| application of imaging flow cytometry for the characterization of intracellular attributes in chinese hamster ovary cell lines at the single-cell level. | biopharmaceutical manufacturing using chinese hamster ovary (cho) cells requires the generation of high-producing clonal cell lines. during cell line development, cell cloning using fluorescence-activated cell sorting (facs) has the potential to combine isolation of single cells with sorting based on specific cellular attributes that correlate with productivity and/or growth, identifying cell lines with desirable phenotypes for manufacturing. this study describes the application of imaging flow ... | 2019 | 30925020 |
| simultaneous detection of nicotinamide adenine nucleotides and adenylate pool to quantify redox and energy states in mab-producing cho cells by capillary electrophoresis. | chinese hamster ovary (cho) cells are predominant in the production of therapeutic proteins to treat various diseases. characterization and investigation of cho cell metabolism in a quick and simple way could boost process and cell line development. therefore, a method to simultaneously detect seven redox- and energy-related metabolites in cho cells by capillary electrophoresis has been developed. an on-line focusing technique was applied to improve the peak shape and resolution by using a 50 μm ... | 2019 | 30923861 |
| crispr/cas9 as a genome editing tool for targeted gene integration in cho cells. | the emergence of crispr/cas9 system as a precise and affordable method for genome editing has prompted its rapid adoption for the targeted integration of transgenes in chinese hamster ovary (cho) cells. targeted gene integration allows the generation of stable cell lines with a controlled and predictable behavior, which is an important feature for the rational design of cell factories aimed at the large-scale production of recombinant proteins. here we present the protocol for crispr/cas9-mediat ... | 2019 | 30912048 |
| bioinformatic analysis of chinese hamster ovary host cell protein lipases. | complete, accurate genome assemblies are necessary to design targets for genetic engineering strategies. successful gene knockdowns and knockouts in chinese hamster ovary (cho) cells may prevent the expression of difficult-to-remove host cell proteins (hcps). hcps, if not removed, can cause problems in stability, safety, and efficacy of the biotherapeutic. a significantly improved chinese hamster (ch) reference genome was used to identify new knockout targets with similar predicted functions and ... | 2018 | 30911190 |
| acute and genetic toxicity of gs-e3d, a new pectin lyase-modified red ginseng extract. | korean red ginseng and its extract have been used as traditional medicines and functional foods in countries worldwide. pectin lyase-modified red ginseng extract (gs-e3d) was newly developed as a dietary supplement for obesity, diabetes-related renal dysfunction, etc. in this study, the safety of gs-e3d on acute toxicity and genotoxicity was evaluated. for acute study, sprague-dawley rats were administrated by oral gavage at a dose of 5000 mg/kg gs-e3d. to evaluate genotoxicity of gs-e3d, we con ... | 2019 | 30904430 |
| identification of mammalian glycoproteins with type-i lacdinac structures synthesized by the glycosyltransferase b3galnt2. | the type-i lacdinac (ldn; galnacβ1-3glcnac) has rarely been observed in mammalian cells except in the o-glycan of α-dystroglycan, in contrast to type-ii ldn structures (galnacβ1-4glcnac) in n- and o-glycans that are present in many mammalian glycoproteins, such as pituitary and hypothalamic hormones. although a β1,3-n-acetylgalactosaminyltransferase 2 (b3galnt2; type-i ldn synthase) has been cloned, the function of type-i ldn in mammalian cells is still unclear, as its carrier protein(s) has not ... | 2019 | 30898876 |
| acaricidal mechanism of scopoletin against tetranychus cinnabarinus. | scopoletin is a promising acaricidal botanical natural compound against tetranychus cinnabarinus, and its acaricidal mechanism maybe involve calcium overload according to our previous study. to seek potential candidate target genes of calcium overload induced by scopoletin in t. cinnabarinus, rna-seq was utilized to detect changes in transcription levels. 24 and 48 h after treatment, 70 and 102 differentially expressed genes were obtained, respectively. target genes included 3 signal transductio ... | 2019 | 30894818 |
| reducing recombinant protein expression during cho pool selection enhances frequency of high-producing cells. | chinese hamster ovary (cho) cells are the most widely used mammalian host for industrial-scale production of monoclonal antibodies (mabs) and other protein biologics. isolation of rare high-producing cho cell lines from heterogeneous populations of stable transfectants is a daunting task and delays the process of manufacturing of novel biologics. a variety of factors that contribute to the low frequency of high-producing clones have been described; however, the impact of metabolic burden and oth ... | 2019 | 30885656 |
| cytoskeletal synchronization of cho cells with polymer functionalized fullerene c60. | recent developments in the field of fullerene c60 and its derivatives suggest its suitability in a wide range of applications ranging from photovoltaic instruments, development of solar based cells, cosmetics to enzyme inhibition treatment, and so on. these innovative applications raised possibilities of intentional or oblivious human-particle contact leading to possible deleterious effects on human health. the current study deals with the interaction of dextran functionalized fullerene c60 (dex ... | 2019 | 30884950 |
| the effects of pertuzumab and its combination with trastuzumab on her2 homodimerization and phosphorylation. | pertuzumab (perjeta) is an anti-her2 monoclonal antibody that is used for treatment of her2-positive breast cancers in combination with trastuzumab (herceptin) and docetaxel and showed promising clinical outcomes. pertuzumab is suggested to block heterodimerization of her2 with egfr and her3 that abolishes canonical function of her2. however, evidence on the exact mode of action of pertuzumab in homodimerization of her2 are limited. in this study, we investigated the effect of pertuzumab and its ... | 2019 | 30884851 |
| reduced apoptosis in chinese hamster ovary cells via optimized crispr interference. | chinese hamster ovary (cho) cells are widely used for biopharmaceutical protein production. one challenge limiting cho cell productivity is apoptosis stemming from cellular stress during protein production. here we applied crispr interference (crispri) to downregulate the endogenous expression of apoptotic genes bak, bax, and casp3 in cho cells. in addition to reduced apoptosis, mitochondrial membrane integrity was improved and the caspase activity was reduced. moreover, we optimized the crispri ... | 2019 | 30883679 |
| impact of depth filtration on disulfide bond reduction during downstream processing of monoclonal antibodies from cho cell cultures. | monoclonal antibody interchain disulfide bond reduction was observed in a chinese hamster ovary manufacturing process that used single-use technologies. a similar reduction has been reported for processes that involved high mechanical shear recovery unit operations, such as continuous flow centrifugation and when the clarified harvest was stored under low dissolved oxygen (do) conditions (trexler-schmidt et al., 2010. biotechnology and bioengineering, 106(3), 452-461). the work described here id ... | 2019 | 30883673 |
