| complexes formed with integrin-α5 and kcnb1 potassium channel wild type or epilepsy-susceptibility variants modulate cellular plasticity via ras and akt signaling. | voltage-gated potassium (k+) channel subfamily b member 1 (kcnb1, kv2.1) and integrin-α5 form macromolecular complexes-named integrin-α5-kcnb1 complexes (ikcs)-in the human brain, but their function was poorly understood. here we report that membrane depolarization triggered ikc intracellular signals mediated by small gtpases of the ras subfamily and protein kinase b (akt) to advance the development of filopodia and lamellipodia in chinese hamster ovary cells, stimulate their motility, and enhan ... | 2019 | 31682765 |
| indomethacin enhances type 1 cannabinoid receptor signaling. | in addition to its known actions as a non-selective cyclooxygenase (cox) 1 and 2 inhibitor, we hypothesized that indomethacin can act as an allosteric modulator of the type 1 cannabinoid receptor (cb1r) because of its shared structural features with the known allosteric modulators of cb1r. indomethacin enhanced the binding of [3h]cp55940 to hcb1r and enhanced aea-dependent [35s]gtpγs binding to hcb1r in chinese hamster ovary (cho) cell membranes. indomethacin (1 μm) also enhanced cp55940-depende ... | 2019 | 31680861 |
| identification and profiling of micrornas expressed in oral buccal mucosa squamous cell carcinoma of chinese hamster. | micrornas are known to play essential role in the gene expression regulation in cancer. in our research, next-generation sequencing technology was applied to explore the abnormal mirna expression of oral squamous cell carcinoma (oscc) in chinese hamster. a total of 3 novel mirnas (novel-117, novel-118, and novel-135) and 11 known mirnas (crg-mir-130b-3p, crg-mir-142-5p, crg-mir-21-3p, crg-mir-21-5p, crg-mir-542-3p, crg-mir-486-3p, crg-mir-499-5p, crg-mir-504, crg-mir-34c-5p, crg-mir-34b-5p and c ... | 2019 | 31666604 |
| an arginase-based system for selection of transfected cho cells without the use of toxic chemicals. | polyamines have essential roles in cell proliferation, dna replication, transcription, and translation processes, with intracellular depletion of putrescine, spermidine, and spermine resulting in cellular growth arrest and eventual death. serum-free media for cho-k1 cells require putrescine supplementation, because these cells lack the first enzyme of the polyamine production pathway, arginase. on the basis of this phenotype, we developed an arginase-based selection system. we transfected cho-k1 ... | 2019 | 31666335 |
| the epitope of pmab-210 is located in platelet aggregation-stimulating domain-3 of pig podoplanin. | podoplanin (pdpn)/t1alpha/aggrus, a small mucin-type transmembrane glycoprotein, has been shown to be expressed on lymphatic endothelial cells and epithelial cells of many organs. pdpn is also upregulated in many cancers, and is involved in cancer metastasis and malignant progression. human pdpn possesses three platelet aggregation-stimulating (plag) domains and the plag-like domain, which bind to c-type lectin-like receptor-2 (clec-2). previously, we reported a novel antipig pdpn (ppdpn) monocl ... | 2019 | 31663836 |
| understanding the mechanism of copurification of "difficult to remove" host cell proteins in rituximab biosimilar products. | host cell proteins (hcps) are considered a critical quality attribute and are linked to safety and efficacy of biotherapeutic products. researchers have identified 10 hcps in chinese hamster ovary (cho) that exhibit common characteristics of product association, coelution, and age-dependent expression and therefore are "difficult to remove" during downstream purification. these include cathepsin d, clusterin, galectin-3-binding protein, g-protein coupled receptor 56, lipoprotein lipase, metallop ... | 2020 | 31661608 |
| effects of different 2a peptides on transgene expression mediated by tricistronic vectors in transfected cho cells. | multicistronic vectors can increase transgene expression and decrease the imbalance of gene expression in the chinese hamster ovary (cho) cell expression system. small, self-cleaving 2a peptides have a high cleavage efficiency and are essential for constructing high-expression multicistronic vectors. in this study, we investigated the effects of two different 2a peptides on transgene expression in cho cells via their mediating action on tricistronic vectors. the enhanced green fluorescent protei ... | 2020 | 31659692 |
| tak-925, an orexin 2 receptor-selective agonist, shows robust wake-promoting effects in mice. | orexin-producing neurons in the lateral hypothalamus are a critical regulator of sleep/wake states, and their loss is associated with narcolepsy type 1 (nt1). orexin peptides act on two g protein-coupled receptors: orexin 1 receptor (ox1r) and orexin 2 receptor (ox2r). ox2r knockout (ko) mice, but not ox1r ko mice, showed clear narcolepsy-like phenotypes, including fragmented sleep-wake cycles. moreover, ox2r-selective antagonists have been shown to induce sleepiness in mice, and activation of o ... | 2019 | 31654653 |
| targeted capture of chinese hamster ovary host cell proteins: peptide ligand binding by proteomic analysis. | the clearance of host cell proteins (hcps) is of crucial importance in biomanufacturing, given their diversity in composition, structure, abundance, and occasional structural homology with the product. the current approach to hcp clearance in the manufacturing of monoclonal antibodies (mabs) relies on product capture with protein a followed by removal of residual hcps in flow-through mode using ion exchange or mixed-mode chromatography. recent studies have highlighted the presence of "problemati ... | 2020 | 31654407 |
| dual role of cofilin in app trafficking and amyloid-β clearance. | the accumulation of amyloid-β (aβ) plays a pivotal early event in the pathogenesis of alzheimer's disease (ad). in the brain, neurons produce aβ by the proteolytic processing of amyloid precursor protein (app) through the endocytic pathway, whereas microglia mediate aβ clearance also via endocytic mechanisms. previous studies have shown the critical importance of cofilin, a filamentous actin-severing protein, in actin dynamics and pathogen-triggered endocytic processes. moreover, the binding of ... | 2019 | 31646885 |
| epitope analysis of an antihorse podoplanin monoclonal antibody pmab-219. | podoplanin (pdpn), which is a mucin-type membrane glycoprotein, is expressed on lymphatic endothelial cells and epithelial cells of many organs. pdpn is also overexpressed in several malignant cancers, and its expression is associated with cancer progression and poor prognosis. human pdpn possesses three platelet aggregation-stimulating (plag) domains and the plag-like domain (pld), which binds to c-type lectin-like receptor-2 (clec-2). previously, we reported a novel antihorse pdpn (horpdpn) mo ... | 2019 | 31638470 |
| determination of the pathological features of npc1 variants in a cellular complementation test. | niemann-pick type c (np-c) is a rare disorder of lipid metabolism caused by mutations within the npc1 and npc2 genes. np-c is a neurovisceral disease leading to a heterogeneous, multisystemic spectrum of symptoms in those affected. until now, there is no investigative tool to demonstrate the significance of single variants within the npc genes. hence, the aim of the study was to establish a test that allows for an objective assessment of the pathological potential of npc1 gene variants. chinese ... | 2019 | 31635081 |
| a critical assessment of the genotoxicity profile of the fungicide tricyclazole. | tricyclazole (8-methyl-[1,2,4]triazolo[3,4-b][1,3]benzothiazole) is a fungicide used globally on rice for treatment of the seasonal rice blast disease. human exposure to this fungicide can occur via dietary and nondietary routes. in a battery of in vitro assays, tricyclazole did not induce gene mutations in bacteria (ames test) or at the hprt locus of cho cells. it was also negative for the induction of micronuclei in human lymphocyte cultures and unscheduled dna synthesis (uds) in primary rat h ... | 2020 | 31633836 |
