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coronavirus interactions with the cellular autophagy machinery.the covid-19 pandemic, caused by the sars-cov-2 virus, is the most recent example of an emergent coronavirus that poses a significant threat to human health. virus-host interactions play a major role in the viral life cycle and disease pathogenesis, and cellular pathways such as macroautophagy/autophagy prove to be either detrimental or beneficial to viral replication and maturation. here, we describe the literature over the past twenty years describing autophagy-coronavirus interactions. there ...202032964796
dynamic multiscale metabolic network modeling of chinese hamster ovary cell metabolism integrating n-linked glycosylation in industrial biopharmaceutical manufacturing.experimental and modeling work, described in this article, is focused on the metabolic pathway of chinese hamster ovary (cho) cells, which are the preferred expression system for monoclonal antibody protein production. cho cells are one of the primary hosts for monoclonal antibodies production, which have extensive applications in multiple fields like biochemistry, biology and medicine. here, an approach to explain cellular metabolism with in silico modeling of a microkinetic reaction network is ...202032970321
electrophoresis-assisted accumulation of conductive nanoparticles for the enhancement of cell electropermeabilization.the use of conductive nanoparticles (nps) was previously proposed as a way to locally amplify the electric field (ef) intensity at the cell membrane to enhance cell electroporation. to achieve this, a close distance between the nps and the cell membrane is mandatory. here, a new method to improve the contact between nps and cell surface using the effects of electric pulses (electrophoretic forces) is explored. the effects of two types of electric pulses are analyzed alone or combined in a two-pu ...202032980738
expanded chinese hamster organ and cell line proteomics profiling reveals tissue-specific functionalities.chinese hamster ovary (cho) cells are the predominant production vehicle for biotherapeutics. quantitative proteomics data were obtained from two cho cell lines (cho-s and cho dg44) and compared with seven chinese hamster (cricetulus griseus) tissues (brain, heart, kidney, liver, lung, ovary and spleen) by tandem mass tag (tmt) labeling followed by mass spectrometry, providing a comprehensive hamster tissue and cell line proteomics atlas. of the 8470 unique proteins identified, high similarity w ...202032985598
constitutive activation and inactivation of mutations inducing cell surface loss of receptor and impairing of signal transduction of agonist-stimulated eel follicle-stimulating hormone receptor.in the present study, we investigated the signal transduction of mutants of the eel follicle-stimulating hormone receptor (eelfshr). specifically, we examined the constitutively activating mutant d540g in the third intracellular loop, and four inactivating mutants (a193v, n195i, r546c, and a548v). to directly assess functional effects, we conducted site-directed mutagenesis to generate mutant receptors. we measured cyclic adenosine monophosphate (camp) accumulation via homogeneous time-resolved ...202032992880
metabolic understanding of disulfide reduction during monoclonal antibody production.the disulfide reduction of intact monoclonal antibodies (mabs) and subsequent formation of low molecular weight (lmw) species pose a direct risk to product stability, potency, and patient safety. although enzymatic mechanisms of reduction are well established, an understanding of the cellular mechanisms during the bioreactor process leading to increased risk of disulfide reduction after harvest remains elusive. in this study, we examined bench, pilot, and manufacturing-scale batches of two mabs ...202032997205
indole-6-carboxaldehyde isolated from sargassum thunbergii (mertens) kuntze prevents oxidative stress-induced cellular damage in v79-4 chinese hamster lung fibroblasts through the activation of the nrf2/ho-1 signaling pathway.the disruption of redox equilibrium by oxidative stress, which is characterized by an overproduction of reactive oxygen species (ros), is considered to be associated with fibroblast death in severe lung diseases. indole-6-carboxaldehyde (i6ca) is a natural indole derivative isolated from sargassum thunbergii, which a type of brown algae. however, the antioxidative effects of i6ca, and their mechanisms, have not been identified. this study was conducted to investigate the potential protective eff ...202032997463
toolbox accelerating glycomics (tag): glycan annotation from maldi-tof ms spectra and mapping expression variation to biosynthetic pathways.glycans present extraordinary structural diversity commensurate with their involvement in numerous fundamental cellular processes including growth, differentiation, and morphogenesis. unlike linear dna and protein sequences, glycans have heterogeneous structures that differ in composition, branching, linkage, and anomericity. these differences pose a challenge to developing useful software for glycomic analysis. to overcome this problem, we developed the novel toolbox accelerating glycomics (tag ...202032998456
