Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| using a dihydrofolate reductase-based strategy for producing the biosimilar version of pertuzumab in cho-s cells. | targeted therapy using monoclonal antibodies (mabs) against epidermal growth factor receptor-2 (erbb2) has been utilized for the treatment of breast cancer, recently. pertuzumab, one of the anti-erbb2 mabs, was approved by fda in 2012 for the treatment of metastatic breast cancer. the aim of this study was to produce biosimilar version of pertuzumab in chinese hamster ovary (cho)-s cell line, and compare its erbb2-binding and biological activities, with commercial drug, perjeta. to this end, a d ... | 2018 | 29437532 |
| strategy for generating sequence-defined aptamer reagent sets for detecting protein contaminants in biotherapeutics. | biologic drugs are typically manufactured in mammalian host cells, and it is critical from a drug safety and efficacy perspective to detect and remove host cell proteins (hcps) during production. this is currently achieved with sets of polyclonal antibodies (pabs), but these suffer from critical shortcomings because their composition is inherently undefined, and they cannot detect nonimmunogenic hcps. in this work, we report a high-throughput screening and array-based binding characterization st ... | 2018 | 29436820 |
| butyrated mannac analog improves protein expression in chinese hamster ovary cells. | the chemical additive sodium butyrate (nabu) has been applied in cell culture media as a direct and convenient method to increase the protein expression in chinese hamster ovary (cho) and other mammalian cells. in this study, we examined an alternative chemical additive, 1,3,4-o-bu3 mannac, for its effect on recombinant protein production in cho. supplementation with 1,3,4-o-bu3 mannac for two stable cho cell lines, expressing human erythropoietin or igg, enhanced protein expression for both pro ... | 2018 | 29427449 |
| tumor targeting with novel pyridyl 6-substituted pyrrolo[2,3- d]pyrimidine antifolates via cellular uptake by folate receptor α and the proton-coupled folate transporter and inhibition of de novo purine nucleotide biosynthesis. | tumor-targeted specificities of 6-substituted pyrrolo[2,3- d]pyrimidine analogues of 1, where the phenyl side-chain is replaced by 3',6' (5, 8), 2',5' (6, 9), and 2',6' (7, 10) pyridyls, were analyzed. proliferation inhibition of isogenic chinese hamster ovary (cho) cells expressing folate receptors (frs) α and β were in rank order, 6 > 9 > 5 > 7 > 8, with 10 showing no activity, and 6 > 9 > 5 > 8, with 10 and 7 being inactive, respectively. antiproliferative effects toward frα- and frβ-expressi ... | 2018 | 29425443 |
| inhibitory effect of bofutsushosan (fang feng tong sheng san) on glucose transporter 5 function in vitro. | bofutsushosan (bts; fang feng tong sheng san in chinese) is a formula in traditional japanese kampo medicine and chinese medicine comprising eighteen crude drugs, and is used to treat obesity and metabolic syndrome. fructose is contained in refreshing beverages as high-fructose corn syrup, and is associated with obesity. fructose is absorbed via glucose transporter 5 (glut5) in the intestine. therefore, the inhibition of glut5 is considered to be a target of obesity drugs. we evaluated the inhib ... | 2018 | 29423591 |
| "antimicrobial and antiproliferative activity of essential oil, aqueous and ethanolic extracts of ocimum micranthum willd leaves". | ocimum micranthum willd is a plant used in traditional medicine practiced in the region of the yucatan peninsula. in particular, it is used for the treatment of cutaneous infections and wound healing, however there are currently no existing scientific studies that support these applications. the aim of the present study was to evaluate the antimicrobial and the in vitro proliferative activity (on healthy mammalian cell lines) of the essential oil and extracts (aqueous and ethanolic) of this plan ... | 2018 | 29422064 |
| improving titer while maintaining quality of final formulated drug substance via optimization of cho cell culture conditions in low-iron chemically defined media. | during biopharmaceutical process development, it is important to improve titer to reduce drug manufacturing costs and to deliver comparable quality attributes of therapeutic proteins, which helps to ensure patient safety and efficacy. we previously reported that relative high-iron concentrations in media increased titer, but caused unacceptable coloration of a fusion protein during early-phase process development. ultimately, the fusion protein with acceptable color was manufactured using low-ir ... | 2018 | 29388872 |
| comparison of the in vitro toxicological activity of various particulate matter. | ultrafine particles (ufps, < 2.5 µm) in air pollutants have been identified as a major cause of respiratory diseases, since they can affect the lung alveoli through the bronchus. in particular, if toxicants such as heavy metals and polycyclic aromatic hydrocarbons (pahs) are present in ufps, they can cause diseases such as asthma, chronic obstructive pulmonary disease, and lung cancer. this study compared in vitro toxicity of various particulate matter including ufps from combustion particles of ... | 2018 | 29415641 |
