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https://www.readbyqxmd.com/read/29441788/expression-purification-and-biochemical-characterization-of-human-afamin
#1
Alessandra Altamirano, Andreas Naschberger, Barbara G Furnrohr, Radka Saldova, Weston B Struwe, Patrick M Jennings, Silvia Millán Martín, Suzana Malic, Immanuel Plangger, Stefan Lechner, Reina Pisano, Nicole Peretti, Bernd Linke, Mario M Aguiar, Friedrich Fresser, Andreas Ritsch, Tihana Lenac Rovis, Christina Goode, Pauline M Rudd, Klaus Scheffzek, Bernhard Rupp, Hans Dieplinger
Afamin is an 87 kDa glycoprotein with five predicted N-glycosylation sites. Afamin's glycan abundance contributes to conformational and chemical inhomogeneity presenting great challenges for molecular structure determination. For the purpose of studying the structure of afamin, various forms of recombinantly expressed human afamin (rhAFM) with different glycosylation patterns were thus created. Wild-type rhAFM and various hypo-glycosylated forms were expressed in CHO, CHO-Lec1 and HEK293T cells. Fully non-glycosylated rhAFM was obtained by transfection of point-mutated cDNA to delete all N-glycosylation sites of afamin...
February 14, 2018: Journal of Proteome Research
https://www.readbyqxmd.com/read/29440289/neo212-inhibits-migration-and-invasion-of-glioma-stem-cells
#2
Nagore I Marín-Ramos, Thu Zan Thein, Hee-Yeon Cho, Stephen D Swenson, Weijun Wang, Axel H Schönthal, Thomas C Chen, Florence M Hofman
Glioblastoma multiforme is a malignant brain tumor noted for its extensive vascularity, aggressiveness, and highly invasive nature, suggesting that cell migration plays an important role in tumor progression. The poor prognosis in GBM is associated with a high rate of tumor recurrence, and resistance to the standard of care chemotherapy, temozolomide (TMZ). The novel compound NEO212, a conjugate of TMZ and perillyl alcohol (POH), has proven to be 10 fold more cytotoxic to glioma stem cells (GSCs) than TMZ, and is active against TMZ-resistant tumor cells...
February 13, 2018: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/29437532/using-a-dihydrofolate-reductase-based-strategy-for-producing-the-biosimilar-version-of-pertuzumab-in-cho-s-cells
#3
Amin Ramezani, Abbas Ghaderi
Targeted therapy using monoclonal antibodies (mAbs) against epidermal growth factor receptor-2 (ErbB2) has been utilized for the treatment of breast cancer, recently. Pertuzumab, one of the anti-ErbB2 mAbs, was approved by FDA in 2012 for the treatment of metastatic breast cancer. The aim of this study was to produce biosimilar version of pertuzumab in Chinese hamster ovary (CHO)-S cell line, and compare its ErbB2-binding and biological activities, with commercial drug, Perjeta. To this end, a dihydrofolate reductase (DHFR)-based strategy was used to produce a CHO-S stable cell pool capable of producing high levels of pertuzumab...
February 13, 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/29436586/amelioration-of-inflammatory-responses-by-socheongryong-tang-a-traditional-herbal-medicine-in-raw-264-7%C3%A2-cells-and-rats
#4
Sang Mi Park, Tae Hoon Lee, Rongjie Zhao, Youn Sook Kim, Ji Yun Jung, Chung A Park, Kyung Hwan Jegal, Sae Kwang Ku, Jae Kwang Kim, Chul Won Lee, Young Woo Kim, Il Je Cho, Won G An, Sang Chan Kim
Socheongryong-Tang (SCRT) is a natural medicine prescription that has been mainly used in East Asia for the treatment of inflammatory disorders, including asthma and allergic rhinitis. The present study evaluated the anti-inflammatory effects of SCRT on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and in a rat model of carrageenan (CA)-induced paw edema. Levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and prostaglandin E2 (PGE2) in the culture supernatant were quantified and nitric oxide (NO) production was monitored...
February 6, 2018: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29431425/droplet-array-based-3d-coculture-system-for-high-throughput-tumor-angiogenesis-assay
#5
Xiaohui Du, Wanming Li, Guan-Sheng Du, Hansang Cho, Min Yu, Qun Fang, Luke P Lee, Jin Fang
Angiogenesis is critical for tumor progression and metastasis, and it progresses through orchestral multicellular interactions. Thus, there is urgent demand for high-throughput tumor angiogenesis assays for concurrent examination of multiple factors. For investigating tumor angiogenesis, we developed a microfluidic droplet array-based cell-coculture system comprising a two-layer polydimethylsiloxane chip featuring 6 × 9 paired-well arrays and an automated droplet-manipulation device. In each droplet-pair unit, tumor cells were cultured in 3D in one droplet by mixing cell suspensions with Matrigel, and in the other droplet, human umbilical vein endothelial cells (HUVECs) were cultured in 2D...
