Publications

TitleAbstractYear(sorted descending)
Filter
PMID
Filter
tbx20 controls the expression of the kcnh2 gene and of herg channels.long qt syndrome (lqts) exhibits great phenotype variability among family members carrying the same mutation, which can be partially attributed to genetic factors. we functionally analyzed the kcnh2 (encoding for kv11.1 or herg channels) and tbx20 (encoding for the transcription factor tbx20) variants found by next-generation sequencing in two siblings with lqts in a spanish family of african ancestry. affected relatives harbor a heterozygous mutation in kcnh2 that encodes for p.t152hfsx180 kv11 ...201728049825
production of human mutant biologically active hepatocyte growth factor in chinese hamster ovary cells.hepatocyte growth factor (hgf) is a potent multifunctional cytokine that affects proliferation, migration, and morphogenesis of various cells. hgf is secreted as an inactive single-chain precursor protein and activated by the cleavage of serine proteases to form heterodimers. in our current study, the cleavage site of hgf was blocked by replaced arg 494 of glu (r494e) that resulted in the single-chain hgf (r494e) unable to be cleaved by serine proteases. we established chinese hamster ovary (cho ...201728045643
asp73-dependent and -independent regulation of the affinity of ligands for human histamine h1 receptors by na().the affinity of ligands for g-protein-coupled receptors (gpcrs) is allosterically regulated by na(+) via a highly conserved aspartate residue (asp(2.50)) in the second transmembrane domain of gpcrs. in the present study, we examined the na(+)-mediated regulation of the affinity of ligands for gq/11-protein-coupled human histamine h1 receptors in chinese hamster ovary cells. the affinities of 3 agonists and 20 antihistamines were evaluated by their displacement curves against the binding of [(3)h ...201728040476
assessing the genotoxicity of two commonly occurring byproducts of water disinfection: chloral hydrate and bromal hydrate.water disinfection treatments result in the formation of disinfection byproducts (dbps) that have been linked to adverse human health outcomes including higher incidence of bladder and colorectal cancer. however, data about the genotoxicity of dbps is limited to only a small fraction of compounds. chloral hydrate (ch) and bromal hydrate (bh) are two trihaloacetaldehydes commonly detected in disinfected waters, but little is known about their genotoxicity, especially bh. we investigated the genot ...201728010927
preferential hydrolysis of truncated oxidized glycerophospholipids by lysosomal phospholipase a2.truncated oxidized glycerophospholipids (ox-pls) are bioactive lipids resulting from oxidative stress. the catabolic pathways for truncated ox-pls are not fully understood. lysosomal phospholipase a2 (lpla2) with phospholipase a and transacylase activities is a key enzyme in phospholipid homeostasis. the present study assessed whether lpla2 could hydrolyze truncated ox-pls. incubation of lpla2 with liposomes consisting of 1,2-o-octadecenyl-sn-glycero-3-phosphocholine (dodpc)/1,2-dioleoyl-sn-glyc ...201727993948
hiv protease inhibitors alter amyloid precursor protein processing via β-site amyloid precursor protein cleaving enzyme-1 translational up-regulation.mounting evidence implicates antiretroviral (arv) drugs as potential contributors to the persistence and evolution of clinical and pathological presentation of hiv-associated neurocognitive disorders in the post-arv era. based on their ability to induce endoplasmic reticulum (er) stress in various cell types, we hypothesized that arv-mediated er stress in the central nervous system resulted in chronic dysregulation of the unfolded protein response and altered amyloid precursor protein (app) proc ...201727993242
high-throughput lipidomic and transcriptomic analysis to compare sp2/0, cho, and hek-293 mammalian cell lines.a combined lipidomics and transcriptomics analysis was performed on mouse myeloma sp2/0, chinese hamster ovary (cho), and human embryonic kidney (hek) cells in order to compare widely used mammalian expression systems. initial thin layer chromatography (tlc) analysis indicated that phosphatidylethanolamine (pe) and phosphatidylcholine (pc) were the major lipid components in all cell lines with lower amounts of sphingomyelin (sm) in sp2/0 compared to cho and hek, which was subsequently confirmed ...201727991764
integrated genome and protein editing swaps α-2,6 sialylation for α-2,3 sialic acid on recombinant antibodies from cho.immunoglobin g with α-2,6 sialylation has been reported to have an impact on antibody-dependent cellular cytotoxicity and anti-inflammatory efficacy. however, production of antibodies with α-2,6 sialylation from chinese hamster ovary cells is challenging due to the inaccessibility of sialyltransferases for the heavy chain n-glycan site and the presence of exclusively α-2,3 sialyltransferases. in this study, combining mutations on the fc regions to allow sialyltransferase accessibility with overe ...201727943633
knockout of a difficult-to-remove cho host cell protein, lipoprotein lipase, for improved polysorbate stability in monoclonal antibody formulations.while the majority of host cell protein (hcp) impurities are effectively removed in typical downstream purification processes, a small population of hcps are particularly challenging. previous studies have identified hcps that are challenging for a variety of reasons. lipoprotein lipase (lpl)-a chinese hamster ovary (cho) hcp that functions to hydrolyze esters in triglycerides-was one of ten hcps identified in previous studies as being susceptible to retention in downstream processing. lpl may d ...201727943242
design, synthesis and biological activity of novel molecules designed to target parp and dna.in order to enhance the cytotoxic potential of poly(adp-ribose) polymerase (parp) inhibitors in brca1 or 2 deficient tumours, we designed a series of molecules containing a 1,2,3-triazene moiety tethered to a parp targeting scaffold. a cell-based selectivity assay involving a brca2-deficient chinese hamster cell line and its corresponding brca2 wild type transfectant, was used to predict the parp targeting potential of the latter agents. the results showed that adding a dna damaging function to ...201728003142
improving the secretory capacity of chinese hamster ovary cells by ectopic expression of effector genes: lessons learned and future directions.chinese hamster ovary (cho) cells are the preferred cell factory for the production of therapeutic glycoproteins. although efforts primarily within bioprocess optimization have led to increased product titers of recombinant proteins (r-proteins) expressed in cho cells, post-transcriptional bottlenecks in the biosynthetic pathway of r-proteins remain to be solved. to this end, the ectopic expression of transgenes (effector genes) offers great engineering potential. however, studies on effector ge ...201727931938
role of a tachykinin-related peptide and its receptor in modulating the olfactory sensitivity in the oriental fruit fly, bactrocera dorsalis (hendel).insect tachykinin-related peptide (trp), an ortholog of tachykinin in vertebrates, has been linked with regulation of diverse physiological processes, such as olfactory perception, locomotion, aggression, lipid metabolism and myotropic activity. in this study, we investigated the function of trp (bdtrp) and its receptor (bdtrpr) in an important agricultural pest, the oriental fruit fly bactrocera dorsalis. bdtrpr is a typical g-protein coupled-receptor (gpcr), and it could be activated by the pu ...201727923683
