| pyruvate kinase muscle-1 expression appears to drive lactogenic behavior in cho cell lines, triggering lower viability and productivity: a case study. | chinese hamster ovary (cho) cell lines are used to express a variety of therapeutic proteins. however, lactogenic behavior displayed by some cho cell lines during manufacturing processes may result in a decline in viability, productivity, and possible alterations in product quality. in cultured cells, lactate is produced during glycolysis through irreversible conversion of phosphoenolpyruvate to pyruvate and then lactate via sequential function of pyruvate kinase and lactate dehydrogenase (ldh) ... | 2019 | 30179303 |
| celludinones, new inhibitors of sterol o-acyltransferase, produced by talaromyces cellulolyticus bf-0307. | new indanones, designated celludinones a ((±)-1) and b (2), were isolated from the culture broth of the fungal strain talaromyces cellulolyticus bf-0307. the structures of celludinones were elucidated by spectroscopic data, including 1d and 2d nmr. celludinone a was found to be a mixture of racemic isomers ((±)-1), which were isolated by a chiral column. compounds (+)-1 and (-)-1 inhibited the sterol o-acyltransferase (soat) 1 and 2 isozymes in a cell-based assay using soat1- and soat2-expressin ... | 2018 | 30177721 |
| potential role of mitochondrial damage and s9 mixture including metabolic enzymes in zno nanoparticles-induced oxidative stress and genotoxicity in chinese hamster lung (chl/iu) cells. | the present study was designed to examine genotoxicity induced by 10-40 nm zinc oxide (zno) nanoparticles using the in vitro system. the frequency of micronuclei was significantly increased in a dose-dependent manner when cultured chinese hamster lung (chl/iu) cells were exposed to zno nanoparticles for 24, 48 and 72 h in the continuous treatment method. the maximal frequency of micronuclei was observed in exposure of chl/iu cells to zno nanoparticles at a concentration of 125 μm. the frequency ... | 2018 | 30173861 |
| direct observation of cargo transfer from hdl particles to the plasma membrane. | exchange of cholesterol between high-density lipoprotein (hdl) particles and cells is a key process for maintaining cellular cholesterol homeostasis. recently, we have shown that amphiphilic cargo derived from hdl can be transferred directly to lipid bilayers. here we pursued this work using a fluorescence-based method to directly follow cargo transfer from hdl particles to the cell membrane. | 2018 | 30173079 |
| model-based identification of cell-cycle-dependent metabolism and putative autocrine effects in antibody producing cho cell culture. | the understanding of cell-cycle-dependent population heterogeneities in mammalian cell culture and their influence on production rates is still limited. furthermore, metabolic regulations arising from self-expressed signaling factors (autocrine/autoinhibitory factors) have been postulated in the past, but no determination of such effects have been made so far for fast-growing production chinese hamster ovary (cho) cells in chemically defined media. in this study, a novel approach combining near- ... | 2018 | 30171773 |
| in vitro genotoxicity studies: n-butyl acrylate l5178y mouse lymphoma (tk+/- locus assay), 2-ethylhexyl acrylate gene mutation assay in chinese hamster v79 cells, and 2-ethylhexyl acrylate micronucleus test in human lymphocytes. | available point mutation tests have shown inconsistent results with various acrylates. most of those tests were performed prior to oecd guidelines and appropriate data regarding cytotoxicity are not given. data from three current oecd guideline compliant experiments conducted under glp are provided. they include (a) an in vitro mouse lymphoma (tk+/-) assay (oecd 490) [3], (b) an in vitro hprt locus gene mutation assay utilizing cultures of chinese hamster v79 cells (oecd 476) [1], and (c) an in ... | 2018 | 30167439 |
| nop receptor pharmacological profile - a dynamic mass redistribution study. | the nociceptin/orphanin fq (n/ofq) peptide nop receptor is coupled to pertussis toxin (ptx)-sensitive g proteins (gi/o) whose activation leads to the inhibition of both camp production and calcium channel activity, and to the stimulation of potassium currents. the label free dynamic mass redistribution (dmr) approach has been demonstrated useful for investigating the pharmacological profile of g protein-coupled receptors. herein, we employ dmr technology to systematically characterize the pharma ... | 2018 | 30161182 |
| affinity maturation and characterization of the ofatumumab monoclonal antibody. | cd20 molecule, a phosphoprotein with 297 amino acids and four transmembrane domains, is a member of ms4a protein family. anti-cd20 antibodies such as ofatumumab, which have been developed for cancer treatment and has demonstrated efficacy in relapsed/refractory chronic lymphocytic leukemia, are among the most successful therapies to date. rational engineering methods can be applied with reasonable success to improve functional characteristics of antibodies. considering the importance of this iss ... | 2019 | 30160776 |
| investigating dependencies of relative biological effectiveness for proton therapy in cancer cells. | relative biological effectiveness (rbe) accounts for the differences in biological effect from different radiation types. the rbe for proton therapy remains uncertain, as it has been shown to vary from the clinically used value of 1.1. in this work we investigated the rbe of protons and correlated the biological differences with the underlying physical quantities. | 2018 | 30159358 |
| differential gene expression of a feed-spiked super-producing cho cell line. | feed supplements are concentrated cell culture media that contain a variety of nutrients, which can be added during a bioprocess. during fed-batch cultivation, feed media are typically added to a growing cell culture to maximize cell and product concentrations. in this study, only a single shot of feed medium was added on day 0 to a basal cell culture medium and compared to non-supplemented basal medium (feed-spiked at day 0 versus control experiments) by cultivation of a recombinant mab express ... | 2018 | 30157452 |
| crispr/cas9 gene editing for the creation of an mgat1-deficient cho cell line to control hiv-1 vaccine glycosylation. | over the last decade, multiple broadly neutralizing monoclonal antibodies (bn-mabs) to the hiv-1 envelope protein (env) gp120 have been described. many of these recognize epitopes consisting of both amino acid and glycan residues. moreover, the glycans required for binding of these bn-mabs are early intermediates in the n-linked glycosylation pathway. this type of glycosylation substantially alters the mass and net charge of envs compared to molecules with the same amino acid sequence but posses ... | 2018 | 30157178 |
| discovery, characterization, and remediation of a c-terminal fc-extension in proteins expressed in cho cells. | protein-based biotherapeutics are produced in engineered cells through complex processes and may contain a wide variety of variants and post-translational modifications that must be monitored or controlled to ensure product quality. recently, a low level (~1-5%) impurity was observed in a number of proteins derived from stably transfected chinese hamster ovary (cho) cells using mass spectrometry. these molecules include antibodies and fc fusion proteins where fc is on the c-terminus of the const ... | 2019 | 30148415 |
| recurring genomic structural variation leads to clonal instability and loss of productivity. | chinese hamster ovary cells, commonly used in the production of therapeutic proteins, are aneuploid. their chromosomes bear structural abnormality and undergo changes in structure and number during cell proliferation. some production cell lines are unstable and lose their productivity over time in the manufacturing process and during the product's life cycle. to better understand the link between genomic structural changes and productivity stability, an immunoglobulin g producing cell line was s ... | 2019 | 30144379 |
