Publications

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mechanistic evaluation of the transfection barriers involved in lipid-mediated gene delivery: interplay between nanostructure and composition.here we present a quantitative mechanism-based investigation aimed at comparing the cell uptake, intracellular trafficking, endosomal escape and final fate of lipoplexes and lipid-protamine/deoxyribonucleic acid (dna) (lpd) nanoparticles (nps) in living chinese hamster ovary (cho) cells. as a model, two lipid formulations were used for comparison. the first formulation is made of the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (dotap) and the zwitterionic lipid dioleoylphosphocholine ...201424296066
aspects of nonviral gene therapy: correlation of molecular parameters with lipoplex structure and transfection efficacy in pyridinium-based cationic lipids.this study seeks correlations between the molecular structures of cationic and neutral lipids, the lipid phase behavior of the mixed-lipid lipoplexes they form with plasmid dna, and the transfection efficacy of the lipoplexes. synthetic cationic pyridinium lipids were co-formulated (1:1) with the cationic lipid 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (epc), and these lipids were co-formulated (3:2) with the neutral lipids 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (dope) or chol ...201424296044
dynamic kisspeptin receptor trafficking modulates kisspeptin-mediated calcium signaling.kisspeptin receptor (kiss1r) signaling plays a critical role in the regulation of reproduction. we investigated the role of kisspeptin-stimulated kiss1r internalization, recycling, and degradation in the modulation of kiss1r signaling. kisspeptin stimulation of chinese hamster ovary or gt1-7 cells expressing kiss1r resulted in a biphasic increase in intracellular ca(2+) ([ca(2+)]i), with a rapid acute increase followed by a more sustained second phase. in contrast, stimulation of the trh recepto ...201424295737
analysis of chinese hamster ovary cell metabolism through a combined computational and experimental approach.optimization of cell culture processes can benefit from the systematic analysis of experimental data and their organization in mathematical models, which can be used to decipher the effect of individual process variables on multiple outputs of interest. towards this goal, a kinetic model of cytosolic glucose metabolism coupled with a population-level model of chinese hamster ovary cells was used to analyse metabolic behavior under batch and fed-batch cell culture conditions. the model was parame ...201424292563
risk assessment of polychlorinated biphenyls and heavy metals in soils of an abandoned e-waste site in china.risk assessment of abandoned e-waste recycling areas received little attention. herein, we report the concentrations of 16 pcbs and 7 heavy metals in soils near an abandoned e-waste recycling plant in taizhou, china. our data showed that levels of tri-, tetra-, penta-, hexa-pcbs were 9.01, 5.56, 12.93, 3.13 mg/kg, and pb, cd, cu were 6082.9, 42.3, 2364.2 mg/kg soil. cd was the most prevalent contaminant with nemerow index value of 44.3. contaminants have been transported from the abandoned site ...201424292442
a novel approach to evaluate the pharmacokinetic biocomparability of a monoclonal antibody derived from two different cell lines using simultaneous crossover design.a parallel study design with a large number of subjects has been a typical path for pharmacokinetic (pk) biocomparability assessment of biotherapeutics with long half-lives and immunogenic propensity, for example, monoclonal antibodies (mab). a recently published innovative bioanalytical method that can quantify mab produced from two different cell lines in the same sample opened an avenue to exploring a simultaneous crossover study design for pk biocomparability assessment of biotherapeutics. s ...201424281691
high-expression β(1) adrenergic receptor/cell membrane chromatography method based on a target receptor to screen active ingredients from traditional chinese medicines.β-adrenergic receptors are important targets for drug discovery. we have developed a new β1 -adrenergic receptor cell membrane chromatography (β1 ar-cmc) with offline ultra-performance lc (uplc) and ms method for screening active ingredients from traditional chinese medicines. in this study, chinese hamster ovary-s cells with high β1 ar expression levels were established and used to prepare a cell membrane stationary phase in a β1 ar-cmc model. the retention fractions were separated and identifi ...201424272943
sex hormone binding globulin and corticosteroid binding globulin as major effectors of steroid action.contrary to the long-held postulate of steroid-hormone binding globulin action, these protein carriers of steroids are major players in steroid actions in the body. this manuscript will focus on our work with sex hormone binding globulin (shbg) and corticosteroid binding globulin (cbg) and demonstrate how they are actively involved in the uptake, intracellular transport, and possibly release of steroids from cells. this manuscript will also discuss our own findings that the steroid estradiol is ...201424269740
insulin-like growth factor ii peptide fusion enables uptake and lysosomal delivery of α-n-acetylglucosaminidase to mucopolysaccharidosis type iiib fibroblasts.enzyme replacement therapy for mps iiib (mucopolysaccharidosis type iiib; also known as sanfilippo b syndrome) has been hindered by inadequate mannose 6 phosphorylation and cellular uptake of rhnaglu (recombinant human α-n-acetylglucosaminidase). we expressed and characterized a modified rhnaglu fused to the receptor-binding motif of igf-ii (insulin-like growth factor 2) (rhnaglu-igf-ii) to enhance its ability to enter cells using the cation-independent mannose 6-phosphate receptor, which is als ...201424266751
nauclea latifolia smith (rubiaceae) exerts antinociceptive effects in neuropathic pain induced by chronic constriction injury of the sciatic nerve.the roots of nauclea latifolia smith (rubiaceae) popularly known as "koumkouma" is used in traditional cameroonian medicine as neuropathic pain remedy and for the treatment of headache, inflammatory pain and convulsion. this study was conducted to evaluate the antinociceptive effects of the alkaloid fraction isolated from nauclea latifolia in neuropathic pain induced by chronic constriction injury (cci) of the sciatic nerve in rat.201424263011
benzothiazole head group based cationic lipids: synthesis and application for gene delivery.a series of benzothiazole based lipids (1-10) containing different derivatives of benzothiazole in the head group region were synthesized to determine the structure-activity relationship for gene delivery. the liposomes formulated were mixed with plasmid dna encoding green fluorescent protein (α5gfp) or β-galactosidase (pcmv-sport-β-gal) and transfected into b16f10 (human melanoma cancer cells), cho (chinese hamster ovary), a-549 (human lung carcinoma cells) and mcf-7 (human breast carcinoma cel ...201424262373
effects of sterilization methods on the physical, chemical, and biological properties of silk fibroin membranes.silk fibroin has been widely explored for many biomedical applications, due to its biocompatibility and biodegradability. sterilization is a fundamental step in biomaterials processing and it must not jeopardize the functionality of medical devices. the aim of this study was to analyze the influence of different sterilization methods in the physical, chemical, and biological characteristics of dense and porous silk fibroin membranes. silk fibroin membranes were treated by several procedures: imm ...201424259492