a defucosylated anti‑cd44 monoclonal antibody 5‑mg2a‑f exerts antitumor effects in mouse xenograft models of oral squamous cell carcinoma.cd44 is widely expressed on the surface of most tissues and all hematopoietic cells, and regulates many genes associated with cell adhesion, migration, proliferation, differentiation, and survival. cd44 has also been studied as a therapeutic target in several cancers. previously, an anti‑cd44 monoclonal antibody (mab), c44mab‑5 (igg1, kappa) was established by immunizing mice with cd44‑overexpressing chinese hamster ovary (cho)-k1 cells. c44mab‑5 recognized all cd44 isoforms, and showed high sen ...202033000243
biological activities of a recombinant fortilin from fenneropenaeus merguiensis.human fortilin, an antiapoptotic protein, has also been implicated in several diseases; however, several potential uses of fortilin have also been proposed. bearing the implications of fortilin in mind, fortilin analog, which has no complication with diseases, is required. since a recombinant full-length fortilin from fenneropenaeus merguiensis (rfm-fortilin (fl)) reported only 44% (3e-27) homologous to human fortilin, therefore the biological activities of the fm-fortilin (fl) and its fragments ...202033002062
pharmacokinetic prediction of an antibody in mice based on an in vitro cell-based approach using target receptor-expressing cells.in vivo pharmacokinetics (pk) studies using mice and monkeys are the main approaches for evaluating and predicting the pk of antibodies, and there is a strong demand for methods that do not require animal experiments. in this work, we focused on quantitatively predicting the nonlinear pk of an antibody based on cell-based assays. an anti-mouse fc gamma receptor iib antibody was used as a model antibody. to determine the pk parameters related to nonspecific elimination in vivo, the plasma concent ...202033004886
toxicity assessment of gryllus bimaculatus (a type of cricket) glycosaminoglycan.we performed general toxicity studies of gryllus bimaculatus (two-spotted cricket) glycosaminoglycan (gbg), including a single, 4-week repeated oral dose toxicity test in icr mice, and short-term genotoxicity tests. the mutagenic potential of the purified gbg was non-genotoxic when it was evaluated using short-term genotoxicity tests, namely ames, chromosome aberration (ca), and micronuclei (mn) tests. in salmonella typhimurium and escherichia coli assays, gbg did not produce any mutagenic respo ...202033005591
animal model and bioinformatics analyses suggest the timp1/mmp9 axis as a potential biomarker in oral squamous cell carcinoma.oral squamous cell carcinoma (oscc) is a common malignant tumor of the head and neck. however, the molecular mechanism underlying its development and progression is yet unclear. genes that are differentially expressed, that is, differentially expressed genes (degs), between normal and diseased tissues are believed to be involved in disease development and progression. to identify the degs in oscc and explore their role in occurrence and progression, we established a chinese hamster oscc model, d ...202033006223
improving the expression of anti-il-2rα monoclonal antibody in the cho cells through optimization of the expression vector and translation efficiency.the growing need for monoclonal antibodies (mabs) necessitates the development of novel and efficient production approaches. regulatory elements like ubiquitous chromatin-opening elements (ucoes) have been employed for improvement of the mab expression in the chinese hamster ovary (cho) cells. sineups are a class of long non-coding rnas, which can improve the translation of partly overlapping mrnas. a combination of these two elements might lead to higher production of mabs. therefore, the curre ...202033007349
a rare hcn4 variant with combined sinus bradycardia, left atrial dilatation, and hypertrabeculation/left ventricular noncompaction phenotype.hcn4 variants have been reported to cause combined sick sinus syndrome (sss) and left ventricular noncompaction (lvnc) cardiomyopathy. this relationship has been proven in few cases and no previous patients have associated left atrial dilatation (lad). our objective was to study a familial disorder characterized by sss, lad, and hypertrabeculation/lvnc and to identify the underlying genetic and electrophysiological characteristics.202033008772
a novel kv7.3 variant in the voltage-sensing s4 segment in a family with benign neonatal epilepsy: functional characterization and in vitro rescue by β-hydroxybutyrate.pathogenic variants in kcnq2 and kcnq3, paralogous genes encoding kv7.2 and kv7.3 voltage-gated k+ channel subunits, are responsible for early-onset developmental/epileptic disorders characterized by heterogeneous clinical phenotypes ranging from benign familial neonatal epilepsy (bfne) to early-onset developmental and epileptic encephalopathy (dee). kcnq2 variants account for the majority of pedigrees with bfne and kcnq3 variants are responsible for a much smaller subgroup, but the reasons for ...202033013448
