TitleAbstractYear(sorted descending)
cell propagation of cholera toxin cta adp-ribosylating factor by exosome mediated this study, we report how the cholera toxin (ct) a subunit (cta), the enzyme moiety responsible for signaling alteration in host cells, enters the exosomal pathway, secretes extracellularly, transmits itself to a cell population. the first evidence for long-term transmission of ct's toxic effect via extracellular vesicles was obtained in chinese hamster ovary (cho) cells. to follow the ct intracellular route towards exosome secretion, we used a novel strategy for generating metabolically-labe ...201829783743
inactivation of deubiquitinase cyld enhances therapeutic antibody production in chinese hamster ovary cells.chinese hamster ovary (cho) cells are promising host engineering cells for industry manufacturing of therapeutic antibodies. however, cell death due to apoptosis remains a huge challenge to augment antibody production, and developing cho cells with enhanced anti-apoptosis and proliferation ability is fundamental for cell line development and high-yielding bioprocesses. deubiquitinase cylindromatosis (cyld) has been proved to be a tumor suppressor by negatively regulating nf-κb and wnt/β-catenin ...201829766242
antibody-assisted target identification reveals afatinib, an egfr covalent inhibitor, down-regulating ribonucleotide reductase.afatinib, used for the first-line treatment of non-small-cell lung carcinoma (nsclc) patients with distinct epidermal growth factor receptor (egfr) mutations, inactivates egfr by mimicking atp structure and forming a covalent adduct with egfr. we developed a method to unravel potential targets of afatinib in nsclc cells through immunoprecipitation of afatinib-labeling proteins with anti-afatinib antiserum and mass spectrometry analysis. ribonucleotide reductase (rnr) is one of target proteins of ...201829765556
zinc supplementation decreases galactosylation of recombinant igg in cho cells.trace element composition of culture medium can be altered to modulate glycoform of recombinant glycoproteins. in this study, we show that zn2+ supplementation at or above 100 μm decreases galactosylation of recombinant igg expressed in chinese hamster ovary cells. this decrease in galactosylation is not due to reduced galactosyltransferase expression. this effect persists upon supplementation of galactose and uridine to the culture, indicating that it may not be due to reduced udp-gal availabil ...201829749563
efficient mab production in cho cells with optimized signal peptide, codon, and utr.antibody drugs have been used to treat a number of diseases successfully. producing antibodies with high yield and quality is necessary for clinical applications of antibodies. for a candidate molecule, optimization of a vector to produce sufficient yield and an accurate primary structure is indispensable in the early stage of the production process development. it is especially important to maintain the fidelity of n-terminal sequence. in order to produce antibodies with a high yield and accura ...201829740673
initial hazard assessment of 4-benzylphenol, a structural analog of bisphenol f: genotoxicity tests in vitro and a 28-day repeated-dose toxicity study in rats.4-benzylphenol (cas no. 101-53-1), a structural analog of bisphenol f, has estrogenic activity in vitro and in vivo, as is the case with bisphenol f. 4-benzylphenol is used in plastics and during organic synthesis. since its safety is largely unknown, we conducted toxicity tests as part of screening risk assessment in an existing chemical safety survey program. based on results of the ames test and the chromosomal aberration test using chinese hamster lung cells (oecd tg 471 and 473), 4-benzylph ...201829705401
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, ...201830017654
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).201832161862
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 ...201829991278
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 ...201830007199
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 ...201830002126
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 ...201830104636
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 ...201830100348
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 ...201830056138
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 ...201830039852
cell culture medium supplemented with taurine decreases basic charge variant levels of a monoclonal explore the impact of taurine on monoclonal antibody (mab) basic charge variants in chinese hamster ovary (cho) cell culture.201830229353
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.201830159358
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 ...201830157452
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 ...201830157178
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 ...201830077924
the expression pattern of the phosphoproteome is significantly changed during the growth phases of recombinant cho cell 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 ...201830076757
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 ...201830076448
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 ...201830069078