cellular responses to individual amino-acid depletion in antibody-expressing and parental cho cell lines.depletion of two nonessential amino acids, asparagine (asn) and glutamine (gln), occurred during a fed-batch production process with a cho cell line expressing a recombinant antibody. this depletion coincided with growth suppression and the onset of the stationary phase. experimental withdrawal of asn led to cell cycle arrest of cell line a in g0/g1 phase. on a mechanistic level, withdrawal of either asn or gln stimulated the amino-acid response (aar) pathway, indicating that depletion of noness ...201424254056
cell line profiling to improve monoclonal antibody production.mammalian cell culture performance is influenced by both intrinsic (genetic) and extrinsic (media and process) factors. in this study, intrinsic capacity of various monoclonal antibody-producing chinese hamster ovary (cho) cell lines was compared by exposing them to the same culture condition. microarray-based transcriptomics and lc-ms/ms shotgun proteomics technologies were utilized to obtain expression landscape of different cell lines. specific transcripts and proteins correlating with produc ...201424249214
microarray platform affords improved product analysis in mammalian cell growth studies.high throughput (ht) platforms serve as a cost-efficient and rapid screening method for evaluating the effect of cell-culture conditions and screening of chemicals. we report the development of a ht cell-based microarray platform to assess the effect of culture conditions on chinese hamster ovary (cho) cells. specifically, growth, transgene expression and metabolism of a gs/methionine sulphoximine (msx) cho cell line, which produces a therapeutic monoclonal antibody, was examined using a microar ...201424227746
in silico and in vitro genotoxicity evaluation of descarboxyl levofloxacin, an impurity in levofloxacin.it is important to establish the safety of impurities in drug substances or drug products. the assessment of genotoxicity of impurities and the determination of acceptable limits for genotoxic impurities was addressed in some recent guidances as a difficult issue. descarboxyl levofloxacin is an impurity isolated from levofloxacin, which may impose a risk without associated benefit. however, there is insufficient toxic information about descarboxyl levofloxacin. this study investigated the genoto ...201424224725
single-cell-based breeding: rational strategy for the establishment of cell lines from a single cell with the most favorable properties.for efficient biomolecule production (e.g., antibodies, recombinant proteins), mammalian cells with high expression rates should be selected from cell libraries, propagated while maintaining a homogenous expression rate, and subsequently stabilized at their high expression rate. clusters of isogenic cells (i.e., colonies) have been used for these processes. however, cellular heterogeneity makes it difficult to obtain cell lines with the highest expression rates by using single-colony-based breed ...201424216459
successful construction and stable expression of an anti-cd45ra scfv-egfp fusion protein in chinese hamster ovary cells.cd45ra has been found highly expressed on leukemia cells and may be a potential target of the disease. in this study, an anti-cd45ra single-chain antibody fragment (scfv3a4) was genetically linked to the n terminus of the enhanced green fluorescent protein (egfp) to generate a scfv3a4-egfp fusion protein. the scfv3a4-egfp with a molecular weight of 57kda was stably expressed and secreted from the transfected cho cells through the er/golgi-dependent pathway. the fusion protein was soluble in the ...201424211776
γ375w fibrinogen-synthesizing cho cells indicate the accumulation of variant fibrinogen within endoplasmic reticulum.hepatic endoplasmic reticulum (er) storage disease (hersd) associated with hypofibrinogenemia has been reported in patients with four types of heterozygous γ-chain variant fibrinogen in the c terminal region. of interest, substitution of γr375w induced hypofibrinogenemia and hersd, whereas γr375g induced dysfibrinogenemia.201424210681
comprehensive, quantitative bioprocess productivity monitoring using fluorescence eem spectroscopy and chemometrics.this study demonstrates the application of fluorescence excitation-emission matrix (eem) spectroscopy to the quantitative predictive analysis of recombinant glycoprotein production cultured in a chinese hamster ovary (cho) cell fed-batch process. the method relies on the fact that eem spectra of complex solutions are very sensitive to compositional change. as the cultivation progressed, changes in the emission properties of various key fluorophores (e.g., tyrosine, tryptophan, and the glycoprote ...201424504094
pecam-1-dependent heme oxygenase-1 regulation via an nrf2-mediated pathway in endothelial cells.the antioxidant enzyme heme oxygenase (ho)-1, which catalyses the first and rate-limiting step of heme degradation, has major anti-inflammatory and immunomodulatory effects via its cell-type-specific functions in the endothelium. in the current study, we investigated whether the key endothelial adhesion and signalling receptor pecam-1 (cd31) might be involved in the regulation of ho-1 gene expression in human endothelial cells (ecs). to this end pecam-1 expression was down-regulated in human umb ...201424500083
controllability analysis of protein glycosylation in cho cells.to function as intended in vivo, a majority of biopharmaceuticals require specific glycan distributions. however, achieving a precise glycan distribution during manufacturing can be challenging because glycosylation is a non-template driven cellular process, with the potential for significant uncontrolled variability in glycan distributions. as important as the glycan distribution is to the end-use performance of biopharmaceuticals, to date, no strategy exists for controlling glycosylation on-li ...201424498415
apolipoprotein e increases cell association of amyloid-β 40 through heparan sulfate and lrp1 dependent pathways.the increased risk of alzheimer's disease (ad) associated with specific apolipoprotein e (apoe) isoforms appears to relate to altered amyloid-β (aβ) homeostasis. clearance of aβ from the brain is reduced in the presence of the ad-associated apoe4 isoform, which may contribute to the accumulation of aβ deposits in the parenchyma and vasculature. the low-density lipoprotein receptor-related protein 1 (lrp1) and heparan sulfate proteoglycans (hspgs), both established apoe receptors, are involved in ...201424491019
analysis of microrna transcription and post-transcriptional processing by dicer in the context of cho cell proliferation.cho cells are the mammalian cell line of choice for recombinant production of therapeutic proteins. however, their low rate of proliferation limits obtainable space-time yields due to inefficient biomass accumulation. we set out to correlate microrna transcription to cell-specific growth-rate by microarray analysis of 5 cho suspension cell lines with low to high specific growth rates. global microrna expression analysis and pearson correlation studies showed that mature microrna transcript level ...201424486028