| carbosilane dendrimers with phosphonium terminal groups are low toxic non-viral transfection vectors for sirna cell delivery. | non-viral gene delivery vectors studied in the gene therapy applications are often designed with the cationic nitrogen containing groups necessary for binding and cell release of nucleic acids. disadvantage is a relatively high toxicity which restricts the in vivo use of such nanoparticles. here we show, that the 3rd generation carbosilane dendrimers possessing (trimethyl)phosphonium (pme3) groups on their periphery were able to effectively deliver the functional sirna into the cells (b14, crice ... | 2019 | 30877030 |
| progression of change in membrane capacitance and cytoplasm conductivity of cells during controlled starvation using dual-frequency dep cytometry. | the dielectric properties of cells are directly related to their morphological and physiological properties and can be used to monitor their status when exposed to stress conditions. in this work, dual-frequency dielectrophoresis (dep) cytometry was employed to measure changes in the membrane capacitance and cytoplasm conductivity of single chinese hamster ovary (cho) cells during the progression of starvation-induced apoptosis. our dual-frequency dep cytometer enables simultaneous measurement o ... | 2019 | 30876633 |
| the effect of green and black tea polyphenols on brca2 deficient chinese hamster cells by synthetic lethality through parp inhibition. | tea polyphenols are known antioxidants presenting health benefits due to their observed cellular activities. in this study, two tea polyphenols, epigallocatechin gallate, which is common in green tea, and theaflavin, which is common in black tea, were investigated for their parp inhibitory activity and selective cytotoxicity to brca2 mutated cells. the observed cytotoxicity of these polyphenols to brca2 deficient cells is believed to be a result of parp inhibition induced synthetic lethality. ch ... | 2019 | 30875717 |
| virus removal filtration of chemically defined chinese hamster ovary cells medium with nanocellulose-based size exclusion filter. | sterility of bioreactors in biotherapeutic processing remains a significant challenge. virus removal size-exclusion filtration is a robust and highly efficient approach to remove viruses. this article investigates the virus removal capacity of nanocellulose-based filter for upstream bioprocessing of chemically defined chinese hamster ovary (cho) cells medium containing pluronic f-68 (powercho™, lonza) and supplemented with insulin-transferrin-selenium (its) at varying process parameters. virus r ... | 2019 | 30871931 |
| host-antibody inductivity of virulent entamoeba histolytica and non-virulent entamoeba moshkovskii in a mouse model. | despite similarities in morphology, gene and protein profiles, entamoeba histolytica and e. moshkovskii show profound differences in pathogenicity. entamoeba histolytica infection might result in amoebic dysentery and liver abscess, while e. moshkovskii causes only mild diarrhea. extensive studies focus on roles of host immune responses to the pathogenic e. histolytica; however, evidence for e. moshkovskii remains scarce. | 2019 | 30867040 |
| a novel led-based 2d-fluorescence spectroscopy system for in-line bioprocess monitoring of chinese hamster ovary cell cultivations-part ii. | this study was performed in order to evaluate a new led-based 2d-fluorescence spectrometer for in-line bioprocess monitoring of chinese hamster ovary (cho) cell culture processes. the new spectrometer used selected excitation wavelengths of 280, 365, and 455 nm to collect spectral data from six 10-l fed-batch processes. the technique provides data on various fluorescent compounds from the cultivation medium as well as from cell metabolism. in addition, scattered light offers information about th ... | 2019 | 32625013 |
| both irradiated and bystander effects link with dna repair capacity and the linear energy transfer. | in comparison with a low linear energy transfer (let) radiation, a high-let radiation induces more complex dna damage. this study wonders whether radiation-induced bystander effect (ribe) is dependent of let. | 2019 | 30858123 |
| selection of vaccinia virus-neutralizing antibody from a phage-display human-antibody library. | although smallpox was eradicated in 1980, it is still considered a potential agent of biowarfare and bioterrorism. smallpox has the potential for high mortality rates along with a major public health impact, eventually causing public panic and social disruption. passive administration of neutralizing monoclonal antibodies (mabs) is an effective intervention for various adverse reactions caused by vaccination and the unpredictable nature of emerging and bioterrorist-related infections. currently, ... | 2019 | 30856707 |
| secreting oviduct epithelial cells of coturnix coturnix japonica (qoec) and changes to their proteome after nonviral transfection. | the quail oviduct (coturnix c. japonica) is a natural candidate avian bioreactor, while the secretive quail oviduct epithelial cells (qoecs) are potential in vitro producers of recombinant proteins and vaccines. in view of the need for highly performing and transformable cell lines, qoec may potentially act as an alternative bioreactor platform to the existing ones, for example, to the chinese hamster ovary. the aim of this work was to characterize qoecs and their response to nucleofection with ... | 2019 | 30854717 |
| transient manipulation of the expression level of selected growth rate correlating micrornas does not increase growth rate in cho-k1 cells. | engineering of chinese hamster ovary cells by manipulating microrna (mirna) expression levels has been shown to induce advantageous, desired phenotypes. most of these studies so far were concerned with increasing productivity or reducing growth rate (with the implied intention of thus freeing cellular resources to also increase productivity). here we evaluated the ability of growth correlating mirnas to increase the growth rate of cho-k1 cells by transient overexpression or knock down, respectiv ... | 2019 | 30853633 |
| rapid establishment of stable retroviral packaging cells and recombinant susceptible target cell lines employing novel transposon vectors derived from sleeping beauty. | viral vector particles derived from murine leukemia virus (mlv) mediate highly efficient stable gene transfer used in gene therapeutic approaches and in the generation of transgenic cell lines. however, the establishment of stable viral packaging cells (vpcs) is a time-consuming challenge. to overcome this limitation, we successfully generated novel sleeping beauty-derived transposon vectors entailing envelope and packaging expression cassettes as well as a transfer vector. upon multiplexed tran ... | 2019 | 30852270 |
| metabolic engineering of chinese hamster ovary cells towards reduced biosynthesis and accumulation of novel growth inhibitors in fed-batch cultures. | chinese hamster ovary (cho) cells in fed-batch cultures are known to consume large amounts of nutrients and divert significant portion of them towards the formation of byproducts, some of which, including lactate and ammonia, are known to be growth inhibitory in nature. a major fraction of these inhibitory metabolites are byproducts or intermediates of amino acid catabolism. limiting the supply of amino acids has been shown to curtail the production of corresponding inhibitory byproducts resulti ... | 2019 | 30851381 |
| chemical, cytotoxic and genotoxic analysis of etidronate in sodium hypochlorite solution. | to test whether the incorporation of a chelation powder, etidronate, marketed for root canal irrigation (dual rinse hedp) into a sodium hypochlorite (naocl) solution induced additional cytotoxic and genotoxic effects not observed with naocl alone. | 2019 | 30848496 |
