Publications

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cellular engineering for therapeutic protein production: product quality, host modification, and process improvement.recombinant proteins offer many therapeutic advantages unavailable in traditional small molecule drugs, but the need for cellular versus chemical synthesis complicates production. avenues for producing therapeutic biologics are continuously expanding, and developments in biochemistry, cell biology, and bioengineering fuel new discoveries that promise safer, more efficient, and cheaper drugs for consumers. numerous approaches to express recombinant proteins exist, but escherichia coli, saccharomy ...201727933725
genotoxicity evaluation of hwanglyeonhaedok-tang, an herbal formula.hwanglyeonhaedok-tang (huanglianjiedu-tang, orengedoku-to), a traditional herbal formula, is used for the treatment of inflammatory, gastrointestinal and cardiovascular diseases.201727916588
identification of human junctional adhesion molecule 1 as a functional receptor for the hom-1 calicivirus on human cells.the hom-1 vesivirus was reported in 1998 following the inadvertent transmission of the animal calicivirus san miguel sea lion virus to a human host in a laboratory. we characterized the hom-1 strain and investigated the mechanism by which human cells could be infected. an expression library of 3,559 human plasma membrane proteins was screened for reactivity with hom-1 virus-like particles, and a single interacting protein, human junctional adhesion molecule 1 (hjam1), was identified. transient e ...201728196955
vesivirus 2117 capsids more closely resemble sapovirus and lagovirus particles than other known vesivirus structures.vesivirus 2117 is an adventitious agent that, in 2009, was identified as a contaminant of chinese hamster ovary cells propagated in bioreactors at a pharmaceutical manufacturing plant belonging to genzyme. the consequent interruption in supply of fabrazyme and cerezyme (drugs used to treat fabry and gaucher diseases, respectively) caused significant economic losses. vesivirus 2117 is a member of the caliciviridae, a family of small icosahedral viruses encoding a positive-sense rna genome. we hav ...201727902397
in vitro protective effects of botryosphaeran, a (1→3;1→6)-β-d-glucan, against mutagens in normal and tumor rodent cells.botryosphaeran (bot) is an exocellular β-d-glucan (carbohydrate biopolymer) of the (1→3;1→6)-linked type produced by botryosphaeria rhodina mamb-05. the cytotoxic, mutagenic, genotoxic, and protective effects of this substance were evaluated in chinese hamster lung fibroblasts (v79) and rat hepatocarcinoma cells (htc) by the micronucleus test (mn) and the comet assay. bot was not genotoxic in either cell line; it decreased the clastogenic effects of doxorubicin, h2o2, and benzo[a]pyrene. these r ...201728137365
genetic engineering of cho cells for viral resistance to minute virus of mice.contamination by the parvovirus minute virus of mice (mvm) remains a challenge in chinese hamster ovary (cho) biopharmaceutical production processes. although infrequent, infection of a bioreactor can be catastrophic for a manufacturer, can impact patient drug supply and safety, and can have regulatory implications. we evaluated engineering a cho parental cell line (chozn(®) gs(-/-) ) to create a new host cell line that is resistant to mvm infection by modifying the major receptors used by the v ...201727642072
murine leukemia virus-derived retroviral vector has differential integration patterns in human cell lines used to produce recombinant factor viii.nowadays recombinant factor viii is produced in murine cells including in chinese hamster ovary (cho) and baby hamster kidney cells (bhk). previous studies, using the murine leukemia virus-derived retroviral vector pmfg-fviii-p140k, modified two recombinant human cell lines, hepg2 and hek293 to produce recombinant factor viii. in order to characterize these cells, the present study aimed to analyze the integration pattern of retroviral vector pmfg-fviii-p140k.201725031062
biogenesis of silver nanoparticles using endophytic fungus pestalotiopsis microspora and evaluation of their antioxidant and anticancer activities.an endophytic fungal strain isolated from the leaves of gymnema sylvestre was identified as pestalotiopsis microspora vj1/vs1 based on nucleotide sequencing of internal transcribed spacer region (its 1-5.8s-its 2) of 18s rrna gene (ncbi accession number kx213894). in this study, an efficient and ecofriendly approach has been reported for the synthesis of silver nanoparticles (agnps) using aqueous culture filtrate of p. microspora. ultraviolet-visible analysis confirmed the synthesis of agnps by ...201727826190
network reconstruction of the mouse secretory pathway applied on cho cell transcriptome data.protein secretion is one of the most important processes in eukaryotes. it is based on a highly complex machinery involving numerous proteins in several cellular compartments. the elucidation of the cell biology of the secretory machinery is of great importance, as it drives protein expression for biopharmaceutical industry, a 140 billion usd global market. however, the complexity of secretory process is difficult to describe using a simple reductionist approach, and therefore a promising avenue ...201728298216
novel citronellyl-based photoprobes designed to identify er proteins interacting with dolichyl phosphate in yeast and mammalian cells.dolichyl phosphate-linked mono- and oligosaccharides (dlo) are essential intermediates in protein n-glycosylation, c- and o-mannosylation and gpi anchor biosynthesis. while many membrane proteins in the endoplasmic reticulum (er) involved in the assembly of dlos are known, essential proteins believed to be required for the transbilayer movement (flip-flopping) and proteins potentially involved in the regulation of dlo synthesis remain to be identified.201727099830
characterization of membrane occupation and recognition nexus repeat containing 3, meiosis expressed gene 1 binding partner, in mouse male germ cells.mammalian spermatogenesis is a well-organized process of cell development and differentiation. meiosis expressed gene 1 (meig1) plays an essential role in the regulation of spermiogenesis. to explore potential mechanisms of meig1's action, a yeast two-hybrid screen was conducted, and several potential binding partners were identified; one of them was membrane occupation and recognition nexus repeat containing 3 (morn3). morn3 mrna is only abundant in mouse testis. in the testis, morn3 mrna is hi ...201725248657
enhancing the toolbox to study il-17a in cattle and sheep.the development of methods to detect cytokine expression by t cell subsets in ruminants is fundamental to strategic development of new livestock vaccines for prevention of infectious diseases. it has been possible to detect t cell expression of ifn-γ, il-4 and il-10 in ruminants for many years but methods to detect expression of il-17a are relatively limited. to address this gap in capability we have cloned bovine and ovine il-17a cdnas and expressed biologically-active recombinant proteins in c ...201728388924
optimizing conditions for calcium phosphate mediated transient transfection.calcium phosphate mediated transfection has been used for delivering dna into mammalian cells in excess of 30 years due to its most low cost for introducing recombinant dna into culture cells. however, multiple factors affecting the transfect efficiency are commonly recognized meanwhile for years, the low transfection efficiency of this approach on higher differentiated and non-tumor cells such as cho and c2c12 limits its application on research.201728386188