mutagenicity of 7-ketocholesterol in cho cells: the role of lipid peroxidation.as an important cholesterol oxide, 7-ketocholesterol plays a deleterious role in the occurrence of cancer. although the fact had been proved that 7-ketocholesterol could induce several biological phenomena, including apoptosis, dna damage, et al., this issue whether 7-ketocholesterol led to mutagenesis in mammalian cells remains largely unexplored. here, we investigated the major role of lipid peroxidation in the genotoxic response to 7-ketocholesterol in chinese hamster ovary (cho) cells. the r ...202033017620
transient multivalent nanobody targeting to cd206-expressing cells via ph-degradable nanogels.to target nanomedicines to specific cells, especially of the immune system, nanobodies can be considered as an attractive tool, as they lack the fc part as compared to traditional antibodies and, thus, prevent unfavorable fc-receptor mediated mistargeting. for that purpose, we have site-specifically conjugated cd206/mmr-targeting nanobodies to three types of dye-labeled nanogel derivatives: non-degradable nanogels, acid-degradable nanogels (with ketal crosslinks), and single polymer chains (also ...202033019594
a novel method to detect autoantibodies against platelets in patients with immune thrombocytopenia.to examine anti-platelet autoantibodies in patients with immune thrombocytopenia (itp) not only provides solid evidence for diagnosis, and also helps to select an individualized strategy for the treatment. the aim of this study is to develop a novel cell-based assay to detect autoantibodies in itp patients.202033031805
development of a new microdosimetric biological weighting function for the rbe10assessment in case of the v79 cell line exposed to ions from1h to238u.an improved biological weighting function (ibwf) is proposed to phenomenologically relate microdosimetric lineal energy probability density distributions with the relative biological effectiveness (rbe) for the in vitro clonogenic cell survival (surviving fraction = 10%) of the most commonly used mammalian cell line, i.e. the chinese hamster lung fibroblasts (v79). the ibwf, intended as a simple and robust tool for a fast rbe assessment to compare different exposure conditions in particle therap ...202033032273
inhibition of poly(adp-ribose) polymerase induces synthetic lethality in brip1 deficient ovarian epithelial cells.pathogenic variations in the homologous recombination (hr) gene, brca1 interacting protein c-terminal helicase 1 (brip1) increase the risk for ovarian cancer. parp inhibitors (parpi) exert a synthetic lethal effect in brca-mutated ovarian cancers. effective hr requires cooperation between brca1 and brip1; therefore, brip1-incompetancy may predict vulnerability to synthetic lethality. here we investigated the response of ovarian epithelial cells with defective brip1 function to parpi, and compare ...202033032822
1-methylpyrene induces chromosome loss and mitotic apparatus damage in a chinese hamster v79-derived cell line expressing both human cyp1a2 and sulfotransferase 1a1.1-methylpyrene (1-mp) is a ubiquitous environmental pollutant and rodent carcinogen. its mutagenic activity depends on sequential activation by various cyp and sulfotransferase (sult) enzymes. previously we have observed induction of micronuclei and mitotic arrest by 1-mp in a chinese hamster (v79)-derived cell line expressing both human cyp1a2 and sult1a1 (v79-hcyp1a2-hsult1a1), however, the mode of chromosome damage and the involvement of mitotic tubulin structures have not been clarified. in ...202033035519
establishment of microbeam radiation therapy at a small-animal irradiator.microbeam radiation therapy is a preclinical concept in radiation oncology. it spares normal tissue more effectively than conventional radiation therapy at equal tumor control. the radiation field consists of peak regions with doses of several hundred gray, whereas doses between the peaks (valleys) are below the tissue tolerance level. widths and distances of the beams are in the submillimeter range for microbeam radiation therapy. a similar alternative concept with beam widths and distances in ...202033038461
systematically gap-filling the genome-scale metabolic model of cho cells.chinese hamster ovary (cho) cells are the leading cell factories for producing recombinant proteins in the biopharmaceutical industry. in this regard, constraint-based metabolic models are useful platforms to perform computational analysis of cell metabolism. these models need to be regularly updated in order to include the latest biochemical data of the cells, and to increase their predictive power. here, we provide an update to icho1766, the metabolic model of cho cells.202033040240
chlorella vulgaris extract as a serum replacement that enhances mammalian cell growth and protein expression.the global cell culture market is experiencing significant growth due to the rapid advancement in antibody-based and cell-based therapies. both rely on the capacity of different living factories, namely prokaryotic and eukaryotic cells, plants or animals for reliable and mass production. the ability to improve production yield is of important concern. among many strategies pursued, optimizing the complex nutritional requirements for cell growth and protein production has been frequently performe ...202033042965
