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bystander effect induced by uvc radiation in chinese hamster v79 cells.in past decades, researches on radiation-induced bystander effect mainly focused on ionizing radiation such as α-particle, β-particle, x-ray and γ-ray. but few researches have been conducted on the ability of ultraviolet (uv) radiation-induced bystander effect, and knowledge of uvc-induced bystander effect is far limited. here, we adopted medium transfer experiment to detect whether uvc could cause bystander effect in chinese hamster v79 cells. we determined the cell viability, apoptosis rate, c ...201624517728
high zinc ion supplementation of more than 30 μm can increase monoclonal antibody production in recombinant chinese hamster ovary dg44 cell culture.effects of high znso4·7h2o supplementation on cell growth and monoclonal antibody (mab) production in chemically defined suspension cultures of recombinant chinese hamster ovary (rcho) dg44 cells were examined. the supplementation of znso4·7h2o up to 120 μm gradually increased specific mab production rate of rcho dg44 cells in the early growth phase (0-4 days of culture). the znso4·7h2o concentration for enhancing mab production without any cytotoxic effects on cell growth was 30-60 μm. in addit ...201626512008
assays for dna double-strand break repair by microhomology-based end-joining repair mechanisms.dna double stranded breaks (dsbs) are one of the most deleterious types of dna lesions. the main pathways responsible for repairing these breaks in eukaryotic cells are homologous recombination (hr) and non-homologous end-joining (nhej). however, a third group of still poorly characterized dsb repair pathways, collectively termed microhomology-mediated end-joining (mmej), relies on short homologies for the end-joining process. here, we constructed gfp reporter assays to characterize and distingu ...201626657630
inactivation of gdp-fucose transporter gene (slc35c1) in cho cells by zfns, talens and crispr-cas9 for production of fucose-free antibodies.removal of core fucose from n-glycans attached to human igg1 significantly enhances its affinity for the receptor fcγriii and thereby dramatically improves its antibody-dependent cellular cytotoxicity activity. while previous works have shown that inactivation of fucosyltransferase 8 results in mutants capable of producing fucose-free antibodies, we report here the use of genome editing techniques, namely zfns, talens and the crispr-cas9, to inactivate the gdp-fucose transporter (slc35c1) in chi ...201626471004
a rapid method for simultaneous evaluation of free light chain content and aggregate content in culture media of chinese hamster ovary cells expressing monoclonal antibodies for cell line screening.the goal of developing a monoclonal antibody (mab) production process is high productivity and high quality. because the productivity and quality of mabs depend on cell line properties, the selection of cell lines suitable for large-scale production is an important stage in process development for mab production. the light chain (lc) is important for antibody folding and assembly in the endoplasmic reticulum; cell lines that secrete a large amount of lcs in the medium secrete high-quality antibo ...201626467692
temperature-sensitive mir-483 is a conserved regulator of recombinant protein and viral vector production in mammalian cells.cell engineering and bioprocess optimizations such as low temperature cultivation represent powerful tools to improve cellular performance and product yields of mammalian production cells. besides monoclonal antibodies (mabs), novel biotherapeutic formats such as viral vectors will gain increasing importance. here, we demonstrate that similar to chinese hamster ovary (cho) cells, product yields of recombinant adeno-associated virus (raav) producing hela cells can be markedly increased by low tem ...201626461143
actn1 rod domain mutation associated with congenital macrothrombocytopenia.mutations in actn1, the gene encoding the actin-crosslinking protein α-actinin-1, cause autosomal dominant macrothrombocytopenia. α-actinin-1 exists as antiparallel dimers, composed of an n-terminal actin-binding domain (abd), four spectrin-like repeats (slrs), which form the spacer rod, and a c-terminal calmodulin-like (cam) domain. all of the previously reported actn1 mutations associated with macrothrombocytopenia reside within the abd and the cam domain and not within the slr domain. in this ...201626453073
quick generation of raman spectroscopy based in-process glucose control to influence biopharmaceutical protein product quality during mammalian cell culture.mitigating risks to biotherapeutic protein production processes and products has driven the development of targeted process analytical technology (pat); however implementing pat during development without significantly increasing program timelines can be difficult. the development of a monoclonal antibody expressed in a chinese hamster ovary (cho) cell line via fed-batch processing presented an opportunity to demonstrate capabilities of altering percent glycated protein product. glycation is cau ...201626587969
slashing the timelines: opting to generate high-titer clonal lines faster via viability-based single cell sorting.chinese hamster ovary (cho) cell line development (cld) is a long and laborious process, which requires up to 5 - 6 months in order to generate and bank cho lines capable of stably expressing therapeutic molecules. additionally, single cell cloning of these production lines is also necessary to confirm clonality of the production lines. here we introduce the utilization of viability staining dye in combination with flow cytometer to isolate high titer clones from a pool of selected cells and sin ...201626587808
hydroxycinnamic acid amides from scopolia tangutica inhibit the activity of m1 muscarinic acetylcholine receptor in vitro.scopolia tangutica maxim (s. tangutica) extracts have been traditionally used as antispasmodic, sedative, and analgesic agents in tibet and in the qinghai province of china. their active compositions are however poorly understood. we have recently isolated five new hydroxycinnamic acid (hca) amides along with two known hca amides, one cinnamic acid amide from these extracts. in this study, we evaluate their abilities to inhibit carbacol-induced activity of m1 muscarinic acetylcholine receptor al ...201626586621
ox2 orexin/hypocretin receptor signal transduction in recombinant chinese hamster ovary cells.there are two subtypes of orexin receptors, ox1 and ox2. signalling pathways have been mapped in much higher detail for ox1 receptors than ox2 receptors. almost all the detailed studies have been performed in chinese hamster ovary cells, and we thus chose the same cell background for the studies on human ox2 receptors to allow comparison to human ox1 receptors. adenylyl cyclase, phospholipase a2, c and d and diacylglycerol lipase activities were assessed by precursor radiolabelling and chromatog ...201626582739
an ultra scale-down approach identifies host cell protein differences across a panel of mab producing cho cell line variants.during the manufacture of biopharmaceutical products, the final product must lie within strict pre-set specifications, for example the host cell protein (hcp) content. a number of specific hcps have been identified in particular products and the interactions between product/hcps have also been recently investigated; however, a comparison of the hcp dynamics between related cell lines and their response to early downstream processing to aid process development and cell line selection has not been ...201626579700