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 ( ...201830063930
dna repair deficient chinese hamster ovary cells exhibiting differential sensitivity to charged particle radiation under aerobic and hypoxic 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 ...201830061540
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 ...201830059122
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 ...201830142424
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 ...201830131369
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 ...201830131171
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 ...201830110655
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 ...201830108268
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 ...201830107380
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 ...201830036451
calibration of pertussis toxin brp batch 1 in a standardised cho cell-based clustering assay.the european pharmacopoeia (ph. eur.) pertussis toxin (pt) biological reference preparation (brp) is used as a working standard for safety testing of acellular pertussis vaccines as prescribed in the ph. eur. monographs 1356 "pertussis vaccine (acellular, component, adsorbed)" and 1595 "pertussis vaccine (acellular, co-purified, adsorbed)". the brp was calibrated in 2006 in the murine histamine sensitisation test (hist) against the world health organization (who) 1st international standard (is) ...201830272557
in vitro and in vivo evaluation of the genotoxicity of eriobotrya japonica leaf extract.eriobotrya japonica leaf is included in the chinese pharmacopoeia, and is widely used as a medicinal material in traditional medicine. the present study investigated the potential genotoxic effects of e. japonica leaf extract (eje) using three standard battery systems. genotoxicity tests were conducted following the test guidelines of the organisation for economic cooperation and development (oecd) and ministry of food and drug safety (mfds), with application of good laboratory practice. the bac ...201830268829
targeting of influenza viral epitopes to antigen-presenting cells by genetically engineered chimeric molecules in a humanized nod scid gamma transfer model.antiviral dna vaccines are a novel strategy in the vaccine development field, which basically consists of the administration of expression vectors coding viral antigen sequences into the host's cells. targeting of conserved viral epitopes by antibody fragments specific to activating cell surface co-receptor molecules on antigen-presenting cells could be an alternative approach for inducing protective immunity. it has been shown that fcγri on human monocytes enhances antigen presentation in vivo. ...201830191743
cell culture media for recombinant protein expression in chinese hamster ovary (cho) cells: history, key components, and optimization strategies.the culture of chinese hamster ovary (cho) cells for modern industrial applications, such as expression of recombinant proteins, requires media that support growth and production. such media must support high viable cell densities while also stimulating the synthesis and extracellular transport of biologic products. early media development efforts in this area yielded basic formulations to sustain growth, viability, and cellular function, albeit comprising animal sourced components, and complex ...201830290072
construction and application of (q)sar models to predict chemical-induced in vitro chromosome drug development, genetic toxicology studies are conducted using in vitro and in vivo assays to identify potential mutagenic and clastogenic effects, as outlined in the international council for harmonisation (ich) s2 regulatory guideline. (quantitative) structure-activity relationship ((q)sar) models that predict assay outcomes can be used as an early screen to prioritize pharmaceutical candidates, or later during product development to evaluate safety when experimental data are unavailable ...201830278198
pharmacological characterization of the orexin/hypocretin receptor agonist nag promising series of small-molecule orexin receptor agonists has been described, but the molecular pharmacological properties, i.e. ability and potency to activate the different orexin receptor-regulated signal pathways have not been reported for any of these ligands. we have thus here assessed these properties for the most potent ligand of the series, 4'-methoxy-n,n-dimethyl-3'-[n-(3-{[2-(3-methylbenzamido)ethyl]amino}phenyl sulfamoyl]-(1,1'-biphenyl)-3-carboxamide (nag 26). chinese hamster ...201830194937
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 ...201830116893
construction strategies for developing expression vectors for recombinant monoclonal antibody production in cho cells.recent years have seen the use of recombinant proteins in the treatment of different diseases. among them, monoclonal antibodies (mabs) are currently the fastest growing class of bio-therapeutic recombinant proteins. chinese hamster ovary (cho) cells are the most commonly used host cells for production of these recombinant mabs. expression vectors determine the expression level and quality of recombinant mabs. currently, few construction strategies for recombinant mabs expression vectors in cho ...201830191354