long-term episomal transgene expression from mitotically stable integration-deficient lentiviral vectors.nonintegrating gene delivery vectors have an improved safety profile compared with integrating vectors, but transgene retention is problematic as nonreplicating episomes are progressively and rapidly diluted out through cell division. we have developed an integration-deficient lentiviral vector (idlv) system generating mitotically stable episomes capable of long-term transgene expression. we found that a transient cell cycle arrest at the time of transduction with idlvs resulted in 13-45% of chi ...201424483952
comparison of clot lysis activity and biochemical properties of originator tenecteplase (metalyse(®)) with those of an alleged biosimilar.the bioengineered tissue plasminogen activator tenecteplase is an important treatment modality of acute myocardial infarction recommended by international guidelines. following introduction of originator tenecteplase (brand names metalyse(®) and tnkase(®)), a "biosimilar" tenecteplase became available for commercial use in india under the brand name elaxim(®) in the absence of indian biosimilar guidelines which came into force from september 15th, 2012. based on a report of biochemical and fibri ...201424550831
human orexin/hypocretin receptors form constitutive homo- and heteromeric complexes with each other and with human cb1 cannabinoid receptors.human ox1 orexin receptors have been shown to homodimerize and they have also been suggested to heterodimerize with cb1 cannabinoid receptors. the latter has been suggested to be important for orexin receptor responses and trafficking. in this study, we wanted to assess the ability of the other combinations of receptors to also form similar complexes. vectors for expression of human ox1, ox2 and cb1 receptors, c-terminally fused with either renilla luciferase or gfp(2) green fluorescent protein ...201424530395
kojic acid protects c57bl/6 mice from gamma-irradiation induced damage.the radioprotective effects of a single administration of kojic acid (ka) against ionizing radiation were evaluated via assessment of 30-day survival and alterations of peripheral blood parameters of adult c57bl/6 male mice. the 30-day survival rate of mice pretreated with ka (75 or 300 mg/kg body weight, ka75 or ka300) subcutaneously 27 h prior to a lethal dose (8 gy, 153.52 cgy/min) of gamma irradiation was higher than that of mice irradiated alone (40% or 60% vs 0%). it was observed that the ...201424528043
effect of brazilian propolis (af-08) on genotoxicity, cytotoxicity and clonogenic death of chinese hamster ovary (cho-k1) cells irradiated with (60)co gamma-radiation.the present study was conducted in order to evaluate the effect of brazilian propolis (af-08; 5, 10, 15, 30, 50, 100, and 200μg/ml) in protecting cho-k1 cells against genotoxic and cytotoxic damage and clonogenic death induced by (60)co gamma-radiation (1.0, 2.0, 4.0, and 6.0gy). for this purpose, three interlinked endpoints were analyzed: induction of dna damage by use of the micronucleus (mn) test (genotoxic damage), cell viability by means of the mts assay, and differential staining (cytotoxi ...201424525380
metabolic control at the cytosol-mitochondria interface in different growth phases of cho cells.metabolism at the cytosol-mitochondria interface and its regulation is of major importance particularly for efficient production of biopharmaceuticals in chinese hamster ovary (cho) cells but also in many diseases. we used a novel systems-oriented approach combining dynamic metabolic flux analysis and determination of compartmental enzyme activities to obtain systems level information with functional, spatial and temporal resolution. integrating these multiple levels of information, we were able ...201424525334
4-hydroxynonenal induces an increase in expression of receptor for activating c kinase 1 (rack1) in chinese hamster v79-4 lung cells.4-hydroxy-trans-2-nonenal (hne) is a cytotoxic α,β-unsaturated aldehyde implicated in the pathology of several diseases that have an oxidative stress mechanism, including atherosclerosis, diabetes, alcohol-induced liver disease, and neurodegenerative disorders. as the most toxic aldehydic product of lipid peroxidation, hne is known to exert a range of biological effects in a concentration-dependent manner. in this study, the effect of hne on the levels of proteins in v79-4 chinese hamster lung c ...201424525193
black pepper constituent piperine: genotoxicity studies in vitro and in vivo.piperine is responsible for the hot taste of black pepper. publications on genotoxicity of piperine are reported: negative ames tests and one in vitro micronucleus test (mnt). in vivo tests were mainly negative. in the majority of the data the administered dose levels did not follow the dose selection requirements of regulatory guidelines of having dose levels up to the maximum tolerated dose (mtd). the only oral high dose studies were a positive in vivo mnt in mice in contrast to a negative in ...201424525095
adap interactions with talin and kindlin promote platelet integrin αiibβ3 activation and stable fibrinogen binding.adap is a hematopoietic-restricted adapter protein that promotes integrin activation and is a carrier for other adapter proteins, src kinase-associated phosphoprotein 1 (skap1) and skap2. in t lymphocytes, skap1 is the adap-associated molecule that activates integrins through direct linkages with rap1 effectors (regulator of cell adhesion and polarization enriched in lymphoid tissues; rap1-interacting adapter molecule). adap also promotes integrin αiibβ3 activation in platelets, which lack skap1 ...201424523237
molecular cloning and characterization of two pig vasoactive intestinal polypeptide receptors (vpac1-r and vpac2-r).we here report the cloning, tissue expression, and functional analyses of the two pig vasoactive intestinal polypeptide (vip) receptors (pvpac1-r and pvpac2-r). the cloned full-length pvpac1-r and pvpac2-r share high structural similarity with their mammalian counterparts. functional assay revealed that the full-length pvpac1-r and pvpac2-r-expressed chinese hamster ovary (cho) cells could be activated by pvip and ppacap38 potently, indicating that pvpac1-r and pvpac2-r are capable of binding vi ...201424520933
mirna expression in cho: nature knows best.improvement of chinese hamster ovary (cho) cells for therapeutic protein production has great potential for the manufacturing of biopharmaceuticals. this commentary by baik and lee discusses the study by klanert et al., which describes an improved tool that will allow greater control over the design of cho cells.201424519876
high-throughput clonepix fl analysis of mab-expressing clones using the ucoe expression system.therapeutic recombinant monoclonal antibodies (mabs) are commonly produced by high-expressing, clonal, mammalian cells. creation of these clones for manufacturing remains heavily reliant on stringent selection and gene amplification, which in turn can lead to genetic instability, variable expression, product heterogeneity and prolonged development timelines. inclusion of cis-acting ubiquitous chromatin opening elements (ucoe™) in mammalian expression vectors has been shown to improve productivit ...201424518824