genetic analysis of the clonal stability of chinese hamster ovary cells for recombinant protein production.chinese hamster ovary (cho) cells are frequently used for the production of recombinant proteins for therapeutical applications. however, the recombinant protein expression level of cho cells may reduce during long-term culture. the physiological changes related to the stability of expression were not well understood. in this study, we performed a series of genetic analysis on stable and unstable clonal derived populations. transcriptome analysis revealed that a large number of differentially ex ...201626563441
chemically defined media modifications to lower tryptophan oxidation of biopharmaceuticals.oxidation of biopharmaceuticals is a major product quality issue with potential impacts on activity and immunogenicity. at eli lilly and company, high tryptophan oxidation was observed for two biopharmaceuticals in development produced in chinese hamster ovary cells. a switch from historical hydrolysate-containing media to chemically defined media with a reformulated basal powder was thought to be responsible, so mitigation efforts focused on media modification. shake flask studies identified th ...201626560440
host cell protein impurities in chromatographic polishing steps for monoclonal antibody purification.downstream purification of monoclonal antibodies (mabs) is normally performed using a platform process that is empirically tuned to optimize impurity removal for each new product. a more fundamental understanding of impurities and the product itself would provide insights into the rational design of efficient downstream processes. this work examines the chromatographic properties of chinese hamster ovary host cell protein (hcp) impurities in non-affinity chromatographic resins commonly used in p ...201626550778
manool, a salvia officinalis diterpene, induces selective cytotoxicity in cancer cells.manool, a diterpene isolated from salvia officinalis, was evaluated by the xtt colorimetric assay for cytotoxicity and selectivity against different cancer cell lines: b16f10 (murine melanoma), mcf-7 (human breast adenocarcinoma), hela (human cervical adenocarcinoma), hepg2 (human hepatocellular carcinoma), and mo59j, u343 and u251 (human glioblastoma). a normal cell line (v79, chinese hamster lung fibroblasts) was used to compare the selectivity of the test substance. manool exhibited higher cy ...201626547581
human cyp2e1-dependent mutagenicity of mono- and dichlorobiphenyls in chinese hamster (v79)-derived cells.polychlorinated biphenyls (pcbs) are a group of persistent organic pollutants with confirmed carcinogenicity to humans. metabolic activation of lower chlorinated pcbs to genotoxic metabolites may involve hydroxylation and further oxidation, and some hydroxylated metabolites may be sulfo-conjugated. however, the genotoxicity of individual pcb compounds is largely unknown. in this study, 15 mono- and dichlorobiphenyls were investigated for genotoxicity using the micronucleus and hprt mutagenicity ...201626547025
cloning and expression of codon-optimized recombinant darbepoetin alfa in leishmania tarentolae t7-tr.darbepoetin alfa is an engineered and hyperglycosylated analog of recombinant human erythropoietin (epo) which is used as a drug in treating anemia in patients with chronic kidney failure and cancer. this study desribes the secretory expression of a codon-optimized recombinant form of darbepoetin alfa in leishmania tarentolae t7-tr. synthetic codon-optimized gene was amplified by pcr and cloned into the plexsy-i-blecherry3 vector. the resultant expression vector, plexsydarbo, was purified, diges ...201626546410
the multidrug transporter mate1 sequesters ocs within an intracellular compartment that has no influence on oc secretion in renal proximal tubules.secretion of organic cations (ocs) across renal proximal tubules (rpts) involves basolateral oc transporter (oct)2-mediated uptake from the blood followed by apical multidrug and toxin extruder (mate)1/2-mediated efflux into the tubule filtrate. whereas oct2 supports electrogenic oc uniport, mate is an oc/h(+) exchanger. as assessed by epifluorescence microscopy, cultured chinese hamster ovary (cho) cells that stably expressed human mate1 accumulated the fluorescent oc n,n,n-trimethyl-2-[methyl( ...201626538438
a markov chain model for n-linked protein glycosylation--towards a low-parameter tool for model-driven glycoengineering.glycosylation is a critical quality attribute of most recombinant biotherapeutics. consequently, drug development requires careful control of glycoforms to meet bioactivity and biosafety requirements. however, glycoengineering can be extraordinarily difficult given the complex reaction networks underlying glycosylation and the vast number of different glycans that can be synthesized in a host cell. computational modeling offers an intriguing option to rationally guide glycoengineering, but the h ...201626537759
improving expression of recombinant human igf-1 using igf-1r knockout cho cell lines.chinese hamster ovary (cho) cells are widely used for the large-scale production of recombinant biopharmaceuticals. however, attempts to express igf-1 (a mutated human insulin-like growth factor 1 ea peptide (higf-1ea mut)) in cho cells resulted in poor cell growth and low productivity (0.1-0.2 g/l). human igf-1 variants negatively impacted cho cell growth via the igf-1 receptor (igf-1r). therefore knockout (ko) of the igf-1r gene in two different cho cell lines as well as knockdown (kd) of igf- ...201626523469
deletion of a telomeric region on chromosome 8 correlates with higher productivity and stability of cho cell lines.chinese hamster ovary (cho) cells are widely used for large scale production of recombinant biopharmaceuticals. although these cells have been extensively used, a demand to further increase the performance, for example, to facilitate the process of clone selection to isolate the highest producing cell lines that maintain stability of production over time is still existing. we compared gene expression profiles of high versus low producing cho clones to identify regulated genes which can be used a ...201626523402
regulation of cell growth and apoptosis through lactate dehydrogenase c over-expression in chinese hamster ovary cells.lactate has long been credited as a by-product, which jeopardizes cell growth and productivity when accumulated over a certain concentration during the manufacturing process of therapeutic recombinant proteins by chinese hamster ovary (cho) cells. a number of efforts to decrease the lactate concentration have been developed; however, the accumulation of lactate is still a critical issue by the late stage of fed-batch culture. therefore, a lactate-tolerant cell line was developed through over-exp ...201626841889
evaluation of the relative biological effectiveness of spot-scanning proton irradiation in vitro.variations in relative biological effectiveness (rbe) from a fixed value of 1.1 are critical in proton beam therapy. to date, studies estimating rbe at multiple positions relative to the spread-out bragg peak (sobp) have been predominantly performed using passive scattering methods, and limited data are available for spot-scanning beams. thus, to investigate the rbe of spot-scanning beams, chinese hamster fibroblast v79 cells were irradiated using the beam line at the hokkaido university hospita ...201626838131