| characterization of glycoengineered anti-her2 monoclonal antibodies produced by using a silkworm-baculovirus expression system. | silkworm-baculovirus expression systems are efficient means for the production of recombinant proteins that provide high expression levels and post-translational modifications. here, we characterized the stability, glycosylation pattern and antibody-dependent cell-mediated cytotoxicity activity of anti-her2 monoclonal antibodies containing native or glycoengineered mammalian-like n-glycans that were produced by using a silkworm-baculovirus expression system. compared with a monoclonal antibody p ... | 2018 | 29415204 |
| debottlenecking protein secretion and reducing protein aggregation in the cellular host. | chinese hamster ovary (cho) cells have been extensively used for industrial production of biotherapeutics. with advances in cell line development and process optimization, production levels of therapeutic proteins using the cho expression system have increased to beyond 10g per liter scale. these high-titer processes could challenge the secretory capacity of cho cells, which can result in degradation and aggregation of the protein of interest. this review discusses bottlenecks in the secretory p ... | 2018 | 29414073 |
| applications of proteomic methods for cho host cell protein characterization in biopharmaceutical manufacturing. | chinese hamster ovary (cho) cells are the most prevalent host organism for production of recombinant therapeutic proteins, including monoclonal antibodies (mabs). regulatory guidance mandates control of the host cell protein (hcp) concentration in the production process, which remains a primary challenge. although hcp concentrations are typically measured by elisa, orthogonal proteomic methods are gaining popularity for identification and quantitation of individual hcp species. recent applicatio ... | 2018 | 29414072 |
| a quantitative pcr-based assay reveals that nucleotide excision repair plays a predominant role in the removal of dna-protein crosslinks from plasmids transfected into mammalian cells. | dna-protein crosslinks (dpcs) are complex dna lesions that induce mutagenesis and cell death. dpcs are created by common antitumor drugs, reactive oxygen species, and endogenous aldehydes. since these agents create other types of dna damage in addition to dpcs, identification of the mechanisms of dpc repair is challenging. in this study, we created plasmid substrates containing site-specific dpc lesions, as well as plasmids harboring lesions that are selectively repaired by the base excision or ... | 2018 | 29413806 |
| grp78 protects cho cells from ribosylation. | d-ribose (rib), a reactive glycation compound that exists in organisms, abnormally increases in the urine of diabetic patients and can yield large amounts of advanced glycation end products (ages), leading to cell dysfunction. however, whether cellular proteins are sensitive to this type of glycation is unknown. in this study, we found that cellular ages accumulate in chinese hamster ovary (cho) cells with increased rib concentration and administration time. mass spectrum analysis of isolated ag ... | 2018 | 29410209 |
| both the ph domain and n-terminal region of oxysterol-binding protein related protein 8s are required for localization to pm-er contact sites. | oxysterol-binding protein-related proteins are implicated in the sensing and transporting lipids at the membrane contact sites. one of the members of the mammalian orp family, orp8, is thought to transport lipids through directly tethering both er and pm membranes. targeting to pm is thought to be mediated by n-terminal pleckstrin homology domain via binding to phosphoinositides. sequence alignments and nmr structural determination revealed that the ph domain of orp8 is atypical and contains an ... | 2018 | 29409900 |
| rimonabant, a potent cb1 cannabinoid receptor antagonist, is a gαi/o protein inhibitor. | rimonabant is a potent and selective cannabinoid cb1 receptor antagonist widely used in animal and clinical studies. besides its antagonistic properties, numerous studies have shown that, at micromolar concentrations rimonabant behaves as an inverse agonist at cb1 receptors. the mechanism underpinning this activity is unclear. here we show that micromolar concentrations of rimonabant inhibited gαi/o-type g proteins, resulting in a receptor-independent block of g protein signaling. accordingly, r ... | 2018 | 29407764 |
| 13 c flux analysis reveals that rebalancing medium amino acid composition can reduce ammonia production while preserving central carbon metabolism of cho cell cultures. | 13 c metabolic flux analysis (mfa) provides a rigorous approach to quantify intracellular metabolism of industrial cell lines. in this study, 13 c mfa was used to characterize the metabolic response of chinese hamster ovary (cho) cells to a novel medium variant designed to reduce ammonia production. ammonia inhibits growth and viability of cho cell cultures, alters glycosylation of recombinant proteins, and enhances product degradation. ammonia production was reduced by manipulating the amino ac ... | 2018 | 29405605 |