solvation dynamics and intermittent oscillation of cell membrane: live chinese hamster ovary cell.dynamics of the exofacial thiols (i.e., cell surface thiol containing membrane proteins) of a live chinese hamster ovary (cho) cell is probed by time-resolved confocal microscopy. for this purpose, a fluorescent probe, 7-(diethylamino)-3-(4-maleimidophenyl)-4-methylcoumarin (cpm) is covalently attached to the exofacial thiols. the emission maximum of cpm bound exofacial thiols indicates a highly exposed and polar environment. using cpm, we studied solvation dynamics, for the first time, at the m ...201424571625
heat exposure enhances radiosensitivity by depressing dna-pk kinase activity during double strand break repair.from the role of double strand dna dependent protein kinase (dna-pkcs) activity of non-homologous end joining (nhej) repair for dna double strand breaks (dsbs), we aim to define possible associations between thermo-sensitisation and the enzyme activities in x-ray irradiated cells.201424571173
mir-7 inhibits colorectal cancer cell proliferation and induces apoptosis by targeting xrcc2.analysis using publicly available algorithms predicts that x-ray repair complementing defective repair in chinese hamster cells 2 (xrcc2), a key component in the homologous recombination repair pathway, is a potential target of micro-ribonucleic acid-7 (mir-7). some studies have shown that both mir-7 and xrcc2 are associated with cancer development. for this purpose, we searched for the possible relationship between mir-7 and xrcc2 in the development of colorectal cancer (crc).201424570594
apoptosis-inducing activity of hplc fraction from voacanga globosa (blanco) merr. on the human colon carcinoma cell.voacanga globosa (blanco), a plant endemic to the philippines, is traditionally used especially by indigenous people of bataan in the treatment of ulcers, wounds and tumorous growths. this study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of hplc fractions from leaves on hct116 human colon carcinoma and a549 human lung carcinoma cell lines. ethanolic extraction was performed on v globosa leaves followed by hexane and ethyl ac ...201424568467
improved expression of recombinant human factor ix by co-expression of ggcx, vkor and furin.recombinant human fix concentrates (rhfix) are essential in the treatment and prevention of bleeding in the bleeding disorder haemophilia b. however, due to the complex nature of fix production yields are low which leads to high treatment costs. here we report the production of rhfix with substantially higher yield by co-expressing human fix with ggcx (γ-glutamyl carboxylase), vkor (vitamin k epoxide reductase) and furin (paired basic amino acid cleaving enzyme) in chinese hamster ovary (cho) ce ...201424567122
[study on the effect of deficient ercc2/xpd gene on the repair of dna damage induced by uvc in cho cell line].to explore the function of ercc2/xpd in the repair of dna damage induced by uvc.201424564106
a phase i/ii, multiple-dose, dose-escalation study of siltuximab, an anti-interleukin-6 monoclonal antibody, in patients with advanced solid tumors.this phase i/ii study evaluated safety, efficacy, and pharmacokinetics of escalating, multiple doses of siltuximab, a chimeric anti-interleukin (il)-6 monoclonal antibody derived from a new chinese hamster ovary (cho) cell line in patients with advanced/refractory solid tumors.201424563479
neonatal genome-wide methylation patterns in relation to birth weight in the norwegian mother and child cohort.although epigenetic regulation plays a critical role in embryonic development, few studies have examined the relationship of epigenome-wide methylation with fetal growth. using the infinium humanmethylation450 beadchip (illumina, inc., san diego, california) in a substudy of 1,046 infants from the norwegian mother and child cohort study (moba) enrolled between 1999 and 2008, we examined epigenome-wide cord blood dna methylation in relation to birth weight. in multivariable-adjusted robust linear ...201424561991
the impact of homologous recombination repair deficiency on depleted uranium clastogenicity in chinese hamster ovary cells: xrcc3 protects cells from chromosome aberrations, but increases chromosome fragmentation.depleted uranium (du) is extensively used in both industry and military applications. the potential for civilian and military personnel exposure to du is rising, but there are limited data on the potential health hazards of du exposure. previous laboratory research indicates du is a potential carcinogen, but epidemiological studies remain inconclusive. du is genotoxic, inducing dna double strand breaks, chromosome damage and mutations, but the mechanisms of genotoxicity or repair pathways involv ...201424561002
long-term activation upon brief exposure to xanomleline is unique to m1 and m4 subtypes of muscarinic acetylcholine receptors.xanomeline is an agonist endowed with functional preference for m1/m4 muscarinic acetylcholine receptors. it also exhibits both reversible and wash-resistant binding to and activation of these receptors. so far the mechanisms of xanomeline selectivity remain unknown. to address this question we employed microfluorometric measurements of intracellular calcium levels and radioligand binding to investigate differences in the short- and long-term effects of xanomeline among muscarinic receptors expr ...201424558448
effect of sodium butyrate on the assembly, charge variants, and galactosylation of antibody produced in recombinant chinese hamster ovary cells.sodium butyrate (nabu) is known to increase the specific productivity of recombinant chinese hamster ovary (rcho) cells. to understand the effects of nabu on the product quality, rcho cells producing monoclonal antibody (mab) were cultivated at various concentrations of nabu (0 to 4 mm). nabu increased correctly assembled mab. in the absence of nabu, the proportions of intact mab (2h2l) and heavy chain dimer (2h) were 81 and 15 %. at 1 mm nabu, the proportion of 2h2l increased to 93 %, whereas t ...201424557571
sodium butyrate enhances the acidic isoform content of recombinant human erythropoietin produced by chinese hamster ovary cells.sodium butyrate is commonly used in mammalian cell cultures to increase the productivity of recombinant proteins. a chinese hamster ovary (cho) cell line producing recombinant human erythropoietin (rhepo) was cultured in commercial medium. addition of 0.5 mm butyrate inhibited the over-growth of the cells after the medium was changed from serum-added medium to serum-free medium. at the 6th day, the addition of butyrate lowered the transcriptional level of sialidases i, ii, and iii compared to th ...201424557070
expression of base excision repair key factors and mir17 in familial and sporadic breast cancer.understanding of brca1/2 interaction with the base excision repair (ber) pathway could improve therapy based on 'synthetic lethality', whose effectiveness is based on homologous recombination deficiency in cells lacking functional brca genes. however, poly (adp-ribose) polymerase (parp) inhibitors failed in some patients and for this reason we explored ber key enzyme expression. in this study, the expression of ber enzymes (redox factor 1/apurinic-apyrimidinic endonuclease 1 (ref1/apex1), nth en ...201424556691
profiling convoluted single-dimension proton nmr spectra: a plackett-burman approach for assessing quantification error of metabolites in complex mixtures with application to cell culture.single-dimension hydrogen, or proton, nuclear magnetic resonance spectroscopy (1d-(1)h nmr) has become an attractive option for characterizing the full range of components in complex mixtures of small molecular weight compounds due to its relative simplicity, speed, spectral reproducibility, and noninvasive sample preparation protocols compared to alternative methods. one challenge associated with this method is the overlap of nmr resonances leading to "convoluted" spectra. while this can be mit ...201424555717