| unraveling what makes a monoclonal antibody difficult-to-express: from intracellular accumulation to incomplete folding and degradation via erad. | although most therapeutic monoclonal antibodies (mabs) can routinely be produced in the multigram per litre range, some mab candidates turn out to be difficult-to-express (dte). in addition, the class of more complex biological formats is permanently increasing and mammalian expression systems like chinese hamster ovary (cho) cell lines can show low performance. hence, there is an urgent need to identify any rate limiting processing step during cellular synthesis. therefore, we assessed the intr ... | 2020 | 31631329 |
| characterization and mutagenesis of chinese hamster ovary cells endogenous retroviruses to inactivate viral particle release. | the chinese hamster ovary (cho) cells used to produce biopharmaceutical proteins are known to contain type-c endogenous retrovirus (erv) sequences in their genome and to release retroviral-like particles. although evidence for their infectivity is missing, this has raised safety concerns. as the genomic origin of these particles remained unclear, we characterized type-c erv elements at the genome, transcriptome, and viral particle rna levels. we identified 173 type-c erv sequences clustering int ... | 2020 | 31631325 |
| awakening dormant glycosyltransferases in cho cells with crispra. | chinese hamster ovary (cho) cells are the preferred workhorse for the biopharmaceutical industry, and crispr/cas9 has proven powerful for generating targeted gene perturbations in cho cells. here, we expand the crispr engineering toolbox with crispr activation (crispra) to increase transcription of endogenous genes. we successfully increased transcription of mgat3 and st6gal1, and verified their activity on a functional level by subsequently detecting that the appropriate glycan structures were ... | 2020 | 31631317 |
| ivabradine for chronic heart rate control in persistent atrial fibrillation. design of the brake-af project. | ivabradine is an inhibitor of the if channel, the main determinant of the pacemaker function of the sinus node. the drug has been approved for the treatment of angina and heart failure. there is some evidence of its role as an inhibitor of atrial-ventricular node (avn) conduction. the aim of the brake-af project is to assess ivabradine use for rate control in atrial fibrillation (af). | 2020 | 31631048 |
| engineering antibodies on the surface of cho cells. | while antibody libraries are traditionally screened in phage, bacterial, or yeast display formats, they are produced in large scale for pharmaceutical and commercial use in mammalian cell lines. the simpler organisms used for screening have significantly different folding and glycosylation machinery than mammalian cells; consequently, clones resulting from these libraries may require further optimization for mammalian cell expression. to streamline the antibody discovery process, we developed a ... | 2020 | 31625108 |
| epitope mapping of pmab-225 an anti-alpaca podoplanin monoclonal antibody using flow cytometry. | podoplanin (pdpn) is a mucin-type membrane glycoprotein, and possesses three platelet aggregation-stimulating (plag) domains: plag1, plag2, and plag3 at the n-terminus of pdpn, and one or two plag-like domains (plds) in the middle of pdpn. pdpn is expressed on normal tissues, such as podocytes of the kidney and type i alveolar cells of the lung, and is also overexpressed in numerous malignant cancers. previously, we reported a novel anti-alpaca podoplanin (apdpn) monoclonal antibody (mab), pmab- ... | 2019 | 31613697 |
| impact of micro and macroporous tff membranes on product sieving and chromatography loading for perfusion cell culture. | bioprocess intensification can be achieved through high cell density perfusion cell culture with continuous protein capture integration. protein passage and cell retention are commonly accomplished using tangential flow filtration systems consisting of microporous membranes. significant challenges, including low efficiency and decaying product sieving over time, are commonly observed in these cell retention devices. here, we demonstrate that a macroporous membrane overcomes the product sieving c ... | 2020 | 31612989 |
| a study on enhanced o-glycosylation strategy for improved production of recombinant human chorionic gonadotropin in chinese hamster ovary cells. | human chorionic gonadotropin (hcg) is a glycoprotein hormone that exists as a heterodimer comprised of an α subunit and β subunit linked with disulfide bridges. the β subunit contains four o-glycosylation sites. previous studies have found that the translation of mrna to polypeptides of the β subunit was a severely limiting step for the expression of recombinant hcg protein in chinese hamster ovary (cho) cells. the effects of o-glycosylation on recombinant hcg protein expression were assessed by ... | 2019 | 31604106 |
| model-driven engineering of n-linked glycosylation in chinese hamster ovary cells. | chinese hamster ovary (cho) cells are used for industrial production of protein-based therapeutics (i.e., "biologics"). here we describe a method for combining systems-level kinetic models with a synthetic biology platform for multigene overexpression to rationally perturb n-linked glycosylation. specifically, we sought to increase galactose incorporation on a secreted immunoglobulin g (igg) protein. we rationally design, build, and test a total of 23 transgenic cell pools that express single or ... | 2019 | 31596566 |
| efficacy of lenvervimab, a recombinant human immunoglobulin, in treatment of chronic hepatitis b virus infection. | the mechanism of action of hepatitis b immunoglobulin (hbig) for chronic hepatitis b (chb) treatment is not fully understood. a clinical trial in japan suggested that hbig therapy might benefit patients with chb.1 lenvervimab, a recombinant monoclonal hbig developed by gc pharma (yongin, south korea), has been screened from an anti-hb-enriched phage-display library cloned from hepatitis b vaccine-boosted human blood cells and was produced in chinese hamster ovary cells.2-4. | 2019 | 31589980 |
| crystal structure of the fab region of a neutralizing antibody against granulocyte-macrophage colony-stimulating factor. | an increased level of granulocyte-macrophage colony-stimulating factor has a potential role in the development of autoimmune diseases, and the neutralization of its activity by monoclonal antibodies is a promising therapy for some diseases. here, the crystal structure of the fab region of ev1007, a fully human antibody expressed in chinese hamster ovary cells that was developed from human peripheral blood mononuclear cells, is described. the structure closely resembles that of mb007, which is th ... | 2019 | 31584011 |
| integration of high-throughput analytics and cell imaging enables direct early productivity and product quality assessment during chinese hamster ovary cell line development for a complex multi-subunit vaccine antigen. | mammalian cell line generation typically includes stable pool generation, single cell cloning and several rounds of clone selection based on cell growth, productivity and product quality criteria. individual clone expansion and phenotype-based ranking is performed initially for hundreds or thousands of mini-scale cultures, representing the major operational challenge during cell line development. automated cell culture and analytics systems have been developed to enable high complexity clone sel ... | 2020 | 31568688 |