integrin αiibβ3-dependent erk signaling is regulated by src and rho kinases in both leu33 and pro33 polymorphic isoforms.platelet integrin αiibβ3 possesses a leu/pro polymorphism at residue 33 (leu33/hpa-1a or pro33/hpa-1b). the pro33 isoform has been suggested to exhibit prothrombotic features. αiibβ3-expressing cho (chinese hamster ovary) cells on immobilized fibrinogen show activation of the map kinase family member erk2, with an enhanced erk2 activity in pro33 cells compared to leu33 cells. in our present work, we examined how the leu/pro polymorphism modulates the erk2 activation stimulated by 2 differently t ...201727923225
a framework to quantify karyotype variation associated with cho cell line instability at a single-cell level.chinese hamster ovary (cho) cells, the major mammalian host cells for biomanufacturing of therapeutic proteins, have been extensively investigated to enhance productivity and product quality. however, cell line instability resulting in unexpected changes in productivity or product quality continues to be a challenge. based on previous reports about causes and characteristics of production instability, we hypothesized that chromosomal rearrangements due to genomic instability are associated with ...201727922175
flavonoids from heliotropium subulatum exudate and their evaluation for antioxidant, antineoplastic and cytotoxic activities ii.the flavonoids are the largest group of phenolic compounds isolated from a wide range of higher plants. these compounds work as antimicrobials, anti-insect agents and protect plants from other types of biotic and abiotic stresses. various researchers have suggested that flavonoids possessed antioxidant, antineoplastic and cytotoxic activities. the main objective of this study was to test dichloromethane fraction of resinous exudate of heliotropium subulatum for their antioxidant, antineoplastic ...201727905025
selection of chemically defined media for cho cell fed-batch culture processes.two cho cell clones derived from the same parental cho(bc®) cell line and producing the same monoclonal antibody (bc-g, a low producing clone; bc-p, a high producing clone) were tested in four basal media in all possible combinations with three feeds (=12 conditions) in fed-batch cultures. higher amino acid feeding did not always lead to higher mab production. the two clones showed differences in cell physiology, metabolism and optimal medium-feed combinations. during the phase transitions of al ...201727900626
rational design of glycoengineered interferon-β analogs with improved aggregation state: experimental validation.recombinant human interferon-β (rhifn-β) used clinically has lower efficacy than expected due to protein instabilities such as aggregation. increasing molecular stability, glycoengineering has been used to improve clinical efficacy for a number of therapeutics; however, often labor-intensive trail-and-error approaches are used to identify additional glycosylation sites. in this study two rhifn-β analogs with one additional glycosylation site, l6t and s75n, identified by a rational in silico appr ...201727881683
segmented linear modeling of cho fed-batch culture and its application to large scale production.we describe a systematic approach to model cho metabolism during biopharmaceutical production across a wide range of cell culture conditions. to this end, we applied the metabolic steady state concept. we analyzed and modeled the production rates of metabolites as a function of the specific growth rate. first, the total number of metabolic steady state phases and the location of the breakpoints were determined by recursive partitioning. for this, the smoothed derivative of the metabolic rates wi ...201727869296
reduced cytotoxicity in pcb-exposed chinese hamster ovary (cho) cells pretreated with vitamin e.the aim of this study was to evaluate protective effects of vitamin e (50 -150 μm) in ovary cells upon cytotoxic effects induced by two structurally distinct pcb congeners - planar "dioxin-like" pcb 77 and non-planar di-ortho-substituted pcb 153 with an emphasis on identifying differences in the mechanism of vitamin e action depending on the structure of congeners. application of three bioassays confirmed that pcbs decrease ovarian cell proliferation with slightly profound effects of pcb 77. pcb ...201727865896
establishment of monoclonal antibodies against cell surface domains of asct2/slc1a5 and their inhibition of glutamine-dependent tumor cell growth.human alanine-serine-cysteine transporter 2 (asct2; slc1a5) is a major transporter of the amino acid glutamine that is known to be overexpressed in certain malignant tumors. in this study, we generated specific monoclonal antibodies (mabs) against asct2 by establishing an asct2-expressing chinese hamster ovary cell line that was used to immunize mice and rats. the mabs km4008, km4012, and km4018 against asct2 were isolated through a cell-based screen; these specifically bound to asct2-positive c ...201727865832
improved recombinant antibody production by cho cells using a production enhancer dna element with repeated transgene integration at a predetermined chromosomal site.chinese hamster ovary (cho) cells are one of the most useful host cell lines for the production of biopharmaceutical proteins. although a series of production processes have been refined to improve protein productivity and cost performance, establishing producer cells is still time-consuming and labor-intensive. recombinase-mediated site-specific gene integration into a predetermined chromosomal locus may enable predictable protein expression, reducing the laborious process of cell screening. we ...201727856232
production and purification of recombinant glucocerebrosidase in transgenic rice cell suspension cultures.gaucher disease, which is caused by deficiency of glucocerebrosidase (gcd), is currently treated by enzyme replacement therapy. plant-based systems produce glycoproteins and can be combined with targeting strategies to generate proteins with terminal mannose structures for macrophage uptake. however, the gliding step for the purification is essential since the produced protein still exists inside cells. in the case of rice-amylase 1a (ramy1a) secretion signal peptide, gcd protein is secreted out ...201727817046
lysosomal alpha-mannosidase and alpha-mannosidosis.lysosomal alpha-mannosidase with acidic ph optimum is ubiquitous in human tissues where is expressed in two major forms, a and b that are the product of a single gene located on chromosome 19. mutations in the gene encoding for alpha-mannosidase cause alpha- mannosidosis, an autosomal recessive disease, resulting in the accumulation of unprocessed mannose containing oligosaccharide material. this rare disease has an estimated incidence of 1/500.0.00 live births and clinically is divided into thr ...201727814608
the effect of particle size on the genotoxicity of gold nanoparticles.despite the increasing biomedical applications of gold nanoparticles (aunps), their toxicological effects need to be thoroughly understood. in the present study, the genotoxic potential of commercially available aunps with varying size (5, 20, and 50 nm) were assessed using a battery of in vitro and in vivo genotoxicity assays. in the comet assay, 20 and 50 nm aunps did not induce obvious dna damage in hepg2 cells at the tested concentrations, whereas 5 nm nps induced a dose-dependent increment ...201727770565
differences in trpc3 and trpc6 channels assembly in mesenteric vascular smooth muscle cells in essential hypertension.canonical transient receptor potential (trpc)3 and trpc6 channels of vascular smooth muscle cells (vsmcs) mediate stretch- or agonist-induced cationic fluxes, contributing to membrane potential and vascular tone. native trpc3/c6 channels can form homo- or heterotetrameric complexes, which can hinder individual trpc channel properties. the possibility that the differences in their association pattern may change their contribution to vascular tone in hypertension is unexplored. functional characte ...201727861908
