubbles (two-stage MBCO2 ) was investigated. Zero and > 5 log reductions of S. pastorianus populations suspended in physiological saline (PS) containing 0 and 10% ethanol, respectively, occurred by the two-stage MBCO2 at a mixing vessel pressure of 1 MPa and a heating coil temperature of 40°C. Conversely, the detected number of surviving S. pastorianus cells in PS containing 5% ethanol was higher in yeast and mould agar (YMA, an optimum agar) than YMA with 2...
October 31, 2017: Biotechnology Progress
Mohammad Golzarijalal, Farzin Zokaee Ashtiani, Bahram Dabir
In this study, shear-induced flocculation modelling of Chlorella sp. microalgae was conducted by combination of Population Balance Modelling (PBM) and CFD. The inhomogeneous Multiple Size Group (MUSIG) and the Euler-Euler two fluid models were coupled via Ansys-CFX-15 software package to achieve both fluid and particle dynamics during the flocculation. For the first time, a detailed model was proposed to calculate the collision frequency and breakage rate during the microalgae flocculation by means of the response surface methodology (RSM) as a tool for optimization...
October 31, 2017: Biotechnology Progress
Jafar Vatandoost, Omid Zarei Sani
The production of recombinant K-proteins (VKD) for therapeutic purposes is an important challenge in the pharmaceutical industry. These proteins are primarily synthesized as precursor molecules and contain prepro-peptide sequences. The pro-peptide is connected to γ-carboxylase enzyme through the γ-carboxylase recognition site for the direct γ-carboxylation of VKD proteins that has a significant impact on their biological activity. Pro-peptides have different attitudes toward γ-carboxylase and certain amino acids in pro-peptide sequences are responsible for the differences in γ-carboxylase affinity...
October 31, 2017: Biotechnology Progress
Frank H Kung, David Sillitti, David I Shreiber, Jeffrey D Zahn, Bonnie L Firestein
The construction of biomaterials with which to limith the growth of cells or to limit the adsorbtion of proteins is essential for understanding biological phenomena. Here, we describe a novel method to simply and easily create thin layers of poly (2-hydroxyethyl methacrylate) (p-HEMA) for protein and cellular patterning via etching with ethanol and microfluidic devices. First, a cell culture surface or glass coverslip is coated with p-HEMA. Next, a polydimethylsiloxane (PDMS) microfluidic is placed onto the p-HEMA surface, and ethanol is aspirated through the device...
October 31, 2017: Biotechnology Progress
Sai Rashmika Velugula-Yellela, Abasha Williams, Nicholas Trunfio, Chih-Jung Hsu, Brittany Chavez, Seongkyu Yoon, Cyrus Agarabi
Monoclonal antibody production in commercial scale cell culture bioprocessing requires a thorough understanding of the engineering process and components used throughout manufacturing. It is important to identify high impact components early on during the lifecycle of a biotechnology derived product. While cell culture media selection is of obvious importance to the health and productivity of mammalian bioreactor operations, other components such as antifoam selection can also play an important role in bioreactor cell culture...
October 31, 2017: Biotechnology Progress
Yu-Chia Wei, Stephanie Braun-Galleani, Maria José Henríquez, Sahan Bandara, Darren Nicholas Nesbeth
Transketolase is a proven biocatalytic tool for asymmetric carbon-carbon bond formation, both as a purified enzyme and within bacterial whole-cell biocatalysts. The performance of Pichia pastoris as a host for transketolase whole-cell biocatalysis was investigated using a transketolase-overexpressing strain to catalyse formation of L-erythrulose from β-hydroxypyruvic acid and glycolaldehyde substrates. Pichia pastoris transketolase coding sequence from the locus PAS_chr1-4_0150 was subcloned downstream of the methanol-inducible AOX1 promoter in a plasmid for transformation of strain GS115, generating strain TK150...
October 31, 2017: Biotechnology Progress
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