| receptor-specific crosstalk between prostanoid e receptor 3 and bombesin receptor subtype 3. | bombesin receptor subtype 3 (brs-3) is a gpcr that is expressed in the cns, peripheral tissues, and tumors. our understanding of brs-3's role in physiology and pathophysiology is limited because its natural ligand is unknown. in an attempt to identify this ligand, we screened toad skin ( bufo bufo gargarizans cantor) extracts and identified prostaglandins as putative ligands. in brs-3-transfected human embryonic kidney (hek) cells, we found that prostaglandins, with prostaglandin e2 (pge2) being ... | 2018 | 29401613 |
| a novel bicistronic gene design couples stable cell line selection with a fucose switch in a designer cho host to produce native and afucosylated glycoform antibodies. | the conserved glycosylation site asn297 of a monoclonal antibody (mab) can be decorated with a variety of sugars that can alter mab pharmacokinetics and recruitment of effector proteins. antibodies lacking the core fucose at asn297 (afucosylated mabs) show enhanced antibody-dependent cell-mediated cytotoxicity (adcc) and increased efficacy. here, we describe the development of a robust platform for the manufacture of afucosylated therapeutic mabs by engineering a chinese hamster ovary (cho) host ... | 2018 | 29400603 |
| impact of cho metabolism on cell growth and protein production: an overview of toxic and inhibiting metabolites and nutrients. | for over three decades, chinese hamster ovary (cho) cells have been the chosen expression platform for the production of therapeutic proteins with complex post-translational modifications. however, the metabolism of these cells is far from perfect and optimized, and requires substantial know how and process optimization and monitoring to perform efficiently. one of the main reasons for this is the production and accumulation of toxic and growth-inhibiting metabolites during culture. lactate and ... | 2018 | 29393587 |
| mild hypothermic culture conditions affect residual host cell protein composition post-protein a chromatography. | host cell proteins (hcps) are endogenous impurities, and their proteolytic and binding properties can compromise the integrity, and, hence, the stability and efficacy of recombinant therapeutic proteins such as monoclonal antibodies (mabs). nonetheless, purification of mabs currently presents a challenge because they often co-elute with certain hcp species during the capture step of protein a affinity chromatography. a quality-by-design (qbd) strategy to overcome this challenge involves identify ... | 2018 | 29381421 |
| exploring the potential application of short non-coding rna-based genetic circuits in chinese hamster ovary cells. | the majority of cell engineering for recombinant protein production to date has relied on traditional genetic engineering strategies, such as gene overexpression and gene knock-outs, to substantially improve the production capabilities of chinese hamster ovary (cho) cells. however, further improvements in cellular productivity or control over product quality is likely to require more sophisticated rational approaches to coordinate and balance cellular pathways. for these strategies to be impleme ... | 2018 | 29377624 |
| analysis of radiation-induced chromosomal aberrations on a cell-by-cell basis after alpha-particle microbeam irradiation: experimental data and simulations. | there is a continued need for further clarification of various aspects of radiation-induced chromosomal aberration, including its correlation with radiation track structure. as part of the emrp joint research project, biologically weighted quantities in radiotherapy (bioquart), we performed experimental and theoretical analyses on chromosomal aberrations in chinese hamster ovary cells (cho-k1) exposed to α particles with final energies of 5.5 and 17.8 mev (absorbed doses: ∼2.3 gy and ∼1.9 gy, re ... | 2018 | 29624483 |
| multiple reaction monitoring targeted lc-ms analysis of potential cell death marker proteins for increased bioprocess control. | the monitoring of protein biomarkers for the early prediction of cell stress and death is a valuable tool for process characterization and efficient biomanufacturing control. a representative set of six proteins, namely gpdh, prdx1, lgals1, cfl1, tagln2 and mdh, which were identified in a previous cho-k1 cell death model using discovery lc-mse was translated into a targeted liquid chromatography multiple reaction monitoring mass spectrometry (lc-mrm-ms) platform and verified. the universality of ... | 2018 | 29607450 |