'click'-xylosides as initiators of the biosynthesis of glycosaminoglycans: comparison of mono-xylosides with xylobiosides.different mono-xylosides and their corresponding xylobiosides obtained by a chemo-enzymatic approach featuring various substituents attached to a triazole ring were probed as priming agents for glycosaminoglycan (gag) biosynthesis in the xylosyltransferase-deficient pgsa-745 chinese hamster ovary cell line. xylosides containing a hydrophobic aglycone moiety were the most efficient priming agents. mono-xylosides induced higher gag biosynthesis in comparison with their corresponding xylobiosides. ...201727618481
detection of a phosphorylated glycine-serine linker in an igg-based fusion protein.molecular mass determination by electrospray ionization mass spectrometry of a recombinant igg-based fusion protein (mab1-f) produced in human embryonic kidney (hek) cells demonstrated the presence of a dominant +79 da product variant. using lc-ms tryptic peptide mapping analysis and collision-induced dissociation (cid) and electron-transfer/higher-energy collision dissociation fragmentations, the modification was localized to the c-terminal serine residue of a glycine-serine linker [(g4s)2] of ...201727661266
identification of an igg cdr sequence contributing to co-purification of the host cell protease cathepsin d.recombinant therapeutic monoclonal antibodies (mabs) must be purified from host cell proteins (hcps), dna, and other impurities present in chinese hamster ovary (cho) cell culture media. hcps can potentially result in adverse clinical responses in patients and, in specific cases, have caused degradation of the final mab product. as reported previously, residual traces of cathepsin d caused particle formation in the final product of mab-1. the current work was focused on identification of a prima ...201727798957
mar-mediated transgene integration into permissive chromatin and increased expression by recombination pathway engineering.untargeted plasmid integration into mammalian cell genomes remains a poorly understood and inefficient process. the formation of plasmid concatemers and their genomic integration has been ascribed either to non-homologous end-joining (nhej) or homologous recombination (hr) dna repair pathways. however, a direct involvement of these pathways has remained unclear. here, we show that the silencing of many hr factors enhanced plasmid concatemer formation and stable expression of the gene of interest ...201727575535
lengthening of high-yield production levels of monoclonal antibody-producing chinese hamster ovary cells by downregulation of breast cancer 1.the establishment process of high-producing chinese hamster ovary (cho) cells for therapeutic protein production is usually laborious and time consuming because of the low probability of obtaining stable, high-producing clones over a long term. thus, development of an efficient approach is required to establish stable, high-producing cells. this study presents a novel method that can efficiently establish sustainably high-producing cell lines by acceleration of transgene amplification and suppre ...201727742176
specific immune response to phospholipase b-like 2 protein, a host cell impurity in lebrikizumab clinical material.host cell proteins are manufacturing process-related impurities that may co-purify with the product despite extensive efforts to optimize the purification process. the risks associated with these impurities can vary and may be patient and/or therapeutic dependent. therefore, it is critical to monitor and control the levels of these impurities in products and their potential impact on safety and efficacy. lebrikizumab is a humanized immunoglobulin g4 monoclonal antibody (mab) that binds specifica ...201727739010
reduction of ammonia and lactate through the coupling of glutamine synthetase selection and downregulation of lactate dehydrogenase-a in cho cells.chinese hamster ovary (cho) cell cultivation for production of therapeutic proteins is accompanied by production of metabolic wastes, mostly ammonia and lactate. to reduce ammonia production, the glutamine synthetase (gs) system was used to develop therapeutic monoclonal antibody (mab)-producing cho cells (sm-0.025). additionally, the lactate dehydrogenase-a (ldh-a) was downregulated with shrna to reduce lactate production in sm-0.025. the resulting mab-producing cell lines (#2, #46, and #52) pr ...201727704181
fish-based analysis of clonally derived cho cell populations reveals high probability for transgene integration in a terminal region of chromosome 1 (1q13).a basic goal in the development of recombinant proteins is the generation of cell lines that express the desired protein stably over many generations. here, we constructed engineered chinese hamster ovary cell lines (cho-s) with a pcho-hvr1 vector that carried an extracellular domain of a vegf receptor (vr) fusion gene. forty-five clones with high hvr1 expression were selected for karyotype analysis. using fluorescence in situ hybridization (fish) and g-banding, we found that pcho-hvr1 was integ ...201727684722
cell-free systems based on cho cell lysates: optimization strategies, synthesis of "difficult-to-express" proteins and future perspectives.nowadays, biotechnological processes play a pivotal role in target protein production. in this context, chinese hamster ovary (cho) cells are one of the most prominent cell lines for the expression of recombinant proteins and revealed as a safe host for nearly 40 years. nevertheless, the major bottleneck of common in vivo protein expression platforms becomes obvious when looking at the production of so called "difficult-to-express" proteins. this class of proteins comprises in particular several ...201727684475
a novel fusion protein, achr-fc, ameliorates myasthenia gravis by neutralizing antiacetylcholine receptor antibodies and suppressing acetylcholine receptor-reactive b cells.most patients with myasthenia gravis (mg) have elevated levels of autoantibodies against the nicotinic acetylcholine receptor (achr) at the neuromuscular junction, which leads to muscle weakness. we developed a fusion protein, achr-fc, as a novel therapeutic biomolecule for patients with mg and examined its efficacy. achr-fc was expressed by chinese hamster ovary cells and purified. we examined the neutralizing activity and cellular cytotoxicity of achr-fc using anti-achr antibody-producing hybr ...201727677608
fam60a plays a role for production stabilities of recombinant cho cell lines.recombinant cho (chinese hamster ovary) cell lines producing therapeutic proteins often lose their production capability during long-term cultivation. to ensure that cho production cell lines can be up-scaled to high-volume bioreactors, labor intensive stability studies of several months have to be performed to deselect clones that are losing productivity over time. the ability to predict whether clones will produce recombinant proteins at constant high levels, for example, through determination ...201727617904
droplet size influences division of mammalian cell factories in droplet microfluidic cultivation.the potential of using droplet microfluidics for screening mammalian cell factories has been limited by the difficulty in achieving continuous cell division during cultivation in droplets. here, we report the influence of droplet size on mammalian cell division and viability during cultivation in droplets. chinese hamster ovary (cho) cells, the most widely used mammalian host cells for biopharmaceuticals production were encapsulated and cultivated in 33, 180 and 320 pl droplets for 3 days. perio ...201727535608