| chemical characterisation and toxicity assessment in vitro and in vivo of the hydroethanolic extract of terminalia argentea mart. leaves. | terminalia argentea mart. (combretaceae), known mainly as "capitão", is a native tree, not endemic, that occurs in the amazon, caatinga, cerrado and atlantic forest in brazil. leaf infusion is popularly mentioned by riverine communities that inhabit the microregion of northern araguaia (mato grosso, brazil) for the treatment of gastric ulcer, bronchitis and haemorrhage. considering the wide medicinal use, lack of studies that evaluate the safety of use and the scarcity of phytochemical studies o ... | 2018 | 30142424 |
| stepwise phosphorylation of leukotriene b4 receptor 1 defines cellular responses to leukotriene b4. | leukotriene b4 (ltb4) receptor type 1 (blt1) is abundant in phagocytic and immune cells and plays crucial roles in various inflammatory diseases. blt1 is phosphorylated at several serine and threonine residues upon stimulation with the inflammatory lipid ltb4 using phos-tag gel electrophoresis to separate differentially phosphorylated forms of blt1, we identified two distinct types of phosphorylation, basal and ligand-induced, in the carboxyl terminus of human blt1. in the absence of ltb4, the b ... | 2018 | 30131369 |
| ligand-induced coupling between oligomers of the m2 receptor and the gi1 protein in live cells. | uncertainty over the mechanism of signaling via g protein-coupled receptors (gpcrs) relates in part to questions regarding their supramolecular structure. gpcrs and heterotrimeric g proteins are known to couple as monomers under various conditions. many gpcrs form oligomers under many of the same conditions, however, and the biological role of those complexes is unclear. we have used dual-color fluorescence correlation spectroscopy to identify oligomers of the m2 muscarinic receptor and of gi1 i ... | 2018 | 30131171 |
| electron donating capacity reduction of dissolved organic matter by solar irradiation reduces the cytotoxicity formation potential during wastewater chlorination. | after treated wastewater is discharged into surface water for unplanned indirect potable reuse, solar irradiation transforms the dissolved organic matter (dom), which would alter the formation of disinfection byproducts (dbps) and change the cytotoxicity formation potential (ctfp) during post-chlorination in drinking water treatment plants. this study investigated the effects of solar irradiation on the ctfp and total organic halogen formation potential (toxfp) of wastewater during post-chlorina ... | 2018 | 30121436 |
| effects of amino acid substitutions on the biological activity of anti-cd20 monoclonal antibody produced by transgenic silkworms (bombyx mori). | recombinant monoclonal antibodies (mabs) have been used in various therapeutic applications including cancer therapy. fc-mediated effector functions play a pivotal role in the tumor-killing activities of some tumor-targeting mabs, and fc-engineering technologies with glyco-engineering or amino acid substitutions at the antibody fc region have been used to enhance cytotoxic activities including antibody-dependent cellular cytotoxicity (adcc). we previously reported that the mabs produced using tr ... | 2018 | 30119885 |
| transcripts of kv7.1 and mink channels and slow delayed rectifier k+ current (iks) are expressed in zebrafish (danio rerio) heart. | zebrafish are increasingly used as a model for human cardiac electrophysiology, arrhythmias, and drug screening. however, k+ ion channels of the zebrafish heart, which determine the rate of repolarization and duration of cardiac action potential (ap) are still incompletely known and characterized. here, we provide the first evidence for the presence of the slow component of the delayed rectifier k+channels in the zebrafish heart and characterize electrophysiological properties of the slow compon ... | 2018 | 30116893 |
| the development and study of recombinant immunoglobulin a to hemagglutinins of the influenza virus. | we obtained recombinant variants of human antibody fi6 broadly specific to hemagglutinins of the influenza a virus. on the basis of a bi-promoter (cmv, hef1-htlv) vector, we developed genetic constructs for the expression of the heavy and light chains of the immunoglobulins of iga1-, iga2m1-, and igg-isotypes. following transfection and selection, stable chinese hamster ovary (cho) cell lines were produced. the antibodies of iga1-, iga2m1-, and igg-isotypes were purified from culture media. we p ... | 2020 | 30116613 |
| high glucose and low specific cell growth but not mild hypothermia improve specific r-protein productivity in chemostat culture of cho cells. | in the biopharmaceutical sector, chinese hamster ovary (cho) cells have become the host of choice to produce recombinant proteins (r-proteins) due to their capacity for correct protein folding, assembly, and posttranslational modification. however, the production of therapeutic r-proteins in cho cells is expensive and presents insufficient production yields for certain proteins. effective culture strategies to increase productivity (qp) include a high glucose concentration in the medium and mild ... | 2018 | 30114204 |
| oligomerization and catalytic parameters of human udp-glucuronosyltransferase 1a10: expression and characterization of the recombinant protein. | udp-glucuronosyltransferase (ugt), as an integral membrane protein localized in the endoplasmic reticulum, has the ability to detoxify potentially hazardous xenobiotic substances. most ugts are expressed in liver, but ugt1a10 has proven to be an extrahepatic enzyme considerably expressed throughout the gastrointestinal tract. earlier studies indicated that different ugt isoforms could exist in higher-order homo-oligomers or at least dimers within the membrane, but the formation of intermolecular ... | 2018 | 30111624 |
| enhanced transgene expression using cis-acting elements combined with the ef1 promoter in a mammalian expression system. | transgene expression in eukaryotic cells suffers from epigenetic effects that result in low or unstable transgene expression and high clonal variability. the use of epigenetic regulators is a promising approach to alleviating such unwanted effects. we investigated the effect of the strong human ef1-α promoter combined with six cis-acting elements on transgene expression in transfected chinese hamster ovary (cho) cells. the six elements included the human cytomegalovirus immediate early core prom ... | 2018 | 30110655 |
| a mixture of atropisomers enhances neutral lipid degradation in mammalian cells with autophagy induction. | atropisomers with a biaryl dihydronaphthopyranone structure, dinapinones a1 (dpa1) (m position) and a2 (dpa2) (p position), were isolated from the fungus culture broth of talaromyces pinophilus fki-3864 as inhibitors of [14c]neutral lipid ([14c]triacylglycerol (tg) and [14c]cholesteryl ester (ce)) synthesis from [14c]oleic acid in chinese hamster ovary-k1 (cho-k1) cells. dpa2 inhibited [14c]tg and [14c]ce synthesis (ic50s, 0.65 and 5.6 μm, respectively), but dpa1 had no inhibitory activity on [1 ... | 2018 | 30108268 |
| isolation and characterization of a chinese hamster ovary heparan sulfate cell mutant defective in both met receptor binding and hepatocyte growth factor nk1/met signaling. | the up-regulation of hepatocyte growth factor/receptor, hgf/met, signal transduction is observed in most of human cancers. specific heparan sulfate structures enhance the hgf/met signaling at both cell and animal-based model systems. biochemical studies indicate that heparan sulfate interacts with hgf and a natural occurring splicing variant nk1 of hgf with similar affinity. however, it is currently unknown if cell surface heparan sulfate binds to met at physiological conditions and if specific ... | 2018 | 30107380 |