| in vitro and in vivo evaluation of 6-o-α-maltosyl-β-cyclodextrin as a potential therapeutic agent against niemann-pick disease type c. | niemann-pick disease type c (npc) is a rare lysosomal storage disease characterized by the dysfunction of intracellular cholesterol trafficking with progressive neurodegeneration and hepatomegaly. we evaluated the potential of 6-o-α-maltosyl-β-cyclodextrin (g2-β-cd) as a drug candidate against npc. the physicochemical properties of g2-β-cd as an injectable agent were assessed, and molecular interactions between g2-β-cd and free cholesterol were studied by solubility analysis and two-dimensional ... | 2019 | 30845767 |
| crispr/cas9 targeted capture of mammalian genomic regions for characterization by ngs. | the robust detection of structural variants in mammalian genomes remains a challenge. it is particularly difficult in the case of genetically unstable chinese hamster ovary (cho) cell lines with only draft genome assemblies available. we explore the potential of the crispr/cas9 system for the targeted capture of genomic loci containing integrated vectors in cho-k1-based cell lines followed by next generation sequencing (ngs), and compare it to popular target-enrichment sequencing methods and to ... | 2019 | 30837529 |
| induction of peripheral lymph node addressin in human nasal mucosa with eosinophilic chronic rhinosinusitis. | eosinophilic chronic rhinosinusitis (ecrs) is characterised by formation of nasal polyps with prominent eosinophilic infiltration; however, how eosinophils are recruited in this pathological setting remains unclear. in the present study, we carried out quantitative immunohistochemical analysis of nasal polyps associated with ecrs (n=30) and non-ecrs (n=30) to evaluate expression of an l-selectin ligand peripheral lymph node addressin (pnad) on vascular endothelial cells. we found that pnad was i ... | 2019 | 30837082 |
| site-specific roles of n-linked oligosaccharides in recombinant eel follicle-stimulating hormone for secretion and signal transduction. | eel follicle-stimulating hormone (eelfsh) is composed of a common α-subunit and a hormone specific β-subunit, both of which contain two n-linked carbohydrate residues. we characterized the biologically active single chains by fusing the α-subunit to the carboxyl terminal region of the eelfsh β-subunit. expression vectors were constructed and the biological activity of the recombinant hormones (rec-hormones) was characterized using chinese hamster ovary (cho) k1 cells expressing the eelfsh recept ... | 2019 | 30836102 |
| electrophysiological and pharmacological characterization of human inwardly rectifying kir2.1 channels on an automated patch-clamp platform. | inwardly rectifying ik1 potassium currents of the heart control the resting membrane potential of ventricular cardiomyocytes during diastole and contribute to their repolarization after each action potential. mutations in the gene encoding kir2.1 channels, which primarily conduct ventricular ik1, are associated with inheritable forms of arrhythmias and sudden cardiac death. therefore, potential iatrogenic inhibition of kir2.1-mediated ik1 currents is a cardiosafety concern during new drug discov ... | 2019 | 30835490 |
| an lc-ms-based lipidomics pre-processing framework underpins rapid hypothesis generation towards cho systems biotechnology. | given a raw lc-ms dataset, it is often required to rapidly generate initial hypotheses, in conjunction with other 'omics' datasets, without time-consuming lipid verifications. furthermore, for meta-analysis of many datasets, it may be impractical to conduct exhaustive confirmatory analyses. in other cases, samples for validation may be difficult to obtain, replicate or maintain. thus, it is critical that the computational identification of lipids is of appropriate accuracy, coverage, and unbiase ... | 2018 | 30830409 |
| mitochondrial aconitase is a key regulator of energy production for growth and protein expression in chinese hamster ovary cells. | mammalian cells like chinese hamster ovary (cho) cells are routinely used for production of recombinant therapeutic proteins. cells require a continuous supply of energy and nutrients to sustain high cell densities whilst expressing high titres of recombinant proteins. cultured mammalian cells are primarily dependent on glucose and glutamine metabolism for energy production. | 2018 | 30830403 |
| in vitro pharmacological profile of ipragliflozin, a sodium glucose co-transporter 2 inhibitor. | ipragliflozin, a selective sodium glucose cotransporter 2 (sglt2) inhibitor, is used for the treatment of type 2 diabetes mellitus. to date, the only known in vitro pharmacological characteristic of ipragliflozin is its selectivity for sglt2 over sglt1, which was previously reported by our group. therefore, in this study, we investigated other in vitro pharmacological characteristics of ipragliflozin and compared them with those of phlorizin, a naturally occurring sglt inhibitor. selectivity of ... | 2019 | 30828082 |
| the fickle cho: a review of the causes, implications, and potential alleviation of the cho cell line instability problem. | chinese hamster ovary (cho) cell-based bioproduction of recombinant proteins can now routinely achieve >5 g/l titers in fed-batches. this progress is partly due to the rapid adaptability of cho cells to various genetic manipulations and changing process conditions. an inherently plastic genome allows for this adaptability; however, it also gives cho cells the propensity for genomic rearrangements. in combination with the genomic and metabolic demand of high producer cells, cho cell plasticity ma ... | 2019 | 30826670 |
| a statistical approach to improve compound screening in cell culture media. | the chinese hamster ovary (cho) cell line is widely used for the production of recombinant proteins due to its high growing capacity and productivity, as well as other cell lines derived later than cho. adapting cell culture media for each specific cell line is a key to exploit these features for cost effective and fast product generation. media supplementation is generally addressed by means of one-factor-at-a-time or classical design of experiments approaches but these techniques may not be ef ... | 2019 | 32625011 |
| production of recombinant human acid β-glucosidase with high mannose-type n-glycans in rice gnt1 mutant for potential treatment of gaucher disease. | gaucher disease is an inherited metabolic disease caused by genetic acid β -glucosidase (gba) deficiency and is currently treated by enzyme replacement therapy. for uptake into macrophages, gba needs to carry terminal mannose residues on their n-glycans. knockout mutant rice of n-acetylglucosaminyltransferase-i (gnt1) have a disrupted n-glycan processing pathway and produce only glycoproteins with high mannose residues. in this study, we introduced a gene encoding recombinant human gba into both ... | 2019 | 30822514 |
| a modifying autoantigen in graves' disease. | the tsh receptor (tshr) is the major autoantigen in graves' disease (gd). bioinformatic analyses predict the existence of several human tshr isoforms from alternative splicing, which can lead to the coexpression of multiple receptor forms. the most abundant of these is tshrv1.3. in silico modeling of tshrv1.3 demonstrated the structural integrity of this truncated receptor isoform and its potential binding of tsh. tissue profiling revealed wide expression of tshrv1.3, with a predominant presence ... | 2019 | 30822352 |
| organic anion‑transporting polypeptides contribute to the uptake of curcumin and its main metabolites by human breast cancer cells: impact on antitumor activity. | curcumin is a natural polyphenolic compound with pronounced anticancer properties, despite its low bioavailability caused by extensive glucuronidation and sulfation. information on the cellular uptake mechanisms and their contribution to the anticancer effects of curcumin and its biotransformation products is limited. the present study, therefore, investigated the role of organic anion‑transporting polypeptides (oatps) in the cellular uptake of curcumin and its major metabolites in oatp‑expressi ... | 2019 | 30816509 |