rational optimization of a monoclonal antibody for simultaneous improvements in its solution properties and biological activity.developability considerations should be integrated with lead engineering of antibody drug candidates in interest of their cost effective translations into medicines. to explore feasibility of this imperative, we have performed rational mutagenesis studies on a monoclonal antibody (mab1) whose development was discontinued owing to manufacturability hurdles. seven computationally designed variants of mab1 containing single point (v44k, e59s, e59t and e59y) and double (v44ke59s, v44ke59t and v44ke5 ...201830189027
application of the crispr/cas9 gene editing method for modulating antibody fucosylation in cho cells.genetic engineering plays an essential role in the development of cell lines for biopharmaceutical manufacturing. advanced gene editing tools can improve both the productivity of recombinant cell lines as well as the quality of therapeutic antibodies. antibody glycosylation is a critical quality attribute for therapeutic biologics because the glycan patterns on the antibody fragment crystallizable (fc) region can alter its clinical efficacy and safety as a therapeutic drug. as an example, recomb ...201830242691
targeting mirnas with crispr/cas9 to improve recombinant protein production of cho cells.micrornas with their unique ability to target hundreds of genes have been highlighted as powerful tools to improve bioprocess behavior of cells. the common approaches to stably deplete mirnas are the use of sponge decoy transcripts or shrna inhibitors, which requires the introduction and expression of extra genetic material. as an alternative, we implemented the crispr/cas9 system in our laboratory to generate chinese hamster ovary (cho) cells which lack the expression of a specific mirna for th ...201830242690
bench-scale stirred-tank bioreactor for recombinant protein production in chinese hamster ovary (cho) cells in suspension.most pharmaceutical biotechnology companies use stirred-tank bioreactors (str) for recombinant protein manufacturing. these bioreactors are used at a variety of different scales ranging from bench to production scales, with working volumes from 10 ml to 25,000 l. bench-scale strs are commonly used to culture mammalian cells for process development, to troubleshoot production scale bioreactors using scale-down models (sdm), or to conduct fundamental research. in this chapter, we describe the oper ...201830242685
cho and hek293 cultivation and transfection in single-use orbitally shaken bioreactors.chinese hamster ovary (cho) and human embryonic kidney 293 (hek293) cells are the two most popular mammalian hosts for the production of recombinant proteins. in this chapter the suspension cultivation and transfection of these cells in small-scale disposable bioreactors is described. the tubespin bioreactor 50 and tubespin bioreactor 600 are designed for the cultivation of suspension cells by orbital shaking and have maximum working volumes of about 15 ml and 400 ml, respectively.201830242684
pei-mediated transient gene expression in cho cells.we describe a method for polyethyleneimine (pei) mediated transient transfection of suspension-adapted chinese hamster ovary (cho-dg44) cells for protein expression applicable at scales from 2 ml to 2 l. the method involves transfection at a high cell density (5 × 106 cells/ml) by direct addition of plasmid dna (pdna) and pei to the culture and subsequent incubation at 31 °c with agitation by orbital shaking. this method requires 0.3 mg/l of coding pdna, 2.7 mg/l of nonspecific (filler) dna and ...201830242678
the interaction of talin with the cell membrane is essential for integrin activation and focal adhesion formation.multicellular organisms have well-defined, tightly regulated mechanisms for cell adhesion. heterodimeric αβ integrin receptors play central roles in this function and regulate processes for normal cell functions, including signaling, cell migration, and development, binding to the extracellular matrix, and senescence. they are involved in hemostasis and the immune response, participate in leukocyte function, and have biological implications in angiogenesis and cancer. proper control of integrin ...201830254158
characterization of cellular states of cho-k1 suspension cell culture through cell cycle and rna-sequencing profiling.chinese hamster ovary (cho) derived cell lines are the preferred host system for the production of therapeutic proteins. the aim of this work was to explore the regulation of suspension-adapted cho-k1 host cell line bioprocesses, especially under a temperature gradient from 37 °c to 31 °c. we analyzed cell cycle behavior through flow cytometry of propidium iodide stained cells and high throughput transcriptome dynamics by rna sequencing. we found a cell culture state characterized by g0/g1 synch ...201830243609
versatile cell-free protein synthesis systems based on chinese hamster ovary cells.we present an alternative production platform for the synthesis of complex proteins. apart from conventionally applied protein production using engineered mammalian cell lines, this protocol describes the preparation and principle of cell-free protein synthesis systems based on cho cell lysates. the cho cell-free system contains endogenous microsomes derived from the endoplasmic reticulum, which enables a direct integration of membrane proteins into a nature like milieu and the introduction of p ...201830242694