novel selective β1-adrenoceptor antagonists for concomitant cardiovascular and respiratory disease.β-blockers reduce mortality and improve symptoms in people with heart disease; however, current clinically available β-blockers have poor selectivity for the cardiac β1-adrenoceptor (ar) over the lung β2-ar. unwanted β2-blockade risks causing life-threatening bronchospasm and reduced efficacy of β2-agonist emergency rescue therapy. thus, current life-prolonging β-blockers are contraindicated in patients with both heart disease and asthma. here, we describe ndd-713 and -825, novel highly β1-selec ...201728400472
impact of anatase titanium dioxide nanoparticles on mutagenic and genotoxic response in chinese hamster lung fibroblast cells (v-79): the role of cellular uptake.the unique physico-chemical properties of nano crystalline anatase titanium dioxide nanoparticles (tio2 nps) render them with different biological and chemical activities. hence, it is widely used in industrial and consumer applications. previous studies have shown the genotoxicity of tio2 nps. however, there is a paucity of data regarding mutagenicity of these nps. in the present study, the cellular uptake, sub-cellular localization, cytotoxicity and short term dna interaction of tio2 nps (1-10 ...201728400324
phic31 integrase can improve the efficiency of different construct designs for monoclonal antibody expression in cho cells.several types of expression vectors have been used for recombinant protein expression in chinese hamster ovary cells (cho) which usually result in variable and unstable levels of expression.201728400295
peroxisome biogenesis: a novel inducible pex19 splicing variant is involved in early stages of peroxisome proliferation.pex19p harbouring a prenylation caax box functions as a chaperone and transporter for peroxisomal membrane proteins in membrane assembly. by functional phenotype-complementation assay using a pex19 chinese hamster ovary cell mutant zp119, we herein cloned a rat cdna encoding a protein similar to pex19p, but with a c-terminal hydrophobic segment in place of the caax box region. the transcript of this gene was highly induced by treatment of rats with a peroxisome proliferator, clofibrate, hence te ...201728391327
the short neuropeptide f modulates olfactory sensitivity of bactrocera dorsalis upon starvation.the insect short neuropeptide f (snpf) family has been shown to modulate diverse physiological processes, such as feeding, appetitive olfactory behavior, locomotion, sleep homeostasis and hormone release. in this study, we identified the snpf (bdsnpf) and its receptor (bdsnpfr) in an important agricultural pest, the oriental fruit fly bactrocera dorsalis (hendel). afterwards, the receptor cdna was functionally expressed in chinese hamster ovary cell lines. activation of bdsnpfr by snpf peptides ...201728377097
arabinosylation of recombinant human immunoglobulin-based protein therapeutics.protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (n-linked), or serine/threonine-linked (o-linked) glycans. in n-linked glycans, fucose levels have b ...201728375048
dna double-strand breaks induced byfractionated neutron beam irradiation for boron neutron capture therapy.to use the 53bp1 foci assay to detect dna double-strand breaks induced by fractionated neutron beam irradiation of normal cells.201728373429
mir-143 targets mapk7 in cho cells and induces a hyperproductive phenotype to enhance production of difficult-to-express proteins.in recent years, the number of complex but clinically effective biologicals such as multi-specific antibody formats and fusion proteins has increased dramatically. however, compared to classical monoclonal antibodies (mabs), these rather artificially designed therapeutic proteins have never undergone millions of years of evolution and thus often turn out to be difficult-to-express using mammalian expression systems such as chinese hamster ovary (cho) cells. in order to provide access to these so ...201728371547
a strategy to accelerate protein production from a pool of clones in chinese hamster ovary cells for toxicology studies.in the biopharmaceutical industry, a clonally derived cell line is typically used to generate material for investigational new drug (ind)-enabling toxicology studies. the same cell line is then used to generate material for clinical studies. if a pool of clones can be used to produce material for ind-enabling toxicology studies ("pool for tox strategy") during the time a lead clone is being selected for clinical material production, the toxicology studies can be accelerated significantly (approx ...201728371489
low glucose concentrations within typical industrial operating conditions have minimal effect on the transcriptome of recombinant cho cells.typically, mammalian cell culture medium contains high glucose concentrations that are analogous to diabetic levels in humans, suggesting that mammalian cells are cultivated in excessive glucose. using rna-seq, this study characterized the chinese hamster ovary (cho) cell transcriptome under two glucose concentrations to assess the genetic effects associated with metabolic pathways, in addition to other global responses. the initial extracellular glucose concentrations used represented high (30 ...201728371311
investigation of toxicity and mutagenicity of cold atmospheric argon plasma.cold atmospheric argon plasma is recognized as a new contact free approach for the decrease of bacterial load on chronic wounds in patients. so far very limited data are available on its toxicity and mutagenicity on eukaryotic cells. thus, the toxic/mutagenic potential of cold atmospheric argon plasma using the microplaster β(®) , which has been used efficiently in humans treating chronic and acute wounds, was investigated using the xtt assay in keratinocytes and fibroblasts and the hgprt (hypox ...201728370324
n-terminal or signal peptide sequence engineering prevents truncation of human monoclonal antibody light chains.monoclonal antibodies (mabs) contain short n-terminal signal peptides on each individual polypeptide that comprises the mature antibody, targeting them for export from the cell in which they are produced. the signal peptide is cleaved from each heavy chain (hc) and light chain (lc) polypeptide after translocation to the er and prior to secretion. this process is generally highly efficient, producing a high proportion of correctly cleaved hc and lc polypeptides. however, mis-cleavage of the signa ...201728369727
segmentation and quantitative analysis of apoptosis of chinese hamster ovary cells from fluorescence microscopy images.accurate and fast quantitative analysis of living cells from fluorescence microscopy images is useful for evaluating experimental outcomes and cell culture protocols. an algorithm is developed in this work to automatically segment and distinguish apoptotic cells from normal cells. the algorithm involves three steps consisting of two segmentation steps and a classification step. the segmentation steps are: (i) a coarse segmentation, combining a range filter with a marching square method, is used ...201728367787
the hybrid search: a mass spectral library search method for discovery of modifications in proteomics.we present a mass spectral library-based method to identify tandem mass spectra of peptides that contain unanticipated modifications and amino acid variants. we describe this as a "hybrid" method because it combines matching both ion m/z and mass losses. the mass loss is the difference between the mass of an ion peak and the mass of its precursor. this difference, termed deltamass, is used to shift the product ions in the library spectrum that contain the modification, thereby allowing library p ...201728367633