February 12, 2018: Analytical Chemistry
https://www.readbyqxmd.com/read/29431186/trogocytosis-of-ligand-receptor-complex-and-its-intracellular-transport-in-cd30-signaling
#6
Makoto Nakashima, Mariko Watanabe, Kaoru Uchimaru, Ryouichi Horie
BACKGROUND INFORMATION: CD30, which is characteristically expressed in classical Hodgkin lymphoma (cHL), is thought to transduce signals by ligation of trimerized CD30 ligand (CD30L) on the surface of surrounding cells and recruitment of downstream molecules. In this report, we propose a new mechanism for CD30 signaling by its ligand. We prepared two stable transformants, CHO cells expressing CD30L fused to mCherry and HeLa cells expressing CD30 fused to GFP. RESULTS: Co-culture of these cells triggered clustering of CD30 and CD30L at the cellular interface, formation of multiple CD30L-CD30 complexes, internalization of these complexes with a portion of the plasma membrane into the HeLa cells, and intracellular transport to the lysosomal compartment...
February 12, 2018: Biology of the Cell
https://www.readbyqxmd.com/read/29429815/long-term-immunogenicity-of-an-initial-booster-dose-of-an-inactivated-vero-cell-culture-derived-japanese-encephalitis-vaccine-je-vc-and-the-safety-and-immunogenicity-of-a-second-je-vc-booster-dose-in-children-previously-vaccinated-with-an-inactivated-mouse
#7
Ki Wook Yun, Hoan Jong Lee, Ji Young Park, Hye-Kyung Cho, Yae-Jean Kim, Kyung-Hyo Kim, Nam Hee Kim, Young Jin Hong, Dong Ho Kim, Hwang Min Kim, Sung-Ho Cha
BACKGROUND: This study was performed with the aim of determining the long-term immunogenicity of an inactivated, Vero cell culture-derived Japanese encephalitis (JE) vaccine (JE-VC) and an inactivated, mouse brain-derived JE vaccine (JE-MB) after the 1st booster dose at 2 years of age, as well as the safety and immunogenicity of the 2nd booster dose of JE-VC at 6 years of age, in children primed and given a 1st booster dose of either JE-VC or JE-MB. METHOD: In this multicenter, open-label clinical trial, the study population consisted of healthy Korean children (aged 6 years) who participated in the previous JE vaccine trial...
February 8, 2018: Vaccine
https://www.readbyqxmd.com/read/29429147/effects-of-several-cosmetic-preservatives-on-ros-dependent-apoptosis-of-rat-neural-progenitor-cells
#8
Onjeon Ryu, Bo Kyung Park, Minji Bang, Kyu Suk Cho, Sung Hoon Lee, Edson Luck T Gonzales, Sung Min Yang, Seonmin Kim, Pyeong Hwa Eun, Joo Young Lee, Kyu-Bong Kim, Chan Young Shin, Kyoung Ja Kwon
Benzalkonium chloride, diazolidinyl urea, and imidazolidinyl urea are commonly used preservatives in cosmetics. Recent reports suggested that these compounds may have cellular and systemic toxicity in high concentration. In addition, diazolidinyl urea and imidazolidinyl urea are known formaldehyde (FA) releasers, raising concerns for these cosmetic preservatives. In this study, we investigated the effects of benzalkonium chloride, diazolidinyl urea, and imidazolidinyl urea on ROS-dependent apoptosis of rat neural progenitor cells (NPCs) in vitro...
February 12, 2018: Biomolecules & Therapeutics
https://www.readbyqxmd.com/read/29427449/butyrated-mannac-analog-improves-protein-expression-in-chinese-hamster-ovary-cells
#9
Bojiao Yin, Qiong Wang, Cheng-Yu Chung, Xiaozhi Ren, Rahul Bhattacharya, Kevin J Yarema, Michael J Betenbaugh
The chemical additive sodium butyrate (NaBu) has been applied in cell culture media as a direct and convenient method to increase the protein expression in Chinese hamster ovary (CHO) and other mammalian cells. In this study, we examined an alternative chemical additive, 1,3,4-O-Bu3 ManNAc, for its effect on recombinant protein production in CHO. Supplementation with 1,3,4-O-Bu3 ManNAc for two stable CHO cell lines, expressing human erythropoietin or IgG, enhanced protein expression for both products with negligible impact on cell growth, viability, glucose utilization, and lactate accumulation...