| functional consequences of a kcnt1 variant associated with status dystonicus and early-onset infantile encephalopathy. | we identified a novel de novo kcnt1 variant in a patient with early-infantile epileptic encephalopathy (eiee) and status dystonicus, a life-threatening movement disorder. we determined the functional consequences of this variant on the encoded kna 1.1 channel to investigate the molecular mechanisms responsible for this disorder. | 2019 | 31560846 |
| trpa1 ion channel determines beneficial and detrimental effects of gyy4137 in murine serum-transfer arthritis. | modulation of nociception and inflammation by sulfide in rheumatoid arthritis and activation of transient receptor potential ankyrin 1 (trpa1) ion channels by sulfide compounds are well documented. the present study aims to investigate trpa1-mediated effects of sulfide donor gyy4137 in k/bxn serum-transfer arthritis, a rodent model of rheumatoid arthritis. trpa1 and somatostatin sst4 receptor wild-type (wt) and knockout mice underwent k/bxn serum transfer and were treated daily with gyy4137. fun ... | 2019 | 31551776 |
| the impact of sialylation linkage-type on the pharmacokinetics of recombinant butyrylcholinesterases. | chinese hamster ovary (cho) cells typically produce glycoproteins with n-glycans terminating in α-2,3 sialylation. human cells produce glycoproteins that include α-2,3 and α-2,6 sialic acids. to examine the impact of altering protein sialylation on pharmacokinetic properties, recombinant human butyrylcholinesterase (bche) was produced in cho cells by knocking out the α-2,3 sialyltransferase genes followed by overexpression of the α-2,6 sialyltransferase (26bche) enzyme. the n-glycan composition ... | 2020 | 31544955 |
| integrated clarification and purification of monoclonal antibodies by membrane based separation of aqueous two-phase systems. | therapeutic monoclonal antibodies (mab) are used for the treatment of numerous serious diseases, which have led to an increasing demand over the last decades. increased cell density and mab titer of the cultivation broth lead to great challenges for the subsequent clarification and capture operations in the downstream process. as an alternative approach to the conventional downstream process, a selective mab extraction via an aqueous two-phase system (atps) directly from the cultivation broth of ... | 2019 | 31544846 |
| evaluating the impact of suramin additive on cho cells producing fc-fusion protein. | to examine the effects of suramin in cho cell cultures in terms of the cell culture performance and quality of the fc-fusion protein. | 2019 | 31541331 |
| endoglin protein interactome profiling identifies trim21 and galectin-3 as new binding partners. | endoglin is a 180-kda glycoprotein receptor primarily expressed by the vascular endothelium and involved in cardiovascular disease and cancer. heterozygous mutations in the endoglin gene (eng) cause hereditary hemorrhagic telangiectasia type 1, a vascular disease that presents with nasal and gastrointestinal bleeding, skin and mucosa telangiectases, and arteriovenous malformations in internal organs. a circulating form of endoglin (alias soluble endoglin, seng), proteolytically released from the ... | 2019 | 31540324 |
| glycan residues balance analysis - greba: a novel model for the n-linked glycosylation of igg produced by cho cells. | the structure of n-linked glycosylation is a very important quality attribute for therapeutic monoclonal antibodies. different carbon sources in cell culture media, such as mannose and galactose, have been reported to have different influences on the glycosylation patterns. accurate prediction and control of the glycosylation profile are important for the process development of mammalian cell cultures. in this study, a mathematical model, that we named glycan residues balance analysis (greba), w ... | 2020 | 31539564 |
| intravenous delivery of a chemically modified sulfamidase efficiently reduces heparan sulfate storage and brain pathology in mucopolysaccharidosis iiia mice. | mucopolysaccharidosis type iiia (mps iiia) is a lysosomal storage disorder (lsd) characterized by severe central nervous system (cns) degeneration. the disease is caused by mutations in the sgsh gene coding for the lysosomal enzyme sulfamidase. sulfamidase deficiency leads to accumulation of heparan sulfate (hs), which triggers aberrant cellular function, inflammation and eventually cell death. there is currently no available treatment against mps iiia. in the present study, a chemically modifie ... | 2019 | 31528541 |
| reprogramming aa catabolism in cho cells with crispr/cas9 genome editing improves cell growth and reduces byproduct secretion. | chinese hamster ovary (cho) cells are the preferred host for producing biopharmaceuticals. amino acids are biologically important precursors for cho metabolism; they serve as building blocks for proteogenesis, including synthesis of biomass and recombinant proteins, and are utilized for growth and cellular maintenance. in this work, we studied the physiological impact of disrupting a range of amino acid catabolic pathways in cho cells. we aimed to reduce secretion of growth inhibiting metabolic ... | 2019 | 31526854 |
| collaborative study for the calibration of the replacement international standard for pertussis toxin for use in histamine sensitisation and cho cell clustering assays. | pertussis toxin (pt) in its detoxified form is one of the major protective antigens in vaccines against bordetella pertussis (whooping cough). reference preparations of native pt are required for the quality control of pertussis vaccines. stocks of the first who international standard (is) for pt (jnih-5) were low and a replacement was required. one candidate material was donated by a vaccine manufacturer to nibsc. it was formulated, lyophilised into sealed glass ampoules and coded 15/126. an in ... | 2019 | 31526657 |
| gpr40 full agonism exerts feeding suppression and weight loss through afferent vagal nerve. | gpr40/ffar1 is a gq protein-coupled receptor expressed in pancreatic β cells and enteroendocrine cells, and mediates insulin and incretin secretion to regulate feeding behavior. several gpr40 full agonists have been reported to reduce food intake in rodents by regulating gut hormone secretion in addition to their potent glucose-lowering effects; however, detailed mechanisms of feeding suppression are still unknown. in the present study, we characterized t-3601386, a novel compound with potent fu ... | 2019 | 31525244 |
| advancing biologics development programs with legacy cell lines: advantages and limitations of genetic testing for addressing clonality concerns prior to availability of late stage process and product consistency data. | the bioprocessing industry uses recombinant mammalian cell lines to generate therapeutic biologic drugs. to ensure consistent product quality of the therapeutic proteins, it is imperative to have a controlled production process. regulatory agencies and the biotechnology industry consider cell line "clonal origin" an important aspect of maintaining process control. demonstration of clonal origin of the cell substrate, or production cell line, has received considerable attention in the past few ye ... | 2020 | 31519780 |
| enhanced transient expression of an anti-cd52 monoclonal antibody in cho cells through utilization of mirna sponge technology. | chinese hamster ovary (cho) cells are the dominant mammalian host system for the production of recombinant therapeutic proteins. improving the viable cell density during culture of recombinant cho cells can greatly affect the production yield. micrornas (mirs) -15a and 16-1 are known as negative regulators of multiple genes involved in cell cycle progression and apoptotic inhibition. mir sponges, which act as decoy targets, are transcripts which contain complementary binding sites to the seed re ... | 2019 | 31516510 |