distinct functions of acyl/alkyl dihydroxyacetonephosphate reductase in peroxisomes and endoplasmic reticulum.plasmalogens are a subclass of ether glycerophospholipids characterized by a vinyl-ether bond at the sn-1 position of the glycerol backbone. plasmalogen biosynthesis is initiated in peroxisomes. at the third step of plasmalogen synthesis, alkyl-dihydroxyacetonephosphate (dhap) is enzymatically reduced to 1-alkyl-sn-glycero-3-phospate by acyl/alkyl dhap reductase (adhapr), whose activity is found in both peroxisomes and microsomes. we herein show that knockdown of adhapr in hela cells reduced the ...202033042986
systematic identification of safe harbor regions in the cho genome through a comprehensive epigenome analysis.the chinese hamster ovary (cho) cell lines that are used to produce commercial quantities of therapeutic proteins commonly exhibit a decrease in productivity over time in culture, a phenomenon termed production instability. random integration of the transgenes encoding the protein of interest into locations in the cho genome that are vulnerable to genetic and epigenetic instability often causes production instability through copy number loss and silencing of expression. several recent publicatio ...202033049068
hypericin, a medicinal compound from st. john's wort, inhibits genotoxicity induced by mutagenic agents in v79 cells.this study evaluated the cytotoxic, genotoxic, and the modulatory effects on dna damage of hypericin in chinese hamster lung fibroblasts (v79 cells). the hypericin is a natural polycyclic quinone, mainly extracted from st. john's wort (hypericum perforatum l.). along with hyperforin, the hypericins are responsible for the antidepressant activity of st. john's wort. cytotoxicity was assessed by the xtt colorimetric assay and the nuclear division index (ndi). the genotoxic activity was studied by ...202033050761
polymer fraction including exosomes derived from chinese hamster ovary cells promoted their growth during serum-free repeated batch culture.while continuous (perfusion) culture of mammalian cells might reduce the reactor size owing to the high cell density, there is the problem of higher medium cost; however, this problem is expected to be solved by the reuse of growth-promoting components in the culture supernatant. the polymer fraction (pf, 10 kda-220 nm) collected from the supernatant of serum-free repeated-batch culture of chinese hamster ovary (cho) cells in not only adhesion but also suspension promoted the cell growth in resp ...202033051156
the quantitative analysis of the mechanism involved in pertussis toxin-mediated cell clustering and its implications in the in vitro quality control of diphtheria tetanus and whole cell pertussis vaccines.some of the adverse side-effects such as leukocytosis, hyperinsulinemia, hypoglycemia and sensitization to histamine, caused by diphtheria, tetanus and whole cell pertussis (dtwp) vaccines are related to the presence of non-inactivated pertussis toxin residues (niptxr). the chinese hamster ovary (cho) cell clustering assay is an in vitro assay to measure niptxr in dtwp vaccines based on the ability of active pertussis toxin (ptx) to cause cellular clustering. to study the biochemical mechanism i ...202033059000
raman spectra-based deep learning: a tool to identify microbial contamination.deep learning has the potential to enhance the output of in-line, on-line, and at-line instrumentation used for process analytical technology in the pharmaceutical industry. here, we used raman spectroscopy-based deep learning strategies to develop a tool for detecting microbial contamination. we built a raman dataset for microorganisms that are common contaminants in the pharmaceutical industry for chinese hamster ovary (cho) cells, which are often used in the production of biologics. using a c ...202033063423
biological activities of a new crotamine-like peptide from crotalus oreganus helleri on c2c12 and cho cell lines, and ultrastructural changes on motor endplate and striated muscle.crotamine and crotamine-like peptides are non-enzymatic polypeptides, belonging to the family of myotoxins, which are found in high concentration in the venom of the crotalus genus. helleramine was isolated and purified from the venom of the southern pacific rattlesnake, crotalus oreganus helleri. this peptide had a similar, but unique, identity to crotamine and crotamine-like proteins isolated from other rattlesnakes species. the variability of crotamine-like protein amino acid sequences may al ...202033065200
development of a high yielding bioprocess for a pre-fusion rsv subunit vaccine.respiratory syncytial virus (rsv) is a highly contagious virus causing severe infection in infants and the elderly. various approaches are being used to develop an effective rsv vaccine. the rsv fusion (f) subunit, particularly the cleaved trimeric pre-fusion f, is one of the most promising vaccine candidates under development. the pre-fusion conformation elicits the majority of neutralizing antibodies during natural infection. however, this pre-fusion conformation is metastable and prone to con ...202033068697