celludinones, new inhibitors of sterol o-acyltransferase, produced by talaromyces cellulolyticus 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 ...201830177721
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 ...201830173861
direct observation of cargo transfer from hdl particles to the plasma 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.201830173079
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- ...201830171773
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 ...201830167439
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 ...201830111624
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 ...201830161182
rap1 binding to the talin 1 f0 domain makes a minimal contribution to murine platelet gpiib-iiia activation.activation of platelet glycoprotein iib-iiia (gpiib-iiia; integrin αiibβ3) leads to high-affinity fibrinogen binding and platelet aggregation during hemostasis. whereas gtp-bound rap1 gtpase promotes talin 1 binding to the β3 cytoplasmic domain to activate platelet gpiib-iiia, the rap1 effector that regulates talin association with β3 in platelets is unknown. rap1 binding to the talin 1 f0 subdomain was proposed to forge the talin 1-rap1 link in platelets. here, we report a talin 1 point mutant ...201830242097
car-t cells based on novel bcma monoclonal antibody block multiple myeloma cell growth.the cell-surface protein b cell maturation antigen (bcma, cd269) has emerged as a promising target for car-t cell therapy for multiple myeloma. in order to create a novel bcma car, we generated a new bcma monoclonal antibody, clone 4c8a. this antibody exhibited strong and selective binding to human bcma. bcma car-t cells containing the 4c8a scfv were readily detected with recombinant bcma protein by flow cytometry. the cells were cytolytic for rpmi8226, h929, and mm1s multiple myeloma cells and ...201830208593
high glucose and low specific cell growth but not mild hypothermia improve specific r-protein productivity in chemostat culture of cho 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 ...201830114204
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 ...201830121436
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 ...201830119885
evaluation of toxicological and teratogenic effects of carbosilane glucose glycodendrimers in zebrafish embryos and model rodent cell lines.glycodendrimers (glyco-ddms) represent a rapidly growing class of nanoparticles with promising properties for biomedical applications but concerns regarding the impact on human health and environment are still justified. here we report, for the first time, the comparative study of in vivo developmental toxicity of carbosilane glyco-ddms and their cytotoxicity in vitro. carbosilane glyco-ddms (generation 1-3) containing 4, 8, and 16 β-d-glucopyranosyl units at the periphery (ddm1glu, ddm2glu, and ...201830182770
point-of-care production of therapeutic proteins of good-manufacturing-practice quality.manufacturing technologies for biologics rely on large, centralized, good-manufacturing-practice (gmp) production facilities and on a cumbersome product-distribution network. here, we report the development of an automated and portable medicines-on-demand device that enables consistent, small-scale gmp manufacturing of therapeutic-grade biologics on a timescale of hours. the device couples the in vitro translation of target proteins from ribosomal dna, using extracts from reconstituted lyophiliz ...201831015674
bioinformatic analysis of chinese hamster ovary host cell protein lipases.complete, accurate genome assemblies are necessary to design targets for genetic engineering strategies. successful gene knockdowns and knockouts in chinese hamster ovary (cho) cells may prevent the expression of difficult-to-remove host cell proteins (hcps). hcps, if not removed, can cause problems in stability, safety, and efficacy of the biotherapeutic. a significantly improved chinese hamster (ch) reference genome was used to identify new knockout targets with similar predicted functions and ...201830911190
site-specific integration ushers in a new era of precise cho cell line engineering.chinese hamster ovary (cho) cells are widely used for the production of therapeutic proteins. customarily, cho production cell lines are established through random integration, which requires laborious screening of many clones to isolate suitable producers. in contrast, site-specific integration (ssi) accelerates cell line development by targeting integration of transgenes to pre-validated genomic loci capable of supporting high and stable expression. to date, a relatively small number of these ...201831086757