| recombinant glycoprotein e of varicella zoster virus contains glycan-peptide motifs that modulate b cell epitopes into discrete immunological signatures. | a recombinant subunit vaccine (shingrix®) was recently licensed for use against herpes zoster. this vaccine is based on glycoprotein e (ge) of varicella zoster virus (vzv), the most abundantly expressed protein of vzv, harboring sites for n- and o-linked glycosylation. the subunit vaccine elicits stronger virus-specific cd4+ t cell response as well as antibody b cell response to ge, compared to the currently used live attenuated vaccine (zostavax®). this situation is at variance with the current ... | 2019 | 30813247 |
| maximum entropy and population heterogeneity in continuous cell cultures. | continuous cultures of mammalian cells are complex systems displaying hallmark phenomena of nonlinear dynamics, such as multi-stability, hysteresis, as well as sharp transitions between different metabolic states. in this context mathematical models may suggest control strategies to steer the system towards desired states. although even clonal populations are known to exhibit cell-to-cell variability, most of the currently studied models assume that the population is homogeneous. to overcome thi ... | 2019 | 30811392 |
| chromatographic purification of recombinant human erythropoietin. | recombinant human erythropoietin is a valuable therapeutic protein used in the treatment of several serious diseases. it exists in different isoforms and is produced by genetically modified mammalian cells such as chinese hamster ovary or human embryonic kidney cells. as for other biopharmaceutical drugs, a key factor for its successful industrial production is to achieve a high degree of purity and to decrease the content of critical impurities to trace amounts. this goal is achieved in the sep ... | 2019 | 30810853 |
| novel column generation-based optimization approach for poly-pathway kinetic model applied to cho cell culture. | mathematical modelling can provide precious tools for bioprocess simulation, prediction, control and optimization of mammalian cell-based cultures. in this paper we present a novel method to generate kinetic models of such cultures, rendering complex metabolic networks in a poly-pathway kinetic model. the model is based on subsets of elementary flux modes (efms) to generate macro-reactions. thanks to our column generation-based optimization algorithm, the experimental data are used to identify t ... | 2019 | 30809468 |
| systematic evaluation of site-specific recombinant gene expression for programmable mammalian cell engineering. | many branches of biology depend on stable and predictable recombinant gene expression, which has been achieved in recent years through targeted integration of the recombinant gene into defined integration sites. however, transcriptional levels of recombinant genes in characterized integration sites are controlled by multiple components of the integrated expression cassette. lack of readily available tools has inhibited meaningful experimental investigation of the interplay between the integratio ... | 2019 | 30807689 |
| model-assisted design of experiments as a concept for knowledge-based bioprocess development. | design of experiments methods offer systematic tools for bioprocess development in quality by design, but their major drawback is the user-defined choice of factor boundary values. this can lead to several iterative rounds of time-consuming and costly experiments. in this study, a model-assisted design of experiments concept is introduced for the knowledge-based reduction of boundary values. first, the parameters of a mathematical process model are estimated. second, the investigated factor comb ... | 2019 | 30806781 |
| crispr/cas9-mediated knockout of microrna-744 improves antibody titer of cho production cell lines. | micrornas (mirnas) are noncoding rnas that serve as versatile molecular engineering tools to improve production cells by overexpression or knockdown of mirnas showing beneficial or adverse effects on cell-culture performance. the genomic knockout (ko) of noncoding rnas in chinese hamster ovary (cho) production cells has not been reported. however, given the significant number of mirnas showing negative effects on cho-bioprocess performance and the development of clustered regularly interspaced s ... | 2019 | 30802343 |
| transcriptomic analysis of igg4 fc-fusion protein degradation in a panel of clonally-derived cho cell lines using rnaseq. | in this study, we report an investigation of a panel of clonally-derived chinese hamster ovary (cho) cell lines exhibiting variability in the proportion of full-length igg4 fc-fusion protein produced. the recombinant protein was found to be degraded during cell culture into four shorter "clipped" species (three of the four cleavage sites occurred at arginine residues) and preliminary analyses suggested that a host cell enzyme was responsible for proteolysis. to identify the specific enzyme respo ... | 2019 | 30802296 |
| model-based optimization of antibody galactosylation in cho cell culture. | exerting control over the glycan moieties of antibody therapeutics is highly desirable from a product safety and batch-to-batch consistency perspective. strategies to improve antibody productivity may compromise quality, while interventions for improving glycoform distribution can adversely affect cell growth and productivity. process design therefore needs to consider the trade-off between preserving cellular health and productivity while enhancing antibody quality. in this work, we present a m ... | 2019 | 30802295 |
| pmab-213: a monoclonal antibody for immunohistochemical analysis against pig podoplanin. | podoplanin (pdpn) is known to be expressed in normal tissues, including lymphatic endothelial cells, renal podocytes, and type i lung alveolar cells. monoclonal antibodies (mabs) against human, mouse, rat, rabbit, dog, cat, and bovine pdpn have already been established; however, mabs against pig pdpn (ppdpn) are lacking. in the present study, mice were immunized with ppdpn-overexpressing chinese hamster ovary (cho)-k1 cells (cho/ppdpn), and hybridomas producing mabs against ppdpn were identified ... | 2019 | 30802179 |
| enhanced cho clone screening: application of targeted locus amplification and next-generation sequencing technologies for cell line development. | early analytical clone screening is important during chinese hamster ovary (cho) cell line development of biotherapeutic proteins to select a clonally derived cell line with most favorable stability and product quality. sensitive sequence confirmation methods using mass spectrometry have limitations in throughput and turnaround time. next-generation sequencing (ngs) technologies emerged as alternatives for cho clone analytics. we report an efficient ngs workflow applying the targeted locus ampli ... | 2019 | 30793505 |
| pmab-219: a monoclonal antibody for the immunohistochemical analysis of horse podoplanin. | monoclonal antibodies (mabs) against human, mouse, rat, rabbit, dog, cat, and bovine podoplanin (pdpn), a lymphatic endothelial cell marker, have been established in our previous studies. however, mabs against horse pdpn (horpdpn), which are useful for immunohistochemical analysis, remain to be developed. in the present study, mice were immunized with horpdpn-overexpressing chinese hamster ovary (cho)-k1 cells (cho/horpdpn), and hybridomas producing mabs against horpdpn were screened using flow ... | 2019 | 30766925 |
| a proteomic profiling dataset of recombinant chinese hamster ovary cells showing enhanced cellular growth following mir-378 depletion. | the proteomic data presented in this article provide supporting information to the related research article "depletion of endogenous mirna-378-3p increases peak cell density of cho dp12 cells and is correlated with elevated levels of ubiquitin carboxyl-terminal hydrolase 14" (costello et al., in press) [1]. control and microrna-378 depleted cho dp12 cells were profiled using label-free quantitative proteomic profiling. cho dp12 cells were collected on day 4 and 8 of batch culture, subcellular pr ... | 2018 | 30761351 |