fx knockout cho hosts can express desired ratios of fucosylated or afucosylated antibodies with high titers and comparable product quality.during antibody dependent cell cytotoxicity (adcc) the target cells are killed by monocytes and natural killer cells. adcc is enhanced when the antibody heavy chain's core n-linked glycan lacks the fucose molecule(s). several strategies have been utilized to generate fully afucosylated antibodies. a commonly used and efficient approach has been knocking out the fut8 gene of the chinese hamster ovary (cho) host cells, which results in expression of antibody molecules with fully afucosylated glyca ...201727666939
cationic lipids bearing succinic-based, acyclic and macrocyclic hydrophobic domains: synthetic studies and in vitro gene transfer.in this communication we describe the construction of four succinic-based cationic lipids, their formulation with plasmid dna (pdna), and an evaluation of their in vitro gene delivery into chinese hamster ovarian (cho-k1) cells. the cationic lipids employed in this work possess either a dimethylamine or trimethylamine headgroup, and a macrocyclic or an acyclic hydrophobic domain composed of, or derived from two 16-atom, succinic-based acyl chains. the synthesized lipids and a co-lipid of neutral ...201727662033
label-free live cell imaging by confocal raman microscopy identifies cho host and producer cell lines.as a possible viable and non-invasive method to identify high producing cells, confocal raman microscopy was shown to be able to differentiate cho host cell lines and derivative production clones. cluster analysis of spectra and their derivatives was able to differentiate between different producer cell lines and a host, and also distinguished between an intracellular region of high lipid and protein content that in structure resembles the endoplasmic reticulum. this ability to identify the er m ...201727440252
increased in vivo effector function of human igg4 isotype antibodies through afucosylation.for some antibodies intended for use as human therapeutics, reduced effector function is desired to avoid toxicities that might be associated with depletion of target cells. since effector function(s), including antibody-dependent cell-mediated cytotoxicity (adcc), require the fc portion to be glycosylated, reduced adcc activity antibodies can be obtained through aglycosylation of the human igg1 isotype. an alternative is to switch to an igg4 isotype in which the glycosylated antibody is known t ...201727216702
use of in vitro assays to assess the potential cytotoxic, genotoxic and antigenotoxic effects of vanillic and cinnamic acid.vanillic acid (va) found in vanilla and cinnamic acid (ca) the precursor of flavonoids and found in cinnamon oil, are natural plant phenolic acids which are secondary aromatic plant products suggested to possess many physiological and pharmacological functions. in vitro and in vivo experiments have shown that phenolic acids exhibit powerful effects on biological responses by scavenging free radicals and eliciting antioxidant capacity. in the present study, we investigated the antioxidant capacit ...201727309403
polymorphisms in dna repair gene and susceptibility to glioma: a systematic review and meta-analysis based on 33 studies with 15 snps in 9 genes.at present, many publications have evaluated the correlation between the dna repair gene polymorphisms and glioma susceptibility. however, the results remain inconclusive. the aim of this research is to exhaustively assess the association of genetic polymorphisms in dna repair genes with glioma risk in human. meta-analysis method was conducted, and 33 studies with 15 snps in 9 genes were included (12553 glioma cases and 17178 controls). correlation strength was evaluated by odds ratio with a 95  ...201727055523
xgfr*, a novel affinity-matured bispecific antibody targeting igf-1r and egfr with combined signaling inhibition and enhanced immune activation for the treatment of pancreatic cancer.the epidermal growth factor receptor (egfr) and the insulin-like growth factor-1 receptor (igf-1r) play critical roles in tumor growth, providing a strong rationale for the combined inhibition of igf-1r and egfr signaling in cancer therapy. we describe the design, affinity maturation, in vitro and in vivo characterization of the bispecific anti-igf-1r/egfr antibody xgfr*. xgfr* is based on the bispecific igg antibody xgfr, which enabled heterodimerization of an igf-1r binding scfab heavy chain w ...201726984378
charge heterogeneity: basic antibody charge variants with increased binding to fc receptors.we identified active isoforms of the chimeric anti-gd2 antibody, ch14.18, a recombinant antibody produced in chinese hamster ovary cells, which is already used in clinical trials. (1,2,3) we separated the antibody by high resolution ion-exchange chromatography with linear ph gradient elution into acidic, main and basic charge variants on a preparative scale yielding enough material for an in-depth study of the sources and the effects of microheterogeneity. the binding affinity of the charge vari ...201727559765
the use of transcription terminators to generate transgenic lines of chinese hamster ovary cells (cho) with stable and high level of reporter gene expression.mammalian cell lines are widely used to produce recombinant proteins. stable transgenic cell lines usually contain many insertions of the expression vector in one genomic region. transcription through transgene can be one of the reasons for target gene repression after prolonged cultivation of cell lines. in the present work, we used the known transcription terminators from the sv40 virus, as well as the human β- and γ-globin genes, to prevent transcription through transgene. the transcription t ...201726483962
understanding the cholesterol metabolism-perturbing effects of docosahexaenoic acid by gas chromatography-mass spectrometry targeted metabonomic profiling.over the past few decades, docosahexaenoic acid (dha) has gained special attention for management of cholesterol-associated metabolic disorders and neurodegenerative diseases such as alzheimer's disease (ad) owing to its neuroprotective, anti-inflammatory and hypolipidemic properties. several epidemiological studies have reported the effect of dha in reducing the risk of developing ad by lowering cholesterol. hypercholesterolemia is a pro-amyloidogenic factor influencing the enzymatic processing ...201726428672
reciprocal translocation observed in end-of-production cells of a commercial cho-based process.during the validation of an additional working cell bank derived from a validated master cell bank to support the commercial production continuum of a recombinant protein, we observed an unexpected chromosomal location of the gene of interest in some end-of-production cells. this event-identified by fluorescence in situ hybridization and multicolour chromosome painting as a reciprocal translocation involving a chromosome region containing the gene of interest with its integral coding and flankin ...201726242789
chinese hamster ovary (cho) host cell engineering to increase sialylation of recombinant therapeutic proteins by modulating sialyltransferase expression.n-glycans of human proteins possess both α2,6- and α2,3-linked terminal sialic acid (sa). recombinant glycoproteins produced in chinese hamster overy (cho) only have α2,3-linkage due to the absence of α2,6-sialyltransferase (st6gal1) expression. the chinese hamster st6gal1 was successfully overexpressed using a plasmid expression vector in three recombinant immunoglobulin g (igg)-producing cho cell lines. the stably transfected cell lines were enriched for st6gal1 overexpression using fitc-sambu ...201725641927