aquaporin-9 facilitates membrane transport of hydrogen peroxide in mammalian cells.aquaporin (aqp) 9, a member of the transmembrane water channel family, is defined as a water/glycerol transporting protein. some aqps including aqp3 and aqp8 have been recently found to transport hydrogen peroxide (h2o2). here we show that aqp9 facilitates the membrane transport of h2o2 in human and mice cells. enforced expression of human aqp9 in chinese hamster ovary-k1 potentiated the increase in cellular h2o2 after adding exogenous h2o2. in contrast, aqp9 knockdown by sirna in human hepatoma ...201626837049
metabolic engineering with multi-objective optimization of kinetic models.kinetic models have a great potential for metabolic engineering applications. they can be used for testing which genetic and regulatory modifications can increase the production of metabolites of interest, while simultaneously monitoring other key functions of the host organism. this work presents a methodology for increasing productivity in biotechnological processes exploiting dynamic models. it uses multi-objective dynamic optimization to identify the combination of targets (enzymatic modific ...201626826510
[stable expression of recombinant human podoplanin in chinese hamster ovary (cho) cells].to construct podoplanin (pdpn) eukaryotic expression plasmid pdpn-pegfp-n1, establish chinese hamster ovary (cho) cell line stably expressing recombinant human pdpn and investigate its biological activity.201626728373
targeted transgene insertion into the cho cell genome using cre recombinase-incorporating integrase-defective retroviral vectors.retroviral vectors have served as efficient gene delivery tools in various biotechnology fields. however, viral dna is randomly inserted into the genome, which can cause problems, such as insertional mutagenesis and gene silencing. previously, we reported a site-specific gene integration system, in which a transgene is integrated into a predetermined chromosomal locus of chinese hamster ovary (cho) cells using integrase-defective retroviral vectors (idrvs) and cre recombinase. in this system, a ...201626724679
permeabilization of the nuclear envelope following nanosecond pulsed electric field exposure.permeabilization of cell membranes occurs upon exposure to a threshold absorbed dose (ad) of nanosecond pulsed electric fields (nspef). the ultimate, physiological bioeffect of this exposure depends on the type of cultured cell and environment, indicating that cell-specific pathways and structures are stimulated. here we investigate 10 and 600 ns duration pef effects on chinese hamster ovary (cho) cell nuclei, where our hypothesis is that pulse disruption of the nuclear envelope membrane leads t ...201626721436
in search of acceptable alternatives to the murine histamine sensitisation test (hist): what is possible and practical?the 'international workshop on alternatives to the murine histamine sensitization test for acellular pertussis vaccines: in search of acceptable alternatives to the murine histamine sensitization test (hist): what is possible and practical?' was held on 4 and 5 march 2015 in london, united kingdom. participants discussed the results of the data generated from an international collaborative study (bsp114 phase 2) sponsored by the european directorate for the quality of medicines & health care (ed ...201628279256
overproduction of recombinant human bone morphogenetic protein-7 in chinese hamster ovary cells.bone morphogenetic protein-7 is a multifunctional growth factor involved in various cellular processes such as osteogenesis, kidney and eye development, brown adipogenesis, and bone metastasis, and thus has been considered to have therapeutic potential for treating various diseases. in this study, we established a chinese hamster ovary (cho) cell line stably overexpressing recombinant human bmp-7 (rhbmp-7). over the course of a 14-day fed-batch culture process in a 7.5-l bioreactor (5-l working ...201626711959
doxycycline exerted neuroprotective activity by enhancing the activation of neuropeptide gpcr pac1.doxycycline has significant neuroprotective effect with anti-inflammatory and anti-apoptotic activity. we found for the first time that doxycycline specially promoted the proliferation of chinese hamster ovary (cho) cells with high expression of neuropeptide pituitary adenylate cyclase-activating polypeptide (pacap) preferring g protein-coupled receptor (gpcr), pacap receptor 1(pac1) and induced the internalization of pac1 tagged with yellow fluorescent protein (yfp) indicating doxycycline inter ...201626700245
overexpression of yy1 increases the protein production in mammalian cells.the production of therapeutic antibodies using mammalian cells remains a high-priority in the biopharmaceutical manufacturing industry. bioengineers have targeted different cellular processes, including transcription, translation, secretion and post-translational modifications, to overcome the metabolic bottlenecks limiting production capacity and create high-producing mammalian cell lines. the polycomb group (pcg) proteins belong to a family of chromatin regulators with important roles in multi ...201626686315
dysbindin-1 modifies signaling and cellular localization of recombinant, human d₃ and d₂ receptors.dystrobrevin binding protein-1 (dysbindin-1), a candidate gene for schizophrenia, modulates cognition, synaptic plasticity and frontocortical circuitry and interacts with glutamatergic and dopaminergic transmission. loss of dysbindin-1 modifies cellular trafficking of dopamine (da) d2 receptors to increase cell surface expression, but its influence upon signaling has never been characterized. further, the effects of dysbindin-1 upon closely related d3 receptors remain unexplored. hence, we exami ...201626685100
glycosylphosphatidylinositol anchor modification machinery deficiency is responsible for the formation of pro-prion protein (prp) in bxpc-3 protein and increases cancer cell motility.the normal cellular prion protein (prp) is a glycosylphosphatidylinositol (gpi)-anchored cell surface glycoprotein. however, in pancreatic ductal adenocarcinoma cell lines, such as bxpc-3, prp exists as a pro-prp retaining its glycosylphosphatidylinositol (gpi) peptide signaling sequence. here, we report the identification of another pancreatic ductal adenocarcinoma cell line, aspc-1, which expresses a mature gpi-anchored prp. comparison of the 24 genes involved in the gpi anchor modification pa ...201626683373
importance of interaction between integrin and actin cytoskeleton in suspension adaptation of cho cells.the biopharmaceutical production process relies upon mammalian cell technology where single cells proliferate in suspension in a chemically defined synthetic environment. this environment lacks exogenous growth factors, usually contributing to proliferation of fibroblastic cell types such as chinese hamster ovary (cho) cells. use of cho cells for production hence requires a lengthy 'adaptation' process to select clones capable of proliferation as single cells in suspension. the underlying molecu ...201626679704
cd147 overexpression promotes tumorigenicity in chinese hamster ovary cells.cd147 overexpresses in many epithelium-originated tumors and plays an important role in tumor migration and invasion. most studies aim at the role of cd147 in tumor progression using tumor cell models. however, the influence of abnormal overexpression of cd147 on neoplastic transformation of normal cells is unknown. here, the role of cd147 in malignant phenotype transformation in cho cells was investigated. three cho cell lines that stably overexpressed cd147 (cho-cd147), egfp-cd147 (cho-egfp-cd ...201626676266