| investigating the effects of stove emissions on ocular and cancer cells. | more than a third of the world's population relies on solid fuels for cooking and heating, with major health consequences. although solid fuel combustion emissions are known to increase the prevalence of illnesses such as chronic obstructive pulmonary disease and lung cancer, however, their effect on the eyes is underexplored. this study assesses the acute toxicity of solid fuel combustion emissions on healthy ocular cells and a cancer cell line. three healthy ocular cell lines (corneal, lens, a ... | 2019 | 30755694 |
| tag-on-demand: exploiting amber codon suppression technology for the enrichment of high-expressing membrane protein cell lines. | membrane proteins play key roles in the evolution of numerous diseases and as a result have become the most dominant class of targets for therapeutic intervention. however, their poor expression and detection oftentimes prohibit drug discovery and screening efforts. herein, we have developed an approach, named 'tag-on-demand' that exploits amber suppression to control the expression of 'tagged' membrane proteins for detection and selections, yet can be turned off for expression of the protein in ... | 2018 | 30753634 |
| challenging the workhorse: comparative analysis of eukaryotic micro-organisms for expressing monoclonal antibodies. | for commercial protein therapeutics, chinese hamster ovary (cho) cells have an established history of safety, proven capability to express a wide range of therapeutic proteins and high volumetric productivities. expanding global markets for therapeutic proteins and increasing concerns for broadened access of these medicines has catalyzed consideration of alternative approaches to this platform. reaching these objectives likely will require an order of magnitude increase in volumetric productivit ... | 2019 | 30739333 |
| an integrated platform for mucin-type o-glycosylation network generation and visualization. | mucin-type o-glycans have profound effects on the structure and stability of glycoproteins. o-glycans on the cell surface proteins also modulate the cell's interactions with the surrounding environments and other cells. the synthetic pathway of o-glycans involves a large number of enzymes with diverse substrate specificity. the expression pattern of these enzymes is cell and tissue-specific, thus making the pathway highly diverse. to facilitate pathway analysis in a cell and tissue-specific fash ... | 2019 | 30739313 |
| a comparison of the ability of leu8- and pro8-oxytocin to regulate intracellular ca2+ and ca2+-activated k+ channels at human and marmoset oxytocin receptors. | the neurohypophyseal hormone oxytocin (ot) regulates biologic functions in both peripheral tissues and the central nervous system. in the central nervous system, ot influences social processes, including peer relationships, maternal-infant bonding, and affiliative social relationships. in mammals, the nonapeptide ot structure is highly conserved with leucine in the eighth position (leu8-ot). in marmosets (callithrix), a nonsynonymous nucleotide substitution in the oxt gene codes for proline in t ... | 2019 | 30739093 |
| study of the mechanism for increased protein expression via transcription potency reduction of the selection marker. | stable transfection of mammalian cells using various expression cassettes for exogenous gene expression has been well established. the impact of critical factors in these cassettes, such as promoter and enhancer elements, on recombinant protein production in mammalian cells has been studied extensively to optimize the expression efficiency. however, few studies on the correlation between the strength of selection marker and the expression of gene of interest (goi) have been reported. here we inv ... | 2019 | 30730009 |
| free, unlinked glycosylphosphatidylinositols on mammalian cell surfaces revisited. | glycosylphosphatidylinositols (gpis) are linked to many cell-surface proteins, anchor these proteins in the membrane, and are well characterized. however, gpis that exist in the free form on the mammalian cell surface remain largely unexplored. to investigate free gpis in cultured cell lines and mouse tissues, here we used the t5-4e10 mab (t5 mab), which recognizes unlinked gpis having an n-acetylgalactosamine (galnac) side chain linked to the first mannose at the nonreducing terminus. we detect ... | 2019 | 30728244 |
| selective inhibition of par4 (protease-activated receptor 4)-mediated platelet activation by a synthetic nonanticoagulant heparin analog. | objective- par4 (protease-activated receptor 4), one of the thrombin receptors in human platelets, has emerged as a promising target for the treatment of arterial thrombotic disease. previous studies implied that thrombin exosite ii, known as a binding site for heparin, may be involved in thrombin-induced par4 activation. in the present study, a heparin octasaccharide analog containing the thrombin exosite ii-binding domain of heparin was chemically synthesized and investigated for anti-par4 eff ... | 2019 | 30727756 |
| c3p3-g1: first generation of a eukaryotic artificial cytoplasmic expression system. | most eukaryotic expression systems make use of host-cell nuclear transcriptional and post-transcriptional machineries. here, we present the first generation of the chimeric cytoplasmic capping-prone phage polymerase (c3p3-g1) expression system developed by biological engineering, which generates capped and polyadenylated transcripts in host-cell cytoplasm by means of two components. first, an artificial single-unit chimeric enzyme made by fusing an mrna capping enzyme and a dna-dependent rna pol ... | 2019 | 30726994 |
| enhancing the functionality of a microscale bioreactor system as an industrial process development tool for mammalian perfusion culture. | without a scale-down model for perfusion, high resource demand makes cell line screening or process development challenging, therefore, potentially successful cell lines or perfusion processes are unrealized and their ability untapped. we present here the refunctioning of a high-capacity microscale system that is typically used in fed-batch process development to allow perfusion operation utilizing in situ gravity settling and automated sampling. in this low resource setting, which involved rout ... | 2019 | 30712286 |
| mycobacterium tuberculosis ag85b:hfcγ1 recombinant fusion protein as a selective receptor-dependent delivery system for antigen presentation. | introducing an effective vaccine for tuberculosis (tb) is an urgent need. mycobacterium tuberculosis (mtb) ag85 complex is suggested for making protective immunodominant antigens for design and development of novel tb vaccine. in the present study, a pdr2ef1-fcγ1 vector has been used to make ag85b:hfcγ1 recombinant fusion protein. briefly, specific xbai and noti site incorporated primers were used to amplify mtb-fbpb gene by pcr, ta-cloned and amplified in e.coli dh5α. the resulting vector then ... | 2019 | 30711546 |
| case study: the characterization and implementation of dielectric spectroscopy (biocapacitance) for process control in a commercial gmp cho manufacturing process. | dielectric spectroscopy (biocapacitance) is an up-and-coming technology for real time monitoring of biomass in cell culture processes and has opened the door for next-generation cell culture process control techniques such as automated on-demand nutrient feeding. in this case study we empirically demonstrate the lower limit of quantitation (loq), probe-to-probe consistency, and scalability of in situ biocapacitance probes using data generated from small- and large-scale chinese hamster ovary (ch ... | 2019 | 30707503 |