establishment of fast-growing serum-free immortalised cells from chinese hamster lung tissues for biopharmaceutical production.chinese hamster (cricetulus griseus) ovary-derived chinese hamster ovary (cho) cells are the most commonly used mammalian hosts for the industrial production of recombinant therapeutics because of their ability to fold, assemble, and perform post-translational modifications, such as glycosylation, on proteins. they are also valuable for their ability to grow in serum-free suspension cultures. in this study, we established a cell line derived from lung tissue of chinese hamsters, named chinese ha ...202033077772
improving antibody production in stably transfected cho cells by crispr-cas9-mediated inactivation of genes identified in a large-scale screen with chinese hamster-specific sirnas.the chinese hamster ovary (cho) cell line is commonly used for the production of biotherapeutics. as cell productivity directly affects the cost of production, methods are developed to manipulate the expression of specific genes that are known to be involved in protein synthesis, folding, and secretion to increase productivity. however, there are no large-scale cho-specific functional screens to identify novel gene targets that impact the production of secreted recombinant proteins. here, a larg ...202033079482
high-resolution three-dimensional chromatin profiling of the chinese hamster ovary cell genome.chinese hamster ovary (cho) cell lines are the pillars of a multi-billion dollar biopharmaceutical industry producing recombinant therapeutic proteins. the effects of local chromatin organisation and epigenetic repression within these cell lines result in unpredictable and unstable transgene expression following random integration. limited knowledge of the cho genome and its higher-order chromatin organisation has thus far impeded functional genomics approaches required to tackle these issues. h ...202033095445
impact of polyallylamine hydrochloride on gene expression and karyotypic stability of multidrug resistant transformed cells.the synthetic polymer, polyallylamine hydrochloride (paa), is found in a variety of applications in biotechnology and medicine. it is used in gene and sirna transfer, to form microcapsules for targeted drug delivery to damaged and tumor cells. conventional chemotherapy often does not kill all cancer cells and leads to multidrug resistance (mdr). until recently, studies of the effects of paa on cells have mainly focused on their morphological and genetic characteristics immediately or several hou ...202033096691
reduced culture temperature differentially affects expression and biophysical properties of monoclonal antibody variants.reduced culture temperature is an increasingly popular practice to improve recombinant protein yields in cho cells. recent studies have attributed the enhancement of protein titers at sub-physiological temperatures to increased mrna levels as well as extended stationary phase. we observed that reducing the culture temperature arrested cell growth, prolonged viability, and increased cell size. however, the reduced culture temperature had a differential effect on protein and mrna expression of clo ...201433101721
ptselect™: a post-transcriptional technology that enables rapid establishment of stable cho cell lines and surveillance of clonal variation.currently, stable chinese hamster ovary cell lines producing therapeutic, recombinant proteins are established either by antibiotic and/or metabolic selection. here, we report a novel technology, ptselect™ that utilizes an sirna cloned upstream of the gene of interest (goi) that is processed to produce functional ptselect™-sirnas, which enable cell enrichment. cells with stably integrated goi are selected and separated from cells without goi by transfecting cd4/sirna mrna regulated by ptselect™- ...202033115662
evidence for co-translational misincorporation of non-canonical amino acid hydroxyproline in recombinant antibodies produced in chinese hamster ovary (cho) cell lines.with the advent of highly sensitive technologies such as tandem mass spectrometry and next-generation sequencing, recombinant antibodies are now routinely analyzed for the presence of low-level sequence variants including amino acid misincorporations. during mab cell culture process development, we found that proline was replaced with the non-canonical amino acid, hydroxyproline, in the protein sequence. we investigated the relationship between proline content in the cell culture media and proli ...202033119652
methylthioadenosine (mta) boosts cell-specific productivities of chinese hamster ovary cultures: dosage effects on proliferation, cell cycle and gene expression.a major goal for process and cell engineering in the biopharmaceutical industry is enhancing production through increasing volumetric and cell-specific productivities (csp). here, we present 5'-deoxy-5'-(methylthio)adenosine (mta), the degradation product of s-(5'-adenosyl)-l-methionine (sam), as a highly attractive native additive which can boost csp by 79% when added to exponentially growing cells at a concentration of 250-300 μm. notably, cell viability and cell size remain higher than in non ...202033128321