growth behavior of human adipose tissue-derived stromal/stem cells at small scale: numerical and experimental investigations.human adipose tissue-derived stromal/stem cells (hascs) are a valuable source of cells for clinical applications, especially in the field of regenerative medicine. therefore, it comes as no surprise that the interest in hascs has greatly increased over the last decade. however, in order to use hascs in clinically relevant numbers, in vitro expansion is required. single-use stirred bioreactors in combination with microcarriers (mcs) have shown themselves to be suitable systems for this task. howe ...201830518117
differential regulation of thermodynamic binding forces of levocetirizine and (s)-cetirizine by lys191 in human histamine h₁ receptors.cetirizine is a zwitterionic second-generation antihistamine containing r- and s-enantiomers, levocetirizine, and (s)-cetirizine. levocetirizine is known to have a higher affinity for the histamine h₁ receptors than (s)-cetirizine; ligand-receptor docking simulations have suggested the importance of the formation of a salt bridge (electrostatic interaction) between the carboxylic group of levocetirizine and the lys191 residue at the fifth transmembrane domain of human histamine h₁ receptors. in ...201830558340
a proteomic profiling dataset of recombinant chinese hamster ovary cells showing enhanced cellular growth following mir-378 depletion.the proteomic data presented in this article provide supporting information to the related research article "depletion of endogenous mirna-378-3p increases peak cell density of cho dp12 cells and is correlated with elevated levels of ubiquitin carboxyl-terminal hydrolase 14" (costello et al., in press) [1]. control and microrna-378 depleted cho dp12 cells were profiled using label-free quantitative proteomic profiling. cho dp12 cells were collected on day 4 and 8 of batch culture, subcellular pr ...201830761351
an lc-ms-based lipidomics pre-processing framework underpins rapid hypothesis generation towards cho systems biotechnology.given a raw lc-ms dataset, it is often required to rapidly generate initial hypotheses, in conjunction with other 'omics' datasets, without time-consuming lipid verifications. furthermore, for meta-analysis of many datasets, it may be impractical to conduct exhaustive confirmatory analyses. in other cases, samples for validation may be difficult to obtain, replicate or maintain. thus, it is critical that the computational identification of lipids is of appropriate accuracy, coverage, and unbiase ...201830830409
tag-on-demand: exploiting amber codon suppression technology for the enrichment of high-expressing membrane protein cell lines.membrane proteins play key roles in the evolution of numerous diseases and as a result have become the most dominant class of targets for therapeutic intervention. however, their poor expression and detection oftentimes prohibit drug discovery and screening efforts. herein, we have developed an approach, named 'tag-on-demand' that exploits amber suppression to control the expression of 'tagged' membrane proteins for detection and selections, yet can be turned off for expression of the protein in ...201830753634
mitochondrial aconitase is a key regulator of energy production for growth and protein expression in chinese hamster ovary cells.mammalian cells like chinese hamster ovary (cho) cells are routinely used for production of recombinant therapeutic proteins. cells require a continuous supply of energy and nutrients to sustain high cell densities whilst expressing high titres of recombinant proteins. cultured mammalian cells are primarily dependent on glucose and glutamine metabolism for energy production.201830830403
febrile temperature elevates the expression of phosphatidylserine on plasmodium falciparum (fcr3csa) infected red blood cell surface leading to increased cytoadhesion.during the asexual intra-erythrocytic cycle, plasmodium (p.) falciparum exports parasitic proteins to the surface of infected red blood cells (irbcs) facilitating its cytoadhesion to various endothelial host receptors. this adhesive behavior is a critical contributor towards disease manifestation. however, little is known about the influence of recurring elevated temperature - a common symptom of the malaria infection - on the adhesive properties of irbcs to endothelial receptors. to address thi ...201830302009
ds-8500a, an orally available g protein-coupled receptor 119 agonist, upregulates glucagon-like peptide-1 and enhances glucose-dependent insulin secretion and improves glucose homeostasis in type 2 diabetic rats.g protein-coupled receptor 119 (gpr119) has been shown to be highly expressed in small intestinal l-cells and pancreatic β-cells and mediates intracellular camp concentration, glucagon-like peptide (glp-1) secretion, and glucose-stimulated insulin secretion (gsis). this study examined the pharmacological effects of 4-(5-{(1r)-1-[4-(cyclopropylcarbonyl) phenoxy]propyl}-1,2,4-oxadiazol-3-yl)-2-fluoro-n-[(2r)-1-hydroxypropan-2-yl]benzamide (ds-8500a), a novel, orally available, selective gpr119 ago ...201830217957
high-throughput screening of antibody-expressing cho clones using an automated shaken deep-well system.biopharmaceutical protein manufacturing requires the highest producing cell lines to satisfy current multiple grams per liter requirements. screening more clones increases the probability of identifying the high producers within the pool of available transfectant candidate cell lines. for the predominant industry mammalian host cell line, chinese hamster ovary (cho), traditional static-batch culture screening does not correlate with the suspension fed-batch culture used in manufacturing, and thu ...201830298994