a strategy for fusion expression and preparation of functional glucagon-like peptide-1 (glp-1) analogue by introducing an enterokinase cleavage site.kglp-1, a 31-amino acid glucagon-like peptide-1 (glp-1) analogue, has a great therapeutic potential for anti-diabetes. in this work, a strategy for expression and purification of functional kglp-1 peptide has been established. kglp-1 cdna was fused with glutathione s-transferase (gst), with an enterokinase cleavage site in the fusion junction. the recombinant fusion protein gst-kglp-1 was affinity purified via the gst-tag, and then digested with enterokinase. the resulting gst part as well as th ...201424737080
efficient production and purification of extracellular domain of human fgfr-fc fusion proteins from chinese hamster ovary cells.the family of fibroblast growth factor receptors (fgfrs) plays an important role in cell growth, survival, differentiation and angiogenesis. the three immunoglobulin-like extracellular domains of fgfr (d1, d2, and d3) are critical for ligand binding and specificity towards fibroblast growth factor and heparan sulfate. fibroblast growth factor receptors are overexpressed in a wide variety of tumors, such as breast, bladder, and prostate cancer, and therefore they are attractive targets for differ ...201424727156
optimization of heterologous protein production in chinese hamster ovary cells under overexpression of spliced form of human x-box binding protein.the optimization of protein production is a complex and challenging problem in biotechnology. different techniques for transcription, translation engineering and the optimization of cell culture conditions have been used to improve protein secretion, but there remain many open problems involving post-translational modifications of the secreted protein and cell line stability.201424725707
effect of glutamine substitution by tca cycle intermediates on the production and sialylation of fc-fusion protein in chinese hamster ovary cell culture.in an effort to reduce the accumulation of ammonia in culture medium, three different tca cycle intermediates, (alpha-ketoglutarate (α-kg), citric acid and succinic acid) along with glutamic acid for a comparison, were examined as a substitute for glutamine with rcho cell line producing a fc-fusion glycoprotein. among them, α-kg showed the best production performance. when cells were cultivated with 4 mm α-kg, the final ammonia concentration did not exceed 3 mm, which is less than one fourth of ...201424721212
ultrasound driven assembly of lignin into microcapsules for storage and delivery of hydrophobic molecules.oil-filled microcapsules of kraft lignin were synthesized by first creating an oil in water emulsion followed by a high-intensity, ultrasound-assisted cross-linking of lignin at the water/oil interface. the rationale behind our approach is based on promoting documented lignin hydrophobic interactions within the oil phase, followed by locking the resulting spherical microsystems by covalent cross-linking using a high intensity ultrasound treatment. as further evidence in support of our rationale, ...201424720505
synthesis, spectral characterization, and antiproliferative studies of mixed ligand titanium complexes of adamantylamine.titanium complexes have been synthesized by the reaction between titanium tetrachloride (ticl4), respective bidentate ligand [4,4' -dimethoxy-2,2' -bipyridine (bpome), 6,6'-dimethyl-2,2'-bipyridine (dpme), 1,2-diaminocyclohexane (dach), 1,10-phenanthroline (phen), and benzoylacetone (bzac)], and adamantylamine (ada) in 1 : 2 : 2 molar ratios, respectively. the structure of synthesized complexes was confirmed using elemental analysis, ftir, uv-visible, (1)h nmr, and mass spectrometry techniques. ...201424715822
surfactant uptake dynamics in mammalian cells elucidated with quantitative coherent anti-stokes raman scattering microspectroscopy.the mechanism of surfactant-induced cell lysis has been studied with quantitative coherent anti-stokes raman scattering (cars) microspectroscopy. the dynamics of surfactant molecules as well as intracellular biomolecules in living chinese hamster lung (chl) cells has been examined for a low surfactant concentration (0.01 w%). by using an isotope labeled surfactant having cd bonds, surfactant uptake dynamics in living cells has been traced in detail. the simultaneous cars imaging of the cell itse ...201424710120
modulation of amyloid precursor protein expression reduces β-amyloid deposition in a mouse model.proteolytic cleavage of the amyloid precursor protein (app) generates β-amyloid (aβ) peptides. prolonged accumulation of aβ in the brain underlies the pathogenesis of alzheimer disease (ad) and is regarded as a principal target for development of disease-modifying therapeutics.201424687915
evaluation of anti-inflammatory and mechanism of action of extract of macrosiphonia longiflora (desf.) müll. arg.macrosiphonia longiflora (desf.) müll. arg (apocynaceae), popularly known as 'velame' and 'velame branco', is a native subshrub that grows in the brazilian cerrado. this plant is widely used in traditional medicine in the form of decoction and infusion, particularly as anti-inflammatory, depurative, anti-rheumatic, antisyphilitic and antiulcer remedy. there is no available information in the literature that has addressed its pharmacological activity and phytochemical analysis.201424681039
microrna-9 and microrna-326 regulate human dopamine d2 receptor expression, and the microrna-mediated expression regulation is altered by a genetic variant.the human dopamine receptor d2 (drd2) has been implicated in the pathophysiology of schizophrenia and other neuropsychiatric disorders. most antipsychotic drugs influence dopaminergic transmission through blocking dopamine receptors, primarily drd2. we report here the post-transcriptional regulation of drd2 expression by two brain-expressed micrornas (mirs), mir-326 and mir-9, in an ex vivo mode, and show the relevance of mir-mediated drd2 expression regulation in human dopaminergic neurons and ...201424675081
rorα and ror γ are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin d.rorα and rorγ are expressed in human skin cells that produce the noncalcemic 20-hydroxyvitamin d3 [20(oh)d3] and 20,23-dihydroxyvitamin d3 [20,23(oh)2d3]. chinese hamster ovary (cho) cells stably expressing a tet-on rorα or rorγ expression vector and a ror-responsive element (rore)-luc reporter, and a mammalian 2-hybrid model examining the interaction between the ligand binding domain (lbd) of rorα or rorγ with an lbd-interacting lxxll-peptide, were used to study ror-antagonist activities. these ...201424668754
a high throughput screening assay system for the identification of small molecule inhibitors of gsp.mis-sense mutations in the α-subunit of the g-protein, gsα, cause fibrous dysplasia of bone/mccune-albright syndrome. the biochemical outcome of these mutations is constitutively active gsα and increased levels of camp. the aim of this study was to develop an assay system that would allow the identification of small molecule inhibitors specific for the mutant gsα protein, the so-called gsp oncogene. commercially available chinese hamster ovary cells were stably transfected with either wild-type ...201424667240