label-free protein quantification of sodium butyrate treated cho cells by esi-uhr-tof-ms.effects of butyrate on cho producer cells are contradictory, promoting productivity and at the same time repressing proliferation. though in previous omics studies the background of butyrate impact on producer cells has been investigated, the knowledge about the mechanism is still very limited. as previous proteomic results on this field are mainly based on 2de-gels, we conducted a label-free ms quantification, based on fast high resolution esi-ms and a straight forward software solution, to gai ...201728363874
production of recombinant human acid α-glucosidase with high-mannose glycans in gnt1 rice for the treatment of pompe disease.lysosomal storage diseases are a group of inherited metabolic disorders. patients are treated with enzyme replacement therapy (ert), in which the replacement enzymes are required to carry terminal mannose or mannose 6-phosphate residues to allow efficient uptake into target cells and tissues. n-acetylglucosaminyltransferase-i (gnti) mediates n-glycosylation in the cis cisternae of the golgi apparatus by adding n-acetylglucosamine to the exposed terminal mannose residue of core n-glycan structure ...201728363873
development of a cho-based cell-free platform for synthesis of active monoclonal antibodies.chinese hamster ovary (cho) cells are routinely optimized to stably express monoclonal antibodies (mabs) at high titers. at the early stages of lead isolation and optimization, hundreds of sequences for the target protein of interest are screened. typically, cell-based transient expression technology platforms are used for expression screening, but these can be time- and resource-intensive. here, we have developed a cell-free protein synthesis (cfps) platform utilizing a commercially available c ...201728350472
antioxidant and cytoprotective effects of morin against hydrogen peroxide-induced oxidative stress are associated with the induction of nrf-2‑mediated ho-1 expression in v79-4 chinese hamster lung fibroblasts.natural phytochemicals of plant origin, including flavonoids, have been found to be potent antioxidants providing beneficial effects against oxidative stress-related diseases. the present study was carried out to investigate the antioxidant properties of morin, a flavonoid originally isolated from the flowering plants of the moraceae family. superoxide dismutase (sod)‑like activity and 2,2'‑azino‑bis‑(3‑ethylbenzothiazoline‑6‑sulfonic acid) (abts•+) radical scavenging activity were determined. w ...201728350063
a high density cho-s transient transfection system: comparison of expicho and expi293.chinese hamster ovary (cho) cells are the principal mammalian host used for stable cell line generation and biotherapeutic protein production. until recently, production of milligrams to grams of protein in cho transient systems was challenging. as such, human embryonic kidney (hek293) cells are the most common mammalian cell type used for transient transfection. the post-translational modifications (ptms) of a protein are dictated in part by the cell line used for expression, and changes in ptm ...201728342833
studies to examine potential tolerability differences between the 5-ht2c receptor selective agonists lorcaserin and cp-809101.lorcaserin (lor) is a selective 5-ht2c receptor agonist that has been fda approved as a treatment for obesity. the most frequently reported side-effects of lor include nausea and headache, which can be dose limiting. we have previously reported that in the rat, while lor produced unconditioned signs characteristic of nausea/malaise, the highly selective 5-ht2c agonist cp-809101 (cp) produced fewer equivalent signs. because this may indicate a subclass of 5-ht2c agonists having better tolerabilit ...201728338324
an aromatic amino acid within intracellular loop 2 of the prostaglandin ep2 receptor is a prerequisite for selective association and activation of gαs.we previously demonstrated that the aromatic moiety of tyr(143) within the intracellular loop 2 (icl2) region of the prostaglandin ep2 receptor plays a crucial role in gs coupling. here we investigated whether the icl2 of the ep2 receptor directly binds to gαs and whether an aromatic moiety affects this interaction. in chinese hamster ovary cells, mutations of tyr(143) reduced the ability of the ep2 receptor to interact with g proteins as demonstrated by gtpγs sensitivity, as well as the ability ...201728336329
antitumor activity of chlpmab-2, a human-mouse chimeric cancer-specific antihuman podoplanin antibody, via antibody-dependent cellular cytotoxicity.human podoplanin (hpdpn), a platelet aggregation-inducing transmembrane glycoprotein, is expressed in different types of tumors, and it binds to c-type lectin-like receptor 2 (clec-2). the overexpression of hpdpn is involved in invasion and metastasis. anti-hpdpn monoclonal antibodies (mabs) such as nz-1 have shown antitumor and antimetastatic activities by binding to the platelet aggregation-stimulating (plag) domain of hpdpn. recently, we developed a novel mouse anti-hpdpn mab, lpmab-2, using ...201728332312
transcriptomic changes in cho cells after adaptation to suspension growth in protein-free medium analysed by a species-specific microarray.chinese hamster ovary (cho) cells are the preferred cell line for production of biopharmaceuticals. these cells are capable to grow without serum supplementation, but drastic changes in their phenotype occur during adaptation to protein-free growth, which typically include the change to a suspension phenotype with reduced growth rate. a possible approach to understand this transformation, with the intention to counteract the reduction in growth by targeted supplementation of protein-free media, ...201728302587
evaluation of two public genome references for chinese hamster ovary cells in the context of rna-seq based gene expression analysis.rna-seq is a powerful transcriptomics tool for mammalian cell culture process development. successful rna-seq data analysis requires a high quality reference for read mapping and gene expression quantification. currently, there are two public genome references for chinese hamster ovary (cho) cells, the predominant mammalian cell line in the biopharmaceutical industry. in this study, we compared these two references by analyzing 60 rna-seq samples from a variety of cho cell culture conditions. am ...201728295162
copper(i)-phosphine polypyridyl complexes: synthesis, characterization, dna/hsa binding study, and antiproliferative activity.a series of copper(i)-phosphine polypyridyl complexes have been investigated as potential antitumor agents. the complexes [cu(pph3)2dpq]no3 (2), [cu(pph3)2dppz]no3 (3), [cu(pph3)2dppa]no3 (4), and [cu(pph3)2dppme]no3 (5) were synthesized by the reaction of [cu(pph3)2no3] with the respective planar ligand under mild conditions. these copper complexes were fully characterized by elemental analysis, molar conductivity, fab-ms, and nmr, uv-vis, and ir spectroscopies. interactions between these coppe ...201728290674
dielectrophoresis study of temporal change in internal conductivity of single cho cells after electroporation by pulsed electric fields.applying sufficiently strong pulsed electric fields to a cell can permeabilize the membrane and subsequently affect its dielectric properties. in this study, we employ a microfluidic dielectrophoresis cytometry technique to simultaneously electroporate and measure the time-dependent dielectric response of single chinese hamster ovary cells. using experimental measurements along with numerical simulations, we present quantitative results for the changes in the cytoplasm conductivity of single cel ...201728289483