cellular responses of hyaluronic acid-coated chitosan recent years, nanotechnology has been proven to offer promising biomedical applications for in vivo diagnostics and drug delivery, stressing the importance of thoroughly investigating the biocompatibility of potentially translatable nanoparticles (nps). herein, we report the cellular responses of uncoated chitosan nps (cs nps) and hyaluronic acid-coated chitosan nps (ha-cs nps) when introduced into chinese hamster ovary cells (cho-k1) in a dose-dependent manner (2.5, 0.25, 0.025, 0.0025, and ...201830310671
toxicological assessment of xanthigen® nutraceutical extract combination: mutagenicity, genotoxicity and oral toxicity.xanthigen® is a nutraceutical combination for weight management capable of increasing energy expenditure via uncoupling protein 1 (ucp-1) in white adipose tissue. it consists of brown seaweed undaria pinnatifida extract, rich in the carotenoid fucoxanthin (fx) and pomegranate seed oil (pso), rich in punicic acid. xanthigen was screened to determine its genotoxicity and 90-days repeated oral toxicity. genotoxicity was assessed with the ames test (ta89, ta100, ta1535, ta1537, wp2), chromosomal abe ...201830386730
cytotoxic and genotoxic potential of the type i photoinitiators bapo and tpo on human oral keratinocytes and v79 fibroblasts.phenylbis(acyl) phosphine oxide (bapo) and diphenyl(acyl) phosphine oxide (tpo) are alternative photoinitiators to camphorquinone (cq) in dental resinous materials. aim of this study was to investigate their cytotoxic/genotoxic potential in human oral keratinocytes (okf6/tert2) and chinese hamster lung fibroblasts (v79) in comparison to cq.201830340767
nanosecond range electric pulse application as a non-viral gene delivery method: proof of concept.current electrotransfection protocols are well-established for decades and, as a rule, employ long micro-millisecond range electric field pulses to facilitate dna transfer while application of nanosecond range pulses is limited. the purpose of this paper is to show that the transfection using ultrashort pulses is possible by regulating the pulse repetition frequency. we have used 200 ns pulses (10-18 kv/cm) in bursts of ten with varied repetition frequency (1 hz-1 mhz). the chinese hamster ovary ...201830341389
influence of chromatin remodeling in the removal of uvc-induced damage in tcr proficient and deficient chinese hamster mammalian cells, nucleotide excision repair system is constituted of two sub-pathways, global genomic repair (ggr) and transcription coupled repair (tcr). deficiency of tcr pathway leads to cockyane syndrome (cs) which is a rare human autosomal recessive disorder. owing to the pivotal role of csb gene in tcr, it's mutation causes severe repair and transcriptional defects in csb patients. csb protein belongs to the atp chromatin remodeling complex, hence presumably an improper chromatin remode ...201830389155
revealing key determinants of clonal variation in transgene expression in recombinant cho cells using targeted genome editing.generation of recombinant chinese hamster ovary (rcho) cell lines is critical for the production of therapeutic proteins. however, the high degree of phenotypic heterogeneity among generated clones, referred to as clonal variation, makes the rcho cell line development process inefficient and unpredictable. here, we investigated the major genomic causes of clonal variation. we found the following: (1) consistent with previous studies, a strong variation in rcho clones in response to hypothermia ( ...201830388888
collaborative study of thresholds for mutagens: proposal of a typical protocol for detection of hormetic responses in cytotoxicity tests.according to the linear no-threshold model (lnt), even the smallest amount of radiation is hazardous. although the lnt is not based on solid data, this hypothesis has been applied to mutagens and carcinogens. as a result, it has been postulated that there are no thresholds for these chemicals. to demonstrate negativity by experiments is practically impossible, because negative data may leave behind the possibility that additional data might make the resolution power high enough to change negativ ...201830338768
genotoxicity of three biofuel candidates compared to reference demand for alternative energy sources increases due to concerns regarding energy security and greenhouse gas emissions. however, little is known regarding the impacts of biofuels to the environment and human health even though the identification of such impacts is important to avoid biofuels leading to undesired effects. in this study mutagenicity and genotoxicity of the three biofuel candidates ethyl levulinate (el), 2-methyltetrahydrofuran (2-mthf) and 2-methylfuran (2-mf) were investig ...201830391874