February 10, 2018: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29427433/highly-sensitive-detection-of-mutations-in-cho-cell-recombinant-dna-using-multi-parallel-single-molecule-real-time-dna-sequencing
#10
Joseph F Cartwright, Karin Anderson, Joseph Longworth, Philip Lobb, David C James
High-fidelity replication of biologic-encoding recombinant DNA sequences by engineered mammalian cell cultures is an essential pre-requisite for the development of stable cell lines for the production of biotherapeutics. However, immortalized mammalian cells characteristically exhibit an increased point mutation frequency compared to mammalian cells in vivo, both across their genomes and at specific loci (hotspots). Thus unforeseen mutations in recombinant DNA sequences can arise and be maintained within producer cell populations...
February 10, 2018: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29408933/inducible-transgenic-expression-of-tripeptidyl-peptidase-1-in-a-mouse-model-of-late-infantile-neuronal-ceroid-lipofuscinosis
#11
Yuliya Nemtsova, Jennifer A Wiseman, Mukarram El-Banna, Peter Lobel, David E Sleat
Late-infantile neuronal ceroid lipofuscinosis is a fatal neurodegenerative disease of children caused by mutations resulting in loss of activity of the lysosomal protease, tripeptidyl peptidase 1 (TPP1). While Tpp1-targeted mouse models of LINCL exist, the goal of this study was to create a transgenic mouse with inducible TPP1 to benchmark treatment approaches, evaluate efficacy of treatment at different stages of disease, and to provide insights into the pathobiology of disease. A construct containing a loxP-flanked stop cassette inserted between the chicken-actin promoter and a sequence encoding murine TPP1 (TgLSL-TPP1) was integrated into the ROSA26 locus in mice by homologous recombination...
2018: PloS One
https://www.readbyqxmd.com/read/29405605/13-c-flux-analysis-reveals-that-rebalancing-medium-amino-acid-composition-can-reduce-ammonia-production-while-preserving-central-carbon-metabolism-of-cho-cell-cultures
#12
Allison G McAtee Pereira, Jason L Walther, Myles Hollenbach, Jamey D Young
13 C metabolic flux analysis (MFA) provides a rigorous approach to quantify intracellular metabolism of industrial cell lines. In this study, 13 C MFA was used to characterize the metabolic response of Chinese hamster ovary (CHO) cells to a novel medium variant designed to reduce ammonia production. Ammonia inhibits growth and viability of CHO cell cultures, alters glycosylation of recombinant proteins, and enhances product degradation. Ammonia production was reduced by manipulating the amino acid composition of the culture medium; specifically, glutamine, glutamate, asparagine, aspartate, and serine levels were adjusted...
February 6, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29400603/a-novel-bicistronic-gene-design-couples-stable-cell-line-selection-with-a-fucose-switch-in-a-designer-cho-host-to-produce-native-and-afucosylated-glycoform-antibodies
#13
Gargi Roy, Tom Martin, Arnita Barnes, Jihong Wang, Rod Brian Jimenez, Megan Rice, Lina Li, Hui Feng, Shu Zhang, Raghothama Chaerkady, Herren Wu, Marcello Marelli, Diane Hatton, Jie Zhu, Michael A Bowen
The conserved glycosylation site Asn297 of a monoclonal antibody (mAb) can be decorated with a variety of sugars that can alter mAb pharmacokinetics and recruitment of effector proteins. Antibodies lacking the core fucose at Asn297 (afucosylated mAbs) show enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and increased efficacy. Here, we describe the development of a robust platform for the manufacture of afucosylated therapeutic mAbs by engineering a Chinese hamster ovary (CHO) host cell line to co-express a mAb with GDP-6-deoxy-D-lyxo-4-hexulose reductase (RMD), a prokaryotic enzyme that deflects an intermediate in the de novo synthesis of fucose to a dead-end product, resulting in the production of afucosylated mAb (GlymaxX™ Technology, ProBioGen)...
February 5, 2018: MAbs
https://www.readbyqxmd.com/read/29398668/nardostachys-jatamansi-dc-ethanol-extract-ameliorates-a%C3%AE-42-cytotoxicity
#14
Quan Feng Liu, Youngjae Jeon, Yung-Wei Sung, Jang Ho Lee, Haemin Jeong, Young-Mi Kim, Hye Sup Yun, Young-Won Chin, Songhee Jeon, Kyoung Sang Cho, Byung-Soo Koo
The Nardostachys jatamansi DC (NJ) root has been used as a sedative or analgesic to treat neurological symptoms and pain in traditional Korean medicine. Here, we investigate the potential effects of NJ on Alzheimer's disease (AD) and reveal the molecular mechanism through which NJ exerts its effects. The neuroprotective effect of the NJ root ethanol extract against β amyloid (Aβ) toxicity was examined in vitro using a cell culture system and in vivo using a Drosophila AD model. The NJ extract and chlorogenic acid, a major component of NJ, inhibited Aβ-induced cell death in SH-SY5Y cells...