multiple expression cassette exchange via tp901-1, r4, and bxb1 integrase systems on a mouse artificial chromosome.the site-specific excision of a target dna sequence for genetic knockout or lineage tracing is a powerful tool for investigating biological systems. currently, site-specific recombinases (ssrs), such as cre or flp recombination target cassettes, have been successfully excised or inverted by a single ssr to regulate transgene expression. however, the use of a single ssr might restrict the complex control of gene expression. this study investigated the potential for expanding the multiple regulati ...201728286726
high glucose enhances camp level and extracellular signal-regulated kinase phosphorylation in chinese hamster ovary cell: usage of br-camp in foreign protein β-galactosidase expression.glucose is a carbon source for chinese hamster ovary (cho) cell growth, while low growth rate is considered to enhance the production of recombinant proteins. the present study reveals that glucose concentrations higher than 1 g/l reduce the growth rate and substantially increase in camp (∼300%) at a high glucose concentration (10 g/l). high glucose also enhances the phosphorylation of extracellular signal-regulated kinase (erk) and p27(kip) by western blot analysis. to determine whether the pho ...201728286121
seroprevalence for 2117-like vesiviruses in italian household dogs.in 2003, a novel calicivirus, the vesivirus (vev) strain 2117, was identified incidentally as a contaminant in chinese hamster ovary (cho) cell cultures by a german pharmaceutical company. similar contaminations have been documented in three additional episodes, in bio-reactors used for production of recombinant drugs. more, recently 2117-like vevs have also been identified at high prevalence in the stools from asymptomatic kennel dogs and only sporadically in diarrhoeic and healthy household do ...201728284600
cytotoxicities and genotoxicities of cements based on calcium silicate and of dental formocresol.increasing interest is being paid to the toxicities of dental materials. the purpose of this study was to determine the cytotoxicities and genotoxicities of endodontic compounds to chinese hamster ovary (cho-k1) reproductive cells. cultured cho-k1 cells were treated with dental formocresol, two types of calcium hydroxide paste, and two types of mineral trioxide aggregate cement for 24h. a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was performed on each culture, and the m ...201728283090
efficient expression of stable recombinant human insulin-like growth factor-1 fusion with human serum albumin in chinese hamster ovary cells.insulin-like growth factor-1 (igf-1) plays a crucial role in cell development, differentiation, and metabolism, and has been a potential therapeutic agent for many diseases. chinese hamster ovary (cho) cells are widely used for production of recombinant therapeutic proteins, but the expression level of igf-1 in cho cells is very low (1,500 µg/l) and the half-life of igf-1 in blood circulation is only 4.5 min according to previous studies. therefore, igf-1 was fused to long-circulating serum prot ...201728281882
binding and biologic characterization of recombinant human serum albumin-etgfbr2 fusion protein expressed in cho cells.transforming growth factor-β1 (tgf-β1) signaling is involved in cell metabolism, growth, differentiation, carcinoma invasion and fibrosis development, which suggests tgf-β1 can be treated as a therapeutic target extensively. because tgf-β1 receptor type α(tgfbr2) is the directed and essential mediator for tgf-β1 signals, the extracellular domain of tgfbr2 (etgfbr2), binding partner for tgf-β1, has been produced in a series of expression systems to inhibit tgf-β1 signaling. however, etgfbr2 is un ...201728281868
proteomic analysis of host cell protein dynamics in the culture supernatants of antibody-producing cho cells.chinese hamster ovary (cho) cells are the most common cell line used for the production of therapeutic proteins including monoclonal antibodies (mabs). host cell proteins (hcps), secreted and released from lysed cells, accumulate extracellularly during the cultures of recombinant cho (rcho) cells, potentially impairing product quality. in an effort to maintain good mab quality during the cultures, hcps accumulated extracellularly in batch and fed-batch cultures of a mab-producing rcho cell line ...201728281648
enzyme replacement therapy for farber disease: proof-of-concept studies in cells and mice.a series of studies were carried out in farber disease (omim #228000) cells and mice to evaluate the feasibility of enzyme replacement therapy (ert) for this disorder. media from chinese hamster ovary (cho) cells overexpressing human recombinant acid ceramidase (rhac) was used to treat fibroblasts from a farber disease patient, leading to significantly reduced ceramide. we also found that chondrocytes from farber disease mice had a markedly abnormal chondrogenic phenotype, and this was corrected ...201728275553
antioxidant effect of thiazolidine molecules in cell culture media improves stability and performance.the ability of cell culture media components to generate reactive species as well as their sensitivity to oxidative degradation, affects the overall stability of media and the behavior of cells cultured in vitro. this study investigates the influence of thiazolidine molecules, formed from the condensation between cysteine and alpha-ketoacids, on the stability of these complex mixtures and on the performance of cell culture processes aiming to produce therapeutically relevant monoclonal antibodie ...201728268250
optimizing cell-free protein expression in cho: assessing small molecule mass transfer effects in various reactor configurations.cell-free protein synthesis (cfps) is an ideal platform for rapid and convenient protein production. however, bioreactor design remains a critical consideration in optimizing protein expression. using turbo green fluorescent protein (tgfp) as a model, we tracked small molecule components in a chinese hamster ovary (cho) cfps system to optimize protein production. here, three bioreactors in continuous-exchange cell-free (cecf) format were characterized. a gfp optical sensor was built to monitor t ...201728266026
understanding of decreased sialylation of fc-fusion protein in hyperosmotic recombinant chinese hamster ovary cell culture: n-glycosylation gene expression and n-linked glycan antennary profile.to understand the effects of hyperosmolality on protein glycosylation, recombinant chinese hamster ovary (rcho) cells producing the fc-fusion protein were cultivated in hyperosmolar medium resulting from adding nacl (415 mosm/kg). the hyperosmotic culture showed increased specific fc-fusion protein productivity (qfc ) but a decreased proportion of acidic isoforms and sialic acid content of the fc-fusion protein. the intracellular and extracellular sialidase activities in the hyperosmotic culture ...201728266015
mirna engineering of cho cells facilitates production of difficult-to-express proteins and increases success in cell line development.in recent years, coherent with growing biologics portfolios also the number of complex and thus difficult-to-express (dte) therapeutic proteins has increased considerably. dte proteins challenge bioprocess development and can include various therapeutic protein formats such as monoclonal antibodies (mabs), multi-specific affinity scaffolds (e.g. bispecific antibodies), cytokines or fusion proteins. hence, the availability of robust and versatile chinese hamster ovary (cho) host cell factories is ...201728262952