restoration of mitochondrial gene expression using a cloned human gene in chinese hamster lung cell mutant.gal(-)32 is a chinese hamster lung cell nuclear mutant that is unable to grow in galactose due to a defect in mitochondrial protein synthesis. since the product of the gal(-)32 gene was unknown, it was imperative to use phenotypic complementation to clone a human gene that corrected the gal(-)32 mutation.201726052559
ch14.18 antibody produced in cho cells in relapsed or refractory stage 4 neuroblastoma patients: a siopen phase 1 study.this study aimed to assess the safety, pharmacokinetic and activity profiles of the human-mouse chimeric monoclonal anti-disialoganglioside gd2 antibody ch14.18 produced in chinese hamster ovary (cho) cells (ch14.18/cho).201723924804
a high-throughput media design approach for high performance mammalian fed-batch cultures.an innovative high-throughput medium development method based on media blending was successfully used to improve the performance of a chinese hamster ovary fed-batch medium in shaking 96-deepwell plates. starting from a proprietary chemically-defined medium, 16 formulations testing 43 of 47 components at 3 different levels were designed. media blending was performed following a custom-made mixture design of experiments considering binary blends, resulting in 376 different blends that were tested ...201723563583
very high density of chinese hamster ovary cells in perfusion by alternating tangential flow or tangential flow filtration in wave bioreactor™-part ii: applications for antibody production and cryopreservation.a high cell density perfusion process of monoclonal antibody (mab) producing chinese hamster ovary (cho) cells was developed in disposable wave bioreactor™ using external hollow fiber (hf) filter as cell separation device. tangential flow filtration (tff) and alternating tangential flow (atf) systems were compared and process applications of high cell density perfusion were studied here: mab production and cryopreservation. operations by perfusion using microfiltration (mf) or ultrafiltration (u ...201723436783
host cell protein dynamics in recombinant cho cells: impacts from harvest to purification and beyond.during the production of recombinant protein products, such as monoclonal antibodies, manufacturers must demonstrate clearance of host cell impurities and contaminants to appropriate levels prior to use in the clinic. these include host cell dna and rna, product related contaminants such as aggregates, and importantly host cell proteins (hcps). despite the importance of hcp removal, the identity and dynamics of these proteins during cell culture and downstream processing (dsp) are largely unknow ...201723328085
characterization of monoclonal antibody size variants containing extra light chains.size exclusion chromatography (sec) is the most commonly used method to separate and quantify monoclonal antibody (mab) size variants. mab-a is an igg1 subtype humanized monoclonal antibody recombinantly produced in chinese hamster ovary (cho) cells. sec analysis of mab-a resolved a peak, named peak 1, which elutes between monomer and dimer peaks. mab-a lots produced from different clones and production scales all have 0.2-0.3% of sec peak 1. electron spray ionization--time of flight mass spectr ...201723255003
identification of a single base-pair mutation of taa (stop codon) → gaa (glu) that causes light chain extension in a cho cell derived igg1.we describe here the identification of a stop codon taa (stop) → gaa (glu) = stop221e mutation on the light chain of a recombinant igg1 antibody expressed in a chinese hamster ovary (cho) cell line. the extended light chain variants, which were caused by translation beyond the mutated stop codon to the next alternative in-frame stop codon, were observed by mass spectra analysis. the abnormal peptide peaks present in tryptic and chymotryptic lc-ms peptide mapping were confirmed by n-terminal sequ ...201723018810
toward a bioengineered heparin: challenges and strategies for metabolic engineering of mammalian cells.heparin is the most widely used pharmaceutical to control blood coagulation in modern medicine. a health crisis that took place in 2008 led to a demand for production of heparin from non-animal sources. since chinese hamster ovary (cho) cells are capable of producing heparan sulfate (hs), a related polysaccharide naturally, and heparin and hs share the same biosynthetic pathway, we hypothesized that heparin could be produced in cho cells by metabolic engineering. we developed stable human n-deac ...201722714556
expression and biological characterization of an anti-cd20 biosimilar candidate antibody: a case study.the cd20 molecule is a non-glycosylated protein expressed mainly on the surface of b lymphocytes. in some pathogenic b cells, it shows an increased expression, thus becoming an attractive target for diagnosis and therapy. rituximab is a chimeric antibody that specifically recognizes the human cd20 molecule. this antibody is indicated for the treatment of non-hodgkin lymphomas and autoimmune diseases, such as rheumatoid arthritis and systemic lupus erythematosus. in this work, we describe the sta ...201722647435
genomics in mammalian cell culture bioprocessing.explicitly identifying the genome of a host organism including sequencing, mapping, and annotating its genetic code has become a priority in the field of biotechnology with aims at improving the efficiency and understanding of cell culture bioprocessing. recombinant protein therapeutics, primarily produced in mammalian cells, constitute a $108 billion global market. the most common mammalian cell line used in biologic production processes is the chinese hamster ovary (cho) cell line, and althoug ...201722079893
evidence for a mammalian late-g1 phase inhibitor of replication licensing distinct from geminin or cdk activity.pre-replication complexes (pre-rcs) are assembled onto dna during late mitosis and g1 to license replication origins for use in s phase. in order to prevent re-replication of dna, licensing must be completely shutdown prior to entry into s phase. while mechanisms preventing re-replication during s phase and mitosis have been elucidated, the means by which cells first prevent licensing during late g1 phase are poorly understood. we have employed a hybrid mammalian / xenopus egg extract replicatio ...201721983086
wavelength-dependent backscattering measurements for quantitative real-time monitoring of apoptosis in living cells.apoptosis--programmed cell death--is a cellular process exhibiting distinct biochemical and morphological changes. an understanding of the early morphological changes that a cell undergoes during apoptosis can provide the opportunity to monitor apoptosis in tissue, yielding diagnostic and prognostic information. there is avid interest regarding the involvement of apoptosis in cancer. the initial response of a tumor to successful cancer treatment is often massive apoptosis. current apoptosis dete ...201720059251
characterization of a new muc1 monoclonal antibody (vu-2-g7) directed to the glycosylated pdtr sequence of muc1.a monoclonal antibody (mab), vu-2-g7, was generated against a synthetic 60-mer muc1 triple tandem repeat peptide with n-acetyl-galactosamine (galnac) o-linked to the threonine in the pdtr region of each repeat (3m galnac). vu-2-g7 and 8 muc1 mabs (vu-3-c6, vu-4-h5, 139h2, a76-a/c7, vu-12-e1, bcp9, mf11 and bw835) were tested against various glycosylated and nonglycosylated muc1 tandem repeat peptides. vu-2-g7 showed strong reactivity with its immunogen, 3m galnac, and much lower reactivity with ...201710867613