February 3, 2018: Biological & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/29393587/impact-of-cho-metabolism-on-cell-growth-and-protein-production-an-overview-of-toxic-and-inhibiting-metabolites-and-nutrients
#15
REVIEW
Sara Pereira, Helene F Kildegaard, Mikael R Andersen
For over three decades, Chinese hamster ovary (CHO) cells have been the chosen expression platform for the production of therapeutic proteins with complex post-translational modifications. However, the metabolism of these cells is far from perfect and optimized, and requires substantial knowhow and process optimization and monitoring to perform efficiently. One of the main reasons for this is the production and accumulation of toxic and growth-inhibiting metabolites during culture. Lactate and ammonium are the most known, but many more have been identified...
February 2, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29388872/improving-titer-while-maintaining-quality-of-final-formulated-drug-substance-via-optimization-of-cho-cell-culture-conditions-in-low-iron-chemically-defined-media
#16
Jianlin Xu, Matthew S Rehmann, Xuankuo Xu, Chao Huang, Jun Tian, Nan-Xin Qian, Zheng Jian Li
During biopharmaceutical process development, it is important to improve titer to reduce drug manufacturing costs and to deliver comparable quality attributes of therapeutic proteins, which helps to ensure patient safety and efficacy. We previously reported that relative high-iron concentrations in media increased titer, but caused unacceptable coloration of a fusion protein during early-phase process development. Ultimately, the fusion protein with acceptable color was manufactured using low-iron media, but the titer decreased significantly in the low-iron process...
February 1, 2018: MAbs
https://www.readbyqxmd.com/read/29387348/age-induced-neuronal-cell-death-is-enhanced-in-g2019s-lrrk2-mutation-with-increased-rage-expression
#17
Hyun Jin Cho, Chengsong Xie, Huaibin Cai
Background: Leucine-rich repeat kinase 2 (LRRK2) mutations represent the most common genetic cause of sporadic and familial Parkinson's disease (PD). Especially, LRRK2 G2019S missense mutation has been identified as the most prevalent genetic cause in the late-onset PD. Advanced glycation end products (AGEs) are produced in high amounts in diabetes and diverse aging-related disorders, such as cardiovascular disease, renal disease, and neurological disease. AGEs trigger intracellular signaling pathway associated with oxidative stress and inflammation as well as cell death...
2018: Translational Neurodegeneration
https://www.readbyqxmd.com/read/29382079/a-simple-method-to-reduce-both-lactic-acid-and-ammonium-production-in-industrial-animal-cell-culture
#18
Nathaniel W Freund, Matthew S Croughan
Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production-termed Lactate Supplementation and Adaptation (LSA) technology-through the use of CHO cells adapted to a lactate-supplemented medium...
January 28, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29377798/1024-pixel-cmos-multimodality-joint-cellular-sensor-stimulator-array-for-real-time-holistic-cellular-characterization-and-cell-based-drug-screening
#19
Jong Seok Park, Moez Karim Aziz, Sensen Li, Taiyun Chi, Sandra Ivonne Grijalva, Jung Hoon Sung, Hee Cheol Cho, Hua Wang
This paper presents a fully integrated CMOS multimodality joint sensor/stimulator array with 1024 pixels for real-time holistic cellular characterization and drug screening. The proposed system consists of four pixel groups and four parallel signal-conditioning blocks. Every pixel group contains 16 × 16 pixels, and each pixel includes one gold-plated electrode, four photodiodes, and in-pixel circuits, within a pixel footprint. Each pixel supports real-time extracellular potential recording, optical detection, charge-balanced biphasic current stimulation, and cellular impedance measurement for the same cellular sample...
February 2018: IEEE Transactions on Biomedical Circuits and Systems
https://www.readbyqxmd.com/read/29377430/preclinical-characterization-of-three-transient-receptor-potential-vanilloid-receptor-1-antagonists-for-early-use-in-human-intradermal-microdose-analgesic-studies
#20
E Sjögren, M M Halldin, O Stålberg, A K Sundgren-Andersson
BACKGROUND: The transient receptor potential vanilloid receptor 1 (TRPV1) is a nonselective cation channel involved in the mediation of peripheral pain to the central nervous system. As such, the TRPV1 is an accessible molecular target that lends itself well to the understanding of nociceptive signalling. This study encompasses preclinical investigations of three molecules with the prospect to establish them as suitable analgesic model compounds in human intradermal pain relief studies...
January 26, 2018: European Journal of Pain: EJP
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