a role of corazonin receptor in larval-pupal transition and pupariation in the oriental fruit fly bactrocera dorsalis (hendel) (diptera: tephritidae).corazonin (crz) is a neuropeptide hormone, but also a neuropeptide modulator that is internally released within the cns, and it has a widespread distribution in insects with diverse physiological functions. here, we identified and cloned the cdnas of bactrocera dorsalis that encode crz and its receptor crzr. mature bdcrz has 11 residues with a unique ser(11) substitution (instead of the typical asn) and a his in the evolutionary variable position 7. the bdcrzr cdna encodes a putative protein of ...201728261106
screening for small molecule inhibitors of statin-induced app c-terminal toxic fragment production.alzheimer's disease (ad) is characterized by neuronal and synaptic loss. one process that could contribute to this loss is the intracellular caspase cleavage of the amyloid precursor protein (app) resulting in release of the toxic c-terminal 31-amino acid peptide app-c31 along with the production of appδc31, full-length app minus the c-terminal 31 amino acids. we previously found that a mutation in app that prevents this caspase cleavage ameliorated synaptic loss and cognitive impairment in a mu ...201728261092
gold nanoparticles and electroporation impose both separate and synergistic radiosensitizing effects in ht-29 tumor cells: an in vitro study.radiation therapy (rt) is the gold standard treatment for more than half of known tumors. despite recent improvements in rt efficiency, the side effects of ionizing radiation (ir) in normal tissues are a dose-limiting factor that restricts higher doses in tumor treatment. one approach to enhance the efficiency of rt is the application of radiosensitizers to selectively increase the dose at the tumor site. gold nanoparticles (gnps) and electroporation (ep) have shown good potential as radiosensit ...201728260889
a systematic approach to time-series metabolite profiling and rna-seq analysis of chinese hamster ovary cell culture.chinese hamster ovary (cho) cells are the primary host used for biopharmaceutical protein production. the engineering of cho cells to produce higher amounts of biopharmaceuticals has been highly dependent on empirical approaches, but recent high-throughput "omics" methods are changing the situation in a rational manner. omics data analyses using gene expression or metabolite profiling make it possible to identify key genes and metabolites in antibody production. systematic omics approaches using ...201728252038
the enhancement of antibody concentration and achievement of high cell density cho cell cultivation by adding nucleoside.recently, with the dramatic increase in demand for therapeutic antibodies, chinese hamster ovary (cho) cell culture systems have made significant progress in recombinant antibody production. over the past two decades, recombinant antibody productivity has been improved by more than 100-fold. medium optimization has been identified as an important key approach for increasing product concentrations. in this study, we evaluated the effects of deoxyuridine addition to fed-batch cultures of antibody- ...201728251404
effects of lysosomal biotherapeutic recombinant protein expression on cell stress and protease and general host cell protein release in chinese hamster ovary cells.recombinant human acid alpha glucosidase (gaa) is the therapeutic enzyme used for the treatment of pompe disease, a rare genetic disorder characterized by gaa deficiency in the cell lysosomes (raben et al., curr mol med. 2002; 2:145-166). the manufacturing process for gaa can be challenging, in part due to protease degradation. the overall goal of this study was to understand the effects of gaa overexpression on cell lysosomal phenotype and host cell protein (hcp) release, and any resultant cons ...201728249362
antibody purification from cho cell supernatant using new multimodal membranes.this contribution describes strategies to purify monoclonal antibodies from chinese hamster ovary (cho) cell culture supernatant using newly designed multimodal membranes (mmms). the mmms were used for the capture step purification of human igg1 following a size-exclusion desalting column to remove chaotropic salts that interfere with igg binding. the mmm column attained higher dynamic binding capacity than a protein a resin column at an equivalent residence time of 1 min. the two-step mmm chrom ...201728248007
an automated micro-total immunoassay system for measuring cancer-associated α2,3-linked sialyl n-glycan-carrying prostate-specific antigen may improve the accuracy of prostate cancer diagnosis.the low specificity of the prostate-specific antigen (psa) for early detection of prostate cancer (pca) is a major issue worldwide. the aim of this study to examine whether the serum pca-associated α2,3-linked sialyl n-glycan-carrying psa (s2,3psa) ratio measured by automated micro-total immunoassay systems (μtas system) can be applied as a diagnostic marker of pca. the μtas system can utilize affinity-based separation involving noncovalent interaction between the immunocomplex of s2,3psa and ma ...201728241428
antiglycopeptide mouse monoclonal antibody lpmab-21 exerts antitumor activity against human podoplanin through antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity.the interaction between podoplanin (pdpn) and c-type lectin-like receptor 2 (clec-2) is involved in tumor malignancy. we have established many monoclonal antibodies (mabs) against human podoplanin using the cancer-specific mab (casmab) technology. lpmab-21, one of the mouse antipodoplanin mabs, is of the igg2a subclass, and its minimum epitope was determined to be thr76-arg79 of the human podoplanin. importantly, sialic acid is linked to thr76; therefore, lpmab-21 is an antiglycopeptide mab (gpm ...201728234556
what can mathematical modelling say about cho metabolism and protein glycosylation?chinese hamster ovary cells have been in the spotlight for process optimization in recent years, due to being the major, long established cell factory for the production of recombinant proteins. a deep, quantitative understanding of cho metabolism and mechanisms involved in protein glycosylation has proven to be attainable through the development of high throughput technologies. here we review the most notable accomplishments in the field of modelling cho metabolism and protein glycosylation.201728228925
evaluating the expression profile and stability of different ucoe containing vector combinations in mab-producing cho cells.as the demand for monoclonal antibodies (mab) increases, more efficient expression methods are required for their manufacturing process. transcriptional gene silencing is a common phenomenon in recombinant cell lines which leads to expression reduction and instability. there are reports on improved antibody expression in ubiquitous chromatin opening element (ucoe) containing both heavy and light chain gene constructs. here we investigate the impact of having these elements as part of the light c ...201728228095
how important is thermodynamics for identifying elementary flux modes?we present a method for computing thermodynamically feasible elementary flux modes (tefms) using equilibrium constants without need of internal metabolite concentrations. the method is compared with the method based on a binary distinction between reversible and irreversible reactions. when all reactions are reversible, adding the constraints based on equilibrium constants reduces the number of elementary flux modes (efms) by a factor of two. declaring in advance some reactions as irreversible, ...201728222104