| enhanced biosynthesis performance of heterologous proteins in cho-k1 cells using crispr-cas9. | chinese hamster ovary (cho) cells are the famous expression system for industrial production of recombinant proteins, such as therapeutic antibodies. however, there still remain bottlenecks in protein quality and weakness in expression efficiency because of the intrinsic genetic properties of the cell. here we have enhanced biosynthesis performance of heterologous proteins in cho-k1 cells using crispr-cas9 by editing the genome precisely with two genes for improving er microenvironment and reinf ... | 2018 | 29683658 |
| a polymeric immunoglobulin-antigen fusion protein strategy for enhancing vaccine immunogenicity. | in this study, a strategy based on polymeric immunoglobulin g scaffolds (pigs) was used to produce a vaccine candidate for mycobacterium tuberculosis. a genetic fusion construct comprising genes encoding the mycobacterial ag85b antigen, an immunoglobulin γ-chain fragment and the tailpiece from immunoglobulin μ chain was engineered. expression was attempted in chinese hamster ovary (cho) cells and in nicotiana benthamiana. the recombinant protein assembled into polymeric structures (tb-pigs) in n ... | 2018 | 29682888 |
| quantitative proteomic analysis of cellular responses to a designed amino acid feed in a monoclonal antibody producing chinese hamster ovary cell line | chinese hamster ovary (cho) cell line is considered as the most common cell line in the biopharmaceutical industry because of its capability in performing efficient post-translational modifications and producing the recombinant proteins, which are similar to natural human proteins. the optimization of the upstream process via different feed strategies has a great impact on the target molecule expression and yield. | 2018 | 29678103 |
| toxic hypf-n oligomers selectively bind the plasma membrane to impair cell adhesion capability. | the deposition of fibrillar protein aggregates in human organs is the hallmark of several pathological states, including highly debilitating neurodegenerative disorders and systemic amyloidoses. it is widely accepted that small oligomers arising as intermediates in the aggregation process, released by fibrils, or growing in secondary nucleation steps are the cytotoxic entities in protein-misfolding diseases, notably neurodegenerative conditions. increasing evidence indicates that cytotoxicity is ... | 2018 | 29590593 |
| combination of temperature shift and hydrolysate addition regulates anti-ige monoclonal antibody charge heterogeneity in chinese hamster ovary cell fed-batch culture. | charge heterogeneity has been broadly studied as a critical quality attribute during monoclonal antibody (mab) production that may subsequently affect product stability and biopotency. however, the charge variation distribution is poorly controlled, so methods of more effective control need to be explored. in this study, the combined effects of temperature shift (37-34, 37-32, or 37-30 °c) and hydrolysate addition (0.100 g/l) to culture feed on the charge heterogeneity of anti-ige mab were inves ... | 2018 | 29589263 |
| [impact of neutron radiation on the viability of tumor cells cultured in the presence of boron-10 isotope]. | to investigate the impact of a neutron beam formed with the accelerator-based epithermal neutron source designed at the g.i. budker institute of nuclear physics (inp) on the viability of human and animal tumor cells cultured in the presence of boron-10 isotope. | 2018 | 30241133 |
| analgesia by deletion of spinal neurokinin 1 receptor expressing neurons using a bioengineered substance p-pseudomonas exotoxin conjugate. | cell deletion approaches to pain directed at either the primary nociceptive afferents or second-order neurons are highly effective analgesic manipulations. second-order spinal neurons expressing the neurokinin 1 (nk1) receptor are required for the perception of many types of pain. to delete nk1+ neurons for the purpose of pain control, we generated a toxin–peptide conjugate using dtnb-derivatized (cys0) substance p (sp) and a n-terminally truncated pseudomonas exotoxin (pe35) that retains the en ... | 2018 | 28814145 |
| toxicity studies of lactobacillus plantarum ps128tm isolated from spontaneously fermented mustard greens. | : probiotics are extensively available to consumers; however, the use of probiotics may not always be safe, and there are few reports on their side effects, including those of lactobacillus. lactobacillus plantarum strain ps128tm isolated from spontaneously fermented mustard greens in taiwan was recently reported to exhibit probiotic properties. in this study, we aimed to assess the safety of strain ps128tm for use in humans via examining genotoxic and oral toxic effects using in vitro and in vi ... | 2019 | 31835837 |
| assessment of dna vaccine encoding toxoplasma gondii microneme complete gene and il-12 as adjuvant in balb/c mice. | toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis in humans and animals. micronemes (mics) are effective candidates for dna vaccine. | 2019 | 31579446 |