| dopamine transporter forms stable dimers in the live cell plasma membrane in a phosphatidylinositol 4,5-bisphosphate-independent manner. | the human dopamine transporter (hdat) regulates the level of the neurotransmitter dopamine (da) in the synaptic cleft and recycles da for storage in the presynaptic vesicular pool. many neurotransmitter transporters exist as oligomers, but the physiological role of oligomerization remains unclear; for example, it has been speculated to be a prerequisite for amphetamine-induced release and protein trafficking. previous studies point to an oligomeric quaternary structure of hdat; however, the exac ... | 2019 | 30705091 |
| slc20a2 variants cause dysfunctional phosphate transport activity in endothelial cells induced from idiopathic basal ganglia calcification patients-derived ipscs. | idiopathic basal ganglia calcification (ibgc) is a rare neuropsychiatric illness also known as fahr's disease or primary familial brain calcification (pfbc). ibgc is caused by slc20a2 variants, which encodes the inorganic phosphate (pi) transporter pit-2, a transmembrane protein associated with pi homeostasis. we have reported novel slc20a2 variants in the japanese population and established an induced pluripotent stem cells (ipscs) from an ibgc patient carrying a slc20a2 variant. to investigate ... | 2019 | 30704756 |
| cho genome mining for synthetic promoter design. | synthetic promoters are an attractive alternative for use in mammalian hosts such as cho cells as they can be designed de novo with user-defined functionalities. in this study, we describe and validate a method for bioprocess-directed design of synthetic promoters utilizing cho genomic sequence information. we designed promoters with two objective features, (i) constitutive high-level recombinant gene transcription, and (ii) upregulated transcription under mild hypothermia or late-stage culture. ... | 2019 | 30703471 |
| comparison of data-acquisition methods for the identification and quantification of histone post-translational modifications on a q exactive hf hybrid quadrupole orbitrap mass spectrometer. | histone post-translational modifications (ptms) play key roles in regulating eukaryotic gene expression. mass spectrometry (ms) has emerged as a powerful method to characterize and quantify histone ptms as it allows unbiased identification and quantification of multiple histone ptms including combinations of the modifications present. | 2019 | 30701600 |
| an oral toxicity test in rats and a genotoxicity study of extracts from the stems of opuntia ficus-indica var. saboten. | opuntia ficus-indica var. saboten (ofis) is used widely in korea to treat constipation due to its diuretic effects and its enhancement of bowel function and appetite. however, its safety has not yet been established. the aim of this study was to evaluate the repeated oral toxicity and genotoxicity of ofis extract (oe). | 2019 | 30691445 |
| silk sericin hydrolysate is a potential candidate as a serum-substitute in the culture of chinese hamster ovary and henrietta lacks cells. | the silk sericin hydrolysate (ssh) from the waste of silk processing as a substitute of fetal bovine serum (fbs) was used for the culture of chinese hamster ovary (cho) cells and henrietta lacks (hela) strain of human cervical cancer cells. the survival ratio of these cells cultured in ssh media were similar to or higher than those in fbs media. especially after the serum was replaced by low concentration of ssh at 15.0 μg/ml for 5 d, the proliferation of both cells was also similar to or higher ... | 2019 | 30690536 |
| current state-of-the-art in plant-based antibody production systems. | monoclonal antibodies represent the major class of biopharmaceutical products (for therapeutics and diagnostics) with an increasing demand that reaches several tons per year worldwide. traditional large-scale manufacturing processes are based on stirred tank bioreactors for the growth of chinese hamster ovary cells (cho) which requires high initial investments and production costs. therefore, there is an urgent need for alternative production platforms that can at least act as a complement to th ... | 2019 | 30684155 |
| dissecting n-glycosylation dynamics in chinese hamster ovary cells fed-batch cultures using time course omics analyses. | n-linked glycosylation affects the potency, safety, immunogenicity, and pharmacokinetic clearance of several therapeutic proteins including monoclonal antibodies. a robust control strategy is needed to dial in appropriate glycosylation profile during the course of cell culture processes accurately. however, n-glycosylation dynamics remains insufficiently understood owing to the lack of integrative analyses of factors that influence the dynamics, including sugar nucleotide donors, glycosyltransfe ... | 2019 | 30682623 |
| pmab-210: a monoclonal antibody against pig podoplanin. | podoplanin (pdpn) is a type i transmembrane glycoprotein that is expressed in normal tissues, including renal corpuscles and type i lung alveolar cells. monoclonal antibodies (mabs) against human, mouse, rat, rabbit, dog, cat, and bovine pdpns have already been established; however, antipig pdpn (ppdpn) mabs have not. we therefore immunized mice with ppdpn-overexpressing chinese hamster ovary (cho)-k1 cells (cho/ppdpn), and screened hybridomas, which are producing anti-ppdpn mabs. one of mabs, p ... | 2019 | 30681406 |
| multiple mechanisms underlie increased cardiac late sodium current. | we recently reported a quantitative relationship between the degree of functional perturbation reported in the literature for 356 variants in the cardiac sodium channel gene scn5a and the penetrance of resulting arrhythmia phenotypes. in the course of that work, we identified multiple scn5a variants, including r1193q, that are common in populations but are reported in human embryonic kidney (hek) cells to generate large late sodium current (ina-l). | 2019 | 30677491 |
| histidine-rich cell-penetrating peptide for cancer drug delivery and its uptake mechanism. | in this work, we report a drug delivery system based on the ph-responsive self-assembly and -disassembly behaviors of peptides. here, a systematically designed histidine-rich lipidated peptide (np1) is presented to encapsulate and deliver an anticancer drug ellipticine (ept) into two model cells: non-small-cell lung carcinoma and chinese hamster ovary cells. the mechanism of ph-responsive peptide self-assembly and -disassembly involved in the drug encapsulation and release process are extensivel ... | 2019 | 30673275 |
| chromophore pre-maturation for improved speed and sensitivity of split-gfp monitoring of protein secretion. | complementation-dependent fluorescence is a powerful way to study co-localization or interactions between biomolecules. a split-gfp variant, involving the self-associating gfp 1-10 and gfp 11, has previously provided a convenient approach to measure recombinant protein titers in cell supernatants. a limitation of this approach is the slow chromophore formation after complementation. here, we alleviate this lag in signal generation by allowing the gfp 1-10 chromophore to mature on a solid support ... | 2019 | 30670736 |
| generation of site-distinct n-glycan variants for in vitro bioactivity testing. | glycosylation, a critical product quality attribute, may affect the efficacy and safety of therapeutic proteins in vivo. chinese hamster ovary fed-batch cell culture batches yielded consistent glycoprofiles of a fc-fusion antibody comprizing three different n-glycosylation sites. by adding media supplements at specific concentrations in cell culture and applying enzymatic glycoengineering, a diverse n-glycan variant population was generated, including high mannose, afucosylated, fucosylated, aga ... | 2019 | 30659587 |