setting time affects in vitro biological properties of root canal sealers.biocompatibility of root canal sealers is important because of the long-term contact of their eluates and/or degradation products with periapical tissues. the literature still lacks studies about the genotoxic effects of these materials and the influence of setting time on biological properties. the cytotoxicity and genotoxicity of an epoxy resin-based sealer (ah plus), a single methacrylate-based sealer (endorez), and a silicone-based sealer (roekoseal) were assessed.201424666905
arp2, a novel pro-apoptotic protein expressed in epithelial prostate cancer lncap cells and epithelial ovary cho transformed cells.neoplastic epithelial cells generate the most aggressive types of cancers such as those located in the lung, breast, colon, prostate and ovary. during advanced stages of prostate cancer, epithelial cells are associated to the appearance of androgen-independent tumors, an apoptotic-resistant phenotype that ultimately overgrows and promotes metastatic events. we have previously identified and electrophysiologically characterized a novel ca(2+)-permeable channel activated during apoptosis in the an ...201424465888
high-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control.decreasing the timeframe for cell culture process development has been a key goal toward accelerating biopharmaceutical development. advanced microscale bioreactors (ambr™) is an automated micro-bioreactor system with miniature single-use bioreactors with a 10-15 ml working volume controlled by an automated workstation. this system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody in a recombinant chinese hamster ovary cell li ...201424449637
mutations in pgap3 impair gpi-anchor maturation, causing a subtype of hyperphosphatasia with mental retardation.glycosylphophatidylinositol (gpi)-anchored proteins play important roles in many biological processes, and mutations affecting proteins involved in the synthesis of the gpi anchor are reported to cause a wide spectrum of intellectual disabilities (ids) with characteristic additional phenotypic features. here, we describe a total of five individuals (from three unrelated families) in whom we identified mutations in pgap3, encoding a protein that is involved in gpi-anchor maturation. three sibling ...201424439110
construction of a public cho cell line transcript database using versatile bioinformatics analysis pipelines.chinese hamster ovary (cho) cell lines represent the most commonly used mammalian expression system for the production of therapeutic proteins. in this context, detailed knowledge of the cho cell transcriptome might help to improve biotechnological processes conducted by specific cell lines. nevertheless, very few assembled cdna sequences of cho cells were publicly released until recently, which puts a severe limitation on biotechnological research. two extended annotation systems and web-based ...201424427317
very-long-chain polyunsaturated fatty acids accumulate in phosphatidylcholine of fibroblasts from patients with zellweger syndrome and acyl-coa oxidase1 deficiency.peroxisomes are subcellular organelles that function in multiple anabolic and catabolic processes, including β-oxidation of very-long-chain fatty acids (vlcfa) and biosynthesis of ether phospholipids. peroxisomal disorders caused by defects in peroxisome biogenesis or peroxisomal β-oxidation manifest as severe neural disorders of the central nervous system. abnormal peroxisomal metabolism is thought to be responsible for the clinical symptoms of these diseases, but their molecular pathogenesis r ...201424418004
new melatonin (mt1/mt2) ligands: design and synthesis of (8,9-dihydro-7h-furo[3,2-f]chromen-1-yl) derivatives.herein we describe the synthesis of novel tricyclic analogues issued from the rigidification of the methoxy group of the benzofuranic analogue of melatonin as mt1 and mt2 ligands. most of the synthesized compounds displayed high binding affinities at mt1 and mt2 receptors subtypes. compound 6b (mt1, ki=0.07nm; mt2, ki=0.08nm) exhibited with the vinyl 6c and allyl 6d the most interesting derivatives of this series. functional activity of these compounds showed full agonist activity with ec50 in t ...201424417958
soluble receptor for advanced glycation end products (srage) attenuates haemodynamic changes to chronic hypoxia in the mouse.the calgranulin-like protein mts1/s100a4 and the receptor for advanced glycation end-products (rage) have recently been implicated in mediating pulmonary arterial smooth muscle cell proliferation and vascular remodelling in experimental pulmonary arterial hypertension (ph). here, the effects of rage antagonism upon 2 weeks of hypobaric hypoxia (10% o2)-induced ph in mice were assessed. treatment with srage was protective against hypobaric hypoxia-induced increases in right ventricular pressure b ...201424417910
cytoskeleton modification and cholesterol depletion affect membrane properties and caveolae positioning of cho cells.the formation of protrusions is necessary for numerous biological processes. it involves extension of the plasma membrane, and the force needed for this is provided by the actin cytoskeleton. tether pulling with optical tweezers can mimic the formation of a protrusion, so we used this method to investigate the effects of modifying not only actin (with latrunculin a) but also microtubules (with nocodazole) and the plasma membrane itself (with methyl-β-cyclodextrin) on the chinese hamster ovary ce ...201424413749
analysis of inter-subunit contacts reveals the structural malleability of extracellular domains in platelet glycoprotein ib-ix complex.the glycoprotein (gp)ib-ix complex is critical to hemostasis and thrombosis. its proper assembly is closely correlated with its surface expression level and requires cooperative interactions among extracellular and transmembrane domains of ibα, ibβ and ix subunits. two interfaces have been previously identified between the extracellular domains of ibβ and ix.201424406065
[construction and expression of a eukaryotic expression vector containing human cr2-fc fusion protein].to construct a eukaryotic expression vector containing human complement receptor 2 (cr2)-fc and express the cr2-fc fusion protein in chinese hamster ovary (cho) cells.201424405588
polyamine transport inhibitors: design, synthesis, and combination therapies with difluoromethylornithine.the development of polyamine transport inhibitors (ptis), in combination with the polyamine biosynthesis inhibitor difluoromethylornithine (dfmo), provides a method to target cancers with high polyamine requirements. the dfmo+pti combination therapy results in sustained intracellular polyamine depletion and cell death. a series of substituted benzene derivatives were evaluated for their ability to inhibit the import of spermidine in dfmo-treated chinese hamster ovary (cho) and l3.6pl human pancr ...201424405276
influence of npc1 genotype on the toxicity of hydroxypropyl-β-cyclodextrin, a potentially therapeutic agent, in niemann-pick type c disease models.hydroxypropyl-β-cyclodextrin (hpbcd) is an attractive drug candidate against niemann-pick type c (npc) disease. however, the safety of hpbcd treatment for npc patients remains to be elucidated. in this study, we examined the acute toxicity of hpbcd in npc1-deficient mice. when treated with hpbcd (20,000 mg/kg, subcutaneously), over half of the wild-type (npc1(+/+)) or npc1(+/-) mice died by 72 h after the injection. in contrast, all of the npc1(-/-) mice survived. marked pathophysiological chang ...201427896072