valeric acid induces cell cycle arrest at g1 phase in cho cell cultures and improves recombinant antibody productivity.to find a more effective chemical reagent for improved monoclonal antibody (mab) production, eight chemical reagents (curcumin, quercein, dl-sulforaphane, thymidine, valeric acid, phenyl butyrate, valproic acid, and lithium chloride) known to induce cell cycle arrest were examined individually as chemical additives to recombinant cho (rcho) cell cultures producing mab. among these chemical additives, valeric acid showed the best production performance. valeric acid decreased specific growth rate ...201626663903
a method for specifically targeting two independent genomic integration sites for co-expression of genes in cho cells.stable mammalian production cell lines in suspension culture enable the reproducible expression of target genes in any desired scale using bioreactor technology. targeted integration methods have been developed to cut down timelines for the generation of stable producer cell lines. the powerful flp recombinase mediated cassette exchange (rmce) technique allows fast integration of target genes in preselected and optimized high expression loci in so called master cell lines. up to now, these cells ...201626658354
identification and characterization of a -1 reading frameshift in the heavy chain constant region of an igg1 recombinant monoclonal antibody produced in cho cells.frameshifts lead to complete alteration of the intended amino acid sequences, and therefore may affect the biological activities of protein therapeutics and pose potential immunogenicity risks. we report here the identification and characterization of a novel -1 frameshift variant in a recombinant igg1 therapeutic monoclonal antibody (mab) produced in chinese hamster ovary cells during the cell line selection studies. the variant was initially observed as an atypical post-monomer fragment peak i ...201626652198
discovery and characterization of hydroxylysine in recombinant monoclonal antibodies.tryptic peptide mapping analysis of a chinese hamster ovary (cho)-expressed, recombinant igg1 monoclonal antibody revealed a previously unreported +16 da modification. through a combination of ms(n) experiments, and preparation and analysis of known synthetic peptides, the possibility of a sequence variant (ala to ser) was ruled out and the presence of hydroxylysine was confirmed. post-translational hydroxylation of lysine was found in a consensus sequence (xkg) known to be the site of modificat ...201626651858
ubiquitous chromatin opening elements (ucoes) effect on transgene position and expression stability in cho cells following methotrexate (mtx) amplification.the requirement for complex therapeutic proteins has resulted in mammalian cells, especially cho cells, being the dominant host for recombinant protein manufacturing. in creating recombinant cho cell lines, the expression vectors integrate into various parts of the genome leading to variable levels of expression and stability of protein production. this makes mammalian cell line development a long and laborious process. therefore, with the intention to accelerate process development of recombina ...201626632501
enhanced sialylation of a human chimeric igg1 variant produced in human and rodent cell lines.glycosylation of the igg-fc is essential for optimal binding and activation of fcγ receptors and the c1q component of complement. however, it has been reported that the effector functions are down-regulated when the fc glycans terminate in sialic acid residues and that sialylated igg mediates anti-inflammatory effects of intravenous immunoglobulin (ivig). although recombinant igg is hypo-sialylated, fc sialylation is shown to be markedly increased when a mouse/human chimeric igg3 phe243ala (f243 ...201626627984
development and characterization of a novel long-acting recombinant follicle stimulating hormone agonist by fusing fc to an fsh-β subunit.does a novel long-acting recombinant human fsh, kn015, a heterodimer composed of fshα and fshβ-fc/fc, offer a potential fsh alternative?201626621853
microfluidic assessment of mechanical cell damage by extensional stress.mammalian cells have been widely used in bioreactors to produce biological products such as pharmaceutical materials. the productivity of such bioreactors is vastly affected by flow-induced cell damage in complicated flow environments, such as agitation-driven turbulence and oxygen bubble bursting at the interface between the culturing medium and air. however, there is no systematic approach to diagnose the cell damage caused by the hydrodynamic stress. in this work, we propose a novel microflui ...201626621113
comparison of three transposons for the generation of highly productive recombinant cho cell pools and cell lines.several naturally occurring vertebrate transposable elements have been genetically modified to enable the transposition of recombinant genes in mammalian cells. we compared three transposons-piggybac, tol2, and sleeping beauty-for their ability to generate cell pools (polyclonal cultures of recombinant cells) and clonal cell lines for the large-scale production of recombinant proteins using chinese hamster ovary cells (cho-dg44) as the host. transfection with each of the dual-vector transposon s ...201626616356
g-protein inhibition profile of the reported gq/11 inhibitor ubo-qic.ubo-qic (fr900359) is the only currently available gq/11 protein inhibitor. however, its characterization has not been published, and we thus set out to do this. gi, gs and gq protein-mediated responses were assessed utilizing endogenous or heterologously expressed receptors in chinese hamster ovary cells. ubo-qic, at 1 μm, was an effective inhibitor of the gq-mediated responses, but was inactive at gi- and gs-mediated responses. gq/11 and g16 responses were additionally compared in hel92.1.7 ce ...201626614908
expression of recombinant human bifunctional peptidylglycine α-amidating monooxygenase in cho cells and its use for insulin analogue modification.the availability of catalytically active peptidylglycine α-amidating monooxygenase (pam) should provide the means to examine its potential use for the chemienzymatic synthesis of bioactive peptides for the purpose of pharmacological studies. hypoglycemic activity is one of the most important features of insulin derivatives. insulin glargine amide was found to show a time/effect profile which is distinctly more flat and thus more advantageous than insulin glargine itself. the aim of the study was ...201626614892
7-chlorofolipastatin, an inhibitor of sterol o-acyltransferase, produced by marine-derived aspergillus ungui nkh-007.a new depsidone, named 7-chlorofolipastatin, and five known structurally related depsidones were isolated from the culture broth of the marine-derived fungus aspergillus ungui nkm-007 by solvent extraction and hplc using an octadecylsilyl column. the structure of 7-chlorofolipastatin was elucidated by various spectroscopic data including 1d and 2d nmr spectroscopy. 7-chlorofolipastatin inhibited sterol o-acyltransferase (soat) 1 and 2 isozymes in cell-based and enzyme assays using soat1- and soa ...201626980608
impact of dissolved oxygen during uv-irradiation on the chemical composition and function of cho cell culture media.ultraviolet (uv) irradiation is advantageous as a sterilization technique in the biopharmaceutical industry since it is capable of targeting non-enveloped viruses that are typically challenging to destroy, as well as smaller viruses that can be difficult to remove via conventional separation techniques. in this work, we investigated the influence of oxygen in the media during uv irradiation and characterized the effect on chemical composition using nmr and lc-ms, as well as the ability of the ir ...201626975046