modulative effects of metabolic effectors on benzo(a)pyrene-induced cytotoxicity and mutagenicity in mammalian cells.conjugation and detoxification of mixed function oxidase (mfo)-mediated benzo(a)pyrene [b(a)p] metabolites with glucuronic acid and glutathione (gsh) are major pathways of b(a)p elimination and ultimately excretion in vivo. we have studied the effects of uridine diphosphate alpha-d-glucuronic acid (udpga) and gsh, a cofactor for the synthesis of glucuronide and gsh conjugates, respectively, on b(a)p-induced cytotoxicity and mutagenicity in mammalian cells. the s9-mix used in the chinese hamster ...20177855867
influence of protein/glycan interaction on site-specific glycan heterogeneity.to study how the interaction between n-linked glycans and the surrounding amino acids influences oligosaccharide processing, we used protein disulfide isomerase (pdi), a glycoprotein bearing 5 n-glycosylation sites, as a model system and expressed it transiently in a chinese hamster ovary (cho)-s cell line. pdi was produced as both secreted sec-pdi and endoplasmic reticulum-retained glycoprotein (er)-pdi, to study glycan processing by er and golgi resident enzymes. quantitative site-specific gly ...201728679530
generation of induced pluripotent stem cells from chinese hamster embryonic fibroblasts.we derived a stable cell line from chinese hamster embryonic fibroblasts by transduction of four mouse transcription factors (m3o, sox2, klf4, and n-myc) using a lentiviral vector. the cell line possess all the characteristics of an induced pluripotent stem cell (ipsc) line. given that chinese hamster ovary (cho) cells are the predominant host cells used for therapeutic protein production and no pluripotent stem cell line or other normal cell line has been isolated from chinese hamster, this ips ...201728677528
clinicopathological implications to micropapillary bladder urothelial carcinoma of the presence of sialyl lewis x-decorated mucin 1 in stroma-facing membranes.bladder urothelial carcinoma (uc) comprises more than 90% of all bladder cancers. among several uc variants, micropapillary uc (mpuc) is a rare one with high potential for lymphovascular invasion and subsequent lymph node metastasis. histologically, mpuc is characterized by the presence of small papillary carcinoma cell clusters surrounded by lacunar spaces. immunohistochemically, the outer circumference of these clusters, that is, the stroma-facing membrane of carcinoma cells, is reportedly alm ...201728666720
antipodocalyxin antibody chpcmab-47 exerts antitumor activity in mouse xenograft models of colorectal adenocarcinomas.podocalyxin (podxl) is expressed in several cancers, including brain tumors and colorectal cancers. podxl overexpression is an independent predictor of progression, metastasis, and poor outcome. we recently immunized mice with recombinant human podxl, which was produced using ln229 glioblastoma cells, and produced a clone pcmab-47 that could be used for investigating podxl expression by flow cytometry and immunohistochemical analysis. herein, we produced a human-mouse chimeric pcmab-47 (chpcmab- ...201728665782
acute toxicity and genotoxicity of fermented traditional medicine oyaksungi-san.the traditional medicine oyaksungi-san (oy) has been prescribed in east asia for hundreds of years for the treatment of stroke, paralysis, and ataxia. oy also has therapeutic effects on arthralgia, myalgia, and rheumatoid arthritis, and recent studies have shown its protective effects against apoptosis of hippocampal cells and its anti-inflammatory effects on the peripheral blood cells of patient with cerebral infarction. many studies have explored the use of traditional medicine and herb materi ...201728664144
accumulative scfv-fc antibody gene integration into the hprt chromosomal locus of chinese hamster ovary cells.we have previously developed an accumulative site-specific gene integration system (agis) using cre-recombinase and mutated loxp sites. agis enables repeated transgene integration into a predetermined chromosomal site in mammalian cells. however, the process of establishing cells with multiple integrated copies of the transgene is still time-consuming. in the present study, we describe an improved version of agis that facilitates and accelerates the establishment of high-producer chinese hamster ...201728662917
monoclonal antibodies expression improvement in cho cells by piggybac transposition regarding vectors ratios and design.establishing stable chinese hamster ovary (cho) cells producing monoclonal antibodies (mabs) usually pass through the random integration of vectors to the cell genome, which is sensitive to gene silencing. one approach to overcome this issue is to target a highly transcribed region in the genome. transposons are useful devices to target active parts of genomes, and piggybac (pb) transposon can be considered as a good option. in the present study, three pb transposon donor vectors containing both ...201728662065
a comprehensive model for heat-induced radio-sensitisation.combined radiotherapy (rt) and hyperthermia (ht) treatments may improve treatment outcome by heat induced radio-sensitisation. we propose an empirical cell survival model (alphar model) to describe this multimodality therapy. the model is motivated by the observation that heat induced radio-sensitisation may be explained by a reduction in the dna damage repair capacity of heated cells. we assume that this repair is only possible up to a threshold level above which survival will decrease exponent ...201728641499
impact of the niemann-pick c1 gene mutation on the total cellular glycomics of cho cells.niemann-pick disease type c (npc) is an autosomal recessive lipid storage disorder, and the majority of cases are caused by mutations in the npc1 gene. in this study, we clarified how a single gene mutation in the npc1 gene impacts the cellular glycome by analyzing the total glycomic expression profile of chinese hamster ovary cell mutants defective in the npc1 gene (npc1 ko cho cells). a number of glycomic alterations were identified, including increased expression of lactosylceramide, gm1, gm2 ...201728628327
proteomic analysis of host cell protein dynamics in the supernatant of fc-fusion protein-producing cho dg44 and dukx-b11 cell lines in batch and fed-batch cultures.chinese hamster ovary (cho) cells are the most widely used host cell lines for the commercial production of therapeutic proteins including fc-fusion proteins. during the culture of recombinant cho (rcho) cells, host cell proteins (hcps), secreted from viable cells and released from dead cells, accumulate extracellularly, potentially impairing product quality. in this study, the hcps that accumulated extracellularly in batch and fed-batch cultures of fc-fusion protein-producing rcho cell lines (d ...201728627725
distinct cellular phenotype linked to defective dna interstrand crosslink repair and homologous recombination.repair of dna interstrand crosslinks (icls) predominantly involves the fanconi anemia (fa) pathway and homologous recombination (hr). the hr repair system eliminates dna double strand breaks (dsbs) that emerge during icls removal. the current study presents a novel cell line, cl‑v8b, representing a new complementation group of chinese hamster cell mutants hypersensitive to dna crosslinking factors. cl‑v8b exhibits increased sensitivity to various dna‑damaging agents, including compounds leading ...201728627616