| bmn 250, a fusion of lysosomal alpha-n-acetylglucosaminidase with igf2, exhibits different patterns of cellular uptake into critical cell types of sanfilippo syndrome b disease pathogenesis. | sanfilippo syndrome type b (sanfilippo b; mucopolysaccharidosis type iiib) occurs due to genetic deficiency of lysosomal alpha-n-acetylglucosaminidase (naglu) and subsequent lysosomal accumulation of heparan sulfate (hs), which coincides with devastating neurodegenerative disease. because naglu expressed in chinese hamster ovary cells is not mannose-6-phosphorylated, we developed an insulin-like growth factor 2 (igf2)-tagged naglu molecule (bmn 250; tralesinidase alfa) that binds avidly to the i ... | 2019 | 30657762 |
| use of a branched linker for enhanced biosensing properties in igg detection from mixed chinese hamster ovary cell cultures. | tris(2-aminoethyl)-amine (tren), a branched amine, was coupled to planar surfaces of alkanethiol self-assembled monolayers (sams) to increase the grafting density of igg-binding peptide (hwrgwv or hwrgwvg) on gold surfaces. one of the three primary amine pendant groups of tren anchors onto the sam, while the other two are available for grafting with the c-termini of the peptide. the ellipsometric peptide density on the sam-branched amine was 1.24 molecules nm-2. the surfaces carrying the peptide ... | 2019 | 30653289 |
| a novel methodology to assess linear energy transfer and relative biological effectiveness in proton therapy using pairs of differently doped thermoluminescent detectors. | a new methodology for assessing linear energy transfer (let) and relative biological effectiveness (rbe) in proton therapy beams using thermoluminescent detectors is presented. the method is based on the different let response of two different lithium fluoride thermoluminescent detectors (lif:mg,ti and lif:mg,cu,p) for measuring charged particles. the relative efficiency of the two detector types was predicted using the recently developed microdosimetric d(z) model in combination with the monte ... | 2019 | 30650402 |
| silkworm storage protein 1 inhibits autophagy-mediated apoptosis. | autophagy is a natural physiological process, and it induces the lysosomal degradation of intracellular components in response to environmental stresses, including nutrient starvation. although an adequate autophagy level helps in cell survival, excessive autophagy triggered by stress such as starvation leads to autophagy-mediated apoptosis. chinese hamster ovary (cho) cells are widely used for producing biopharmaceuticals, including monoclonal antibodies. however, apoptosis induced by high stre ... | 2019 | 30646576 |
| increased growth rate and productivity following stable depletion of mir-7 in a mab producing cho cell line causes an increase in proteins associated with the akt pathway and ribosome biogenesis. | cell line engineering using micrornas represents a desirable route for improving the efficiency of recombinant protein production by cho cells. in this study we generated stable cho dp12 cells expressing a mir-7 sponge transcript which sequesters mir-7 from its endogenous targets. depletion of mir-7 results in a 65% increase in cell growth and >3-fold increase in yield of secreted igg protein. quantitative labelfree lc-ms/ms proteomic profiling was carried out to identify the targets of mir-7 an ... | 2019 | 30641232 |
| secretion analysis of intracellular "difficult-to-express" immunoglobulin g (igg) in chinese hamster ovary (cho) cells. | the chinese hamster ovary (cho) cell line is the most widely used host cell for therapeutic antibody production. although its productivity has been improved by various strategies to satisfy the growing global demand, some difficult-to-express (dte) antibodies remain at low secretion levels. to improve the production of various therapeutic antibodies, it is necessary to determine possible rate-limiting steps in dte antibody secretion in comparison with other high igg producers. here, we analyzed ... | 2019 | 30637508 |
| analysis of golgi ph in chinese hamster ovary cells using ratiometric ph-sensitive fluorescent proteins. | acidic golgi ph plays an important role in protein glycosylation, one of the critical quality attributes of therapeutic proteins. to determine the intracellular golgi ph during culture, stable chinese hamster ovary (cho) cell clones expressing phluorin2, a ratiometric ph-sensitive fluorescent protein (fp), in the cis- and trans-golgi, were constructed by fusing phluorin2 with specific targeting proteins, acetylglucosaminyltransferase, and a galactosyltransferase, respectively. stable cho cell cl ... | 2019 | 30636290 |
| optimization of sequence, display, and mode of operation of igg-binding peptide ligands to develop robust, high-capacity affinity adsorbents that afford high igg product quality. | this work presents the use of peptide ligand hwrgwv and its cognate sequences to develop affinity adsorbents that compete with protein a in terms of binding capacity and quality of the eluted product. first, the peptide ligand was conjugated to crosslinked agarose resins (workbeads) at different densities and using different spacer arms. the optimization of ligand density and display resulted in values of static and dynamic binding capacity of 85 mg/ml and 65 mg/ml, respectively. a selected pept ... | 2019 | 30621158 |
| zinc oxide nanoparticles induced gene mutation at the hgprt locus and cell cycle arrest associated with apoptosis in v-79 cells. | in recent years, the large-scale production of zno nanoparticles (nps) for various applications is increasing exponentially and may pose serious health issues when inhaled either during occupational exposure or in consumer settings. the mechanisms underlying the toxicity of nps have recently been studied intensively. despite the existing studies, the mutagenicity of zno nps in the eukaryotic system is still unclear. therefore, the aim of the present study was to investigate the mutagenic potenti ... | 2019 | 30618096 |
| expression in eukaryotic cells and purification of synthetic gene encoding enterocin p: a bacteriocin with broad antimicrobial spectrum. | due to the emergence of multidrug-resistant bacteria, treatment options for infectious diseases are decreasing. bacteriocins are small antimicrobial peptides produced by numerous bacteria that offer alternative therapeutic strategies to combat multidrug-resistant bacterial infections. we evaluated the cloning, functional expression, and antimicrobial activities of enterocin p (entp), a class ii bacteriocin member, in chinese hamster ovary (cho) cells. a synthetic gene matching cho cell codon usa ... | 2019 | 30617751 |
| anti-hyperglycemic and genotoxic studies of 1-o-methyl chrysophanol, a new anthraquinone isolated from amycolatopsis thermoflava strain sfma-103. | the compound 1-o-methyl chrysophanol (omc) which belongs to a class of hydroxyanthraquinones was isolated from amycolatopsis thermoflava strain sfma-103 and studied for their anti-diabetic properties. omc was evaluated as an anti-diabetic agent based on in silico studies which initially predicted the binding energy with α-amylase (-188.81 kj mol-1) and with α-glucosidase (70.53 kj mol-1). further, these results were validated based on enzyme inhibition assays where omc demonstrated enzyme inhibi ... | 2019 | 30614298 |
| analysis of the immunoglobulin g (igg) secretion efficiency in recombinant chinese hamster ovary (cho) cells by using citrine-fusion igg. | biopharmaceuticals represented by immunoglobulin g (igg) are produced by the cultivation of recombinant animal cells, especially chinese hamster ovary (cho) cells. it is thought that the intracellular secretion process of igg is a bottleneck in the production of biopharmaceuticals. many studies on the regulation of endogenous secretory protein expression levels have shown improved productivity. however, these strategies have not universally improved the productivity of various proteins. a more r ... | 2019 | 30610509 |