production of trimeric sars-cov-2 spike protein by cho cells for serological covid-19 testing.we describe scalable and cost-efficient production of full length, his-tagged severe acute respiratory syndrome coronavirus 2 (sars-cov-2) spike glycoprotein trimer by chinese hamster ovary (cho) cells that can be used to detect sars-cov-2 antibodies in patient sera at high specificity and sensitivity. transient production of spike in both human embryonic kidney (hek) and cho cells mediated by polyethyleneimine was increased significantly (up to 10.9-fold) by a reduction in culture temperature t ...202033128388
formation of carbonaceous and nitrogenous iodinated disinfection byproducts from biofilm extracellular polymeric substances by the oxidation of iodide-containing waters with lead dioxide.lead dioxide (pbo2) is an important form of lead mineral scales in drinking water pipes. iodide (i-) widely presents in source waters and can be thermodynamically oxidized by pbo2 to the reactive iodine species (i2/hoi). biofilm extracellular polymeric substances (eps) are nonnegligible precursors of disinfection byproducts (dbps). the aim was to study the oxidation of i- by pbo2 and formation of iodinated dbps (i-dbps) from eps. at a high molar ratio of pbo2 to i- (> 100), the observed rate con ...202033128980
trans-cinnamaldehyde prevents oxidative stress-induced apoptosis in v79-4 chinese hamster lung fibroblasts through the nrf2-mediated ho-1 activation.oxidative stress, which is characterized by overproduction of reactive oxygen species (ros), is considered a major risk factor associated with fibroblast death in severe lung diseases such as idiopathic pulmonary fibrosis. trans-cinnamaldehyde (tca), the major phytochemical constituent in cinnamon, is known to possess strong anti-oxidant activity. however, whether tca can defend lung fibroblasts against oxidative injury remains to be elucidated. therefore, this study was conducted to investigate ...202033132316
bioluminescence as a sensitive electroporation indicator in sub-microsecond and microsecond range of electrical pulses.the cell membrane permeabilization in electroporation studies is usually quantified using fluorescent markers such as propidium iodide (pi) or yo-pro, while chinese hamster ovary cell line frequently serves as a model. in this work, as an alternative, we propose a sensitive methodology for detection and analysis of electroporation phenomenon based on bioluminescence. luminescent mice myeloma sp2/0 cells (transfected using luciferase-pcdna3 plasmid) were used as a cell model. electroporation has ...202033142215
adiponectin forms a complex with atherogenic ldl and inhibits its downstream effects.adiponectin, an adipocyte-derived protein, has anti-atherogenic and anti-diabetic effects, but how it confers the anti-atherogenic effects is not well understood. to study the anti-atherogenic mechanisms of adiponectin, we examined whether it interacts with atherogenic low-density lipoprotein (ldl) to attenuate ldl's atherogenicity. l5, the most electronegative subfraction of ldl, induces atherogenic responses similarly to copper-oxidized ldl (oxldl). unlike native ldl endocytosed via the ldl re ...202033144330
igist-a kinetic bioassay for pertussis toxin based on its effect on inhibitory gpcr signaling.detection of pertussis toxin (ptx) activity is instrumental for the development and manufacturing of pertussis vaccines. these quality and safety measures require thousands of mice annually. here, we describe interference in gαi-mediated signal transduction (igist), an animal-free kinetic bioassay for detection of ptx, by measuring its effect on inhibitory g protein-coupled receptor (gpcr) signaling. ptx adp-ribosylates inhibitory α-subunits of the heterotrimeric g proteins, thereby perturbing t ...202033147407
a novel lamprey antibody sequence to multimerize and increase the immunogenicity of recombinant viral and bacterial vaccine antigens.hemagglutinin, the major surface protein of influenza viruses, was recombinantly expressed in eukaryotic cells as a monomer instead of its native trimer, and was only immunogenic when administered with an adjuvant [pion et al. 2014]. in order to multimerize this antigen to increase its immunogenicity, a cysteine-rich peptide sequence found at the extreme c-terminus of lamprey variable lymphocyte receptor (vlr)-b antibodies was fused to various recombinant hemagglutinin (rha) proteins from a and ...202033153770
the cag promoter maintains high-level transgene expression in hek293 cells.the vast majority of therapeutic recombinant proteins are produced in mammalian cell lines. however, proteins generated in non-human cell lines, such as chinese hamster ovary (cho) cells, are decorated with human-like glycan structures that differ from those of human cells, and these may induce immunogenic responses in human cells. human embryonic kidney cells (hek293f) are also extensively used as hosts for the expression of recombinant therapeutic proteins, but their utility is limited by the ...202033155423
control of igg glycosylation in cho cell perfusion cultures by greba mathematical model supported by a novel targeted feed, tafe.the n-linked glycosylation pattern is an important quality attribute of therapeutic glycoproteins. it has been reported by our group and by others that different carbon sources, such as glucose, mannose and galactose, can differently impact the glycosylation profile of glycoproteins in mammalian cell culture. acting on the sugar feeding is thus an attractive strategy to tune the glycan pattern. however, in case of feeding of more than one carbon source simultaneously, the cells give priority to ...202033161144