rapid protein production from stable cho cell pools using plasmid vector and the cumate gene-switch.to rapidly produce large amounts of recombinant proteins, the generation of stable chinese hamster ovary (cho) cell pools represents a useful alternative to large-scale transient gene expression (tge). we have developed a cell line (chobri/rcta) allowing the inducible expression of recombinant proteins, based on the cumate gene switch. after the identification of optimal plasmid dna topology (supercoiled vs linearized plasmid) for peipro™ mediated transfection and of optimal conditions for methi ...201728625678
short-term cooling increases serum triglycerides and small high-density lipoprotein levels in humans.cold exposure and β3-adrenergic receptor agonism, which both activate brown adipose tissue, markedly influence lipoprotein metabolism by enhancing lipoprotein lipase-mediated catabolism of triglyceride-rich lipoproteins and increasing plasma high-density lipoprotein (hdl) levels and functionality in mice. however, the effect of short-term cooling on human lipid and lipoprotein metabolism remained largely elusive.201728625343
[the establishment and identification of stable cho cell line with canine transferrin receptor expression].to construct eukaryotic expressing recombinant vector of canine transferrin receptor gene (tfr ), then to transfect chinese hamster ovary (cho) cells with the vector for establishment of stable expression of tfr in cho cell line.201728612559
low-density lipoprotein receptor-related protein-1 (lrp1) c4408r mutant promotes amyloid precursor protein (app) α-cleavage in vitro.previous studies have demonstrated that the low-density lipoprotein receptor-related protein-1 (lrp1) plays conflicting roles in alzheimer's disease (ad) pathogenesis, clearing β-amyloid (aβ) from the brain while also enhancing app endocytosis and resultant amyloidogenic processing. we have recently discovered that co-expression of mutant lrp1 c-terminal domain (lrp1-ct c4408r) with swedish mutant amyloid precursor protein (appswe) in chinese hamster ovary (cho) cells decreases aβ production, wh ...201728612181
differential killing and radio-modifying effects of iodoacetate in mammalian normal and cancer cells.to explore possible applications of iodoacetate (ia), a glycolytic inhibitor, in cancer treatment, we screened its cytotoxicity and radioprotective/sensitizing efficacy in three different mammalian cell lines; a549 (human lung carcinoma), mcf7 (human mammary cancer), a non-cancerous cho (chinese hamster ovary) cells and human lymphocytes. experiments were carried out using ia concentrations ranging from 0.01 to 2.5 µg/ml, with or without 60coγ-radiation. in the outcomes, ia was found to exhibit ...201728612110
variability among tsh measurements can be reduced by combining a glycoengineered calibrator to epitope-defined immunoassays.measuring protein markers with variable glycosylation, such as thyroid-stimulating hormone (tsh), with high accuracy is not an easy task. despite highly sensitive third-generation tests, discrepancies among tsh assays still remain unsolved and are the focus of important standardization efforts. earlier work from our group showed that a lack of similarity in epitope expression between standards and samples may account for discordant hormone measurements. in this study, we aimed at producing a gly ...201728611942
danirixin: a reversible and selective antagonist of the cxc chemokine receptor 2.cxc chemokine receptor 2 (cxcr2) is a key receptor in the chemotaxis of neutrophils to sites of inflammation. the studies reported here describe the pharmacological characterization of danirixin, a cxcr2 antagonist in the diaryl urea chemical class. danirixin has high affinity for cxcr2, with a negative log of the 50% inhibitory concentration (pic50) of 7.9 for binding to chinese hamster ovary cell (cho)-expressed human cxcr2, and 78-fold selectivity over binding to cho-expressed cxcr1. danirixi ...201728611093
chomine: an integrated data warehouse for cho systems biology and modeling.the last decade has seen a surge in published genome-scale information for chinese hamster ovary (cho) cells, which are the main production vehicles for therapeutic proteins. while a single access point is available at www.chogenome.org, the primary data is distributed over several databases at different institutions. currently research is frequently hampered by a plethora of gene names and ids that vary between published draft genomes and databases making systems biology analyses cumbersome and ...201728605771
extracellular signal-regulated kinase 5 associates with casein kinase ii to regulate gpib-ix-mediated platelet activation via the pten/pi3k/akt pathway.essentials the mechanisms of extracellular signal-regulated kinase 5 (erk5) in gpib-ix signaling are unclear. function of erk5 in gpib-ix was tested using aggregation, western blotting, and mass spectrometry. the protein interacting with erk5 in human platelets was identified as casein kinase ii (ckii). erk5 associates with ckii to regulate the activation of the pi3k/akt pathway in gpib-ix signaling.201728603902
activation of unfolded protein response pathway is important for valproic acid mediated increase in immunoglobulin g productivity in recombinant chinese hamster ovary cells.process engineering to improve product quality and titers is gaining importance at late-stage cell culture process development. valproic acid, a us food and drug administration-approved histone deacetylase (hdac) inhibitor, has been shown to improve cell culture performance with higher productivities and minimal effect on the product quality. however, the wider physiological impact of valproic acid on recombinant cells has not been investigated till date. in this study, we investigate the role o ...201728601608
investigation of the relative biological effectiveness and uniform isobiological killing effects of irradiation with a clinical carbon sobp beam on dna repair deficient cho cells.spread-out bragg peak (sobp) c ions have been used clinically in charged particle radiation therapy for years. an sobp beam consists of various monoenergetic bragg peaks; however, the biological effect of irradiation with an sobp beam track has not been well-studied. in order to determine the clinically prospective molecular targets, radiosensitivity to the beam track in dna repair deficient cell lines was investigated. a total of four distinct chinese hamster ovary (cho) cell lines, including c ...201728599495
a new fully human recombinant fsh (follitropin epsilon): two phase i randomized placebo and comparator-controlled pharmacokinetic and pharmacodynamic trials.what are the differences and similarities of pharmacokinetic (pk) and pharmacodynamic (pd) characteristics of the novel recombinant human fsh follitropin epsilon expressed in the human cell line glycoexpress compared with a chinese hamster ovary (cho) derived compound and a urinary derived product?201728591833
characterization of a novel modification of a cho-produced mab: evidence for the presence of tyrosine sulfation.herein we describe the investigation of a chinese hamster ovary (cho)-expressed human mab molecule found partially modified by a +80 da adduct. this mass difference, suggestive of a single sulfation or phosphorylation addition, was observed by mass analysis of the intact and reduced molecule by mass spectrometry (ms). the modification was located on tyrosine 31 (y31) of the light chain in the complementarity-determining region 1 by liquid chromatography (lc)-ms peptide mapping and electron trans ...201728590151