| trehalose uptake and dehydration effects on the cryoprotection of cho-k1 cells expressing tret1. | chinese hamster ovary cells (cho-k1 cells) in which the trehalose transporter (tret1) is expressed can have greater cryoprotection than ordinary cho-k1 cells. this study examines the uptake characteristics of trehalose into cells via tret1 and determines the influence of intracellular trehalose on the freeze-thaw viabilities. in our experiments, the intracellular trehalose concentration is controlled by the extracellular trehalose concentration and the immersion time in a freezing solution. in t ... | 2019 | 31499026 |
| mitochondria supply atp to the er through a mechanism antagonized by cytosolic ca2. | the endoplasmic reticulum (er) imports atp and uses energy from atp hydrolysis for protein folding and trafficking. however, little is known about how this vital atp transport occurs across the er membrane. here, using three commonly used cell lines (cho, ins1 and hela), we report that atp enters the er lumen through a cytosolic ca2+-antagonized mechanism, or caatier (ca2+-antagonized transport into er). significantly, we show that mitochondria supply atp to the er and a serca-dependent ca2+ gra ... | 2019 | 31498082 |
| identification of novel phytocannabinoids from ganoderma by label-free dynamic mass redistribution assay. | located throughout the body, cannabinoid receptors (cb1 and cb2) are therapeutic targets for obesity/metabolic diseases, neurological/mental disorders, and immune modulation. phytocannabinoids are greatly important for the development of new medicines with high efficacy and/or minor side effects. plants and fungi are used in traditional medicine for beneficial effects to mental and immune system. the current research studied five fungi from the genus ganoderma and five plants: ganoderma hainanen ... | 2020 | 31494202 |
| four ways to fit an ion channel model. | mathematical models of ionic currents are used to study the electrophysiology of the heart, brain, gut, and several other organs. increasingly, these models are being used predictively in the clinic, for example, to predict the risks and results of genetic mutations, pharmacological treatments, or surgical procedures. these safety-critical applications depend on accurate characterization of the underlying ionic currents. four different methods can be found in the literature to fit voltage-sensit ... | 2019 | 31493859 |
| dynamics of mechanosensitive nascent adhesion formation. | cellular migration is a tightly regulated process that involves actin cytoskeleton, adaptor proteins, and integrin receptors. forces are transmitted extracellularly through protein complexes of these molecules, called adhesions. adhesions anchor the cell to its substrate, allowing it to migrate. in chinese hamster ovary cells, three classes of adhesion can be identified: nascent adhesions (nas), focal complexes, and focal adhesions, ranked here ascendingly based on size and stability. to underst ... | 2019 | 31493858 |
| hplc-esi-qqq based standardization, mutagenic and genotoxic potential of methanol extract of ziziphus mauritiana lam leaves. | the leaves of ziziphus mauritiana lam have been an integral part of the traditional system of medicine for the treatment of inflammation, wounds, fever, asthma and liver disorders. the leaves are utilised as an edible vegetable in rural parts of india and indonesia. despite its pharmacological significance, ziziphus mauritiana lam lacks scientific evidence on its mutagenic and genotoxic potential. | 2020 | 31493461 |
| role of human n-acetyltransferase 2 genetic polymorphism on aromatic amine carcinogen-induced dna damage and mutagenicity in a chinese hamster ovary cell mutation assay. | carcinogenic aromatic amines such as 4-aminobiphenyl (abp) and 2-aminofluorene (af) require metabolic activation to form electrophilic intermediates that mutate dna leading to carcinogenesis. bioactivation of these carcinogens includes n-hydroxylation catalyzed by cyp1a2 followed by o-acetylation catalyzed by arylamine n-acetyltransferase 2 (nat2). to better understand the role of nat2 genetic polymorphism in abp- and af-induced mutagenesis and dna damage, nucleotide excision repair-deficient (u ... | 2020 | 31490564 |
| chloramination of iodide-containing waters: formation of iodinated disinfection byproducts and toxicity correlation with total organic halides of treated waters. | the formation of iodinated disinfection byproducts (i-dbps) in drinking waters is of a concern due to their higher cyto- and genotoxicity than their chlorinated and brominated analogues. this study investigated the formation of i-dbps under chloramination conditions using preformed chloramine and associated cyto- and geno-toxicities obtained with chinese hamster ovary (cho) cell assay. cyto- and geno-toxicity of the samples were also calculated using dbp toxicity index values and correlated with ... | 2019 | 31484087 |
| intracerebroventricular enzyme replacement therapy with β-galactosidase reverses brain pathologies due to gm1 gangliosidosis in mice. | autosomal recessive mutations in the galactosidase β1 (glb1) gene cause lysosomal β-gal deficiency, resulting in accumulation of galactose-containing substrates and onset of the progressive and fatal neurodegenerative lysosomal storage disease, gm1 gangliosidosis. here, an enzyme replacement therapy (ert) approach in fibroblasts from gm1 gangliosidosis patients with recombinant human β-gal (rhβ-gal) produced in chinese hamster ovary cells enabled direct and precise rhβ-gal delivery to acidified ... | 2020 | 31481471 |
| phosphorylation of unique c-terminal sites of the mu-opioid receptor variants 1b2 and 1c1 influences their gs association following chronic morphine. | we recently demonstrated in rat spinal cord that a regimen of escalating doses of systemic morphine, analogous to regimens used clinically for chronic pain management, selectively up-regulates the mu-opioid receptor (mor) splice variants mor-1b2 and mor-1c1 mrna and functional protein. this study investigated the potential relevance of up-regulating mor-1b2 and mor-1c1 to the ability of chronic morphine to shift mor signaling from predominantly gi /go inhibitory to gs stimulatory. specifically, ... | 2020 | 31479519 |
| impacts of intentional mycoplasma contamination on cho cell bioreactor cultures. | mycoplasma contamination events in biomanufacturing facilities can result in loss of production and costly cleanups. mycoplasma may survive in mammalian cell cultures with only subtle changes to the culture and may penetrate the 0.2 µm filters often used in the primary clarification of harvested cell culture fluid. culture cell-based and indicator cell-based assays that are used to detect mycoplasma are highly sensitive but can take up to 28 days to complete and cannot be used for real-time deci ... | 2019 | 31478189 |
| at1aa (angiotensin ii type-1 receptor autoantibodies): cause or consequence of human primary aldosteronism? | at1aa (angiotensin ii type-1 receptor autoantibodies) were first detected in patients with primary aldosteronism (pa) because of aldosterone-producing adenoma (apa) with an in-house developed assay, but it remained unclear if they can be ascertained also with commercially available assays and if they have a functional role. aims of our study were to investigate if (1) commercially available kits allow detection of raised at1aa titer in apa; (2) this titer is normalized by adrenalectomy; and (3) ... | 2019 | 31476908 |
| a role for alternative end-joining factors in homologous recombination and genome editing in chinese hamster ovary cells. | crispr technologies greatly foster genome editing in mammalian cells through site-directed dna double strand breaks (dsbs). however, precise editing outcomes, as mediated by homologous recombination (hr) repair, are typically infrequent and outnumbered by undesired genome alterations. by using knockdown and overexpression studies in chinese hamster ovary (cho) cells as well as characterizing repaired dna junctions, we found that efficient hr-mediated genome editing depends on alternative end-joi ... | 2019 | 31476574 |