| comparison of integrases identifies bxb1-ga mutant as the most efficient site-specific integrase system in mammalian cells. | phage-derived integrases can catalyze irreversible, site-specific integration of transgenic payloads into a chromosomal locus, resulting in mammalian cells that stably express transgenes or circuits of interest. previous studies have demonstrated high-efficiency integration by the bxb1 integrase in mammalian cells. here, we show that a point mutation (bxb1-ga) in bxb1 target sites significantly increases bxb1-mediated integration efficiency at the rosa26 locus in chinese hamster ovary cells, res ... | 2019 | 30609349 |
| a pathogenic tau fragment compromises microtubules, disrupts insulin signaling and induces the unfolded protein response. | human tauopathies including alzheimer's disease, progressive supranuclear palsy and related disorders, are characterized by deposition of pathological forms of tau, synaptic dysfunction and neuronal loss. we have previously identified a pathogenic c-terminal tau fragment (tau35) that is associated with human tauopathy. however, it is not known how tau fragmentation affects critical molecular processes in cells and contributes to impaired physiological function. chinese hamster ovary (cho) cells ... | 2019 | 30606258 |
| fmoc-ff and hexapeptide-based multicomponent hydrogels as scaffold materials. | short peptides or single amino acids are interesting building blocks for fabrication of hydrogels, frequently used as extracellular matrix-mimicking scaffolds for cell growth in tissue engineering. the combination of two or more peptide hydrogelators could allow obtaining different materials exhibiting new architectures, tunable mechanical properties, high stability and improved biofunctionality. here we report on the synthesis, formulation and multi-scale characterization of peptide-based mixed ... | 2019 | 30601569 |
| interaction of cell culture process parameters for modulating mab afucosylation. | the extent of afucosylation, which refers to the absence of core fucose on fc glycans, can correlate positively with the antibody-dependent cellular cytotoxicity (adcc) activity of a monoclonal antibody (mab). therefore, it is important to maintain consistent afucosylation during cell culture process scale-up in bioreactors for a mab with adcc activity. however, there is currently a lack of understanding about the impact of partial pressure of carbon dioxide (pco2 )-a parameter that can vary wit ... | 2019 | 30597531 |
| c-c motif chemokine ligand (ccl) production in equine peripheral blood mononuclear cells identified by newly generated monoclonal antibodies. | chemokines are soluble molecules directing immune cell trafficking and homing, mediating inflammation, and initiating immune responses to infection. in horses, the analysis of chemokines has been limited by the lack of specific antibodies. we generated mabs specific for the equine c-c motif chemokine ligands (ccl) ccl2 (mcp-1), ccl3 (mip-1α), ccl5 (rantes) and ccl11 (eotaxin) using hybridoma technology. antibody specificity was confirmed by intracellular staining of chinese hamster ovary cells t ... | 2018 | 30596378 |
| cxcl10 production in equine monocytes is stimulated by interferon-gamma. | c-x-c motif ligand 10 (cxcl10) is a pro-inflammatory chemokine and has been extensively evaluated in people and mice. in horses, cxcl10 and its involvement in host immunity has rarely been analyzed due to the lack of specific antibodies. we generated a mab specific for the equine chemokine cxcl10 using hybridoma technology. antibody specificity was confirmed by intracellular staining and flow cytometric analysis of chinese hamster ovary (cho) cells expressing equine rcxcl10, while cho cells expr ... | 2019 | 30593347 |
| utilizing a regulated targeted integration cell line development approach to systematically investigate what makes an antibody difficult to express. | chinese hamster ovary (cho) cells are conventionally used to generate therapeutic cell lines via random integration (ri), where desired transgenes are stably integrated into the genome. targeted integration (ti) approaches, which involve integration of a transgene into a specific locus in the genome, are increasingly utilized for cho cell line development (cld) in recent years. none of these cld approaches, however, are suitable for expression of toxic or difficult-to-express molecules, or for d ... | 2019 | 30592567 |
| secretory leakage of igg1 aggregates from recombinant chinese hamster ovary cells. | aggregation of therapeutic antibodies is one of the most important issues to be resolved in manufacturing processes because of reduced efficacy and immunogenicity. despite aggregation studies in vitro, little is known about the aggregation mechanism in cell culture processes. in this study, we investigated the process of aggregate formation of igg1 antibodies during the culture of chinese hamster ovary (cho) cells to determine how aggregation occurs. a recombinant cho cell line was cultivated in ... | 2019 | 30580968 |
| tak-442, a direct factor xa inhibitor, inhibits monocyte chemoattractant protein 1 production in endothelial cells via involvement of protease-activated receptor 1. | oral blood coagulation inhibitors and their receptors, such as factor xa (fxa), thrombin, and the thrombin receptor protease-activated receptor 1 (par1), are entered into clinical trials for acute coronary syndrome therapy; however, the results obtained so far are different for each drug. the underlying mechanisms of the results have not been fully investigated. we studied the in vitro anti-inflammatory effects of the selective fxa inhibitor tak-442 on human endothelial cells, with comparing tho ... | 2018 | 30568593 |
| structural and epitope analysis (t- and b-cell epitopes) of hepatitis c virus (hcv) glycoproteins: an in silico approach. | chronic infection with hepatitis c virus (hcv) poses a major risk for liver disease like cirrhosis, liver failure and hepatocellular carcinoma. in terms of percentage, the prevalence of hcv in india was found to be low to moderate (1-1.5%), but in terms of sheer numbers, india has a significant number of global hcv patients. presently, hcv can be treated with direct acting-antibody drugs but there is no prophylactic or therapeutic vaccine available against it. in hcv infection, t- and b-cell imm ... | 2018 | 30568344 |
| differential regulation of thermodynamic binding forces of levocetirizine and (s)-cetirizine by lys191 in human histamine h₁ receptors. | cetirizine is a zwitterionic second-generation antihistamine containing r- and s-enantiomers, levocetirizine, and (s)-cetirizine. levocetirizine is known to have a higher affinity for the histamine h₁ receptors than (s)-cetirizine; ligand-receptor docking simulations have suggested the importance of the formation of a salt bridge (electrostatic interaction) between the carboxylic group of levocetirizine and the lys191 residue at the fifth transmembrane domain of human histamine h₁ receptors. in ... | 2018 | 30558340 |
| multiple modes of action of hydrophobic amines and their guanidine analogues on asic1a. | hydrophobic monoamines containing only a hydrophobic/aromatic moiety and protonated amino group are a recently described class of acid-sensing ion channel (asic) modulators. intensive studies have revealed a number of active compounds including endogenous amines and pharmacological agents and shown that these compounds potentiate and inhibit asics depending on their specific structure and on subunit composition of the target channel. the action of monoamines also depends on the application proto ... | 2019 | 30557561 |