| functional analysis of ldlr (low-density lipoprotein receptor) variants in patient lymphocytes to assess the effect of evinacumab in homozygous familial hypercholesterolemia patients with a spectrum of ldlr activity. | homozygous familial hypercholesterolemia is a rare disease usually caused by ldlr (low-density lipoprotein receptor) mutations. homozygous familial hypercholesterolemia is characterized by markedly elevated ldl-c (low-density lipoprotein cholesterol) levels and an extremely high risk of premature atherosclerotic cardiovascular disease. a phase 2, proof-of-concept study (nct02265952) demonstrated that evinacumab, a fully human monoclonal antibody to angptl3 (angiopoietin-like 3 protein), reduced ... | 2019 | 31578082 |
| identification of cp77 as the third orthopoxvirus samd9 and samd9l inhibitor with unique specificity for a rodent samd9l. | orthopoxviruses (opxvs) have a broad host range in mammalian cells, but chinese hamster ovary (cho) cells are nonpermissive for vaccinia virus (vacv). here, we revealed a species-specific difference in host restriction factor samd9l as the cause for the restriction and identified orthopoxvirus cp77 as a unique inhibitor capable of antagonizing chinese hamster samd9l (chsamd9l). two known vacv inhibitors of samd9 and samd9l (samd9&l), k1 and c7, can bind human and mouse samd9&l, but neither can b ... | 2019 | 30918078 |
| development of an anti-bear podoplanin monoclonal antibody pmab-247 for immunohistochemical analysis. | sensitive and specific monoclonal antibodies (mabs) targeting podoplanin (pdpn) are needed for immunohistochemical analyses using formalin-fixed paraffin-embedded tissues because pdpn is known as a lymphatic endothelial cell maker in pathology. recently, we established anti-pdpn mabs against many species, such as human, mouse, rat, rabbit, dog, cat, bovine, pig, horse, goat, tiger, alpaca, and tasmanian devil. however, anti-bear pdpn (bpdpn) has not been established yet. in this study, we immuni ... | 2019 | 31061899 |
| establishment of a monoclonal antibody pmab-233 for immunohistochemical analysis against tasmanian devil podoplanin. | monoclonal antibodies (mabs) against not only human, mouse, and rat but also rabbit, dog, cat, bovine, pig, and horse podoplanins (pdpns) have been established in our previous studies. pdpn is used as a lymphatic endothelial cell marker in pathological diagnoses. however, mabs against tasmanian devil pdpn (taspdpn), which are useful for immunohistochemical analysis, remain to be developed. herein, mice were immunized with taspdpn-overexpressing chinese hamster ovary (cho)-k1 (cho/taspdpn) cells, ... | 2019 | 30984883 |
| intact bioactivities and improved pharmacokinetic of the sl335-ifn-β-1a fusion protein that created by genetic fusion of sl335, a human anti-serum albumin fab, and human interferon-β. | recombinant human interferon beta (rifn-β) has long been used as a first-line treatment for multiple sclerosis (ms), and any attempt to develop a long-acting rifn-β is desirable since only one pegylated version of long-acting rifn-β-1a (plegridy) is currently available in clinics. previously, we reported that sl335, a human fab molecule specific to serum albumin, exhibits an extended serum half-life via utilizing the fcrn recycling mechanism. with the ultimate goal of developing a long-acting ri ... | 2019 | 30684504 |
| establishment of a monoclonal antibody pmab-231 for tiger podoplanin. | podoplanin (pdpn), also known as t1alpha, has been used as a lung type i alveolar cell marker in the pathophysiological condition. although we have established several monoclonal antibodies (mabs) against mammalian pdpns, mabs against tiger pdpn (tigpdpn), which are useful for immunohistochemical analysis, remain to be developed. in this study, we immunized mice with tigpdpn-overexpressing chinese hamster ovary (cho)-k1 cells (cho/tigpdpn) and screened hybridomas producing mabs against tigpdpn u ... | 2019 | 31009336 |
| increased mab production in amplified cho cell lines is associated with increased interaction of creb1 with transgene promoter. | most therapeutic monoclonal antibodies in biopharmaceutical processes are produced in chinese hamster ovary (cho) cells. technological advances have rendered the selection procedure for higher producers a robust protocol. however, information on molecular mechanisms that impart the property of hyper-productivity in the final selected clones is currently lacking. in this study, an igg-producing industrial cell line and its methotrexate (mtx)-amplified progeny cell line were analyzed using transcr ... | 2019 | 32577618 |
| na+ /ca2+ exchanger 1 is a key mechanosensitive molecule of the esophageal myenteric neurons. | our earlier studies showed that mechanical stretch activates inhibitory motor neurons of the oesophagus; however, the underlying molecular mechanisms are unclear. here, we sought to examine if na+ /ca2+ exchanger 1 (ncx1) is responsible for the mechanosensitivity in the esophageal myenteric neurons (emn) of rats and humans. | 2019 | 30466198 |