role of extracellular cysteine residues in the adenosine a2a receptor.the g protein-coupled a2a adenosine receptor represents an important drug target. crystal structures and modeling studies indicated that three disulfide bonds are formed between ecl1 and ecl2 (i, cys71(2.69)-cys159(45.43); ii, cys74(3.22)-cys146(45.30), and iii, cys77(3.25)-cys166(45.50)). however, the a2bar subtype appears to require only disulfide bond iii for proper function. in this study, each of the three disulfide bonds in the a2aar was disrupted by mutation of one of the cysteine residue ...201626969588
generation and characterization of a human/mouse chimeric gd2-mimicking anti-idiotype antibody ganglidiximab for active immunotherapy against neuroblastoma.vaccination with proteins mimicking gd2 that is highly expressed on neuroblastoma (nb) cells is a promising strategy in treatment of nb, a pediatric malignancy with poor prognosis. we previously showed efficacy of ganglidiomab in vivo, a murine anti-idiotype (anti-id) igg1. in order to tailor immune responses to variable regions, we generated a new human/mouse chimeric anti-id antibody (ab) ganglidiximab by replacing murine constant fragments with corresponding human igg1 regions. dna sequences ...201626967324
12-heter1/gpr31, a high-affinity 12(s)-hydroxyeicosatetraenoic acid receptor, is significantly up-regulated in prostate cancer and plays a critical role in prostate cancer progression.previously we identified and deorphaned g-protein-coupled receptor 31 (gpr31) as the high-affinity 12(s)-hydroxyeicosatetraenoic acid [12(s)-hete] receptor (12-heter1). here we have determined its distribution in prostate cancer tissue and its role in prostate tumorigenesis using in vitro and in vivo assays. data-mining studies strongly suggest that 12-heter1 expression positively correlates with the aggressiveness and progression of prostate tumors. this was corroborated with real-time pcr anal ...201626965684
enhanced translation initiation factor 4g levels correlate with production levels of monoclonal antibodies in recombinant cho cell lines.using cells to manufacture protein-based therapeutics or biopharmaceuticals is a rapidly expanding industrial activity. chinese hamster ovary (cho) cells are the most frequently used mammalian host-expression system for the manufacture of biopharmaceuticals. over the past ∼30 years academic and industrial researchers have studied cell expression characteristics with aims to improve product yield, quality, scalability and reproducibility. although many steps in the gene expression and secretion p ...201626965386
thyroid stimulating antibodies are highly prevalent in hashimoto's thyroiditis and associated orbitopathy.thyroid-associated orbitopathy (tao) rarely occurs in patients with hashimoto's thyroiditis (ht).201626964732
a synthetic biology standard for chinese hamster ovary cell genome monitoring and contaminant detection by polymerase chain reaction.chinese hamster ovary (cho) cells are the current industry standard for production of therapeutic monoclonal antibodies at commercial scales. production optimisation in cho cells hinges on analytical technologies such as the use of the polymerase chain reaction (pcr) to quantify genetic factors within the cho genome and to detect the presence of contaminant organisms. pcr-based assays, whilst sensitive and accurate, are limited by (i) requiring lengthy sample preparation and (ii) a lack of stand ...201627652083
investigation of microbubble cavitation-induced calcein release from cells in vitro.in the present study, microbubble (mb) cavitation signal analysis was performed together with calcein release evaluation in both pressure and exposure duration domains of the acoustic field. a passive cavitation detection system was used to simultaneously measure mb scattering and attenuation signals for subsequent extraction efficiency relative to mb cavitation activity. the results indicate that the decrease in the efficiency of extraction of calcein molecules from chinese hamster ovary cells, ...201627637933
role of exposure/recovery schedule in micronuclei induction by several promutagens in v79-derived cells expressing human cyp2e1 and sult1a1.the standard procedure for the micronucleus test in cell lines requires a short exposure (≤0.5 cell cycle) to the test compounds followed by a long recovery (≥1.5 cell cycle), and in case of negative or equivocal results, a second test with extended exposure (≥2 cell cycles) without or with a recovery time. in general the two procedures are advantageous for detecting clastogens and aneugens, respectively. however, whether the recommended procedures apply to micronucleus tests with promutagens in ...201627637483
endoplasmic reticulum-directed recombinant mrna displays subcellular localization equal to endogenous mrna during transient expression in cho cells.when expressing pharmaceutical recombinant proteins in mammalian cells, the protein is commonly directed through the secretory pathway, in a signal peptide-dependent manner, to acquire specific post-translational modifications and to facilitate secretion into the culture medium. one key premise for this is the direction of the mrna encoding the recombinant protein to the surface of the endoplasmic reticulum (er) for subsequent protein translocation into the secretory pathway. to evaluate the eff ...201627624596
identifying model error in metabolic flux analysis - a generalized least squares approach.the estimation of intracellular flux through traditional metabolic flux analysis (mfa) using an overdetermined system of equations is a well established practice in metabolic engineering. despite the continued evolution of the methodology since its introduction, there has been little focus on validation and identification of poor model fit outside of identifying "gross measurement error". the growing complexity of metabolic models, which are increasingly generated from genome-level data, has nec ...201627619919
granular activated carbon treatment may result in higher predicted genotoxicity in the presence of bromide.certain unregulated disinfection byproducts (dbps) are more of a health concern than regulated dbps. brominated species are typically more cytotoxic and genotoxic than their chlorinated analogs. the impact of granular activated carbon (gac) on controlling the formation of regulated and selected unregulated dbps following chlorine disinfection was evaluated. the predicted cyto- and genotoxicity of dbps was calculated using published potencies based on the comet assay for chinese hamster ovary cel ...201627467860
tumor targeting with novel 6-substituted pyrrolo [2,3-d] pyrimidine antifolates with heteroatom bridge substitutions via cellular uptake by folate receptor α and the proton-coupled folate transporter and inhibition of de novo purine nucleotide biosynthesis.targeted antifolates with heteroatom replacements of the carbon vicinal to the phenyl ring in 1 by n (4), o (8), or s (9), or with n-substituted formyl (5), acetyl (6), or trifluoroacetyl (7) moieties, were synthesized and tested for selective cellular uptake by folate receptor (fr) α and β or the proton-coupled folate transporter. results show increased in vitro antiproliferative activity toward engineered chinese hamster ovary cells expressing frs by 4-9 over the ch2 analogue 1. compounds 4-9 ...201627458733
high expression of xrcc6 promotes human osteosarcoma cell proliferation through the β-catenin/wnt signaling pathway and is associated with poor prognosis.increasing evidences show that xrcc6 (x-ray repair complementing defective repair in chinese hamster cells 6) was upregulated and involved in tumor growth in several tumor types. however, the correlation of xrcc6 and human osteosarcoma (os) is still unknown. this study was conducted with the aim to reveal the expression and biological function of xrcc6 in os and elucidate the potential mechanism. the mrna expression level of xrcc6 was measured in osteosarcoma cells and os samples by quantitative ...201627455247