| mrna transfection into cho-cells reveals production bottlenecks. | obtaining highly productive chinese hamster ovary (cho)-cell clones for the production of therapeutic proteins relies on multiple time-consuming selection steps. several cho-cell strains with high degrees of genomic and epigenetic variation are available. each harbor potential advantages and disadvantages for any given product, particularly those considered difficult to express. a simple test system to quickly assess compatibility of cell line and product may therefore prove useful. transient pl ... | 2020 | 31475780 |
| bcat1 and bcat2 disruption in cho cells has cell line-dependent effects. | in recombinant protein expression using chinese hamster ovary (cho) cells, chemically defined media contain essential amino acids such as branched chain amino acids (bcaas) leucine, isoleucine and valine. availability of amino acids is critical as these are building blocks for protein synthesis. however, breakdown of amino acids can lead to build up of toxic intermediates and metabolites that decrease cell growth, productivity and product quality. bcaa catabolism also hampers the usage of bcaas ... | 2019 | 31465797 |
| novel single-chain variant of antibody against mesothelin established by phage library. | mesothelin (msln) shows increased expression in various cancer cells. for clinical application of antibodies as a positron emission tomography (pet) imaging reagent, a human shortened antibody is essential both for avoiding redundant immune responses and for providing rapid imaging. therefore, we cloned a single-chain fragment of variable regions (scfv) from a human-derived gene sequence. this was achieved through the construction of a naïve phage library derived from human tonsil lymphocytes. u ... | 2019 | 31461572 |
| rapid characterization of herg channel kinetics ii: temperature dependence. | ion channel behavior can depend strongly on temperature, with faster kinetics at physiological temperatures leading to considerable changes in currents relative to room temperature. these temperature-dependent changes in voltage-dependent ion channel kinetics (rates of opening, closing, inactivating, and recovery) are commonly represented with q10 coefficients or an eyring relationship. in this article, we assess the validity of these representations by characterizing channel kinetics at multipl ... | 2019 | 31451180 |
| rapid characterization of herg channel kinetics i: using an automated high-throughput system. | predicting how pharmaceuticals may affect heart rhythm is a crucial step in drug development and requires a deep understanding of a compound's action on ion channels. in vitro herg channel current recordings are an important step in evaluating the proarrhythmic potential of small molecules and are now routinely performed using automated high-throughput patch-clamp platforms. these machines can execute traditional voltage-clamp protocols aimed at specific gating processes, but the array of protoc ... | 2019 | 31447109 |
| comparing two automated high throughput viable-cell counting systems for cell culture applications. | cell counting and viability assessment is an integral part of mammalian cell line development. while manual counting with a hemocytometer is still the gold standard method, its subjectivity and high labor intensity has resulted in its reduced use in favor of automated systems. in addition, some of these automated systems offer multiwell plate based high throughput cell count, which is an asset for biopharmaceutical companies generating hundreds of high-performance cell lines per year. in this st ... | 2019 | 31442502 |
| cathepsin d: removal strategy on protein a chromatography, near real time monitoring and characterisation during monoclonal antibody production. | monoclonal antibody (mab) fragmentation is a well-known degradation pathway that results in product loss and can significantly impact product quality, efficacy, or even cause immunogenic reactions, thus potentially endangering patients' health. it is recognised that residual proteases present among host cell proteins (hcps) such as those expressed by chinese hamster ovary (cho) can induce fragmentation, and failure of their complete removal during downstream processing could cause fragmentation ... | 2019 | 31442501 |
| functional and structural characterization of hspb1/hsp27 from chinese hamster ovary cells. | small heat shock proteins (shsps) endow cells with stress tolerance. of the various shsps in mammals, hspb1, also known as hsp27, is the most ubiquitous. to examine the structure and function of hspb1, we expressed, purified, and characterized hspb1 from chinese hamster (cricetulus griseus) ovary cells (cghspb1). cghspb1 forms a large oligomeric structure. we observed a monodisperse 16-mer with an elongated sphere, but this is affected by changes in various conditions, including temperature. und ... | 2019 | 31441240 |
| gatifloxacin-1,2,3-triazole-isatin hybrids and their antimycobacterial activities. | a series of gatifloxacin-1,2,3-triazole-isatin hybrids 8a-l were designed, synthesized, and screened for their in vitro antimycobacterial activity as well as cytotoxicity toward chinese hamster ovary (cho) cells. the synthesized hybrids showed considerable activity against mtb h37 rv and two mdr-mtb strains with minimal inhibitory concentration (mic) of 0.25-8 μg/ml, and the hybrid 8a (micmtb h37rv : 0.25 μg/ml and micmdr-mtb : 0.5 and 1 μg/ml) was found to be most active against the tested stra ... | 2019 | 31441087 |
| genotoxicity and glucose tolerance induction by acetyltriethylcitrate, substitute plasticizer compared to di(2-ethylhexyl)phthalate. | as di(2-ethylhexyl) phthalate (dehp), one of phthalates, is classified as probable human carcinogens in epa, acetyltriethyl citrate(atec), one of aliphatic esters, could be applied to dehp substitute. atec is used as plasticizers in cosmetics and nail products. here, we studied whether atec might have genotoxic potential and induce glucose tolerance as compared to dehp. genotoxicity was determined by ames test with histidine-requiring salmonella typhimurium (ta98, ta100, ta1535 and ta1537) and t ... | 2019 | 31439862 |
| structure-activity relationship study: mechanism of cyto-genotoxicity of nitropyrazole-derived high energy density materials family. | stringent toxicological tests have to be performed prior to the industrial development of alternative chemicals particularly high energy dense materials (hedms) such as explosives. the properties (e.g., power, stability) of these compounds are constantly being improved, the current axis of research being the nitration of nitrogen heterocycles leading to hedms such as nitropyrazole-derived molecules. however, except for 3,4,5-trinitropyrazole (3,4,5-tnp), which was shown to be highly toxic in mic ... | 2019 | 31437493 |
| volatile organic compound (voc) emissions of cho and t cells correlate to their expansion in bioreactors. | volatile organic compound (voc) emissions were measured from chinese hamster ovary (cho) cell and t cell bioreactor gas exhaust lines with the goal of non-invasively metabolically profiling the expansion process. measurements of cellular 'breath' were made directly from the gas exhaust lines using polydimethylsiloxane (pdms)-coated magnetic stir bars, which underwent subsequent thermal desorption-gas chromatography-mass spectrometry (td-gc-ms) analysis. baseline voc profiles were observed from b ... | 2019 | 31430743 |