single intense microsecond electric pulse induces radiosensitization to ionizing radiation: effects of time intervals between electric pulse and ionizing irradiation.background and objective: recent studies have shown the potential of electroporation (ep) as a physical radiosensitizer for ionizing radiation (ir). the amount of sensitizing effect depends on some factors the most important of them is the time interval between the ep and ir. this experimental in vitro study aims to investigate the radiosensitizing effect of ep exposure prior to ir and also evaluate the effects of ep-ir time intervals on the amount of radiosensitizing effects. methods: chinese h ...201830319980
γh2ax prefers late replicating metaphase chromosome regions.dna damage response (ddr) constitutes a protein pathway to handle eukaryotic dna lesions in the context of chromatin. ddr engages the recruitment of signaling, transducer, effector, chromatin modifiers and remodeling proteins, allowing cell cycle delay, dna repair or induction of senescence or apoptosis. an early ddr-event includes the epigenetic phosphorylation of the histone variant h2ax on serine 139 of the c-termini, so-called gammah2ax. gammah2ax foci detected by immunolabeling on interphas ...201830442336
breast cancer: an examination of the potential of ackr3 to modify the response of cxcr4 to cxcl12.upon binding with the chemokine cxcl12, the chemokine receptor cxcr4 has been shown to promote breast cancer progression. this process, however, can be affected by the expression of the atypical chemokine receptor ackr3. given ackr3's ability to form heterodimers with cxcr4, we investigated how dual expression of both receptors differed from their lone expression in terms of their signalling pathways. we created single and double cxcr4 and/or ackr3 chinese hamster ovary (cho) cell transfectants. ...201830441765
histone tails decrease n7-methyl-2'-deoxyguanosine depurination and yield dna-protein cross-links in nucleosome core particles and cells.monofunctional alkylating agents preferentially react at the n7 position of 2'-deoxyguanosine in duplex dna. methylated dna, such as that produced by methyl methanesulfonate (mms) and temozolomide, exists for days in organisms. the predominant consequence of n7-methyl-2'-deoxyguanosine (mdg) is widely believed to be abasic site (ap) formation via hydrolysis, a process that is slow in free dna. examination of mdg reactivity within nucleosome core particles (ncps) provided two general observations ...201830429328
initial hazard assessment of benzyl salicylate: in vitro genotoxicity test and combined repeated-dose and reproductive/developmental toxicity screening test in rats.benzyl salicylate is used as a fragrance ingredient and an ultraviolet light absorber, but its toxicity is unknown. therefore, toxicity tests and hazard classification were conducted for screening assessment under the japanese chemical substances control law. benzyl salicylate was found to be non-genotoxic in vitro based on the chromosomal aberration test using chinese hamster lung cells. however, the combined repeated-dose and reproductive/developmental screening toxicity test, in which male an ...201830359701
bioprocess development of antibody-drug conjugate production for cancer treatment.antibody-drug conjugate (adc) is a class of targeted cancer therapies that combine the advantages of monoclonal antibody (mab)'s specific targeting and chemotherapy's potent cytotoxicity. the therapeutic effect of adc is significantly affected by its bioproduction process. this study aims to develop an effective adc production process using anti-her2 mab-drug as a model therapeutic. first, a high titer (>2 g/l) of mab was produced by chinese hamster ovary cells from fed-batch cell culture. both ...201830352095
establishment of p38bf, a core-fucose-deficient mouse-canine chimeric antibody against dog podoplanin.podoplanin (pdpn), a type i transmembrane sialoglycoprotein, is expressed in normal tissues, including lymphatic endothelial cells, pulmonary type i alveolar cells, and renal podocytes. the overexpression of pdpn in cancers is associated with hematogenous metastasis by interactions with the c-type lectin-like receptor 2 (clec-2). we have previously reported the development of a mouse monoclonal antibody (mab) clone, pmab-38 (igg1, kappa), against dog pdpn (dpdpn). pmab-38 reacted strongly with c ...201830362926
genotoxicity evaluation of an ethanol extract mixture of astragali radix and salviae miltiorrhizae radix.myelophil, a combination of astragali radix and salviae radix, is one of the most commonly used remedies for disorders of qi and blood in traditional chinese medicine. based on the clinical applications of these plants, in particular to pregnant woman, this study aimed to evaluate the genotoxic potential of an ethanol extract mixture of the above two herbs, called myelophil. following the organization for economic cooperation and development (oecd) guideline methods, a genotoxicity test was cond ...201830402128
the treatment effect of novel hghrh homodimer to male infertility hamster.extra-hypothalamic growth hormone-releasing hormone (ghrh) plays an important role in reproduction. to study the treatment effect of grin (a novel hghrh homodimer), the infertility models of 85 male chinese hamsters were established by intraperitoneally injecting 20 mg/kg of cyclophosphamide once in a week for 5 weeks and the treatment with grin or human menopausal gonadotropin (hmg) as positive control was evaluated by performing a 3-week mating experiment. 2-8 mg/kg of grin and 200 u/kg of hmg ...201830402024