screening and subcloning of high producer transfectomas using semisolid media and automated colony picker.generation of high-producing clones is a perquisite for achieving recombinant protein yields suitable for biopharmaceutical production. however, in many industrially important cell lines used to produce recombinant proteins such as chinese hamster ovary, mouse myeloma line (ns0), and hybridomas, only a minority of clones show significantly above-average productivity. thus, in order to have a reasonable probability of finding rare high-producing clones, a large number of clones need to be screene ...201424515462
quantitative estimation of dna damage by photon irradiation based on the microdosimetric-kinetic model.the microdosimetric-kinetic (mk) model is one of the models that can describe the fraction of cells surviving after exposure to ionizing radiation. in the mk model, there are specific parameters, k and yd, where k is an inherent parameter to represent the number of potentially lethal lesions (plls) and yd indicates the dose-mean lineal energy in kev/μm. assuming the plls to be dna double-strand breaks (dsbs), the rate equations are derived for evaluating the dsb number in the cell nucleus. in th ...201424515253
fibs-enabled noninvasive metabolic profiling.in the era of computational biology, new high throughput experimental systems are necessary in order to populate and refine models so that they can be validated for predictive purposes. ideally such systems would be low volume, which precludes sampling and destructive analyses when time course data are to be obtained. what is needed is an in situ monitoring tool which can report the necessary information in real-time and noninvasively. an interesting option is the use of fluorescent, protein-bas ...201424513729
photoluminescent green carbon nanodots from food-waste-derived sources: large-scale synthesis, properties, and biomedical applications.we have developed a simple approach for the large-scale synthesis of water-soluble green carbon nanodots (g-dots) from many kinds of large food waste-derived sources. about 120 g of g-dots per 100 kg of food waste can be synthesized using our simple and environmentally friendly synthesis approach. the g-dots exhibit a high degree of solubility in water because of the abundant oxygen-containing functional groups around their surface. the narrow band of photoluminescence emission (400-470 nm) conf ...201424512145
micromolar concentrations of rimonabant directly inhibits delta opioid receptor specific ligand binding and agonist-induced g-protein activity.there is a growing number of evidence showing, that the cannabinoid receptor 1 (cb1) antagonist rimonabant has many non-cannabimimetic actions, such as affecting the opioid system. the direct effect of rimonabant on opioid receptors has been studied so far mainly on μ-opioid receptors. however recently the δ-opioid receptor (dor) receives much more attention as before, due to its potential therapeutic applications, such as nociception or treatment for psychiatric disorders.201424508403
potential theranostics application of bio-synthesized silver nanoparticles (4-in-1 system).in this report, we have designed a simple and efficient green chemistry approach for the synthesis of colloidal silver nanoparticles (b-agnps) that is formed by the reduction of silver nitrate (agno3) solution using olax scandens leaf extract. the colloidal b-agnps, characterized by various physico-chemical techniques exhibit multifunctional biological activities (4-in-1 system). firstly, bio-synthesized silver nanoparticles (b-agnps) shows enhanced antibacterial activity compared to chemically ...201424505239
protective activity of a novel resveratrol analogue, hs-1793, against dna damage in 137cs-irradiated cho-k1 cells.resveratrol has received considerable attention as a polyphenol with anti-oxidant, anti-carcinogenic, and anti-inflammatory effects. radiation is an important component of therapy for a wide range of malignant conditions. however, it causes damage to normal cells and, hence, can result in adverse side effects. this study was conducted to examine whether hs-1793, a novel resveratrol analogue free from the restriction of metabolic instability and the high dose requirement of resveratrol, induces a ...201424403520
in-silico modeling of glycosylation modulation dynamics in herg ion channels and cardiac electrical signals.cardiac action potentials (ap) are produced by the orchestrated functions of ion channels. a slight change in ion channel activity may affect the ap waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. however, little is known about how reduced glycosylation impacts the gating of herg (human ether-a-go-go related gene) channel, which is partiall ...201424403418
effects of copper on cho cells: insights from gene expression analyses.copper concentration can impact lactate metabolism in chinese hamster ovary (cho) cells. in our previous study, a 20-fold increase in initial copper concentration enabled cho cultures to shift from net lactate production to net lactate consumption, and achieve higher cell growth and productivity. in this follow-up study, we used transcriptomics to investigate the mechanism of action (moa) of copper that mediates this beneficial metabolism shift. from microarray profiling (days 0-7), the number o ...201424403277
gain-of-function kcnh2 mutations in patients with brugada syndrome.brugada syndrome (brs) is an inherited disease characterized by right precordial st segment elevation on electrocardiograms (ecgs) that predisposes patients to sudden cardiac death as a result of polymorphic ventricular tachyarrhythmia or ventricular fibrillation (vf). in brs patients, except for scn5a, mutations in other responsible genes are poorly elucidated.201424400717
scaffold/matrix attachment regions from cho cell chromosome enhanced the stable transfection efficiency and the expression of transgene in cho cells.two scaffold/matrix attachment regions (s/mars) from chinese hamster ovary (cho) cell chromosomes were identified computationally and cloned as the domain boundaries regions. as partition chromosomes in independently regulated units to increase the exogenous gene expression, these two s/mars (mar1 and mar6) flanked expression vectors were respectively transfected into cho cells. the results showed that mar1 and mar6 were much more potent to increase the stable transfection efficiency than the ch ...201424397363
isolating and characterizing three alternatively spliced mu opioid receptor variants: mmor-1a, mmor-1o, and mmor-1p.extensive alternative pre-mrna splicing of the mu opioid receptor gene, oprm1, has demonstrated an array of splice variants in mice, rats and humans. three classes of splice variants have been identified: full-length seven transmembrane (tm) domain variants with c-terminal splicing, truncated 6tm variants and single tm variants. the current studies isolates and characterizes an additional three full-length c-terminal splice variants generated from the mouse oprm1 gene: mmor-1a, mmor-1o, and mmor ...201424375714