| identification of verapamil binding sites within human kv1.5 channel using mutagenesis and docking simulation. | the phenylalkylamine class of l-type ca2+ channel antagonist verapamil prolongs the effective refractory period (erp) of human atrium, which appears to contribute to the efficacy of verapamil in preventing reentrant-based atrial arrhythmias including atrial fibrillation. this study was designed to investigate the molecular and electrophysiological mechanism underlying the action of verapamil on human kv1.5 (hkv1.5) channel that determines action potential duration and erp in human atrium. | 2019 | 30816676 |
| kcnq1 rescues tmc1 plasma membrane expression but not mechanosensitive channel activity. | transmembrane channel-like protein isoform 1 (tmc1) is essential for the generation of mechano-electrical transducer currents in hair cells of the inner ear. tmc1 disruption causes hair cell degeneration and deafness in mice and humans. although thought to be expressed at the cell surface in vivo, tmc1 remains in the endoplasmic reticulum when heterologously expressed in standard cell lines, precluding determination of its roles in mechanosensing and pore formation. here, we report that the kcnq ... | 2019 | 30613966 |
| exposure to the fungicide captan induces dna base alterations and replicative stress in mammalian cells. | the classification of the fungicide captan (cas number: 133-06-2) as a carcinogen agent is presently under discussion. despite the mutagenic effect detected by the ames test and carcinogenic properties observed in mice, the genotoxicity of this pesticide in humans is still unclear. new information is needed about its mechanism of action in mammalian cells. here, we show that chinese hamster ovary (cho) cells exposed to captan accumulate fpg-sensitive dna base alterations. in cho and hela cells, ... | 2019 | 30471166 |
| lion/web: a web-based ontology enrichment tool for lipidomic data analysis. | a major challenge for lipidomic analyses is the handling of the large amounts of data and the translation of results to interpret the involvement of lipids in biological systems. | 2019 | 31141612 |
| early porcine sapovirus infection disrupts tight junctions and uses occludin as a coreceptor. | the genus sapovirus belongs to the family caliciviridae, and its members are common causative agents of severe acute gastroenteritis in both humans and animals. some caliciviruses are known to use either terminal sialic acids or histo-blood group antigens as attachment factors and/or cell surface proteins, such as cd300lf, cd300ld, and junctional adhesion molecule 1 of tight junctions (tjs), as receptors. however, the roles of tjs and their proteins in sapovirus entry have not been examined. in ... | 2019 | 30463963 |
| the comet assay: assessment of in vitro and in vivo dna damage. | anthropogenic activities, indiscriminate and rapid industrialization as well as pursuance of a better life has led to an increase in the concentration of chemicals, like pesticides, automobile exhausts, and new chemical entities, in the environment, which have an adverse effect on all living organisms including humans. sensitive and robust test systems are thus required for accurate hazard identification and risk assessment. the comet assay has been used widely as a simple, rapid, and sensitive ... | 2019 | 31473963 |
| identification of lark as a novel and conserved g-quadruplex binding protein in invertebrates and vertebrates. | double-stranded dnas are usually present in the form of linear b-form double-helix with the base pairs of adenine (a) and thymine (t) or cytosine (c) and guanine (g), but g-rich dna can form four-stranded g-quadruplex (g4) structures, which plays important roles in transcription, replication, translation and protection of telomeres. in this study, a rna recognition motif (rrm)-containing protein, bmlark, was identified and demonstrated to bind g4 structures in the promoters of a transcription fa ... | 2019 | 31165881 |
| new insight into the classification and evolution of glucose transporters in the metazoa. | because glucose is an essential energy source for living organisms, glucose transporters (gluts) are present in all species worldwide. encoded by the solute carrier family 2 gene family, the glut proteins generally have 12 transmembrane helices (tmhs). in total, 14 glut proteins have been identified in humans (hgluts), and they are divided into 3 classes on the basis of their transport characteristics and sequence similarities. herein, we report the use of protein sequence similarity networks (s ... | 2019 | 30888203 |
| simultaneous expression of clophensor and slc26a3 reveals the nature of endogenous oxalate transport in cho cells. | clophensor, a fluorescent fusion protein, is a dual function biosensor that has been utilized as a tool for the simultaneous measurement of intracellular chloride and ph in cells. clophensor has traditionally been used in conjunction with fluorescence microscopy for single cell measurements. here, we present a promising multi-well format advancement for the use of clophensor as a potential high-throughput method capable of measuring fluorescence signal intensity across a well of confluent cells ... | 2019 | 30837228 |