interaction of approved drugs with synaptic vesicle protein 2a.levetiracetam (lev) and its recently approved derivative brivaracetam are anti-epileptic drugs with a unique mechanism of action. the synaptic vesicle protein 2a (sv2a) was previously identified as their main target. in the current study, we tested a collection of 500 approved drugs for interaction with the human sv2a protein expressed in chinese hamster ovary cells. competition binding studies were performed using cell lysates with high sv2a expression and [(3) h]brivaracetam as a radioligand. ...201728220535
glycoengineering of esterase activity through metabolic flux-based modulation of sialic acid.this report describes the metabolic glycoengineering (mge) of intracellular esterase activity in human colon cancer (ls174t) and chinese hamster ovary (cho) cells. in silico analysis of the carboxylesterases ces1 and ces2 suggested that these enzymes are modified with sialylated n-glycans, which are proposed to stabilize the active multimeric forms of these enzymes. this premise was supported by treating cells with butanolylated mannac to increase sialylation, which in turn increased esterase ac ...201728218815
unveiling gene trait relationship by cross-platform meta-analysis on chinese hamster ovary cell transcriptome.in the past few years, transcriptome analysis has been increasingly employed to better understand the physiology of chinese hamster ovary (cho) cells at a global level. as more transcriptome data accumulated, meta-analysis on data sets collected from various sources can potentially provide better insights on common properties of those cells. here, we performed meta-analysis on transcriptome data of different cho cell lines obtained using nimblegen or affymetrix microarray platforms. hierarchical ...201728218403
manipulating cholesterol status within cells.cellular cholesterol levels are intricately controlled to maintain homeostasis. here, we describe ways in which cellular cholesterol status can be manipulated for the study of cholesterol homeostasis, including sterol starvation (by culturing cells in lipoprotein-deficient serum and pretreating/treating with the cholesterol-lowering drug, statin) and sterol enrichment (using cholesterol complexed to cyclodextrin, and low-density lipoprotein). we also describe how to prepare lipoprotein-deficient ...201728205165
antioxidant and cytoprotective effects of morin against hydrogen peroxide-induced oxidative stress are associated with the induction of nrf-2‑mediated ho-1 expression in v79-4 chinese hamster lung fibroblasts.natural phytochemicals of plant origin, including flavonoids, have been found to be potent antioxidants providing beneficial effects against oxidative stress-related diseases. the present study was carried out to investigate the antioxidant properties of morin, a flavonoid originally isolated from the flowering plants of the moraceae family. superoxide dismutase (sod)‑like activity and 2,2'‑azino‑bis‑(3‑ethylbenzothiazoline‑6‑sulfonic acid) (abts•+) radical scavenging activity were determined. w ...201728204816
distinct c-mannosylation of netrin receptor thrombospondin type 1 repeats by mammalian dpy19l1 and dpy19l3.thrombospondin type 1 repeats (tsrs) occur in diverse proteins involved in adhesion and signaling. the two extracellular tsrs of the netrin receptor unc5a contain wxxwxxwxxc motifs that can be c-mannosylated on all tryptophans. a single c-mannosyltransferase (dumpy-19, dpy-19), modifying the first two tryptophans, occurs in caenorhabditis elegans, but four putative enzymes (dpy-19-like 1-4, dpy19l1-4) exist in mammals. single and triple crispr-cas9 knockouts of the three homologs that are expres ...201728202721
multiple blood-brain barrier transport mechanisms limit bumetanide accumulation, and therapeutic potential, in the mammalian brain.there is accumulating evidence that bumetanide, which has been used over decades as a potent loop diuretic, also exerts effects on brain disorders, including autism, neonatal seizures, and epilepsy, which are not related to its effects on the kidney but rather mediated by inhibition of the neuronal na-k-cl cotransporter isoform nkcc1. however, following systemic administration, brain levels of bumetanide are typically below those needed to inhibit nkcc1, which critically limits its clinical use ...201728192112
magnetic sorting of membrane associated igg for phenotype-based selection of stable antibody producing cells.to establish a simple and widely accessible technique for rapidly selecting high producing chinese hamster ovary (cho) cells engineered to express a monoclonal antibody (mab), we have exploited the transient display of recombinant protein on their cell surface. in combination with magnetic bead-based methods, we demonstrate the ability to select for cells of high productivity in the absence of any metabolic-based selection method. this technique is sufficient to obtain genetically stable enginee ...201728189705
fatty-acyl chain profiles of cellular phosphoinositides.phosphoinositides are rapidly turning-over phospholipids that play key roles in intracellular signaling and modulation of membrane effectors. through technical refinements we have improved sensitivity in the analysis of the phosphoinositide pi, pip, and pip2 pools from living cells using mass spectrometry. this has permitted further resolution in phosphoinositide lipidomics from cell cultures and small samples of tissue. the technique includes butanol extraction, derivatization of the lipids, po ...201728189644
ultra-deep next generation mitochondrial genome sequencing reveals widespread heteroplasmy in chinese hamster ovary cells.recent sequencing of the chinese hamster ovary (cho) cell and chinese hamster genomes has dramatically advanced our ability to understand the biology of these mammalian cell factories. in this study, we focus on the powerhouse of the cho cell, the mitochondrion. utilizing a high-resolution next generation sequencing approach we sequenced the chinese hamster mitochondrial genome for the first time and surveyed the mutational landscape of cho cell mitochondrial dna (mtdna). depths of coverage rang ...201728188893
bioreactor scale up and protein product quality characterization of piggybac transposon derived cho pools.chinese hamster ovary (cho) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mabs), bispecific antibodies, and fc-fusion proteins. creating and characterizing the stable cho clonally-derived cell lines (cdcls) needed to manufacture these therapeutic proteins is a lengthy and laborious process. therefore, cho pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development. we recen ...201728188692
evaluation of heavy chain c-terminal deletions on productivity and product quality of monoclonal antibodies in chinese hamster ovary (cho) cells.monoclonal antibodies (mabs) have been well established as potent therapeutic agents and are used to treat many different diseases. during cell culture production, antibody charge variants can be generated by cleavage of heavy chain (hc) c-terminal lysine and proline amidation. differences in levels of charge variants during manufacturing process changes make it challenging to demonstrate process comparability. in order to reduce heterogeneity and achieve consistent product quality, we generated ...201728188688
proteomics profiling of chimeric-truncated tissue plasminogen activator producing- chinese hamster ovary cells cultivated in a chemically defined medium supplemented with protein hydrolysatesculture media enrichment through the addition of protein hydrolysates is beneficial for achieving higher protein expression.201728187683