tracking dipeptides at work-uptake and intracellular fate in cho culture.market demands for monoclonal antibodies (mabs) are steadily increasing worldwide. as a result, production processes using chinese hamster ovary cells (cho) are in the focus of ongoing intensification studies for maximizing cell-specific and volumetric productivities. this includes the optimization of animal-derived component free (adcf) cultivation media as part of good cell culture practice. dipeptides are known to improve cho culture performance. however, little or even conflicting assumption ...201627447702
unexpected potency differences between b-cell-activating factor (baff) antagonist antibodies against various forms of baff: trimer, 60-mer, and membrane-bound.therapeutic agents antagonizing b-cell-activating factor/b-lymphocyte stimulator (baff/blys) are currently in clinical development for autoimmune diseases; belimumab is the first food and drug administration-approved drug in more than 50 years for the treatment of lupus. as a member of the tumor necrosis factor superfamily, baff promotes b-cell survival and homeostasis and is overexpressed in patients with systemic lupus erythematosus and other autoimmune diseases. baff exists in three recognize ...201627440419
emerging disinfection byproducts, halobenzoquinones: effects of isomeric structure and halogen substitution on cytotoxicity, formation of reactive oxygen species, and genotoxicity.halobenzoquinones (hbqs) are a structurally diverse class of water disinfection byproducts. here, we report a systematic study on the effects of isomeric structure and the type and number of halogen substitutions of hbqs on their cytotoxicity, formation of reactive oxygen species (ros), and genotoxicity. dynamic responses and ic50 histograms were obtained using real-time cell analysis, clearly ranking the cytotoxicity of the hbqs in chinese hamster ovary (cho-k1) cells. strong isomeric structure ...201626812484
effect of fc-glycan structure on the conformational stability of igg revealed by hydrogen/deuterium exchange and limited proteolysis.human therapeutic immunoglobulin gamma (igg) molecules contain an n-glycan on each of their fc ch2 domains. these glycans include high-mannose, hybrid, and complex types. recombinant igg molecules containing high-mannose glycans have been shown to clear faster in human blood, and exhibit decreased thermal stability. the molecular mechanism behind these observations, however, is not well understood. in this work, we used hydrogen/deuterium exchange combined with mass spectrometry (hdx ms), as wel ...201626812426
comparative assessment of glycosylation of a recombinant human fsh and a highly purified fsh extracted from human urine.glycosylation is an important ptm and is critical for the manufacture and efficacy of therapeutic glycoproteins. glycan significantly influences the biological properties of human follicle-stimulating hormone (hfsh). using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified fsh (uhfsh) obtained from human urine with that of a recombinant human fsh (rhfsh) obtained from chinese hamster ovary (cho) cells. intact and subunit masses, n-glycans, n-glycosylat ...201626812091
mechanisms of beauvericin toxicity and antioxidant cellular defense.beauvericin (bea) is a secondary metabolite produced by many species of fungus fusarium. this study determines the injury (cell viability, cell proliferation, mitochondrial membrane potential, cell death and dna damage) and the intracellular defense mechanisms (catalase and superoxide dismutase) in chinese hamster ovary (cho-k1) cells after bea exposure. the results obtained in this study demonstrated that bea induces cytotoxicity in a dose- and time-dependent manner in cho-k1 cells. moreover, d ...201626809139
blockade of kv1.5 by paroxetine, an antidepressant drug.paroxetine, a selective serotonin reuptake inhibitor (ssri), has been reported to have an effect on several ion channels including human ether-a-go-go-related gene in a ssri-independent manner. these results suggest that paroxetine may cause side effects on cardiac system. in this study, we investigated the effect of paroxetine on kv1.5, which is one of cardiac ion channels. the action of paroxetine on the cloned neuronal rat kv1.5 channels stably expressed in chinese hamster ovary cells was inv ...201626807026
metabolic analysis of antibody producing chinese hamster ovary cell culture under different stresses conditions.chinese hamster ovary (cho) cells are commonly used as the host cell lines concerning their ability to produce therapeutic proteins with complex post-translational modifications. in this study, we have investigated the time course extra- and intracellular metabolome data of the cho-k1 cell line, under a control and stress conditions. the addition of nacl and trehalose greatly suppressed cell growth, where the maximum viable cell density of nacl and trehalose cultures were 2.2-fold and 2.8-fold l ...201626803706
amygdala-hippocampal phospholipase d (pld) signaling as novel mechanism of cocaine-environment maladaptive conditioned responses.drug-environment associative memory mechanisms and the resulting conditioned behaviors are key contributors in relapse to cocaine dependence. recently, we reported rat amygdala phospholipase d as a key convergent downstream signaling partner in the expression of cocaine-conditioned behaviors mediated by glutamatergic and dopaminergic pathways. in the present study, 1 of the 2 known upstream serotonergic targets of phospholipase d, the serotonin (5-hydroxytryptamine) 2c receptor, was investigated ...201626802567
mechanisms underlying epigenetic and transcriptional heterogeneity in chinese hamster ovary (cho) cell lines.recombinant cell lines developed for therapeutic antibody production often suffer instability or lose recombinant protein expression during long-term culture. heterogeneous gene expression among cell line subclones may result from epigenetic modifications of dna or histones, the protein component of chromatin. we thus investigated in such cell lines, dna methylation and the chromatin environment along the human eukaryotic translation elongation factor 1 alpha 1 (eef1a1) promoter in an antibody p ...201626800878
protein purification and cloning of diacylglycerol lipase from rat brain.diacylglycerol (dg) lipase, which hydrolyses 1-stearoyl-2-arachidonyl-sn-glycerol to produce an endocannabinoid, 2-arachidonoylglycerol, was purified from the soluble fraction of rat brain lysates. dg lipase was purified about 1,200-fold by a sequential column chromatographic procedure. among proteins identified by mass spectrometry analysis in the partially purified dg lipase sample, only ddhd domain containing two (ddhd2), which was formerly regarded as a phospholipase a1, exhibited significan ...201626790472
the effect of leonurus sibiricus plant extracts on stimulating repair and protective activity against oxidative dna damage in cho cells and content of phenolic compounds.leonurus sibiricus l. has been used as a traditional and medicinal herb for many years in asia and europe. this species is known to have antibacterial, anti-inflammatory, and antioxidant activity and has demonstrated a reduction of intracellular reactive oxygen species. all tested extracts of l. sibiricus showed protective and dna repair stimulating effects in chinese hamster ovary (cho) cells exposed to h2o2. preincubation of the cho cells with 0.5 mg/ml of plant extracts showed increased expre ...201626788249
nav1.5 n-terminal domain binding to α1-syntrophin increases membrane density of human kir2.1, kir2.2 and nav1.5 channels.cardiac excitability and refractoriness are largely determined by the function and number of inward rectifier k⁺ channels (kir2.1-2.3), which are differentially expressed in the atria and ventricles, and nav1.5 channels. we have focused on how nav1.5 and kir2.x function within a macromolecular complex by elucidating the molecular determinants that govern nav1.5/kir2.x reciprocal modulation.201626786162
pharmacokinetics and pharmacodynamics of ch14.18/cho in relapsed/refractory high-risk neuroblastoma patients treated by long-term infusion in combination with il-2.ch14.18 manufactured in chinese hamster ovary (cho) cells is currently being evaluated in clinical trials. short-term infusion (sti) (8-20 h/day; 4-5 days) of 100 mg/m2 ch14.18/cho (dinutiximab β) per cycle in combination with cytokines is standard treatment of neuroblastoma (nb) patients. as pain is a limiting factor, we investigated a novel delivery method by continuous long-term infusion (lti) of 100 mg/m2 over 10 days. 53 nb patients were treated with 5-6 cycles of 6 × 106 iu/m2 subcutaneous ...201626785755
emulsions made of oils from seeds of gm flax protect v79 cells against oxidative stress.polyunsaturated fatty acids, sterols, and hydrophilic phenolic compounds are components of flax oil that act as antioxidants. we investigated the impact of flax oil from transgenic flax in the form of emulsions on stressed chinese hamster pulmonary fibroblasts. we found that the emulsions protect v79 cells against the h2o2 and the effect is dose dependent. they reduced the level of intracellular reactive oxygen species and protected genomic dna against damage. the rate of cell proliferation incr ...201626779302
multi-walled carbon nanotubes (nm401) induce ros-mediated hprt mutations in chinese hamster lung fibroblasts.although there is an important set of data showing potential genotoxic effects of nanomaterials (nms) at the dna (comet assay) and chromosome (micronucleus test) levels, few studies have been conducted to analyze their potential mutagenic effects at gene level. we have determined the ability of multi-walled carbon nanotubes (mwcnt, nm401), to induce mutations in the hprt gene in chinese hamster lung (v79) fibroblasts. nm401, characterized in the eu nanogenotox project, were further studied withi ...201626774957
lovastatin induces platelet apoptosis.statins are widely used in the prevention of atherosclerosis and treatment of coronary artery disease because of pleiotropic effects on thrombosis. thrombocytopenia and hemorrhage occurred in some statin-treated patients, but the reason remains unclear. in the current study, we show that lovastatin dose-dependently induces depolarization of mitochondrial inner transmembrane potential, leading to up-regulation of bak, down-regulation of bcl-xl, and activation of caspase-3/8/9. lovastatin treatmen ...201626773364
sox3 binds to 11β-hydroxysteroid dehydrogenase gene promoter suggesting transcriptional interaction in catfish.in fishes, the expression of steroidogenic enzyme genes and their related transcription factors (tfs) are critical for the regulation of steroidogenesis and gonadal development. 11-kt is the potent androgen and hence, 11β-hsd, enzyme involved in 11-kt production is important. regulation of 11β-hsd gene was never studied in any fishes. at first 11β-hsd was cloned and recombinant protein was tested for enzyme activity prior to expression and promoter motif analysis. expression changes revealed sta ...201626772480
chinese hamster ovary-sphingomyelin synthase2 biospecific extraction and liquid chromatography with tandem mass spectrometry analysis for the prediction of bioactive components of rhizoma polygoni cuspidati.a novel strategy for predicting bioactive components in traditional chinese medicines using chinese hamster ovary-sphingomyelin synthase2 (cho-sms2 ) cell biospecific extraction and high-performance liquid chromatography with diode array detection and tandem mass spectrometry analysis was proposed. the hypothesis is that when cells are incubated with the extract of traditional chinese medicines, the potential bioactive components in the traditional chinese medicines should selectively combine wi ...201626763406
a novel regulatory element (e77) isolated from cho-k1 genomic dna enhances stable gene expression in chinese hamster ovary cells.vectors flanked by regulatory dna elements have been used to generate stable cell lines with high productivity and transgene stability; however, regulatory elements in chinese hamster ovary (cho) cells, which are the most widely used mammalian cells in biopharmaceutical production, are still poorly understood. we isolated a novel gene regulatory element from cho-k1 cells, designated e77, which was found to enhance the stable expression of a transgene. a genomic library was constructed by combini ...201626762773
molecular design and downstream processing of turoctocog alfa (novoeight), a b-domain truncated factor viii molecule.turoctocog alfa (novoeight) is a third-generation recombinant factor viii (rfviii) with a truncated b-domain that is manufactured in chinese hamster ovary cells. no human or animal-derived materials are used in the process. the aim of this study is to describe the molecular design and purification process for turoctocog alfa. a five-step purification process is applied to turoctocog alfa: protein capture on mixed-mode resin; immunoaffinity chromatography using a unique, recombinantly produced an ...201626761578
a dual near-infrared and dielectric spectroscopies strategy to monitor populations of chinese hamster ovary cells in bioreactor.to develop a new strategy combining near-infrared (nir) and dielectric spectroscopies for real-time monitoring and in-depth characterizing populations of chinese hamster ovary cells throughout cultures performed in bioreactors.201626758724
association between single-nucleotide polymorphisms in dna double-strand break repair genes and prostate cancer aggressiveness in the spanish population.novel predictors of prognosis and treatment response for prostate cancer (pca) are required to better individualize treatment. single-nucleotide polymorphisms (snps) in four genes directly (xrcc5 (x-ray repair complementing defective repair in chinese hamster cells 5) and xrcc6 (x-ray repair complementing defective repair in chinese hamster cells 6)) or indirectly (parp1 and major vault protein (mvp)) involved in non-homologous end joining were examined in 494 spanish pca patients.201626754263
robustness analysis of elementary flux modes generated by column generation.elementary flux modes (efms) are vectors defined from a metabolic reaction network, giving the connections between substrates and products. efms-based metabolic flux analysis (mfa) estimates the flux over each efm from external flux measurements through least-squares data fitting. the measurements used in the data fitting are subject to errors. a robust optimization problem includes information on errors and gives a way to examine the sensitivity of the solution of the efms-based mfa to these er ...201626748294
structural and functional analysis of four non-coding y rnas from chinese hamster cells: identification, molecular dynamics simulations and dna replication initiation assays.the genes coding for y rnas are evolutionarily conserved in vertebrates. these non-coding rnas are essential for the initiation of chromosomal dna replication in vertebrate cells. however thus far, no information is available about y rnas in chinese hamster cells, which have already been used to detect replication origins and alternative dna structures around these sites. here, we report the gene sequences and predicted structural characteristics of the chinese hamster y rnas, and analyze their ...201626733090
expanding the 3-o-sulfate proteome--enhanced binding of neuropilin-1 to 3-o-sulfated heparan sulfate modulates its activity.binding of proteins to heparan sulfate is driven predominantly by electrostatic interactions between positively charged amino acid residues in the protein and negatively charged sulfate groups located at various positions along the polysaccharide chain. although many heparin/heparan-sulfate-binding proteins have been described, few exhibit preferential binding for heparan sulfates containing relatively rare 3-o-sulfated glucosamine residues. to expand the "3-o-sulfate proteome," affinity matrice ...201626731579
rapid amplification system for recombinant protein production in chinese hamster ovary (cho) cells.recombinant therapeutic proteins have changed the face of modern medicine in the present trend and they continue to provide innovative therapies for deadly diseases. this study describes the development of a novel stable expression system for rapid amplification of genes in chinese hamster ovary (cho) cells. the expression system consists of a host cho cell line and an expression vector (pub-pyori-d-c) which encodes for polyomavirus (py) origin of replication (pyori) for amplification of integra ...201626950459
quantitative feature extraction from the chinese hamster ovary bioprocess bibliome using a novel meta-analysis workflow.the scientific literature concerning chinese hamster ovary (cho) cells grows annually due to the importance of cho cells in industrial bioprocessing of therapeutics. in an effort to start to catalogue the breadth of cho phenotypes, or phenome, we present the cho bibliome. this bibliographic compilation covers all published cho cell studies from 1995 to 2015, and each study is classified by the types of phenotypic and bioprocess data contained therein. using data from selected studies, we also pr ...201626948029
high throughput gene complementation screening permits identification of a mammalian mitochondrial protein synthesis (ρ(-)) mutant.to identify nuclear dna (ndna) oxidative phosphorylation (oxphos) gene mutations using cultured cells, we have developed a complementation system based on retroviral transduction with a full length cdna expression library and selection for oxhos function by growth in galactose. we have used this system to transduce the chinese hamster v79-g7 oxphos mutant cell line with a defect in mitochondrial protein synthesis. the complemented cells were found to have acquired the cdna for the bs6m polypepti ...201626946086
mir-122 targets the vanin 1 gene to regulate its expression in chickens.as the most abundant microrna (mirna) in the liver, mir-122 plays important roles in the growth and development of liver, lipid metabolism, and liver diseases. vanin 1 (vnn1) plays an important role in hepatic lipid metabolism, and vnn1 may serve as a potential therapeutic target for the treatment of metabolic diseases caused by overactivated gluconeogenesis. in our previous rna-seq study, we found the expression of vnn1 increased significantly when the expression of mir-122 (gga-mir-122-5p) was ...201626944978
effect of cholesterol on cellular uptake of cancer drugs pirarubicin and ellipticine.the cell membrane is a major barrier for drug transport. given that many cancer drugs must passively cross the cell membrane, understanding drug-membrane interactions is crucial. we used fluorescence-activated cell sorting to investigate how cholesterol influences the transport of the cancer drugs ellipticine and pirarubicin across cell membranes. we showed that cholesterol depletion helped pirarubicin cross the membranes of nonsmall cell lung carcinoma and chinese hamster ovary cells. in contra ...201626937690
blockade of kv1.5 channels by the antidepressant drug sertraline.sertraline, a selective serotonin reuptake inhibitor (ssri), has been reported to lead to cardiac toxicity even at therapeutic doses including sudden cardiac death and ventricular arrhythmia. and in a ssri-independent manner, sertraline has been known to inhibit various voltage-dependent channels, which play an important role in regulation of cardiovascular system. in the present study, we investigated the action of sertraline on kv1.5, which is one of cardiac ion channels. the eff ect of sertra ...201626937216
advances in recombinant antibody manufacturing.since the first use of chinese hamster ovary (cho) cells for recombinant protein expression, production processes have steadily improved through numerous advances. in this review, we have highlighted several key milestones that have contributed to the success of cho cells from the beginning of their use for monoclonal antibody (mab) expression until today. the main factors influencing the yield of a production process are the time to accumulate a desired amount of biomass, the process duration, ...201626936774
analysis of possible genotoxicity of the herbicide flurochloridone and its commercial formulations: endo iii and fpg alkaline comet assays in chinese hamster ovary (cho-k1) cells.cytotoxic and genotoxic effects of flurochloridone (flc) and its formulations twin pack gold(®) and rainbow(®) were evaluated in cho-k1 cells. using the alkaline single-cell gel electrophoresis (scge) assay, we observed that flc (15 μg/ml), twin pack gold(®) or rainbow(®) induced primary dna damage, increasing the frequency of damaged nucleoids. vitamin e pretreatment did not modify the effect. decreased cell viability was observed only in twin pack gold(®)-treated cultures and was significantly ...201626921020
fevipiprant (qaw039), a slowly dissociating crth2 antagonist with the potential for improved clinical efficacy.here we describe the pharmacologic properties of a series of clinically relevant chemoattractant receptor-homologous molecules expressed on t-helper type 2 (crth2) receptor antagonists, including fevipiprant (nvp-qaw039 or qaw039), which is currently in development for the treatment of allergic diseases. [(3)h]-qaw039 displayed high affinity for the human crth2 receptor (1.14 ± 0.44 nm) expressed in chinese hamster ovary cells, the binding being reversible and competitive with the native agonist ...201626916831
performance optimization of continuous countercurrent tangential chromatography for antibody capture.recent studies have demonstrated that continuous countercurrent tangential chromatography (cctc) can effectively purify monoclonal antibodies from clarified cell culture fluid. cctc has the potential to overcome many of the limitations of conventional packed bed protein a chromatography. this paper explores the optimization of cctc in terms of product yield, impurity removal, overall productivity, and buffer usage. modeling was based on data from bench-scale process development and cctc experime ...201626914276
chemical inhibition of autophagy: examining its potential to increase the specific productivity of recombinant cho cell lines.chinese hamster ovary (cho) cells activate and undergo apoptosis and autophagy for various environmental stresses. unlike apoptosis, studies on increasing the production of therapeutic proteins in cho cells by targeting the autophagy pathway are limited. in order to identify the effects of chemical autophagy inhibitors on the specific productivity (qp ), nine chemical inhibitors that had been reported to target three different phases of autophagy (metformin, dorsomorphin, resveratrol, and sp6001 ...201626914152
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