metabolic profiling of cho cell cultures at different working volumes and agitation speeds using spin tube reactors.culture systems based on spin tube reactors have been consolidated in the development of manufacturing processes based on chinese hamster ovary (cho) cells. despite their widespread use, there is little information about the consequences of varying operational setting parameters on the culture performance of recombinant cho cell lines. here we investigated the effect of varying working volumes and agitation speeds on cell growth, protein production and cell metabolism of two clonally-derived cho ...202033169492
surface antifouling modification on polyethylene filtration membranes by plasma polymerization.surface modification on microporous polyethylene (pe) membranes was facilitated by plasma polymerizing with two hydrophilic precursors: ethylene oxide vinyl ether (eo1v) and diethylene oxide vinyl ether (eo2v) to effectively improve the fouling against mammalian cells (chinese hamster ovary, cho cells) and proteins (bovine serum albumin, bsa). the plasma polymerization procedure incorporated uniform and pin-hole free ethylene oxide-containing moieties on the filtration membrane in a dry single-s ...202033172217
single-cell transcriptome analyses reveal heterogeneity in suspension cultures and clonal markers of cho-k1 cells.cell-to-cell variability in cell populations arises from a combination of intrinsic factors and extrinsic factors related to the milieu. however, the heterogeneity of high cell density suspension cultures for therapeutic protein production remains unknown. here, we illustrate the increasing heterogeneity in the cellular transcriptome of serum-free adapted cho k1 cells during high cell density suspension culture over time without concomitant changes in the genomic sequence. cell cycle-dependent s ...202033179258
high-level expression of the monomeric sars-cov-2 s protein rbd 320-537 in stably transfected cho cells by the eef1a1-based plasmid vector.the spike (s) protein is one of the three proteins forming the coronaviruses' viral envelope. the s protein of the severe acute respiratory syndrome coronavirus 2 (sars-cov-2) has a spatial structure similar to the s proteins of other mammalian coronaviruses, except for a unique receptor-binding domain (rbd), which is a significant inducer of host immune response. recombinant sars-cov-2 rbd is widely used as a highly specific minimal antigen for serological tests. correct exposure of antigenic d ...202133529230
development of a colloidal gold-based immunochromatographic strip for rapid detection of severe acute respiratory syndrome coronavirus 2 spike protein.the outbreak and worldwide pandemic of the novel severe acute respiratory syndrome coronavirus 2 (sars-cov-2) have a significant impact on global economy and human health. in order to reduce the disease spread, 16 monoclonal antibodies (mcabs) again sars-cov-2 were generated by immunized mice with the spike protein receptor binding domain (rbd), which was expressed in chinese hamster ovary cell (cho). a colloidal gold-based immunochromatographic strip was developed with two mcabs to detect sars- ...202133777026
preclinical development of a molecular clamp-stabilised subunit vaccine for severe acute respiratory syndrome coronavirus 2.efforts to develop and deploy effective vaccines against severe acute respiratory syndrome coronavirus 2 (sars-cov-2) continue at pace. here, we describe rational antigen design through to manufacturability and vaccine efficacy of a prefusion-stabilised spike (s) protein, sclamp, in combination with the licensed adjuvant mf59 'mf59c.1' (seqirus, parkville, australia).202133841880
the cryo-em structure of vesivirus 2117 highlights functional variations in entry pathways for viruses in different clades of the vesivirus genus.vesivirus 2117 is an adventitious agent that has been responsible for lost productivity in biopharmaceutical production following contamination of chinese hamster ovary cell cultures in commercial bioreactors. a member of the caliciviridae, 2117 is classified within the vesivirus genus in a clade that includes canine and mink caliciviruses but is distinct from the vesicular exanthema of swine virus (vesv) clade, which includes the extensively studied feline calicivirus (fcv). we have used cryoge ...202133853966
structural and functional characterization of sars-cov-2 rbd domains produced in mammalian cells.as the severe acute respiratory syndrome coronavirus 2 (sars-cov-2) pandemic is still ongoing and dramatically influences our life, the need for recombinant viral proteins for diagnostics, vaccine development, and research is very high. the spike (s) protein, and particularly its receptor-binding domain (rbd), mediates the interaction with the angiotensin-converting enzyme 2 (ace2) receptor on host cells and may be modulated by its structural features. therefore, well-characterized recombinant r ...202133871970