| a negative feedback loop regulates integrin inactivation and promotes neutrophil recruitment to inflammatory sites. | neutrophils are abundant circulating leukocytes that are rapidly recruited to sites of inflammation in an integrin-dependent fashion. contrasting with the well-characterized regulation of integrin activation, mechanisms regulating integrin inactivation remain largely obscure. using mouse neutrophils, we demonstrate in this study that the gtpase activating protein arap3 is a critical regulator of integrin inactivation; experiments with chinese hamster ovary cells indicate that this is not restric ... | 2019 | 31427445 |
| safety evaluation of pq birch allergy immunotherapy to support product development. | pq birch represents an allergen-specific immunotherapy for the treatment of birch pollinosis. it consists of native birch pollen extract chemically modified with glutaldehyde adsorbed to l-tyrosine in its microcrystalline form with addition of the adjuvant monophosphoryl lipid a (mpl®). a nonclinical safety testing strategy was designed based upon interpretation of current legislation and regulatory intelligence and comprised genotoxicity studies (bacterial reverse mutation and chinese hamster o ... | 2019 | 31425728 |
| uveitis sarcoidosis presumably initiated after administration of shingrix vaccine. | uveitis is a form of intraocular inflammation that can occur as a result of infection, injury, vaccination, or as a systemic manifestation of autoimmune or inflammatory diseases including sarcoidosis. sarcoidosis is an inflammatory disease that involves the formation of abnormal granulomas in multiple organ systems. the hallmark of sarcoidosis is a non-caseating granuloma seen on biopsy. here, we present a case report of a healthy 56-year-old caucasian female who presented with uveitis sarcoidos ... | 2019 | 31423396 |
| a comparison of the oligosaccharide structures of antithrombin derived from plasma and recombinant using potelligent® technology. | human antithrombin (at) has two isoforms of which the predominant α-form is glycosylated on all four possible glycosylation sites and the lower abundant β-isoform lacks the oligosaccharide on asn135. the main oligosaccharide structure of human at consists of biantennary complex-type oligosaccharides lacking a core fucose. generally, chinese hamster ovary (cho) cells produce recombinant human at (rhat) with core-fucosylated oligosaccharides. however, rhat lacking core-fucose oligosaccharides can ... | 2019 | 31423004 |
| clonal variations in cho igf signaling investigated by silac-based phosphoproteomics and lfq-ms. | chinese hamster ovary (cho) cells are commonly used for the production of monoclonal antibodies. omics technologies have been used to elucidate cellular switch points which result in higher monoclonal antibody (mab) productivity and process yields in cho and other biopharmaceutical production cell lines such as human or mouse. currently, investigations of the phosphoproteome in cho cell lines are rare yet could provide further insights into cellular mechanisms related to target product expressio ... | 2019 | 31420692 |
| impact of fc n-glycan sialylation on igg structure. | human igg antibodies containing terminal alpha 2,6-linked sialic acid on their fc n-glycans have been shown to reduce antibody-dependent cell-mediated cytotoxicity and possess anti-inflammatory properties. although terminal sialylation on complex n-glycans can happen via either an alpha 2,3-linkage or an alpha 2,6-linkage, sialic acids on human serum igg fc are almost exclusively alpha 2,6-linked. recombinant iggs expressed in chinese hamster ovary (cho) cells, however, have sialic acids through ... | 2020 | 31411531 |
| peptide-driven targeted drug-delivery system comprising turn-on near-infrared fluorescent xanthene-cyanine reporter for real-time monitoring of drug release. | targeted drug delivery (tdd) is an efficient strategy for cancer treatment. however, the real-time monitoring of drug delivery is still challenging because of a pronounced lack of tdd systems capable of providing a near-infrared (nir) fluorescence signal for the detection of drug-release events. herein, a new tdd system, comprising a turn-on nir fluorescent reporter attached to an anticancer drug and targeting peptide, is reported. this system provides both tdd and nir fluorescence monitoring of ... | 2019 | 31403246 |
| consequences of trace metal variability and supplementation on chinese hamster ovary (cho) cell culture performance: a review of key mechanisms and considerations. | trace metals are supplied to chemically-defined media (cdm) for optimal chinese hamster ovary (cho) cell culture performance during the production of monoclonal antibodies and other therapeutic proteins. however, lot-to-lot and vendor-to-vendor variability in raw materials consequently leads to an imbalance of trace metals that are supplied to cdm. this imbalance can yield detrimental effects rooted in several primary mechanisms and pathways including oxidative stress, apoptosis, lactate accumul ... | 2019 | 31403183 |
| zinc supplementation increases protein titer of recombinant cho cells. | in order to study the impact of zinc and copper on the titer levels of mab and recombinant protein in cho cells, the igg-expressing (dp12) and epo-expressing (sk15) cell lines were cultured in chemically defined media with increasing concentrations of either metal. supplementation with 25 mg/l in cdm media resulted in a significant increase in epo (1.7-fold) and igg (2.6-fold) titers compared to control (no added zinc). titers at this zn concentration in cdm containing the insulin replacing agen ... | 2019 | 31396753 |
| interphase cytogenetic analysis of micronucleated and multinucleated cells supports the premature chromosome condensation hypothesis as the mechanistic origin of chromothripsis. | the discovery of chromothripsis in cancer genomes challenges the long-standing concept of carcinogenesis as the result of progressive genetic events. despite recent advances in describing chromothripsis, its mechanistic origin remains elusive. the prevailing conception is that it arises from a massive accumulation of fragmented dna inside micronuclei (mn), whose defective nuclear envelope ruptures or leads to aberrant dna replication, before main nuclei enter mitosis. an alternative hypothesis i ... | 2019 | 31390832 |
| recombinant hemagglutinin produced from chinese hamster ovary (cho) stable cell clones and a pelc/cpg combination adjuvant for h7n9 subunit vaccine development. | the novel h7n9 avian influenza a virus has caused human infections in china since 2013; some isolates from the fifth wave of infections have emerged as highly pathogenic avian influenza viruses. recombinant hemagglutinin proteins of h7n9 viruses can be rapidly and efficiently produced with low-level biocontainment facilities. in this study, recombinant h7 antigen was obtained from engineered stable clones of chinese hamster ovary (cho) cells for subsequent large-scale production. the stable cho ... | 2019 | 31383491 |
| treatment with 3-aminobenzamide negates the radiofrequency-induced adaptive response in two cell models. | in previous investigations, we demonstrated that pre-exposure of different cell cultures to radiofrequency fields can reduce the damage induced by genotoxic agents, an effect resembling the so-called adaptive response. in this study, we pre-exposed human peripheral blood lymphocytes and chinese hamster lung fibroblast cell line to 1950 mhz, umts (universal mobile telecommunication system) signal, for 20 h, and then treated cultures with mitomycin-c. after confirming the induction of an adaptive ... | 2019 | 31382475 |
| process intensification in fed-batch production bioreactors using non-perfusion seed cultures. | although process intensification by continuous operation has been successfully applied in the chemical industry, the biopharmaceutical industry primarily uses fed-batch, rather than continuous or perfusion methods, to produce stable monoclonal antibodies (mabs) from chinese hamster ovary (cho) cells. conventional fed-batch bioreactors may start with an inoculation viable cell density (vcd) of ~0.5 × 106 cells/ml. increasing the inoculation vcd in the fed-batch production bioreactor (referred to ... | 2020 | 31379298 |