site-specific monitoring of n-glycosylation profiles of a ctla4-fc-fusion protein from the secretory pathway to the extracellular environment.glycosylation often plays a key role in the safety and efficacy of therapeutic proteins to patients, thus underlying the need for consistent control of this important post-translational modification during biologics production. in this study, we profiled the site-specific evolution of n-glycans on a ctla4-fc-fusion protein, from the intracellular secretory pathway to the conditioned medium (cm) in fed-batch cell culture. for this, we developed an approach that combined sub-cellular fractionation ...201728186328
fabrication of electrospun hpgl scaffolds via glycidyl methacrylate cross-linker: morphology, mechanical and biological properties.electrospinning is a suitable method to produce scaffolds composed of nanoscale to microscale fibers, which are comparable to the extracellular matrix (ecm). hyperbranched polyglycerol (hpgl) is a highly biocompatible polyether polyol potentially useful for the design of fibrous scaffolds mimicking the ecm architecture. however, scaffolds developed from hpgl have poor mechanical properties and morphological stability in the aqueous environments required for tissue engineering applications. this ...201728183666
method comparison for analyzing wound healing rates.wound healing scratch assay is a frequently used method to characterize cell migration, which is an important biological process in the course of development, tissue repair, and immune response for example. the measurement of wound healing rate, however, varies among different studies. here we summarized these measurements into three types: i) direct rate average; ii) regression rate average; and iii) average distance regression rate. using chinese hamster ovary (cho) cells as a model, we compar ...201728177756
the vdac2-bak axis regulates peroxisomal membrane permeability.peroxisomal biogenesis disorders (pbds) are fatal genetic diseases consisting of 14 complementation groups (cgs). we previously isolated a peroxisome-deficient chinese hamster ovary cell mutant, zp114, which belongs to none of these cgs. using a functional screening strategy, vdac2 was identified as rescuing the peroxisomal deficiency of zp114 where vdac2 expression was not detected. interestingly, knockdown of bak or overexpression of the bak inhibitors bcl-xl and mcl-1 restored peroxisomal bio ...201728174205
small-molecule-biased formyl peptide receptor agonist compound 17b protects against myocardial ischaemia-reperfusion injury in mice.effective treatment for managing myocardial infarction (mi) remains an urgent, unmet clinical need. formyl peptide receptors (fpr) regulate inflammation, a major contributing mechanism to cardiac injury following mi. here we demonstrate that fpr1/fpr2-biased agonism may represent a novel therapeutic strategy for the treatment of mi. the small-molecule fpr1/fpr2 agonist, compound 17b (cmpd17b), exhibits a distinct signalling fingerprint to the conventional fpr1/fpr2 agonist, compound-43 (cmpd43). ...201728169296
high-resolution imaging and quantification of plasma membrane cholesterol by nanosims.cholesterol is a crucial lipid within the plasma membrane of mammalian cells. recent biochemical studies showed that one pool of cholesterol in the plasma membrane is "accessible" to binding by a modified version of the cytolysin perfringolysin o (pfo*), whereas another pool is sequestered by sphingomyelin and cannot be bound by pfo* unless the sphingomyelin is destroyed with sphingomyelinase (smase). thus far, it has been unclear whether pfo* and related cholesterol-binding proteins bind unifor ...201728167768
atf6β-based fine-tuning of the unfolded protein response enhances therapeutic antibody productivity of chinese hamster ovary cells.the dynamics of protein folding and secretion are key issues in improving the productivity and robustness of chinese hamster ovary (cho) producer cells. high recombinant protein secretion in cho producer clones triggers the activation of the unfolded protein response (upr), an intracellular response to the accumulation of unfolded and misfolded proteins in the endoplasmic reticulum (er). we previously reported that the human microrna (mirna) mir-1287 enhances productivity in igg expressing cho c ...201728165157
the fibrous form of intracellular inclusion bodies in recombinant variant fibrinogen-producing cells is specific to the hepatic fibrinogen storage disease-inducible variant fibrinogen.fibrinogen storage disease (fsd) is a rare disorder that is characterized by the accumulation of fibrinogen in hepatocytes and induces liver injury. six mutations in the γc domain (γg284r, γt314p, γd316n, the deletion of γg346-q350, γg366s, and γr375w) have been identified for fsd. our group previously established γ375w fibrinogen-producing chinese hamster ovary (cho) cells and observed aberrant large granular and fibrous forms of intracellular inclusion bodies. the aim of this study was to inve ...201728161763
a vector based on the chicken hypersensitive site 4 insulator element replicates episomally in mammalian cell.gene therapy in mammalian cells requires vectors exhibiting long-term stability and high expression. episomal gene expression vectors offer a safe and attractive alternative to those that integrate into the host cell genome. in the present study, we developed a new episomal vector based on the insulator, chicken hypersensitive site 4 (chs4). the chs4 element was artificially synthesized, cloned into the pegfp-c1 vector, and used to transfect chinese hamster ovary (cho) and human chang liver cell ...201728155604
dedicated high dose rate (192)ir brachytherapy radiation fields for in vitro cell exposures at variable source-target cell distances: killing of mammalian cells depends on temporal dose rate fluctuation.afterloading brachytherapy is conducted by the stepwise movement of a radioactive source through surgically implanted applicator tubes where at predefined dwell positions calculated dwell times optimize spatial dose delivery with respect to a planned dose level. the temporal exposure pattern exhibits drastic fluctuations in dose rate at a given coordinate and within a single treatment session because of the discontinuous and repeated source movement into the target volume. this could potentially ...201728145285
engineering the expression of an anti-interleukin-13 antibody through rational design and mutagenesis.high levels of protein expression are key to the successful development and manufacture of a therapeutic antibody. here, we describe two related antibodies, ab001 and ab008, where ab001 shows a markedly lower level of expression relative to ab008 when stably expressed in chinese hamster ovary cells. we use single-gene expression vectors and structural analysis to show that the reduced titer is associated with the vl cdr2 of ab001. we adopted two approaches to improve the expression of ab001. fir ...201728130326
cell-controlled hybrid perfusion fed-batch cho cell process provides significant productivity improvement over conventional fed-batch cultures.a simple method originally designed to control lactate accumulation in fed-batch cultures of chinese hamster ovary (cho) cells has been modified and extended to allow cells in culture to control their own rate of perfusion to precisely deliver nutritional requirements. the method allows for very fast expansion of cells to high density while using a minimal volume of concentrated perfusion medium. when the short-duration cell-controlled perfusion is performed in the production bioreactor and is i ...201728128436