culture media optimization for chinese hamster ovary cell growth and expression of recombinant varicella-zoster virus glycoprotein e.herpes zoster (hz) is caused by reactivation of varicella-zoster virus (vzv) latent in the sensory ganglia and causes severe pain, often leading to postherpetic neuralgia (phn). two prophylactic vaccines against hz are currently licensed for human use, a live attenuated vaccine and a subunit vaccine containing recombinant vzv glycoprotein e (ge) as antigen. the latter has superior protective efficacy against hz and phn. during hz subunit vaccine development, we obtained chinese hamster ovary (ch ...202133897103
development of anti-human t cell immunoreceptor with ig and itim domains (tigit) monoclonal antibodies for flow cytometry.immune checkpoint inhibitors targeting programmed cell death-ligand 1 (pd-l1), programmed cell death-1 (pd-1), and cytotoxic t lymphocyte-associated antigen 4 (ctla-4) recently made a significant survival rate improvement in cancer treatment. t cell immunoreceptor with ig and itim domains (tigit) is expressed in t and nk cells related to their activities. it has a single extracellular immunoglobulin domain, a type 1 transmembrane domain, and a single intracellular itim. tigit binds with poliovir ...202133900817
culturing chinese hamster ovary cells on cyclo olefin polymer triggers epithelial-mesenchymal transition and spheroid formation, which increases the foreign gene expression driven by the moloney murine leukemia virus long terminal repeat promoter.chinese hamster ovary (cho) cells are frequently used for recombinant protein production (rpp) as a host. while the rpp has been proven successful, there is still a compelling need for further improvement. cyclo olefin polymer (cop) is a plastic material widely utilized due to its properties including its low protein absorption. we applied this as a raw material for rpp cell culture to see if the cop is suitable. a recombinant cho cell line expressing the human erythropoietin (hepo) gene under t ...202133913259
immunogenicity and safety of hepatitis b vaccination in patients with type 2 diabetes in china: an open-label randomized controlled trial.hepatitis b virus (hbv) infection remains a global public health challenge. patients with diabetes are at greater risk of hbv infection than healthy people. the immunogenicity and safety of two major hepatitis b vaccines were evaluated in chinese patients with diabetes.202133992438
identification of a new cell-penetrating peptide derived from the african swine fever virus cd2v protein.the african swine fever virus (asfv) is a huge and complex dna virus that can lead to the acute death of pigs and cause huge losses to the global swine industry. the cd2v protein is a transmembrane protein encoded by the asfv's ep402r gene, which can effectively inhibit the bystander lymphocyte proliferation in response to mitogens and mediate the absorption of red blood cells to asfv-infected cells. the cd2v protein contains repetitive amino acid sequences ([kpcppp]3 labeled as raas), which is ...202134006158
production of monoclonal antibodies targeting aminopeptidase n in the porcine intestinal mucosal epithelium.porcine aminopeptidase n (apn), a membrane-bound metallopeptidase abundantly present in small intestinal mucosa, can initiate a mucosal immune response without any interference such as low protein expression, enzyme inactivity, or structural changes. this makes apn an attractive candidate in the development of vaccines that selectively target the mucosal epithelium. previous studies have shown that apn is a receptor protein for both enterotoxigenic escherichia coli (e. coli) f4 and transmissible ...202134096912
efficacy of minute virus of mice (mvm) inactivation utilizing high temperature short time (htst) pasteurization and suitability assessment of pasteurized, concentrated glucose feeds in chinese hamster ovary (cho) cell expression systems.there is a growing need to provide effective adventitious agent mitigation for high risk upstream cell culture raw materials used for the production of biologics. it is also highly important in the growing fields of cell and gene therapies. glucose is a critical raw material necessary for effective cell growth and productivity; however, glucose is the highest risk animal-origin-free raw material for viral contamination, and often the highest risk raw material in the upstream process as more comp ...202134257631
previously published ethno-pharmacological reports reveal the potentiality of plants and plant-derived products used as traditional home remedies by bangladeshi covid-19 patients to combat sars-cov-2.several plants have traditionally been used since antiquity to treat various gastroenteritis and respiratory symptoms similar to covid-19 outcomes. the common symptoms of covid-19 include fever or chills, cold, cough, flu, headache, diarrhoea, tiredness/fatigue, sore throat, loss of taste or smell, asthma, shortness of breath, or difficulty breathing, etc. this study aims to find out the plants and plant-derived products which are being used by the covid-19 infected patients in bangladesh and ho ...202134305428
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