cell surface proteoglycan-mediated uptake and accumulation of the alzheimer's disease peptide aβ(1-42).proteoglycans (pgs) have been found in alzheimer's disease amyloid-β (aβ) plaques and their glycosaminoglycan chains reportedly influence aβ aggregation, neurotoxicity and intracellular accumulation in cell and animal models, but their exact pathophysiological role(s) remain unclear. we have studied the cellular uptake of fluorescently labelled aβ(1-42) and aβ(1-40) peptides in normal cho cells (k1) and the mutant cell line (pgsa-745) which lacks all protein-attached heparan and chondroitin sulf ...201830409516
glucagon-related peptides from phylogenetically ancient fish reveal new approaches to the development of dual gcgr and glp1r agonists for type 2 diabetes therapy.the insulinotropic and antihyperglycaemic properties of glucagons from the sea lamprey (petromyzontiformes), paddlefish (acipenseriformes) and trout (teleostei) and oxyntomodulin from dogfish (elasmobranchii) and ratfish (holocephali) were compared with those of human glucagon and glp-1 in mammalian test systems. all fish peptides produced concentration-dependent stimulation of insulin release from brin-bd11 rat and 1.1 b4 human clonal β-cells and isolated mouse islets. paddlefish glucagon was t ...201830391422
establishment of monoclonal antibody pmab-202 against horse podoplanin.podoplanin (pdpn), a type i transmembrane glycoprotein, is expressed in several body tissues, including podocytes of renal glomerulus, type i alveolar cells of lung, and lymphatic endothelial cells. pdpn activates platelet aggregation by binding to c-type lectin-like receptor-2 (clec-2) presented on platelets. monoclonal antibodies (mabs) against human-, mouse-, rat-, rabbit-, dog-, bovine-, and cat-pdpn have already been established. however, anti-horse pdpn mabs have not yet been developed. in ...201830362932
depletion of endogenous mirna-378-3p increases peak cell density of cho dp12 cells and is correlated with elevated levels of ubiquitin carboxyl-terminal hydrolase 14.mirnas are potent molecular regulators of cellular behaviour. the manipulation of these small non-coding rnas has been used to enhance industrially relevant phenotypes in chinese hamster ovary (cho) cells. we investigated the stable depletion of six mirnas; mir-204-5p, 338-3p, 378-3p, 409-3p, 455-3p and 505-3p, robustly associated with cell growth rate from a previous profiling study. inhibition of endogenous mir-378-3p function by mirna-sponge-decoy improved peak cell density by 59%. quantitati ...201830389639
purification of recombinant human butyrylcholinesterase on hupresin®.affinity chromatography on procainamide-sepharose has been an important step in the purification of butyrylcholinesterase (bche) and acetylcholinesterase (ache) since its introduction in 1978. the procainamide affinity gel has limitations. in the present report a new affinity gel called hupresin® was evaluated for its ability to purify truncated, recombinant human butyrylcholinesterase (rhubche) expressed in a stably transfected chinese hamster ovary cell line. we present a detailed example of t ...201830384187
proline 36 of the factor xiii activation peptide plays a crucial role in substrate recognition and zymogen activation.the activation peptide of blood coagulation factor xiii (ap-fxiii) has important functions in stabilizing the fxiii-a2 dimer and regulating fxiii activation. contributions of many of its 37 amino acids to these functions have been described. however, the role of proline 36, which is adjacent to the thrombin cleavage site at arg37, has not yet been studied in detail. we approached this question when we came across a patient with congenital fxiii deficiency in whom we detected a novel pro36ser mut ...201830419598
domain unfolding of monoclonal antibody fragments revealed by non-reducing sds-page.monoclonal antibodies and derived fragments are used extensively both experimentally and therapeutically. thorough characterization of such antibodies is necessary and includes assessment of their thermal and storage stabilities. thus, assessment of the underlying conformational stabilities of the antibodies is also important. we recently documented that non-reducing sds-page can be used to assess both monoclonal and polyclonal igg domain thermal unfolding in sds. utilizing this same h2e2 anti-c ...201830417132
cytotoxic and membrane cholesterol effects of ultraviolet irradiation and zinc oxide nanoparticles on chinese hamster ovary cells.zinc oxide (zno) nanoparticles are suspected to produce toxic effects toward mammalian cells; however, discrepancies in the extent of this effect have been reported between different cell lines. simultaneously, high levels of ultraviolet (uv-c) radiation can have carcinogenic effects. the mechanism of this effect is also not well understood. due to similarities in phenotype morphology after cell exposure to zno nanoparticles and uv-c irradiation, we emit the hypothesis that the toxicity of both ...201830445712
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