| bell-shaped agonist activation of 5-ht1a receptor-coupled gαi3 g-proteins: receptor density-dependent switch in receptor signaling. | a previous study observed bell-shaped concentration-response isotherms for activation of gαi3 g-protein subunits by high efficacy 5-ht1a receptor agonists in a chinese hamster ovary (cho) cell line expressing high levels of these receptors. this suggested that a signaling switch took place in that cell line (from gαi3 to activation of other g-proteins) but it was unclear if such effects are observed for 5-ht1a receptors in other cellular environments. here, using an antibody capture-based [35s]g ... | 2019 | 31376526 |
| clostridium perfringens epsilon toxin vaccine candidate lacking toxicity to cells expressing myelin and lymphocyte protein. | a variant form of clostridium perfringens epsilon toxin (y30a-y196a) with mutations, which shows reduced binding to madin-darby canine kidney (mdck) cells and reduced toxicity in mice, has been proposed as the next-generation enterotoxaemia vaccine. here we show that, unexpectedly, the y30a-y196a variant does not show a reduction in toxicity towards chinese hamster ovary (cho) cells engineered to express the putative receptor for the toxin (myelin and lymphocyte protein; mal). the further additi ... | 2019 | 31372245 |
| exploring the n-glycosylation profile of glycoprotein b from human cytomegalovirus expressed in cho and nicotiana tabacum by-2 cells. | the ability to control the glycosylation pattern of recombinant viral glycoproteins represents a major prerequisite before their use as vaccines. the aim of this study consisted of expressing the large soluble ectodomain of glycoprotein b (gb) from human cytomegalovirus (hmcv) in nicotiana tabacum bright yellow-2 (by-2) suspension cells and of comparing its glycosylation profile with that of gb produced in chinese hamster ovary (cho) cells. gb was secreted in the by-2 culture medium at a concent ... | 2019 | 31370181 |
| cellular targeting of bispecific antibody-functionalized poly(ethylene glycol) capsules: do shape and size matter? | in the present study, a capsule system that consists of a stealth carrier based on poly(ethylene glycol) (peg) and functionalized with bispecific antibodies (bsabs) is introduced to examine the influence of the capsule shape and size on cellular targeting. hollow spherical and rod-shaped peg capsules with tunable aspect ratios (ars) of 1, 7, and 18 were synthesized and subsequently functionalized with bsabs that exhibit dual specificities to peg and epidermal growth factor receptor (egfr). dosim ... | 2019 | 31369234 |
| gut microbial metabolites of linoleic acid are metabolized by accelerated peroxisomal β-oxidation in mammalian cells. | microorganisms in animal gut produce unusual fatty acids from the ingested diet. two types of hydroxy fatty acids (hfas), 10-hydroxy-cis-12-octadecenoic acid (hya) and 10-hydroxy-octadecanoic acid (hyb), are linoleic acid (la) metabolites produced by lactobacillus plantarum. in this study, we investigated the metabolism of these hfas in mammalian cells. when chinese hamster ovary (cho) cells were cultured with hya, approximately 50% of the supplemented hya disappeared from the dish within 24 h. ... | 2019 | 31351225 |
| germinality does not necessarily define mab expression and thermal stability. | the production potential of recombinant monoclonal antibody (mab) expressing cell lines depends, among other factors, on the intrinsic antibody structure determined by the amino acid sequence. in this study, we investigated the influence of somatic mutations in the v(d)j sequence of four individual, mature model mabs on the expression potential. therefore, we defined four couples, each consisting of one naturally occurring mab (2g12, ustekinumab, 4b3, and 2f5) and the corresponding germline-deri ... | 2019 | 31350616 |
| predicting industrial-scale cell culture seed trains-a bayesian framework for model fitting and parameter estimation, dealing with uncertainty in measurements and model parameters, applied to a nonlinear kinetic cell culture model, using an mcmc method. | for production of biopharmaceuticals in suspension cell culture, seed trains are required to increase cell number from cell thawing up to production scale. because cultivation conditions during the seed train have a significant impact on cell performance in production scale, seed train design, monitoring, and development of optimization strategies is important. this can be facilitated by model-assisted prediction methods, whereby the performance depends on the prediction accuracy, which can be i ... | 2019 | 31347693 |
| an unconventional uptake rate objective function approach enhances applicability of genome-scale models for mammalian cells. | constraint-based modeling has been applied to analyze metabolism of numerous organisms via flux balance analysis and genome-scale metabolic models, including mammalian cells such as the chinese hamster ovary (cho) cells-the principal cell factory platform for therapeutic protein production. unfortunately, the application of genome-scale model methodologies using the conventional biomass objective function is challenged by the presence of overly-restrictive constraints, including essential amino ... | 2019 | 31341637 |
| pmab-235: a monoclonal antibody for immunohistochemical analysis against goat podoplanin. | sensitive and specific monoclonal antibodies (mabs) against not only human but also mouse, rat, rabbit, dog, cat, bovine, pig, and horse podoplanins (pdpns) have been established in our previous studies. however, anti-goat pdpn (gpdpn) has not been established yet. pdpn has been utilized as a lymphatic endothelial cell marker especially in pathological diagnoses; therefore, mabs for immunohistochemical analyses using formalin-fixed paraffin-embedded tissues are needed. although we recently demon ... | 2019 | 31338471 |
| fabrication of amyloid-β-secreting alginate microbeads for use in modelling alzheimer's disease. | according to the amyloid cascade hypothesis, the earliest trigger in the development of alzheimer's disease (ad) is the accumulation of toxic amyloid-β (aβ) fragments, eventually leading to the classical features of the disease: amyloid plaques, neurofibrillary tangles and synaptic and neuronal loss. the lack of relevant non-transgenic preclinical models reflective of disease progression is one of the main factors hindering the discovery of effective drug treatments. to this end, we have develop ... | 2019 | 31329175 |
| egf receptor stalls upon activation as evidenced by complementary fluorescence correlation spectroscopy and fluorescence recovery after photobleaching measurements. | to elucidate the molecular details of the activation-associated clustering of epidermal growth factor receptors (egfrs), the time course of the mobility and aggregation states of egfp tagged egfr in the membranes of chinese hamster ovary (cho) cells was assessed by in situ mobility assays. fluorescence correlation spectroscopy (fcs) was used to probe molecular movements of small ensembles of molecules over short distances and time scales, and to report on the state of aggregation. the diffusion ... | 2019 | 31323980 |
| activin a-induced cachectic wasting is attenuated by systemic delivery of its cognate propeptide in male mice. | in cancer, elevated activin levels promote cachectic wasting of muscle, irrespective of tumor progression. in excess, activins a and b use the myostatin signaling pathway in muscle, triggering a decrease in protein synthesis and an increase in protein degradation, which ultimately leads to atrophy. recently, we demonstrated that local delivery of engineered activin and myostatin propeptides (natural inhibitors of these growth factors) could induce profound muscle hypertrophy in healthy mice. add ... | 2019 | 31322699 |