| incorporation of cd40 ligand or granulocyte-macrophage colony stimulating factor into hantaan virus (htnv) virus-like particles significantly enhances the long-term immunity potency against htnv infection. | hantavirus infections cause severe haemorrhagic fever with renal syndrome (hfrs) in humans and are associated with high fatality rates. in 2017, numerous outbreaks were reported in china and germany. this represents a significant public-healthcare issue with no effective hfrs vaccines that offer a long-term immune response. in this study, we investigated the long-term humoral and cellular immune responses and protective immunity of hantaan virus (htnv) granulocyte-macrophage colony stimulating f ... | 2019 | 30657443 |
| glyco-engineered cho cell lines producing alpha-1-antitrypsin and c1 esterase inhibitor with fully humanized n-glycosylation profiles. | recombinant chinese hamster ovary (cho) cells are able to provide biopharmaceuticals that are essentially free of human viruses and have n-glycosylation profiles similar, but not identical, to humans. due to differences in n-glycan moieties, two members of the serpin superfamily, alpha-1-antitrypsin (a1at) and plasma protease c1 inhibitor (c1inh), are currently derived from human plasma for treating a1at and c1inh deficiency. deriving therapeutic proteins from human plasma is generally a cost-in ... | 2019 | 30513349 |
| production of soluble pmhc-i molecules in mammalian cells using the molecular chaperone tapbpr. | current approaches for generating major histocompatibility complex (mhc) class-i proteins with desired bound peptides (pmhc-i) for research, diagnostic and therapeutic applications are limited by the inherent instability of empty mhc-i molecules. using the properties of the chaperone tap-binding protein related (tapbpr), we have developed a robust method to produce soluble, peptide-receptive mhc-i molecules in chinese hamster ovary cells at high yield, completely bypassing the requirement for la ... | 2019 | 32725167 |
| host cell protein removal from biopharmaceutical preparations: towards the implementation of quality by design. | downstream processing of protein products of mammalian cell culture currently accounts for the largest fraction of the total production cost. a major challenge is the removal of host cell proteins, which are cell-derived impurities. host cell proteins are potentially immunogenic and can compromise product integrity during processing and hold-up steps. there is an increasing body of evidence that the type of host cell proteins present in recombinant protein preparations is a function of cell cult ... | 2019 | 29654903 |
| a different perspective: how much innovation is really needed for monoclonal antibody production using mammalian cell technology? | as biopharmaceutical companies have optimized cell line and production culture process development, titers of recombinant antibodies have risen steadily to 3-8 g/l for fed-batch mammalian cultures at production scales of 10 kl or larger. most new antibody products are produced from chinese hamster ovary (cho) cell lines, and there are relatively few alternative production hosts under active evaluation. many companies have adopted a strategy of using the same production cell line for early clinic ... | 2019 | 29721583 |
| bioprocessing of recombinant cho-k1, cho-dg44, and cho-s: cho expression hosts favor either mab production or biomass synthesis. | chinese hamster ovary (cho) cells comprise a variety of lineages including cho-dxb11, cho-k1, cho-dg44, and cho-s. despite all cho cell lines sharing a common ancestor, extensive mutagenesis, and clonal selection has resulted in substantial genetic heterogeneity among them. data from sequencing show that different genes are missing in individual cho cell lines and each cell line harbors a unique set of mutations with relevance to the bioprocess. however, not much literature is available about th ... | 2019 | 29701329 |
| digenic heterozigosity in scn5a and cacna1c explains the variable expressivity of the long qt phenotype in a spanish family. | a known long qt syndrome-related mutation in nav1.5 cardiac channels (p.r1644h) was found in 4 members of a spanish family but only 1 of them showed prolongation of the qt interval. in the other 3 relatives, a novel missense mutation in cav1.2 cardiac channels was found (p.s1961n). here, we functionally analyzed p.s1961n cav1.2 channels to elucidate whether this mutation regulates the expressivity of the long qt syndrome phenotype in this family. | 2019 | 29691127 |
| epigenetic regulation of gene expression in chinese hamster ovary cells in response to the changing environment of a batch culture. | the existence of dynamic cellular phenotypes in changing environmental conditions is of major interest for cell biologists who aim to understand the mechanism and sequence of regulation of gene expression. in the context of therapeutic protein production by chinese hamster ovary (cho) cells, a detailed temporal understanding of cell-line behavior and control is necessary to achieve a more predictable and reliable process performance. of particular interest are data on dynamic, temporally resolve ... | 2019 | 30512195 |