| the gagome: a cell-based library of displayed glycosaminoglycans. | glycosaminoglycans (gags) are essential polysaccharides in normal physiology and disease. however, understanding of the contribution of specific gag structures to specific biological functions is limited, largely because of the great structural heterogeneity among gags themselves, as well as technical limitations in the structural characterization and chemical synthesis of gags. here we describe a cell-based method to produce and display distinct gags with a broad repertoire of modifications, a ... | 2018 | 30104636 |
| detecting and imaging o-glcnac sites using glycosyltransferases: a systematic approach to study o-glcnac. | o-glcnacylation is a reversible serine/threonine glycosylation for regulating protein activity and availability inside cells. in a given protein, o-glcnacylated and unoccupied o-linked β-n-acetylglucosamine (o-glcnac) sites are referred to as closed and open sites, respectively. the balance between open and closed sites is believed to be dynamically regulated. in this report, closed sites are detected using in vitro incorporation of galnaz by b3galnt2, and open sites are detected by in vitro inc ... | 2018 | 30100348 |
| suitable hepatitis b vaccine for adult immunization in china: a systematic review and meta-analysis. | hepatitis b virus (hbv) infection remains an important public health problem in china, and adults need to be vaccinated. this systematic review and meta-analysis assessed the appropriate immunization of adults in china. only randomized controlled trials (rcts) were eligible, and seroprotection was defined as anti-hbs≥ 10 miu/ml; 18,308 participants in 27 studies were included. relative risk (rr) and random effects models were used. twenty micrograms of hbv vaccine resulted in a better response t ... | 2019 | 30089437 |
| chromawizard: an open source image analysis software for multicolor fluorescence in situ hybridization analysis. | multicolor image analysis finds its applications in a broad range of biological studies. specifically, multiplex fluorescence in situ hybridization (m-fish) for chromosome painting facilitates the analysis of individual chromosomes in complex metaphase spreads and is widely used to detect both numerical and structural aberrations. while this is well established for human and mouse karyotypes, for which species sophisticated software and analysis tools are available, other organisms and species a ... | 2018 | 30089195 |
| ph excursions impact cho cell culture performance and antibody n-linked glycosylation. | ph excursions exist due to frequent base addition and environmental heterogeneity in large-scale bioreactors. such excursions could lead to suboptimal culture performance. here we investigated the impact of ph excursions on cell culture performance and n-linked glycosylation for three mab-producing chinese hamster ovary cell lines. frequent ph excursions were introduced by bolus base addition (in total 2-6% of initial volume, fixed bolus addition distributed from day 2 to 8) into small-scale bio ... | 2018 | 30088083 |
| glycan-masking hemagglutinin antigens from stable cho cell clones for h5n1 avian influenza vaccine development. | refocusing of b-cell responses can be achieved by preserving the overall fold of the antigen structure but selectively mutating the undesired antigenic sites with additional n-linked glycosylation motifs for glycan masking the vaccine antigen. we previously reported that glycan-masking recombinant h5 hemagglutinin (rh5ha) antigens on residues 83, 127, and 138 (g127 + g138 or g83 + g127 + 138 rh5ha) elicited broader neutralizing antibodies and protection against heterologous clades/subclades of h ... | 2019 | 30080931 |
| isolation of recombinant human antithrombin isoforms by cellufine sulfate affinity chromatography. | human antithrombin (hat) is a major serine protease inhibitor that regulates blood coagulation in human plasma, and it has been applied for the treatment of antithrombin (at) deficiency and disseminated intravascular coagulation (dic). in the past, hat for therapeutic use has been obtained from human plasma; however, hat can now be sourced from transgenic animals and chinese hamster ovary (cho) cells by recombinant technology. the dominant form of hat in plasma is the α form, which is glycosylat ... | 2018 | 30077924 |
| the expression pattern of the phosphoproteome is significantly changed during the growth phases of recombinant cho cell culture. | post-translational modification of proteins by reversible phosphorylation plays a pivotal role in regulating key cellular processes including transcription, translation, cell proliferation, differentiation, apoptosis, and signal transduction. despite the importance of the phosphorylation level of regulation, little work has been carried out on the phosphoproteomic characterization of chinese hamster ovary (cho) cells in bioprocess-relevant conditions. growth control strategies are often used to ... | 2018 | 30076757 |
| pharmacological properties of asp7657, a novel, potent, and selective prostaglandin ep4 receptor antagonist. | we determined the pharmacologic profile of asp7657, trans-4-[({[1-(quinolin-2-ylmethyl)-5-(trifluoromethyl)-1h-indol-7 yl] carbonyl} amino) methyl] cyclohexanecarboxylic acid methanesulfonate (1:1), a newly synthesized selective e-type prostaglandin (ep)4 receptor antagonist using several in vitro and in vivo experiments. asp7657 exhibited high affinity for rat and human ep4 receptors, with ki values of 6.02 nm and 2.21 nm, respectively. in addition, asp7657 potently inhibited the pge2-induced c ... | 2018 | 30076448 |
| robotic selection for the rapid development of stable cho cell lines for hiv vaccine production. | the production of envelope glycoproteins (envs) for use as hiv vaccines is challenging. the yield of envs expressed in stable chinese hamster ovary (cho) cell lines is typically 10-100 fold lower than other glycoproteins of pharmaceutical interest. moreover, envs produced in cho cells are typically enriched for sialic acid containing glycans compared to virus associated envs that possess mainly high-mannose carbohydrates. this difference alters the net charge and biophysical properties of envs a ... | 2018 | 30071025 |
| egcg improves recombinant protein productivity in chinese hamster ovary cell cultures via cell proliferation control. | chinese hamster ovary cell lines are good manufacturing practice-certified host cells and are widely used in the field of biotechnology to produce therapeutic antibodies. recombinant protein productivity in cells is strongly associated with cell growth. to control cell proliferation, many approaches have previously been tested including: genetic engineering, chemical additives such as cell cycle inhibitors, and temperature shift of the culture. to be widely adopted in the biopharmaceutical indus ... | 2018 | 30069612 |
| media photo-degradation in pharmaceutical biotechnology - impact of ambient light on media quality, cell physiology, and igg production in cho cultures. | many vital components in bioprocess media are prone to photo-conversion or photo-degradation upon exposure to ambient light, with severe negative consequences for biomass yield and overall productivity. however, there is only limited awareness of light irradiation as a potential risk factor when working in transparent glass bioreactors, storage vessels or disposable bag systems. the chemical complexity of most media renders a root-cause analysis difficult. this study investigated in a novel, hol ... | 2018 | 30069078 |
| perspectives on progressive strategies and recent trends in the production of recombinant human factor viii. | factor viii (fviii), a glycoprotein cofactor, plays a crucial role in the intrinsic blood coagulation pathway. as one of the most essential blood clotting factors known today, fviii is the largest and most complex commercialized therapeutic protein used in the treatment of hemophilia a, an x-linked recessive disorder. two lyophilized fviii concentrates (viz., plasma fractionated and recombinant) are in use to treat hemorrhagic episodes in patients suffering with hemophilia a. recombinant fviii ( ... | 2018 | 30063930 |
| dna repair deficient chinese hamster ovary cells exhibiting differential sensitivity to charged particle radiation under aerobic and hypoxic conditions. | it has been well established that hypoxia significantly increases both cellular and tumor resistance to ionizing radiation. hypoxia associated radiation resistance has been known for some time but there has been limited success in sensitizing cells to radiation under hypoxic conditions. these studies show that, when irradiated with low linear energy transfer (let) gamma-rays, poly (adp-ribose), polymerase (parp), fanconi anemia (fanc), and mutant chinese hamster ovary (cho) cells respond similar ... | 2018 | 30061540 |
| activatable magnetic resonance nanosensor as a potential imaging agent for detecting and discriminating thrombosis. | the early detection and accurate characterization of life-threatening diseases such as cardiovascular disease and cancer are critical to the design of treatment. knowing whether or not a thrombus in a blood vessel is new (fresh) or old (constituted) is very important for physicians to decide a treatment protocol. we have designed smart mri nano-sensors that can detect, sense and report the stage or progression of cardiovascular diseases such as thrombosis. the nanosensors were functionalized wit ... | 2018 | 30059122 |
| brassica rapa hairy root based expression system leads to the production of highly homogenous and reproducible profiles of recombinant human alpha-l-iduronidase. | the brassica rapa hairy root based expression platform, a turnip hairy root based expression system, is able to produce human complex glycoproteins such as the alpha-l-iduronidase (idua) with an activity similar to the one produced by chinese hamster ovary (cho) cells. in this article, a particular attention has been paid to the n- and o-glycosylation that characterize the alpha-l-iduronidase produced using this hairy root based system. this analysis showed that the recombinant protein is charac ... | 2019 | 30058762 |
| site specificity of eel luteinizing hormone n-linked oligosaccharides in signal transduction. | eel luteinizing hormone (eellh) is composed of a common α-subunit and hormone specific β-subunit, both of which contain asparagine-linked carbohydrate residues, located at positions 56 and 79 on the α-subunit and position 10 on the β-subunit. the specific roles of the individual carbohydrate chains are poorly defined in eel. thus, we characterized the biologically active single chains by fusing the α-subunit to the carboxyl terminal region of the eellh β-subunit. site-directed mutagenesis of the ... | 2018 | 30056138 |
| glyco-engineered cell line and computational docking studies reveals enterotoxigenic escherichia coli cfa/i fimbriae bind to lewis a glycans. | we have previously reported clinical data to suggest that colonization factor i (cfa/i) fimbriae of enterotoxigenic escherichia coli (etec) can bind to lewis a (lea), a glycan epitope ubiquitous in the small intestinal mucosa of young children (<2 years of age), and individuals with a genetic mutation of fut2. to further elucidate the physiological binding properties of this interaction, we engineered chinese hamster ovary (cho-k1) cells to express lea or leb determinants on both n- and o-glycan ... | 2018 | 30050155 |
| host cell protein profiling in biopharmaceutical harvests. | biopharmaceuticals contain residual host cell protein (hcp) impurities, a complex mixture of endogenous proteins from production cell lines such as chinese hamster ovary (cho) cells. the composition of hcp impurities at harvest hinges on multiple factors, e.g., identity of cell line, cell density and viability at harvest, or other process parameters. two-dimensional differential gel electrophoresis (2-d dige) was used to compare hcp in 15 null cell culture harvest supernatants, which are represe ... | 2018 | 30048127 |
| fluorescence "off" and "on" signalling of esculetin in the presence of copper and thiol: a possible implication in cellular thiol sensing. | the interaction of the cupric ion with esculetin, a dihydroxy coumarin derivative, was studied by absorption and fluorescence spectroscopic methods in aqueous medium. esculetin formed a complex in the presence of the cupric ion which was characterised by the shift in its absorption band from 350 nm to 389 nm and the quenching of its fluorescence intensity at 466 nm. from job's plot and fluorescence quenching studies, the stoichiometry of the copper ion and esculetin in the complex was estimated ... | 2018 | 30043019 |
| chlorotyrosines versus volatile byproducts from chlorine disinfection during washing of spinach and lettuce. | following the food safety modernization act of 2011 in the u.s., guidelines for disinfection washes in food packaging facilities are under consideration to control pathogen risks. however, disinfectant exposures may need optimization because the high concentrations of chlorine disinfectant promote the formation of high levels of disinfection byproducts (dbps). when chlorine doses up through the 200 mg/l as cl2 range relevant to the current practice were applied to spinach and lettuce, significan ... | 2018 | 30040386 |
| development of a shake tube-based scale-down model for perfusion cultures. | the use of benchtop bioreactors (brs) for the development of mammalian cell perfusion cultures is expensive and time consuming, given its complexity in equipment and operation. scale-down models, going from liter to milliliter scale, are needed to support the rapid determination of suitable operating conditions in terms of viable cell density (vcd), perfusion rate, and medium composition. in this study, we compare the performance of steady-state perfusion cultures in orbitally shaken tube and br ... | 2018 | 30039852 |
| culture temperature modulates half antibody and aggregate formation in a chinese hamster ovary cell line expressing a bispecific antibody. | therapeutic bispecific antibodies are formed by assembly of multichain polypeptides. in general, a bispecific antibody has two different light chains and two different heavy chains that fold and correctly pair via engineered interchain interactions. because of some incorrect assembly, product-related impurities can be prevalent (e.g., half molecules, mispaired light chains, homodimers), requiring its removal during subsequent purification. in this study, we investigated the modulation of impurit ... | 2018 | 30036451 |
| transcriptome analysis of cho cell size increase during a fed-batch process. | in a chinese hamster ovary (cho) cell fed-batch process, arrest of cell proliferation and an almost threefold increase in cell size occurred, which is associated with an increase in cell-specific productivity. in this study, transcriptome analysis is performed to identify the molecular mechanisms associated with this. cell cycle analysis reveals that the cells are arrested mainly in the g0 /g1 phase. the cell cycle arrest is associated with significant up-regulation of cyclin-dependent kinases i ... | 2019 | 30024106 |
| single copy transgene integration in a transcriptionally active site for recombinant protein synthesis. | for the biomanufacturing of protein biologics, establishing stable cell lines with high transgene transcription is critical for high productivity. modern genome engineering tools can direct transgene insertion to a specified genomic locus and can potentially become a valuable tool for cell line generation. in this study, the authors survey transgene integration sites and their transcriptional activity to identify characteristics of desirable regions. a lentivirus containing destabilized green fl ... | 2018 | 30024101 |
| toxicity and antigenotoxic effect of hispolon derivatives: role of structure in modulating cellular redox state and thioredoxin reductase. | hispolon (hs), a bioactive polyphenol, and its derivatives such as hispolon monomethyl ether (hme), hispolon pyrazole (hp), and hispolon monomethyl ether pyrazole (hmep) were evaluated for comparative toxicity and antigenotoxic effects. the stability of hs derivatives in biological matrices followed the order hs < hp ≈ hme < hmep. the cytotoxicity analysis of hs derivatives indicated that hp and hmep were less toxic than hs and hme, respectively, in both normal and tumor cell types. the mechanis ... | 2018 | 30023935 |
| laccase-mediated enhancement of the antioxidant activity of propolis and poplar bud exudates. | the treatment of propolis and poplar bud exudates with laccase from trametes versicolor and 2,2,6,6-tetramethyl-1-piperidinyloxy free radical increased the antioxidant activity, as evaluated by the 2,2'-diphenyl picrylhydrazyl (dpph)- and t-butyl-ooh-induced dna breakage comet assay analyses. the effect was highest for shorter reaction times. propolis showed the highest antioxidant activity in the dpph test, whereas poplar bud exudates were more active in reducing the t-butyl-ooh-induced lesions ... | 2017 | 30023668 |
| analysis of intracellular igg secretion in chinese hamster ovary cells to improve igg production. | the production of biopharmaceutical immunoglobulin g (igg) using cultured mammalian cells, especially chinese hamster ovary (cho) cells is well established and has been markedly improved through the modification of cells and cell culture engineering technologies. the establishment of high-production cell lines remains a challenge. the intracellular secretion of igg has been investigated to identify and solve the rate-limiting steps in antibody production. however, strategies that regulate the ex ... | 2019 | 30017708 |
| the contributions of individual galactosyltransferases to protein specific n-glycan processing in chinese hamster ovary cells. | galactosylation as part of n-glycan processing is conducted by a set of beta-1,4-galactosyltransferases (b4galts), with b4galt1 as the dominant isoenzyme for this reaction. nevertheless, the exact contributions of this key-player as well as of the other isoenzymes involved in n-glycosylation, b4galt2, b4galt3 and b4galt4, have not been studied in-depth. to increase the understanding of the protein- and site-specific activities of individual galactosyltransferases in chinese hamster ovary cells, ... | 2018 | 30017654 |
| genotoxicity and hemocompatibility of a novel calcium aluminate-based cement. | the aim of this in vitro study was to evaluate the genotoxicity and hemocompatibility of a novel calcium aluminate-based cement, endobinder (eb) (binderware, são carlos, sp, brazil) and compare it with angelus white mineral trioxide aggregate (mta) (awmta) (angelus, soluções odontológicas, londrina, pr, brazil). | 2018 | 32161862 |
| simulation and experimental study on the mechanism of the chlorination of azithromycin. | azithromycin (azi) has been listed as an emerging contaminant by the us epa since 2009 because it is frequently detected in wastewater, surface water, and drinking water. in this paper, the chlorination of azi in drinking water was simulated and studied. the results indicated that new compounds were generated in the chlorination of azi. the byproducts were identified by liquid chromatography-quadrupole-time-of-flight mass spectrometry (lc-q-tof), and four of the byproducts were detected in real ... | 2018 | 30014912 |
| generation of flp-intm-ready dg44 and lec 3.2.8.1 cho cell lines for quick and easy constitutive protein expression. | the well-characterized cell line chinese hamster ovary (cho) has been used to produce numerous biopharmaceuticals and is an important tool for basic research. however, introducing foreign dna into specially modified cho cells such as dg44 and lec 3.2.8.1 can sometimes be an arduous process. here we show that the flp-intm plasmid can be modified to produce a fluorescent tracer protein tag (mcherrytm) as a fusion reporter, to allow for the rapid selection of single-cell sorted, isogenic flp-intm-r ... | 2018 | 30014730 |
| differential expression of micrornas in recombinant chinese hamster ovary cells treated with sodium butyrate using digital rna counting. | sodium butyrate (nabu) is an efficient supplement for increasing recombinant protein production in chinese hamster ovary (cho) cell culture. to elucidate the effects of nabu on mirna expression profile in recombinant cho (rcho) cells, differentially expressed mirnas in nabu-treated rcho cells were assessed by nanostring ncounter analysis. this result showed that eight mature mouse mirnas (let-7b, let-7d, mir-15b, mir-25, mir-27a, mir-99a, mir-125a-5p, and mir-125b-5p) were differentially express ... | 2018 | 30012463 |
| improving the production of recombinant human bone morphogenetic protein-4 in chinese hamster ovary cell cultures by inhibition of undesirable endocytosis. | endocytic regulation serves a critical role in modulating the extracellular level of signaling molecules, such as bone morphogenetic proteins (bmps). unfortunately, endocytosis may result in poor yields of recombinant human bmp-4 (rhbmp-4) from chinese hamster ovary (cho) cell cultures. when rhbmp-4 was incubated with cho cells, rhbmp-4 was actively internalized into cells. cell surface bound heparan sulfate proteoglycans (hspgs) served as the major receptors for rhbmp-4 internalization. removal ... | 2018 | 30011067 |
| a novel mammalian cell line development platform utilizing nanofluidics and optoelectro positioning technology. | generating a highly productive cell line is resource intensive and typically involves long timelines because of the need to screen large numbers of candidates in protein production studies. this has led to miniaturization and automation strategies to allow for reductions in resources and higher throughput. current approaches rely on the use of standard cell culture vessels and bulky liquid handling equipment. new nanofludic technologies offer novel solutions to surpass these limits, further mini ... | 2018 | 30009534 |
| cardiovascular pharmacology of k2p17.1 (task-4, talk-2) two-pore-domain k+ channels. | k2p17.1 (task-4, talk-2) potassium channels are expressed in the heart and represent potential targets for pharmacological management of atrial and ventricular arrhythmias. reduced k2p17.1 expression was found in atria and ventricles of heart failure (hf) patients. modulation of k2p17.1 currents by antiarrhythmic compounds has not been comprehensively studied to date. the objective of this study was to investigate acute effects of clinically relevant antiarrhythmic drugs on human k2p17.1 channel ... | 2018 | 30008082 |
| single cell dielectrophoresis study of apoptosis progression induced by controlled starvation. | nutrient depletion in fed-batch cultures and at the end of batch cultures is among the main causes of stress on cells and a trigger of apoptosis. in this study, we investigated changes in the cytoplasm conductivity of chinese hamster ovary (cho) cells under controlled starvation. employing a single-cell dielectrophoresis (dep) cytometer, we measured the dep response of cho cells incubated in a medium without glucose and glutamine over a 48-h period. using the measured data in conjunction with nu ... | 2018 | 30007208 |
| carbon dots-decorated na2w4o13 composite with wo3 for highly efficient photocatalytic antibacterial activity. | photodisinfection by semiconductors has been proven to be an effective method for achieving antibacterial or antifungal activity. however, the toxicity of the nanomaterial to the environment and organisms is a major concern. herein, a highly efficient and environmentally friendly photodisinfection material of a carbon dots (cds) decorated na2w4o13 composite with wo3 photocatalyst was fabricated via a facile hydrothermal-calcination approach. the tem (transmission electron microscopy) images show ... | 2018 | 30007199 |
| n-linked glycosylation modulates the immunogenicity of recombinant human factor viii in hemophilia a mice. | immune responses to factor viii remain the greatest complication in the treatment of severe hemophilia a. recent epidemiological evidence has highlighted that recombinant factor viii produced in baby hamster kidney cells is more immunogenic than factor viii produced in chinese hamster ovary cells. glycosylation differences have been hypothesized to influence the immunogenicity of these synthetic concentrates. in two hemophilia a mouse models, baby hamster kidney cell-derived factor viii elicited ... | 2018 | 30002126 |
| novel bivalent and d-peptide ligands of cxcr4 mobilize hematopoietic progenitor cells to the blood in c3h/hej mice. | the interaction of sdf-1α (also known as cxcl12) with the cxcr4 receptor plays a critical role in the retention of hematopoietic stem cells (hscs) in bone marrow. the viral macrophage inflammatory protein-ii (vmip-ii), a human herpesvirus-8 (hhv-8)-encoded viral chemokine, can bind the cxcr4 receptor and inhibit endogenous ligand-induced calcium responses and cell migration. previously, we used the bivalent ligand approach to link synthetically two unnatural d-amino acid peptides derived from th ... | 2018 | 29991278 |
| leaky expression of the tet-on system hinders control of endogenous mirna abundance. | with the ability to affect multiple genes and fundamental pathways simultaneously, mirna engineering of chinese hamster ovary (cho) cells has significant advantages over single gene expression or repression. tight control of these molecular triggers is desirable as it could in theory allow on/off or even tunable regulation of desirable cellular phenotypes. the present study investigated the potential of employing a tetracycline inducible (tet-on) system for conditional knockdown of specific mirn ... | 2019 | 29989353 |
| control of amino acid transport into chinese hamster ovary cells. | amino acid transporters (aats) represent a key interface between the cell and its environment, critical for all cellular processes: energy generation, redox control, and synthesis of cell and product biomass. however, very little is known about the activity of different functional classes of aats in chinese hamster ovary (cho) cells, how they support cell growth and productivity, and the potential for engineering their activity and/or the composition of amino acids in growth media to improve cho ... | 2018 | 29987891 |
| genome sequence comparison between chinese hamster ovary (cho) dg44 cells and mouse using end sequences of cho bac clones based on bac-fish results. | chinese hamster ovary (cho) cells have frequently been used in biotechnology as a mammalian host cell platform for expressing genes of interest. previously, we constructed a detailed physical chromosomal map of the cho dg44 cell line by fluorescence in situ hybridization (fish) imaging using 303 bacterial artificial chromosome (bac) clones as hybridization probes (bac-fish). bac-fish results revealed that the two longest chromosomes were completely paired. however, other chromosomes featured par ... | 2018 | 29987698 |
| application of microrna targeted 3'utrs to repress dhfr selection marker expression for development of recombinant antibody expressing cho cell pools. | the dihydrofolate reductase (dhfr) system is used for the selection of recombinant chinese hamster ovary (cho) cell lines using the inhibitor methotrexate (mtx). during clonal selection, endogenous dhfr expression, and resistance to mtx allows the selection of cells expressing sufficient dhfr to survive. here, the authors describe a novel vector platform for the dhfr system, whereby addition of a synthetic 3'utr destabilizes dhfr expression. mirs ability to negatively regulate gene expression by ... | 2018 | 29981248 |
| paxillin phosphorylation at serine 273 and its effects on rac, rho and adhesion dynamics. | focal adhesions are protein complexes that anchor cells to the extracellular matrix. during migration, the growth and disassembly of these structures are spatiotemporally regulated, with new adhesions forming at the leading edge of the cell and mature adhesions disassembling at the rear. signalling proteins and structural cytoskeletal components tightly regulate adhesion dynamics. paxillin, an adaptor protein within adhesions, is one of these proteins. its phosphorylation at serine 273 (s273) is ... | 2018 | 29975690 |
| construction of anti-her2 recombinants as targeting modules for a drug-delivery system against her2-positive cells. | recombinant antibodies have been investigated and used in applications such as targeting modules of drug-delivery systems (dds) against cancers. this study aimed to prepare recombinant antibodies against her2, containing sortase a (srta) recognition sequence, that are applicable as targeting modules in dds after linkage with the drug-carrier containing oligoglycine-acceptor peptide by srta transpeptidation. | 2018 | 29970568 |
| crispr/cas9-mediated knockout of six glycosyltransferase genes in nicotiana benthamiana for the production of recombinant proteins lacking β-1,2-xylose and core α-1,3-fucose. | plants offer fast, flexible and easily scalable alternative platforms for the production of pharmaceutical proteins, but differences between plant and mammalian n-linked glycans, including the presence of β-1,2-xylose and core α-1,3-fucose residues in plants, can affect the activity, potency and immunogenicity of plant-derived proteins. nicotiana benthamiana is widely used for the transient expression of recombinant proteins so it is desirable to modify the endogenous n-glycosylation machinery t ... | 2019 | 29969180 |
| effects of silver sulfide quantum dots coated with 2-mercaptopropionic acid on genotoxic and apoptotic pathways in vitro. | quantum dots (qds) are highly promising nanomaterials in bioimaging system because of their bright fluorescence, broad uv excitation, narrow emission band, and high photostability. recently, there is a great activity on ag2s quantum dots for both imaging and drug/gene delivery due to the potential of having a better cytocompatability and near infrared luminescence. 2-mercaptopropionic acid (2 mp a)-coated silver sulfide (ag2s) qds were reported as the most luminescent, stable, anionic ag2s qds i ... | 2018 | 29958870 |
| high cell density perfusion culture has a maintained exoproteome and metabolome. | the optimization of bioprocesses for biopharmaceutical manufacturing by chinese hamster ovary (cho) cells can be a challenging endeavor and, today, heavily relies on empirical methods treating the bioreactor process and the cells as black boxes. multi-omics approaches have the potential to reveal otherwise unknown characteristics of these systems and identify culture parameters to more rationally optimize the cultivation process. here, the authors have applied both metabolomic and proteomic prof ... | 2018 | 29957873 |
| photosensitization of chinese hamster v79 cells to photoactivated alpha-terthienyl involving membrane damage and oxidative stress. | photosensitization of v79 chinese hamster lung fibroblasts was tested to investigate if the cells can fit the photoactive effect of alpha-terthienyl for safety application. using 15-min photoirradiation of a black light (320-400 nm, 40 w), alpha-terthienyl was significantly photoactivated and caused v79 cells to be shrinkage, detachment and necrosis. the photoactivated alpha-terthienyl played a concentration-dependent stress to decrease cell survival and to induce cell death with median inhibito ... | 2018 | 29957498 |
| a biotinylated and endoplasmic reticulum-targeted glutathione-responsive zinc(ii) phthalocyanine for targeted photodynamic therapy. | a biotinylated glutathione (gsh)-responsive zinc(ii) phthalocyanine has been prepared and characterized. with a 2,4-dinitrobenzenesulfonyl moiety, its fluorescence emission and singlet oxygen generation were silenced in its intact state. upon exposure to high concentration of gsh, its photosensitizing properties were restored in phosphate buffered saline and inside tumor cells. it also showed preferential uptake on hepg2 human hepatocarcinoma cells (with higher biotin receptor expression) rather ... | 2018 | 29956487 |
| automated single-cell analysis and isolation system: a paradigm shift in cell screening methods for bio-medicines. | we have developed an automated robot that facilitates non-invasive isolation of a single cell with the most favorable properties from arrays containing >105 cells, thus allowing the establishment of new cell screening methods for bio-medicines. in this chapter, an outline of the proposed automated single-cell analysis and isolation system (hereafter called 'single-cell robot') is reviewed by comparison with a conventional fluorescence-activated cell sorter (facs). the single-cell robot could per ... | 2018 | 29943292 |
| untangling the mechanism of 3-methyladenine in enhancing the specific productivity: transcriptome analysis of recombinant chinese hamster ovary cells treated with 3-methyladenine. | 3-methyladenine (3-ma) is a chemical additive that enhances the specific productivity (q p ) in recombinant chinese hamster ovary (rcho) cell lines. different from its widely known function of inhibiting autophagy, 3-ma has instead shown to increase autophagic flux in various rcho cell lines. thus, the mechanism by which 3-ma enhances the qp requires investigation. to evaluate the effect of 3-ma on transcriptome dynamics in rcho cells, rna-seq was performed with fc-fusion protein-producing rcho ... | 2018 | 29940077 |
| luminescence from lanthanide(iii) ions bound to the glycocalyx of chinese hamster ovary cells. | lanthanide(iii) ions bind to the glycocalyx of chinese hamster ovary (cho) cells and give rise to a unique luminescent fingerprint. following direct excitation of terbium(iii) and europium(iii) ions in the visible part of the spectrum, we are able to collect emission spectra pixel-by-pixel in images of cho cells. following data analysis that removes the background signal, the fine structure of the europium(iii) luminescence indicate that the lanthanide(iii) ions are bound to a single structure o ... | 2018 | 29939467 |
| sonochemical synthesis of small boron oxide nanoparticles. | the synthesis of small boron oxide nanoparticles (nps) is reported. a sonochemical approach in the presence of a capping agent was employed to produce approximately 4-5-nm-sized b2o3 nps, including the 10b isotopically enriched form. the morphology and composition of the nps were established using transmission electron microscopy and diffraction, respectively. x-ray photoelectron and fourier transform infrared spectroscopies provided information about surface functionalization of the b2o3 nps, w ... | 2018 | 29939022 |
| functional comparison of high and low molecular weight basic fibroblast growth factors. | acid-electrolyzed functional water (fw) is obtained through the electrolysis of sodium chloride solution. stimulation of the human fibroblastic cell line hela by fw led to the augmented secretion of basic fibroblast growth factor (bfgf). immunoprecipitation followed by western blot analysis revealed that both high and low molecular weight isoforms of bfgf were secreted in response to fw treatment. to explore intracellular bfgf localization, a cell fractionation assay was performed. despite the p ... | 2018 | 29923210 |
| ultra-low carbon dioxide partial pressure improves the galactosylation of a monoclonal antibody produced in chinese hamster ovary cells in a bioreactor. | to explore the influence of ultra-low carbon dioxide partial pressure (pco2) on the monoclonal antibody (mab) n-glycosylation profile in chinese hamster ovary (cho) cell culture. | 2018 | 29923054 |
| meta-analysis of publicly available chinese hamster ovary (cho) cell transcriptomic datasets for identifying engineering targets to enhance recombinant protein yields. | transcriptomics has been extensively applied to the investigation of the cho cell platform for the production of recombinant biotherapeutic proteins to identify transcripts whose expression is regulated and correlated to (non)desirable cho cell attributes. however, there have been few attempts to analyze the findings across these studies to identify conserved changes and generic targets for cho cell platform engineering. here, the authors have undertaken a meta-analysis of cho cell transcriptomi ... | 2018 | 29917317 |
| stable expression of infliximab in crispr/cas9-mediated bak1-deficient cho cells. | to establish stable infliximab-expressing chinese hamster ovary (cho) cells with high tolerance to serum-free culture. | 2018 | 29915899 |
| mpo (myeloperoxidase) reduces endothelial glycocalyx thickness dependent on its cationic charge. | objective- the leukocyte heme-enzyme mpo (myeloperoxidase) exerts proinflammatory effects on the vascular system primarily linked to its catalytic properties. recent studies have shown that mpo, depending on its cationic charge, mediates neutrophil recruitment and activation. here, we further investigated mpo's extracatalytic properties and its effect on endothelial glycocalyx (eg) integrity. approach and results- in vivo staining of murine cremaster muscle vessels with alcian blue 8gx provided ... | 2018 | 29903730 |
| mechanisms of precipitate formation during the purification of an fc-fusion protein. | protein precipitates that arise during bioprocessing can cause manufacturing challenges, but they can also aid in clearance of host-cell protein (hcp) and dna impurities. such precipitates differ from many protein precipitates that have been studied previously in their heterogeneous composition, particularly in the presence of high concentrations of the product protein. here, we characterize the precipitates that form after neutralization of protein a purified and viral-inactivated material of a ... | 2018 | 29896879 |
| heterologous expression of ctp:phosphocholine cytidylyltransferase from plasmodium falciparum rescues chinese hamster ovary cells deficient in the kennedy phosphatidylcholine biosynthesis pathway. | the plasmodial ctp:phosphocholine cytidylyltransferase (pfcct) is a promising antimalarial target, which can be inhibited to exploit the need for increased lipid biosynthesis during the erythrocytic life stage of plasmodium falciparum. notable structural and regulatory differences of plasmodial and mammalian ccts offer the possibility to develop species-specific inhibitors. the aim of this study was to use cho-mt58 cells expressing a temperature-sensitive mutant cct for the functional characteri ... | 2018 | 29895950 |
| resveratrol ameliorates microcystin-lr-induced testis germ cell apoptosis in rats via sirt1 signaling pathway activation. | microcystin-leucine arginine (mc-lr), a cyclic heptapeptide produced by cyanobacteria, is a strong reproductive toxin. studies performed in rat sertoli cells and chinese hamster ovary cells have demonstrated typical apoptosis after mc-lr exposure. however, little is known on how to protect against the reproductive toxicity induced by mc-lr. the present study aimed to explore the possible molecular mechanism underlying the anti-apoptosis and protective effects of resveratrol (res) on the co-cultu ... | 2018 | 29890735 |
| safety assessment of the cistanche tubulosa health food product memoregain®: genotoxicity and 28-day repeated dose toxicity test. | the pharmacological effects of cistanches herba, known as "ginseng of the desert", have been extensively studied. in this study, we aimed to assess the genotoxic and oral toxic effects of the cistanche tubulosa health food product memoregain® using in vitro and in vivo tests. ames tests using five strains of salmonella typhimurium showed no signs of increased reverse mutation upon exposure to memoregain® up to a concentration of 5 mg/plate. exposure of chinese hamster ovary (cho-k1) cells to mem ... | 2018 | 29886232 |
| optimization of a high-cell-density polyethylenimine transfection method for rapid protein production in cho-ebna1 cells. | for pre-clinical evaluation of biotherapeutic candidates, protein production by transient gene expression (tge) in chinese hamster ovary (cho) cells offers important advantages, including the capability of rapidly and cost-effectively generating recombinant proteins that are highly similar to those produced in stable cho clones. we have established a novel cho clone (cho-3e7) expressing a form of the epstein-barr virus nuclear antigen-1 (ebna-1) with improved tge productivity relative to parenta ... | 2018 | 29886030 |
| effective and efficient characterization of chinese hamster ovary production cell lines using automated intracellular staining and statistical modeling. | mammalian cell line development is critical to bioproduct manufacturing. success requires selecting a line with desirable performance characteristics, including consistent expression throughout the proposed manufacturing window. given the genetic and phenotypic flux inherent to immortalized lines such as chinese hamster ovary cells, clonally-derived cell line characterization is vital. we describe here the development and implementation of a novel addition to our characterization approach to ens ... | 2018 | 29882242 |
| the binding of captopril to angiotensin i-converting enzyme triggers activation of signaling pathways. | hypertension is a global health problem, and angiotensin i (ang i)-converting enzyme (ace) inhibitors are largely used to control this pathology. recently, it has been shown that ace can also act as a transducer signal molecule when its inhibitors or substrates bind to it. this new role of ace could contribute to understanding some of the effects not explained by its catalytic activity only. in this study, we investigated signaling pathway activation in chinese hamster ovary (cho) cells stably e ... | 2018 | 29874111 |
| a gold nanoparticle bio-optical transponder to dynamically monitor intracellular ph. | a ph-sensitive bio-optical transponder (ph-bot) capable of simultaneously reporting the timing of intracellular dna cargo release from a gold nanoparticle (aunp) and the evolving intracellular ph (ph i) during endosomal maturation is demonstrated. the ph-bot is designed with a triple-dye-labeled duplex dna appended to a 6.6 nm aunp, utilizing ph-responsive fluorescein paired with dylight405 as a surface energy transfer (set) coupled dye pair to ratiometrically report the ph at and after cargo re ... | 2018 | 29874043 |
| lactoferrin prevents susceptibility of wehi 231 cells to anti-ig-induced cell death promoting cell differentiation. | immature b cells are susceptible to apoptosis due to ligation of surface immunoglobulin receptors. the wehi 231 cell line represents a useful model to study the mode of action of factors preventing apoptosis. in this work we investigated the protective effects of multi-species lactoferrins in anti-mouse ig-induced wehi 231 cell death. bovine milk-derived lactoferrin (blf), recombinant human lactoferrin expressed in chinese hamster ovary cells - rhlf(cho) or in human endothelial kidney cells - rh ... | 2018 | 29871734 |
| amp-activated protein kinase mediates the effect of leptin on avian autophagy in a tissue-specific manner. | autophagy, a highly conserved intracellular self-digestion process, plays an integral role in maintaining cellular homeostasis. although emerging evidence indicate that the endocrine system regulates autophagy in mammals, there is still a scarcity of information on autophagy in avian (non-mammalian) species. here, we show that intracerebroventricular administration of leptin reduces feed intake, modulates the expression of feeding-related hypothalamic neuropeptides, activates leptin receptor and ... | 2018 | 29867578 |
| crispr/cas9-multiplexed editing of chinese hamster ovary b4gal-t1, 2, 3, and 4 tailors n-glycan profiles of therapeutics and secreted host cell proteins. | in production of recombinant proteins for biopharmaceuticals, n-glycosylation is often important for protein efficacy and patient safety. igg with agalactosylated (g0)-n-glycans can improve the activation of the lectin-binding complement system and be advantageous in the therapy of lupus and virus diseases. in this study, the authors aimed to engineer cho-s cells for the production of proteins with g0-n-glycans by targeting b4gal-t isoform genes with crispr/cas9. indel mutations in genes encodin ... | 2018 | 29862652 |
| clonal variation in productivity and proteolytic clipping of an fc-fusion protein in cho cells: proteomic analysis suggests a role for defective protein folding and the upr. | product degradation, such as clipping, is a common quality issue in the production of fc-fusion proteins from chinese hamster ovary (cho) cells. degradation of proteins is mainly due to the action of either intracellular or extracellular host cell proteases. this study was carried out to understand more fundamentally the intracellular events that may play a role in determining why cell lines from the same cell line development project can vary with regards to the extent of fc-fusion protein clip ... | 2018 | 29860056 |
| a new cho (chinese hamster ovary)-derived cell line expressing anti-tnfα monoclonal antibody with biosimilar potential. | tumor necrosis factor alpha (tnfα) is a pro-inflammatory cytokine that mediates the homeostasis of immune responses; its exacerbated production is associated with the pathogenesis of autoimmune and chronic inflammatory diseases. anti-tnfα drugs have revolutionized the treatment of inflammatory conditions such as rheumatoid arthritis and crohn's disease. currently, a worldwide race is on stage for the production of biosimilars moved by patent expiration of monoclonal antibodies (mabs), such as an ... | 2018 | 29855993 |
| physiological responses of chinese hamster ovary cells to a productivity-enhancing yeast extract. | hydrolysates play important roles in enhancing the productivity of recombinant proteins in mammalian cell cultures. lacking of detailed understanding of the mechanisms, hydrolysate is commonly regarded as an unstable factor which should be used with cautions. a yeast extract (ye) was approved to improve the fc-fusion protein productivity in a recombinant chinese hamster ovary (cho) cell line. to elucidate the responses of cells to hydrolysates, we further elaborate their physiological changes du ... | 2018 | 29853300 |
| a crispr/cas9 based engineering strategy for overexpression of multiple genes in chinese hamster ovary cells. | manipulation of multiple genes to engineer chinese hamster ovary (cho) cells for better performance in production processes of biopharmaceuticals has recently become more and more popular. yet, identification of useful genes and the unequivocally assessment of their effect alone and in combination(s) on the cellular phenotype is difficult due to high variation between subclones. here, we present development and proof-of-concept of a novel engineering strategy using multiplexable activation of ar ... | 2018 | 29852271 |