| influence of environmental conditions on the fusion of cationic liposomes with living mammalian cells. | lipid-based nanoparticles, also called vesicles or liposomes, can be used as carriers for drugs or many types of biological macromolecules, including dna and proteins. efficiency and speed of cargo delivery are especially high for carrier vesicles that fuse with the cellular plasma membrane. this occurs for lipid mixture containing equal amounts of the cationic lipid dotap and a neutral lipid with an additional few percents of an aromatic substance. the fusion ability of such particles depends o ... | 2019 | 31319557 |
| expression, purification, and characterization of a human complement component c3 analog that lacks the c-terminal c345c domain. | the complement system consists of a series of soluble and cell-surface proteins that serve numerous roles in innate immunity, development, and homeostasis. despite its many functions, the central event in the complement system is the proteolytic activation of the 185 kda complement component 3 (c3) into its opsonin and anaphylatoxin fragments known as c3b (175 kda) and c3a (10 kda), respectively. the c3 protein is comprised of thirteen separate structural domains, several of which undergo extens ... | 2019 | 31319063 |
| coexpression of cmp-sialic acid transporter reduces n-glycolylneuraminic acid levels of recombinant glycoproteins in chinese hamster ovary cells. | recombinant glycoproteins expressed in chinese hamster ovary (cho) cells contain two forms of sialic acids; n-acetylneuraminic acid (neu5ac) as a major type and n-glycolylneuraminic acid (neu5gc) as a minor type. the neu5gc glycan moieties in therapeutic glycoproteins can elicit immune responses because they do not exist in human. in the present work, to reduce neu5gc levels of recombinant glycoproteins from cho cell cultures, we coexpressed cytidine-5'-monophosphate-sialic acid transporter (cmp ... | 2019 | 31317538 |
| glyconanoparticles for targeted tumor therapy of platinum anticancer drug. | an important requirement to decrease the side effects of chemotherapy drugs is to develop nanocarriers with precise biological functions. in this work, a set of glyconanoparticles was prepared via self-assembly of amphiphilic glycoblock copolymers for the targeted delivery of a hydrophobic chemotherapy drug. well-defined glycoblock copolymers that consist of 1,1-di-tert-butyl 3-(2-(metyloyloxy)ethyl)-butane-1,1,3-tricarboxylate (maetc) together with three different protected-sugar moieties (β-d- ... | 2019 | 31314508 |
| kinetic basis of metformin-mpp interactions with organic cation transporter oct2. | organic cation transporter 2 (oct2) clears the blood of cationic drugs. efforts to understand oct2 selectivity as a means to predict the potential of new molecular entities (nmes) to produce unwanted drug-drug interactions typically assess the influence of the nmes on inhibition of transport. however, the identity of the substrate used to assess transport activity can influence the quantitative profile of inhibition. metformin and 1-methyl-4-phenylpyridinium (mpp), in particular, display markedl ... | 2019 | 31313952 |
| characterization of chinese hamster ovary cells with disparate chromosome numbers: reduction of the amount of mrna relative to total protein. | chromosomes in chinese hamster ovary (cho) cells are labile. we have shown that high-chromosome-number cho cells have greater potential to become robust producers of recombinant proteins. one explanation being the increase in transgene integration sites. however, high-chromosome-number cell clones produce more igg3 following culture of single-cell clones, even under conditions that yield the same number of integrations as cells with normal chromosome numbers. here, we characterized high-chromoso ... | 2020 | 31303495 |
| disinfection byproducts and their toxicity in wastewater effluents treated by the mixing oxidant of clo2/cl2. | mixing oxidant of chlorine dioxide (clo2) and chlorine (cl2) often applied in water disinfection. two secondary wastewater effluents at different ammonium-n levels (0.1 and 1.6 mg n l-1) were treated with the mixing oxidant (clo2/cl2) to evaluate the formation of disinfection byproducts (dbps) and the associated cytotoxicity of treated wastewaters. the total chlorine concentrations of clo2 and cl2 were maintained at 10 mg l-1 as cl2 with varied mixing ratios of clo2 to cl2. the formation of 37 h ... | 2019 | 31302364 |
| bioisosteric ferrocenyl aminoquinoline-benzimidazole hybrids: antimicrobial evaluation and mechanistic insights. | phenyl- and bioisosteric ferrocenyl-derived aminoquinoline-benzimidazole hybrid compounds were synthesised and evaluated for their in vitro antiplasmodial activity against the chloroquine-sensitive nf54 and multi-drug resistant k1 strains of the human malaria parasite, plasmodium falciparum. all compounds were active against the two strains, generally showing enhanced activity in the k1 strain, with resistance indices less than 1. cytotoxicity studies using chinese hamster ovarian cells revealed ... | 2019 | 31301563 |
| development of a new promoter to avoid the silencing of genes in the production of recombinant antibodies in chinese hamster ovary cells. | the production of recombinant proteins in mammalian cell lines is one of the most important areas in biopharmaceutical industry. viral transcriptional promoters are widely used to express recombinant proteins in mammalian cell lines. however, these promoters are susceptible to silencing, thus limiting protein productivity. some cpg islands can avoid the silencing of housekeeping genes; for that reason, they have been used to increase the production of recombinant genes in cells of animal origin. ... | 2019 | 31297150 |
| mode-of-action analysis of the effects induced by nicotine in the in vitro micronucleus assay. | nicotine's genotoxic potential has been extensively studied in vitro. while the results of mammalian cell-based studies have inferred that it can potentially damage chromosomes, in general and with few exceptions, adverse dna effects have been observed primarily at supraphysiological concentrations in nonregulatory assays that provide little information on its mode-of-action (moa). in this study, a modern-day regulatory genotoxicity assessment was conducted using a flow cytometry-based in vitro ... | 2019 | 31294873 |
| mir-106b promotes therapeutic antibody expression in cho cells by targeting deubiquitinase cyld. | micrornas (mirnas) function as important regulators of major cellular processes, such as cell cycle, proliferation, development, and apoptosis. recently, mirna engineering of chinese hamster ovary (cho) cells has emerged as a promising strategy for enhancing therapeutic antibody production. previously, we have reported that inhibition of deubiquitinase cylindromatosis (cyld) remarkably enhanced the therapeutic antibody production in cho cells. however, the mechanisms regulating cyld in cho cells ... | 2019 | 31292678 |
| cytoplasmic conductivity as a marker for bioprocess monitoring: study of chinese hamster ovary cells under nutrient deprivation and reintroduction. | the ability to monitor the status of cells during nutrient limitation is important for optimizing bioprocess growth conditions in batch and fed-batch cultures. the activity level of na+ /k+ atpase pumps and cytoplasm ionic concentrations are directly influenced by the nutrient level, and thus, cytoplasm conductivity can be used as a markerless indicator of cell status. in this work, we monitored the change in cytoplasm conductivity of chinese hamster ovary (cho) cells during nutrient deprivation ... | 2019 | 31286489 |