the β4 subunit of the voltage-gated calcium channel (cacnb4) regulates the rate of cell proliferation in chinese hamster ovary cells.the β subunits of voltage-gated calcium channel (vgcc) are cytosolic proteins that interact with the vgcc pore -forming subunit and participate in the trafficking of the channel to the cell membrane and in ion influx regulation. β subunits also exert functions independently of their binding to vgcc by translocation to the cell nucleus including the control of gene expression. mutations of the neuronal cacnb4 (β4) subunit are linked to human neuropsychiatric disorders including epilepsy and intel ...201728587927
new mammalian expression systems.there are an increasing number of recombinant antibodies and proteins in preclinical and clinical development for therapeutic applications. mammalian expression systems are key to enabling the production of these molecules, and chinese hamster ovary (cho) cell platforms continue to be central to delivery of the stable cell lines required for large-scale production. increasing pressure on timelines and efficiency, further innovation of molecular formats and the shift to new production systems are ...201728585079
impact of s-sulfocysteine on fragments and trisulfide bond linkages in monoclonal antibodies.the quality of recombinant proteins such as monoclonal antibodies produced using chinese hamster ovary cell-based mammalian systems is dependent on many factors, including cell line, process and cell culture media. due to these factors, the generated product is heterogeneous and may have chemically-induced modifications or post-translational modifications that affect antibody stability, functionality and, in some cases, patient safety. this study demonstrates that s-sulfocysteine, a cysteine der ...201728581887
identification of fc gamma receptor glycoforms that produce differential binding kinetics for rituximab.fc gamma receptors (fcγr) bind the fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as adcc, cdc and adcp. the immune effector cell activity is directly linked to a productive molecular engagement of fcγrs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the fcγr glycans in this interaction. we provide a complete description of the glycan ...201728576848
orexin receptor agonist yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity.two promising lead structures of small molecular orexin receptor agonist have been reported, but without detailed analyses of the pharmacological properties. one of them, 1-(3,4-dichlorophenyl)-2-[2-imino-3-(4-methylbenzyl)-2,3-dihydro-1h-benzo[d]imidazol-1-yl]ethan-1-ol (yan 7874), is commercially available, and we set out to analyze its properties. as test system we utilized human ox1 and ox2 orexin receptor-expressing chinese hamster ovary (cho) k1 cells as well as control cho-k1 and neuro-2a ...201728575023
myeloma-derived light chain paired with a diagnostic monoclonal antibody hinders immunoassay performance.monoclonal antibodies are widely used as the capture and detection reagents in diagnostic immunoassays. in the past, myeloma fusion partners expressing endogenous heavy and/or light chains were often used to generate hybridoma cell lines. as a result, mixed populations of antibodies were produced that can cause inaccurate test results, poor antibody stability, and significant lot-to-lot variability. we describe one such scenario where the p3u1 (p3x63ag8u.1) myeloma fusion partner was used in the ...201728557609
the ef-1α promoter maintains high-level transgene expression from episomal vectors in transfected cho-k1 cells.in our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (mars) and containing the cytomegalovirus (cmv) promoter allow transgenes to be maintained episomally in chinese hamster ovary (cho) cells. however, the transgene expression was unstable and the number of copies was low. in this study, we focused on enhancers, various promoters and promoter variants that could improve the transgene expression stability, expression magnit ...201728557288
development of a 3-step straight-through purification strategy combining membrane adsorbers and resins.the pressures to efficiently produce complex biopharmaceuticals at reduced costs are driving the development of novel techniques, such as in downstream processing with straight-through processing (stp). this method involves directly and sequentially purifying a particular target with minimal holding steps. this work developed and compared six different 3-step stp strategies, combining membrane adsorbers, monoliths, and resins, to purify a large, complex, and labile glycoprotein from chinese hams ...201728556541
co-overexpression of mgat1 and mgat4 in cho cells for production of highly sialylated albumin-erythropoietin.terminal sialic acids on n-glycan of recombinant human erythropoietin are very important for in vivo half-life, as this glycoprotein has three n-glycosylation sites. n-acetylglucosaminyltransferases i, ii, iv, and v (i.e. mgat1, mgat2, mgat4, and mgat5) catalyze the formation of a glycan antennary structure. these enzymes display different reaction kinetics for a common substrate and generally show low expression in chinese hamster ovary (cho) cells. therefore, genetic control of mgat expression ...201728554385
mammalian systems biotechnology reveals global cellular adaptations in a recombinant cho cell line.effective development of host cells for therapeutic protein production is hampered by the poor characterization of cellular transfection. here, we employed a multi-omics-based systems biotechnology approach to elucidate the genotypic and phenotypic differences between a wild-type and recombinant antibody-producing chinese hamster ovary (cho) cell line. at the genomic level, we observed extensive rearrangements in specific targeted loci linked to transgene integration sites. transcriptional re-wi ...201728544881
kir2.1 and k2p1 channels reconstitute two levels of resting membrane potential in cardiomyocytes.outward and inward background currents across the cell membrane balance, determining resting membrane potential. inward rectifier k+ channel subfamily 2 (kir2) channels primarily maintain the resting membrane potential of cardiomyocytes. human cardiomyocytes exhibit two levels of resting membrane potential at subphysiological extracellular k+ concentrations or pathological hypokalaemia, however, the underlying mechanism is unclear. in the present study, we show that human cardiomyocytes derived ...201728543529
the ik1/kir2.1 channel agonist zacopride prevents and cures acute ischemic arrhythmias in the rat.arrhythmogenesis in acute myocardial infarction (mi) is associated with depolarization of resting membraine potential (rmp) and decrease of inward rectifier potassium current (ik1) in cardiomyocytes. however, clinical anti-arrhythmic agents that primarily act on rmp by enhancing the ik1 channel are not currently available. we hypothesized that zacopride, a selective and moderate agonist of the ik1/kir2.1 channels, prevents and cures acute ischemic arrhythmias. to test this viewpoint, adult sprag ...201728542320
cellular response to exponentially increasing and decreasing dose rates: implications for treatment planning in targeted radionuclide therapy.the treatment of cancer using targeted radionuclide therapy is of interest to nuclear medicine and radiation oncology because of its potential for killing tumor cells while minimizing dose-limiting toxicities to normal tissue. the ionizing radiations emitted by radiopharmaceuticals deliver radiation absorbed doses over protracted periods of time with continuously varying dose rates. as targeted radionuclide therapy becomes a more prominent part of cancer therapy, accurate models for estimating t ...201728541775
Refresh
firstprev
...11121314151617181920...
nextlast
Displaying items 1201 - 1300 of 26566