taming hyperactive hdnase i: stable inducible expression of a hyperactive salt- and actin-resistant variant of human deoxyribonuclease i in cho cells.while the most common causes of clonal instability are dna copy number loss and silencing, toxicity of the expressed protein(s) may also induce clonal instability. human dnase i (hdnase i) is used therapeutically for the treatment of cystic fibrosis (cf) and may have potential benefit for use in systemic lupus erythematosus (sle). hdnase i is an endonuclease that catalyzes degradation of extracellular dna and is inhibited by both salt and g-actin. engineered versions of hdnase i, bearing multipl ...201728127892
application of (13)c flux analysis to identify high-productivity cho metabolic phenotypes.industrial bioprocesses place high demands on the energy metabolism of host cells to meet biosynthetic requirements for maximal protein expression. identifying metabolic phenotypes that promote high expression is therefore a major goal of the biotech industry. we conducted a series of (13)c flux analysis studies to examine the metabolic response to igg expression during early stationary phase of cho cell cultures grown in 3l fed-batch bioreactors. we examined eight clones expressing four differe ...201728122259
a novel approach to characterize host cell proteins associated with therapeutic monoclonal antibodies.recombinant monoclonal antibody (mab) products are widely produced in the pharmaceutical industry using chinese hamster ovary (cho) cells. host cell proteins (hcps) are one of many process-related impurities generated during the production of mab products. the multi-analyte hcp enzyme linked immunosorbent assay (elisa) is the industry standard accepted assay to measure clearance of hcps from recombinant protein therapeutics. while similar platform processes are used for expression and purificati ...201728112396
expression and characterization of a codon-optimized blood coagulation factor viii.essentials recombinant factor viii (fviii) is known to be expressed at a low level in cell culture. to increase expression, we used codon-optimization of a b-domain deleted fviii (bdd-fviii). this resulted in 7-fold increase of the expression level in cell culture. the biochemical properties of codon-optimized bdd-fviii were similar to the wild-type protein.201728109042
from the cover: an investigation of the genotoxicity and interference of gold nanoparticles in commonly used in vitro mutagenicity and genotoxicity assays.the suitability of 4 in vitro assays, commonly used for mutagenicity and genotoxicity assessment, was investigated in relation to treatment with 14 nm citrate-stabilized gold nanoparticles (aunps). specifically, the ames test was conducted without metabolic activation, where no mutagenic effects were observed. high resolution transmission electron microscopy and cytoviva dark-field image analysis showed that aunps did not enter the bacterial cells, thus confirming the unreliability of the ames t ...201728108664
a highly efficient modified human serum albumin signal peptide to secrete proteins in cells derived from different mammalian species.signal peptides (sps) are key elements in the production of recombinant proteins; however, little information is available concerning different sp in mammalian cells other than cho. in order to study the efficiency of different sps to direct the traffic along the secretory pathway of the green fluorescence protein (gfp) and a scfv-fc fusion protein; cho-k1, hek293 and ns0 cell lines were transfected in a transient and stable way. sp of human azurocidin (az), modified human albumin (msa), modifie ...201728089882
secretory pathway optimization of cho producer cells by co-engineering of the mitosrna-1978 target genes cers2 and tbc1d20.chinese hamster ovary (cho) cells are the most commonly used host for the production of biopharmaceuticals. although transcription and translation engineering strategies have been employed to generate high-producer cell clones, the secretory pathway still remains a bottleneck in cellular productivity. in this study we show that ectopic expression of a human mitochondrial genome-encoded small rna (mitosrna-1978) in an igg expressing cho cell line strongly improved specific productivity by functio ...201728088541
adverse and beneficial effects in chinese hamster lung fibroblast cells following radiofrequency exposure.in this study, the effect of radiofrequency (rf) exposure to 1950 mhz, universal mobile telecommunication system signal, was investigated in chinese hamster lung fibroblast cell line (v79). genotoxic and cytotoxic effects of 20-h exposure at specific absorption rate (sar) values from 0.15 w/kg to 1.25 w/kg were measured by means of cytokinesis-block micronucleus (mn) assay. exposure was carried out blinded under strictly controlled conditions of dosimetry and temperature. the effect of rf exposu ...201728072461
development, qualification, validation and application of the neutral red uptake assay in chinese hamster ovary (cho) cells using a vitrocell® vc10® smoke exposure system.cytotoxicity assessment of combustible tobacco products by neutral red uptake (nru) has historically used total particulate matter (tpm) or solvent captured gas vapor phase (gvp), rather than fresh whole smoke. here, the development, validation and application of the nru assay in chinese hamster ovary (cho) cells, following exposure to fresh whole smoke generated with the vitrocell® vc10® system is described. whole smoke exposure is particularly important as both particulate and vapor phases of ...201728062357
molecular characterizations of natalisin and its roles in modulating mating in the oriental fruit fly, bactrocera dorsalis (hendel).initially, natalisin (ntl) was identified from three holometabolous insect species, drosophila melanogaster, tribolium castaneum and bombyx mori, and was documented to regulate reproductive behaviours in d. melanogaster and t. castaneum. in this study, we report the sequences of the ntl precursor and its receptor (ntlr) from an important agricultural pest, bactrocera dorsalis (hendel). ntlr is a typical g-protein coupled receptor and phylogenetic analysis showed that b. dorsalis ntlr was closely ...201727862548
a semi-empirical glycosylation model of a camelid monoclonal antibody under hypothermia cell culture conditions.the impact of cell culture environment on the glycan distribution of a monoclonal antibody (mab) has been investigated through a combination of experiments and modeling. a newly developed cho duxb cell line was cultivated at two levels of initial glutamine (gln) concentrations (0, 4 mm) and incubation temperatures of (33 and 37 °c) in batch operation mode. hypothermia was applied either through the entire culture duration or only during the post-exponential phase. beyond reducing cell growth and ...201728285402
inhibition of glycosylation on a camelid antibody uniquely affects its fcγri binding activity.glycoengineering of mabs has become common practice in attempts to generate the ideal mab candidate for a wide range of therapeutic applications. the effects of these glycan modifications on the binding affinity of igg mabs for fcγriiia and their cytotoxicity are well known. however, little is understood about the effect that these modifications have on binding to the high affinity fcγri receptor. this study analyzed the effect of variable n-glycosylation on a human-llama hybrid mab (eg2-hfc, 80 ...201727721040
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