genetic variation in the two-pore domain potassium channel, task-1, may contribute to an atrial substrate for arrhythmogenesis.the two-pore domain potassium channel, k2p3.1 (task-1) modulates background conductance in isolated human atrial cardiomyocytes and has been proposed as a potential drug target for atrial fibrillation (af). task-1 knockout mice have a predominantly ventricular phenotype however, and effects of task-1 inactivation on atrial structure and function have yet to be demonstrated in vivo. the extent to which genetic variation in kcnk3, that encodes task-1, might be a determinant of susceptibility to af ...201424374141
inhibition of cardiac two-pore-domain k+ (k2p) channels by the antiarrhythmic drug vernakalant--comparison with flecainide.the mixed ion channel blocker, vernakalant (rsd1235), is effective in rapid conversion of atrial fibrillation (af) to sinus rhythm (sr). suppression of cardiac two-pore-domain potassium (k2p) channels causes action potential prolongation and has recently been proposed as a novel antiarrhythmic strategy. the objective of this study was to investigate acute effects of vernakalant on human k2p2.1 (trek-1) and k2p3.1 (task-1) channels to provide a more complete picture of its antiarrhythmic mechanis ...201424374008
geneoptimizer program-assisted cdna reengineering enhances srage autologous expression in chinese hamster ovary cells.soluble receptor for advanced glycation end products (srage) is a secreted mammalian protein that functions as a decoy to counter-react rage signaling-resultant pathological conditions, and has high therapeutic potentials. our prior studies showed that recombinant human srage expressed in chinese hamster, ceanothus griseus, ovary (cho) cells is modified by specific n-glycosylation, and exhibits higher bioactivity than that expressed in other host systems including insect spodoptera frugiperda ce ...201424373844
signaling regulates activity of dhcr24, the final enzyme in cholesterol synthesis.the role of signaling in regulating cholesterol homeostasis is gradually becoming more widely recognized. here, we explored how kinases and phosphorylation sites regulate the activity of the enzyme involved in the final step of cholesterol synthesis, 3β-hydroxysterol δ24-reductase (dhcr24). many factors are known to regulate dhcr24 transcriptionally, but little is known about its posttranslational regulation. we developed a system to specifically test human ectopic dhcr24 activity in a model cel ...201424363437
dynamics of growth and metabolism controlled by glutamine availability in chinese hamster ovary cells.the physiology of animal cells is characterized by constantly changing environmental conditions and adapting cellular responses. applied dynamic metabolic flux analysis captures metabolic dynamics and can be applied to industrially relevant cultivation conditions. we investigated the impact of glutamine availability or limitation on the physiology of cho k1 cells in eight different batch and fed-batch cultivations. varying glutamine availability resulted in global metabolic changes. we observed ...201424362913
neutralization of leukotriene c4 and d4 activity by monoclonal and single-chain antibodies.cysteinyl leukotrienes (lts) are key mediators in inflammation. to explore the structure of the antigen-recognition site of a monoclonal antibody against ltc4 (mabltc), we previously isolated full-length cdnas for heavy and light chains of the antibody and prepared a single-chain antibody comprising variable regions of these two chains (scfvltc).201424361619
allosteric modulators of human a2b adenosine receptor.among adenosine receptors (ars) the a2b subtype exhibits low affinity for the endogenous agonist compared with the a1, a2a, and a3 subtypes and is therefore activated when concentrations of adenosine increase to a large extent following tissue damages (e.g. ischemia, inflammation). for this reason, a2b ar represents an important pharmacological target.201424361612
dose-rate effect was observed in t98g glioma cells following bnct.it is generally said that low let radiation produce high dose-rate effect, on the other hand, no significant dose rate effect is observed in high let radiation. although high let radiations are produced in bnct, little is known about dose-rate effect of bnct.201424360864
tubb1 mutation disrupting microtubule assembly impairs proplatelet formation and results in congenital macrothrombocytopenia.this report describes a family with tubb1-associated macrothrombocytopenia diagnosed based on abnormal platelet β1-tubulin distribution. a circumferential marginal microtubule band was undetectable, whereas microtubules were frayed and disorganized in every platelet from the affected individuals. patients were heterozygous for novel tubb1 p.f260s that locates at the α- and β-tubulin intradimer interface. mutant β1-tubulin was not incorporated into microtubules with endogenous α-tubulin, and α-tu ...201424344610
calcium electroporation in three cell lines: a comparison of bleomycin and calcium, calcium compounds, and pulsing conditions.electroporation with calcium (calcium electroporation) can induce atp depletion-associated cellular death. in the clinical setting, the cytotoxic drug bleomycin is currently used with electroporation (electrochemotherapy) for palliative treatment of tumors. calcium electroporation offers several advantages over standard treatment options: calcium is inexpensive and may readily be applied without special precautions, as is the case with cytostatic drugs. therefore, details on the use of calcium e ...201424342489
low ph inactivation for xenotropic gamma retrovirus in recombinant human tnf-α receptor immunoglobulin g and mechanism of inactivation.cho-derived recombinant proteins for human therapeutic are used commonly. there are noninfectious endogenous retroviruses in cho cells. validation study for inactivation process is required. murine xenotropic gamma retrovirus (x-mulv) is a model virus in validation study. in our previous study, optimum conditions for x-mulv inactivation were sifted. in this study, we performed a further research on low ph inactivation for evaluation of x-mulv clearance in manufacturing of recombinant human tnf-α ...201424341971
asparagine-linked glycosylation of bone morphogenetic protein-2 is required for secretion and osteoblast differentiation.bone morphogenetic protein-2 (bmp-2), a glycosylated protein, has been demonstrated to play a key role in osteoblast differentiation. however, the function of its glycosylation is incompletely understood. in this study, we examined the role that n-linked glycans (nlg) play in the secretion of bmp-2. blocking the addition of nlgs or inhibiting initial glycan processing prevented the secretion of bmp-2. to identify the specific glycosylation sites, we abolished potential sites of n-linked glycosyl ...201424336949
digital mrna profiling of n-glycosylation gene expression in recombinant chinese hamster ovary cells treated with sodium butyrate.to understand the effects of sodium butyrate (nabu) on protein glycosylation, recombinant chinese hamster ovary (rcho) cells producing fc-fusion glycoprotein were subjected to 3mm nabu. the addition of nabu to the cultures reduced the relative proportion of acidic isoforms and sialic acid content of the glycoprotein. fifty-two n-glycosylation-related gene expressions were also assessed by the nanostring ncounter system, which can provide a direct digital readout using custom-designed color-coded ...201424333461
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