| the mouse-canine chimeric anti-dog podoplanin antibody p38b exerts antitumor activity in mouse xenograft models. | podoplanin (pdpn) is a type i transmembrane heavily glycosylated sialoglycoprotein that is expressed in normal tissues such as pulmonary type i alveolar cells, renal podocytes, and lymphatic endothelial cells. pdpn overexpression in cancerous tissue is associated with hematogenous metastasis through interactions with the c-type lectin-like receptor 2 (clec-2). previously, we have reported the development of a mouse monoclonal antibody (mab), pmab-38 (igg1, kappa) against dog pdpn (dpdpn). pmab-3 ... | 2019 | 30519645 |
| 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 |
| impact of cell culture media additives on igg glycosylation produced in chinese hamster ovary cells. | glycosylation is a key critical quality attribute for monoclonal antibodies and other recombinant proteins because of its impact on effector mechanisms and half-life. in this study, a variety of compounds were evaluated for their ability to modulate glycosylation profiles of recombinant monoclonal antibodies produced in chinese hamster ovary cells. compounds were supplemented into the cell culture feed of fed-batch experiments performed with a cho k1 and a cho dg44 cell line expressing a recombi ... | 2019 | 30552760 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| multistage antiplasmodium activity of astemizole analogues and inhibition of hemozoin formation as a contributor to their mode of action. | a drug repositioning approach was leveraged to derivatize astemizole (ast), an antihistamine drug whose antimalarial activity was previously identified in a high-throughput screen. the multistage activity potential against the plasmodium parasite's life cycle of the subsequent analogues was examined by evaluating against the parasite asexual blood, liver, and sexual gametocytic stages. in addition, the previously reported contribution of heme detoxification to the compound's mode of action was i ... | 2019 | 30525439 |
| a novel muc1 aptamer-modified plga-epirubicin-pβae-antimir-21 nanocomplex platform for targeted co-delivery of anticancer agents in vitro and in vivo. | conventional chemotherapy suffers from several drawbacks, including toxic side effects together with the development of resistance to the chemical agents. therefore, exploring alternative therapeutic approaches as well as developing targeted delivery systems are in demand. oligonucleotide-based therapy has emerged as a promising and alternative procedure for treating malignancies involving gene-related diseases. in the current study, a targeted delivery system was designed to target cancer cells ... | 2019 | 30537619 |
| amber suppression coupled with inducible surface display identifies cells with high recombinant protein productivity. | cell line development (cld) for biotherapeutics is a time- and resource-intensive process requiring the isolation and screening of large numbers of clones to identify high producers. novel methods aimed at enhancing cell line screening efficiency using markers predictive of productivity early in the cld process are needed to reliably generate high-yielding cell lines. to enable efficient and selective isolation of antibody expressing chinese hamster ovary cells by fluorescence-activated cell sor ... | 2019 | 30536645 |
| 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 |
| evaluation of pta-derived ruthenium(ii) and iridium(iii) quinoline complexes against chloroquine-sensitive and resistant strains of the plasmodium falciparum malaria parasite. | cationic 1,3,5‑triaza‑phosphaadamantane (pta) quinoline ruthenium(ii) and iridium(iii) complexes were successfully synthesized and characterized using standard spectroscopic and analytical techniques. the complexes were evaluated for their in vitro antiplasmodial activities against the chloroquine-sensitive (cqs) nf54 and chloroquine-resistant (cqr) k1 strains of the plasmodium falciparum species of the malaria parasite and were found to exhibit good activities in the sensitive strain but modera ... | 2019 | 30529881 |
| efficient interleukin-21 production by optimization of codon and signal peptide in chinese hamster ovarian cells. | interleukin-21 is a common γ-chain cytokine that controls the immune responses of b cells, t cells, and natural killer cells. targeting il-21 to strengthen the immune system is promising for the development of vaccines as well as anti-infection and anti-tumor therapies. however, the practical application of il-21 is limited by the high production cost. in this study, we improved il-21 production by codon optimization and selection of appropriate signal peptide in cho-k1 cells. codon-optimized or ... | 2019 | 30544288 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |