Publications

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expression of recombinant classical swine fever virus e2 glycoprotein by endogenous txnip promoter in stable transgenic cho cells.as the main immunogen that could stimulate neutralized antibody in pigs, recombinant e2 protein of csfv was expressed in cho-dhfr-cells driven by endogenous txnip promoter from chinese hamster. different fragments of txnip promoter were amplified by pcr from isolated genomic dna of cho cells and cloned into different expression vectors. compared with cmv promoter, cho-ptxnip-4-re2 (f12) cell clone with the highest yield of re2 protein was established by random insertion of the expression cassett ...202032774204
development and characterization of anti-sheep podoplanin monoclonal antibodies pmab-253 and pmab-260.anti-podoplanin (pdpn) monoclonal antibodies (mabs) are needed as markers for lymphatic endothelial cells or type i alveolar cells in immunohistochemical analyses. we have developed anti-pdpn mabs for many species, including humans, mice, rats, rabbits, dogs, cats, bovines, pigs, tasmanian devils, alpacas, tigers, whales, goats, horses, and bears. this study develops and characterizes anti-sheep pdpn (spdpn) mabs using cell-based immunization and screening (cbis) method. a rap14 tag was added to ...202032679010
osmolality as a lever to modulate the n-glycolylneuraminicacid (neu5gc) level of a recombinant glycoprotein produced in chinese hamster ovary cells.glycoproteins could be highly sialylated, and controlling the sialic acid levels for some therapeutic proteins is critical to ensure product consistency and efficacy. n-acetylneuraminic acid (neu5ac, or nana) and n-glycolylneuraminic acid (neu5gc, or ngna) are the two most common forms of sialic acids produced in mammalian cells. as neu5gc is not produced in humans and can elicit immune responses, minimizing neu5gc formation is important in controlling this quality attribute for complex glycopro ...202032542945
high throughput generation of a resource of the human secretome in mammalian cells.the proteins secreted by human tissues and blood cells, the secretome, are important both for the basic understanding of human biology and for identification of potential targets for future diagnosis and therapy. here, a high-throughput mammalian cell factory is presented that was established to create a resource of recombinant full-length proteins covering the majority of those annotated as 'secreted' in humans. the full-length dna sequences of each of the predicted secreted proteins were gener ...202032502629
vanadium: risks and possible benefits in the light of a comprehensive overview of its pharmacotoxicological mechanisms and multi-applications with a summary of further research trends.vanadium (v) is an element with a wide range of effects on the mammalian organism. the ability of this metal to form organometallic compounds has contributed to the increase in the number of studies on the multidirectional biological activity of its various organic complexes in view of their application in medicine.202032305626
lack of acute toxicity and mutagenicity from recombinant epinephelus lanceolatus piscidin expressed in pichia pastoris.the antimicrobial peptide (amp) piscidin was identified from epinephelus lanceolatus and demonstrated to possess antimicrobial and immune-related functions. supplementation of feed with recombinant epinephelus lanceolatus piscidin (rep)-expressing yeast pellets may minimize the excessive use of antibiotics and control pathogens in aquaculture or animal husbandry. however, before implementing rep as a supplement, it is necessary to understand whether it harbors any toxicity. since toxicological i ...202032290449
gene (hprt) and chromosomal (mn) mutations of nickel metal powder in v79 chinese hamster cells.nickel metal is a naturally occurring element used in many industrial and consumer applications. human epidemiological data and animal cancer bioassays indicate that nickel metal is not likely to be a human carcinogen. yet, nickel metal is classified as a suspected human carcinogen (clp) and possibly carcinogenic to humans (iarc). there are no reliable studies on the potential for nickel metal to induce gene and micronucleus (mn) mutations. to fill these datagaps and increase our understanding o ...202032014793
hamster neogenin, a host-cell protein contained in a respiratory syncytial virus candidate vaccine, induces antibody responses in rabbits but not in clinical trial participants.a recombinant respiratory syncytial virus (rsv) fusion glycoprotein candidate vaccine (rsv-pref) manufactured in chinese hamster ovary cells was developed for immunization of pregnant women, to protect newborns against rsv disease through trans-placental antibody transfer. traces of a host-cell protein, hamster neogenin (haneo1), were identified in purified rsv-pref antigen material. given the high amino-acid sequence homology between haneo1 and human neogenin (huneo1), there was a risk that pot ...202031951765
development of an anti-sheep podoplanin monoclonal antibody pmab-256 for immunohistochemical analysis of lymphatic endothelial cells.sensitive and specific monoclonal antibodies (mabs) targeting podoplanin (pdpn) are needed for immunohistochemical analyses as a marker for lymphatic endothelial cells. we recently have developed anti-pdpn mabs against many species, including human, mouse, rat, rabbit, dog, cat, bovine, pig, tasmanian devil, alpaca, tiger, whale, goat, horse, and bear. however, anti-sheep pdpn (spdpn) has not yet been established. in this study, we used the cell-based immunization and screening method for the de ...202032349619
towards a new avenue for producing therapeutic proteins: microalgae as a tempting green biofactory.most of the recent approved therapeutic proteins are multi-subunit biologics, which need glycosylation and disulfide bridges for their correct conformation and biological functions. currently, there exist many protein-based drugs that are mostly produced in the chinese hamster ovary (cho) cells. however, this expression system appears to associate with some limitations both in upstream and downstream processing steps, including low growth rate, sensitivity to different stresses and pathogens, an ...202031862234
recombinant epstein-barr virus glycoprotein 350 as a serological antigen.epstein-barr virus (ebv) glycoprotein 350 (gp350) is the most abundant glycoprotein expressed on the ebv envelope, the major target for neutralizing antibodies and also essential for virion attachment to b lymphocytes. several studies have addressed ebv gp350 as a vaccine candidate, but less commonly as a potential antigen for serological assays. the aim of the current study was to develop a diagnostic tool to quantify ebv gp350-specific igg in previously ebv-infected individuals. a construct en ...202032650039
fam83g is a novel inducer of apoptosis.the family with sequence similarity 83 (fam83) protein family g (fam83g) possesses a predicted consensus phosphorylation motif for serine/threonine-protein kinase d1/protein kinase c mu (pkd1/pkcμ) at serine residue 356 (s356). in this study, overexpressed wild-type fam83g coimmunoprecipitated with pkd1/pkcμ in chinese hamster ovary (cho) cells inhibited heat shock protein 27 (hsp27) phosphorylation at s82 and reduced the living cell number. the expression of a fam83g phosphorylation-resistant m ...202032570757
quinoline-triazole half-sandwich iridium(iii) complexes: synthesis, antiplasmodial activity and preliminary transfer hydrogenation studies.iridium(iii) half-sandwich complexes containing 7-chloroquinoline-1,2,3-triazole hybrid ligands were synthesised and their inhibitory activities evaluated against the plasmodium falciparum malaria parasite. supporting computational analysis revealed that metal coordination to the quinoline nitrogen occurs first, forming a kinetic product that, upon heating over time, forms a more stable cyclometallated thermodynamic product. single crystal x-ray diffraction confirmed the proposed molecular struc ...202032697227
low density lipoprotein receptor variants in the beta-propeller subdomain and their functional impact.background: pathogenic variants in the low density lipoprotein receptor gene are associated with familial hypercholesterolemia. some of these variants can result in incorrect folding of the ldlr protein, which is then accumulated inside the cell and cannot fulfill its function to internalize ldl particles. we analyzed the functional impact of 10 ldlr variants localized in the beta-propeller of epidermal growth factor precursor homology domain. the experimental part of the work was complemented b ...202032695144
in situ fucosylation of the wnt co-receptor lrp6 increases its endocytosis and reduces wnt/β-catenin signaling.wnt/β-catenin signaling regulates critical, context-dependent transcription in numerous physiological events. among the well-documented mechanisms affecting wnt/β-catenin activity, modification of n-glycans by l-fucose is the newest and the least understood. using a combination of chinese hamster ovary cell mutants with different fucosylation levels and cell-surface fucose editing (in situ fucosylation [isf]), we report that α(1-3)-fucosylation of n-acetylglucosamine (glcnac) in the galβ(1-4)-gl ...202032649905
antibodies to the muscarinic acetylcholine receptor m3 in primary biliary cholangitis inhibit receptor function on cholangiocytes.background and aims: in primary biliary cholangitis (pbc), antibodies to a peptide of the muscarinic acetylcholine receptor 3 (machr3) have been described. since the machr3 is expressed on cholangiocytes and machr3-signaling is involved in the pathogenesis of chronic inflammatory biliary diseases, we wanted to investigate whether anti-machr3-antibodies influence the function of the receptor and the proliferative response of cholangiocytes. methods: immunoglobulins were isolated by ammonium sulfa ...202032695096
lc-ms/ms-based quantitative proteomic and phosphoproteomic analysis of cho-k1 cells adapted to growth in glutamine-free media.this study aims to provide insights into the molecular mechanisms underlying adaptation of cho-k1 cells to growth in glutamine-free media and potentially identifying critical signalling proteins and pathways involved in this phenotype.202032648187
s-adenosylmethionine and methylthioadenosine boost cellular productivities of antibody forming chinese hamster ovary cells.the improvement of cell specific productivities for the formation of therapeutic proteins is an important step towards intensified production processes. among others, the induction of the desired production phenotype via proper media additives is a feasible solution provided that said compounds adequately trigger metabolic and regulatory programs inside the cells. in this study, s-(5'-adenosyl)- l-methionine (sam) and 5'-deoxy-5'-(methylthio)adenosine (mta) were found to stimulate cell specific ...202032644191
media supplementation for targeted manipulation of monoclonal antibody galactosylation and fucosylation.monoclonal antibodies are critically important biologics as the largest class of molecules used to treat cancers, rheumatoid arthritis, and other chronic diseases. antibody glycosylation is a critical quality attribute that has ramifications for patient safety and physiological efficacy-one that can be modified by such factors as media formulation and process conditions during production. using a design-of-experiments approach, we examined the effect of 2-f-peracetyl fucose (2fp), uridine, and g ...202032662879
malignant tissues produce divergent antibody glycosylation of relevance for cancer gene therapy effectiveness.gene therapy approaches now allow for the production of therapeutic antibodies by healthy or cancerous human tissues directly in vivo, and, with an increasing number of gene delivery methods available, the cell type for expression can be chosen. yet, little is known about the biophysical changes introduced by expressing antibodies from producer cells or tissues targeted by gene therapy approaches, nor about the consequences for the type of glycosylation. the effects of different glycosylation on ...202032643525
random epigenetic modulation of cho cells by repeated knockdown of dna methyltransferases increases population diversity and enables sorting of cells with higher production capacities.chinese hamster ovary (cho) cells produce a large share of today's biopharmaceuticals. still, the generation of satisfactory producer cell lines is a tedious undertaking. recently, it was found that cho cells, when exposed to new environmental conditions, modify their epigenome, suggesting that cells adapt their gene expression pattern to handle new challenges. the major aim of the present study was to employ artificially induced, random changes in the dna-methylation pattern of cho cells to div ...202032662873
improving the reaction mix of a pichia pastoris cell-free system using a design of experiments approach to minimise experimental effort.a renaissance in cell-free protein synthesis (cfps) is underway, enabled by the acceleration and adoption of synthetic biology methods. cfps has emerged as a powerful platform technology for synthetic gene network design, biosensing and on-demand biomanufacturing. whilst primarily of bacterial origin, cell-free extracts derived from a variety of host organisms have been explored, aiming to capitalise on cellular diversity and the advantageous properties associated with those organisms. however, ...202032637667
what cho is made of: variations in the biomass composition of chinese hamster ovary cell lines.cell line-specific, genome-scale metabolic models enable rigorous and systematic in silico investigation of cellular metabolism. such models have recently become available for chinese hamster ovary (cho) cells. however, a key ingredient, namely an experimentally validated biomass function that summarizes the cellular composition, was so far missing. here, we close this gap by providing extensive experimental data on the biomass composition of 13 parental and producer cho cell lines under various ...202032619503
improving chinese hamster ovary host cell protein elisa using ella®: an automated microfluidic platform.chinese hamster ovary (cho) cells are a mammalian cell line used in the production of therapeutic proteins. host cell proteins (hcps) are process-related impurities that are derived from the host cell expression system. during biopharmaceutical drug development, removal of hcps is required. enzyme-linked immunosorbent assay (elisa) is a common technique to quantitate hcps, but is a labor-intensive process that takes up to 7 h. ella® is an automated instrument that utilizes microfluidics and glas ...202032615786
metabolic engineering challenges of extending n-glycan pathways in chinese hamster ovary cells.in mammalian cells, n-glycans may include multiple n-acetyllactosamine (poly-lacnac) units that can play roles in various cellular functions and properties of therapeutic recombinant proteins. previous studies indicated that β-1,3-n-acetylglucosaminyltransferase 2 (b3gnt2) and β-1,4-galactotransferase 1 (b4galt1) are two of the primary glycosyltransferases involved in generating lacnac units. in the current study, knocking out sialyltransferase genes slightly enhanced the lacnac content (≥4 repe ...202032663509
directed evolution approach to enhance efficiency and speed of outgrowth during single cell subcloning of chinese hamster ovary cells.chinese hamster ovary (cho) cells are the working horse of the pharmaceutical industry. to obtain high producing cell clones and to satisfy regulatory requirements single cell cloning is a necessary step in cell line development. however, it is also a tedious, labor intensive and expensive process. here we show an easy way to enhance subclonability using subcloning by single cell sorting itself as the selection pressure, resulting in improved subcloning performance of three different host cell l ...202032612755
the p.p888l sap97 polymorphism increases the transient outward current (ito,f) and abbreviates the action potential duration and the qt interval.synapse-associated protein 97 (sap97) is an anchoring protein that in cardiomyocytes targets to the membrane and regulates na+ and k+ channels. here we compared the electrophysiological effects of native (wt) and p.p888l sap97, a common polymorphism. currents were recorded in cardiomyocytes from mice trans-expressing human wt or p.p888l sap97 and in chinese hamster ovary (cho)-transfected cells. the duration of the action potentials and the qt interval were significantly shorter in p.p888l-sap97 ...202032612162
functional pasta consumption in healthy volunteers modulates abcg1-mediated cholesterol efflux capacity of hdl.prevention of cardiovascular (cv) disease is considered a central issue in public health and great attention is payed to nutritional approaches, including consumption of functional foods to reduce cv risk in individuals without indications for anti-atherosclerotic drugs. cholesterol efflux capacity (cec) is an important anti-atherogenic property of hdl and a marker of cv risk. we evaluated the effect of a daily consumption of an innovative whole-wheat synbiotic pasta, compared to a control whole ...202032605885
evaluation of osteogenic/cementogenic modulating potential of pai-1 transfected media for stem cells.in vitro evaluation of the effects of plasminogen activator inhibitor-1 (pai-1) transfected-conditioned media (p-cm) on the differentiation of human periodontal ligament stem cells (hpdlscs) and human periapical follicular stem cells (hpafscs).202032603295
design, construction, and expression of recombinant human interferon beta gene in cho-s cell line using ebv-based expression system.codon optimization has been considered as a powerful strategy to increase the expression level of protein therapeutics in mammalian cells. as an empirical approach to study the effects of the codon usage and gc content on heterologous gene expression in suspension adapted chinese hamster ovary (cho-s) cells, we redesigned the recombinant human interferon beta (rhifn- β) gene based on the codon preference of the cho cell in a way to increase the gc content in the third position of each codon.202032582354
expanding the chinese hamster ovary cell long noncoding rna transcriptome using rnaseq.our ability to study chinese hamster ovary (cho) cell biology has been revolutionised over the last decade following the development of next generation sequencing technology and publication of reference dna sequences for cho cells and the chinese hamster. rna sequencing has not only enabled the association of transcript expression with bioreactor conditions and desirable bioprocess phenotypes but played a key role in the characterisation of protein coding and small noncoding rnas. the annotation ...202032558938
lycopene alleviates oxidative stress via the pi3k/akt/nrf2pathway in a cell model of alzheimer's disease.oxidative stress (os) plays an important role in neurodegenerative diseases such as alzheimer's disease (ad). lycopene is a pigment with potent antioxidant and anti-tumor effects. however, its potential role in central nervous system is not well-defined. the aim of this study was to investigate the effect of lycopene on the cell model of ad and determine its underlying mechanisms.202032551202
cytotoxicity and mutagenicity of narrowband uvb to mammalian cells.phototherapy using narrowband ultraviolet-b (nb-uvb) has been shown to be more effective than conventional broadband uvb (bb-uvb) in treating a variety of skin diseases. to assess the difference in carcinogenic potential between nb-uvb and bb-uvb, we investigated the cytotoxicity via colony formation assay, genotoxicity via sister chromatid exchange (sce) assay, mutagenicity via hypoxanthine phosphoribosyltransferase (hprt) mutation assay, as well as cyclobutane pyrimidine dimer (cpd) formation ...202032545288
using metabolomics to identify cell line-independent indicators of growth inhibition for chinese hamster ovary cell-based bioprocesses.chinese hamster ovary (cho) cells are widely used for the production of biopharmaceuticals. efforts to improve productivity through medium design and feeding strategy optimization have focused on preventing the depletion of essential nutrients and managing the accumulation of lactate and ammonia. in addition to ammonia and lactate, many other metabolites accumulate in cho cell cultures, although their effects remain largely unknown. elucidating these effects has the potential to further improve ...202032429145
small intestine proteomics coupled with serum metabolomics reveal disruption of amino acid metabolism in chinese hamsters with type 2 diabetes mellitus.type 2 diabetes mellitus (t2dm) is a metabolic disorder characterized by hyperglycemia, with metabolic disturbances resulting from defects in insulin secretion, insulin resistance (ir), or both. chinese hamsters have potential value as non-obese animal models of spontaneous t2dm for studying the pathogenesis and molecular characteristics of diabetes. in this study, the molecular characteristics of the chinese hamster diabetes animal model were investigated through small intestine proteomics and ...202032428569
applications of small molecules in modulating productivity and product quality of recombinant proteins produced using cell cultures.mammalian cell cultures have been used extensively for production of recombinant protein therapeutics such as monoclonal antibodies, fusion proteins and enzymes for decades. small molecules have been investigated as media supplements to improve process productivity and reduce cost of goods. those chemicals can lead to significant yield improvement through different mechanisms such as cell cycle modulation, cellular redox regulation, etc. in addition to productivity, small molecules have also bee ...202032540474
endogenous p21-dependent transgene control for cho cell engineering.numerous engineering efforts have been made in chinese hamster ovary (cho) cells for high level production of therapeutic proteins. however, the dynamic regulation of transgene expression is limited in current systems. here, we investigated the effective regulation of transgene expression in cho cells via targeted integration-based endogenous gene tagging with engineering target genes. targeted integration of egfp-human bcl-2 into the p21 locus effectively reduced the apoptosis, compared with ra ...202032539343
putative phospholipase b-like 2 is not responsible for polysorbate degradation in monoclonal antibody drug products.polysorbates (ps) are surfactants commonly added in a therapeutic protein drug product as excipients to protect proteins from denaturation and aggregation during storage, transportation, and delivery. significant degradation of ps in drug products could lead to shortened drug shelf lives and ps-degrading activity in drug products must be minimized. identification of lipases that degrade ps could lead to better process control in drug manufacturing. in 2016, phospholipase b-like 2 (plbd2) was pro ...202032534029
insulin degrading enzyme (ide) expressed by chinese hamster ovary (cho) cells is responsible for degradation of insulin in culture media.chinese hamster ovary (cho) cells cultured in serum-free chemically-defined media (cdm) are used for manufacturing of therapeutic proteins. growth factors, such as insulin are commonly utilized in manufacturing platforms to enhance cho cell viability and growth. here we report that insulin is degraded in the culture media over time mainly due to the activity of the insulin degrading enzyme (ide). insulin degradation was faster in cell lines that released more ide, which negatively impacted cell ...202032526262
functional production of human antibody by the filamentous fungus aspergillus oryzae.monoclonal antibodies (mabs) as biopharmaceuticals take a pivotal role in the current therapeutic applications. generally mammalian cell lines, such as those derived from chinese hamster ovaries (cho), are used to produce the recombinant antibody. however, there are still concerns about the high cost and the risk of pathogenic contamination when using mammalian cells. aspergillus oryzae, a filamentous fungus recognized as a gras (generally regarded as safe) organism, has an ability to secrete a ...202032514366
a site-specific integration reporter system that enables rapid evaluation of crispr/cas9-mediated genome editing strategies in cho cells.targeted gene knockout and site-specific integration (ssi) are powerful genome editing techniques to improve the development of industrially relevant chinese hamster ovary (cho) cell lines. however, past efforts to perform ssi in cho cells are characterized by low efficiencies. moreover, numerous strategies proposed to boost ssi efficiency in mammalian cell types have yet to be evaluated head to head or in combination to appreciably boost efficiencies in cho. to enable systematic and rapid optim ...202032500600
process optimization using high throughput automated micro-bioreactors in chinese hamster ovary cell cultivation.optimization of bioprocesses to increase the yield of desired products is of importance in the biopharmaceutical industry. this can be achieved by strain selection and by developing bioprocess parameters. shake flasks have been used for this purpose. they, however, lack the capability to control the process parameters such as ph and dissolved oxygen (do). this limitation can be overcome with the help of an automated micro-bioreactor. these bioreactors mimic cultivation at a larger scale. one of ...202032478715
activation of oxytocin neurons improves cardiac function in a pressure-overload model of heart failure.this work shows long-term restoration of the hypothalamic oxytocin (oxt) network preserves oxt release, reduces mortality, cardiac inflammation, fibrosis, and improves autonomic tone and cardiac function in a model of heart failure. intranasal administration of oxt in patients mimics the short-term changes seen in animals by increasing parasympathetic-and decreasing sympathetic-cardiac activity. this work provides the essential translational foundation to determine if approaches that mimic parav ...202032478209
modulation of acid-sensing ion channels by hydrogen sulfide.acid-sensing ion channels (asics) have been implicated in many physiological and patho-physiological processes like synaptic plasticity, inflammation, pain perception, stroke-induced brain damage and, drug-seeking behaviour. although asics have been shown to be modulated by gasotransmitters like nitric oxide (no), their regulation by hydrogen sulfide (h2s) is not known. here, we present strong evidence that h2s potentiates asics-mediated currents. low ph-induced current in chinese hamster ovary ...202032446393
production of integrin αiibβ3 in stably transfected and clonal chinese hamster ovary cells for functional and structural studies. 202032445462
impacts on product quality attributes of monoclonal antibodies produced in cho cell bioreactor cultures during intentional mycoplasma contamination events.a mycoplasma contamination event in a biomanufacturing facility can result in costly cleanups and potential drug shortages. mycoplasma may survive in mammalian cell cultures with only subtle changes to the culture and penetrate the standard 0.2-µm filters used in the clarification of harvested cell culture fluid. previously, we reported a study regarding the ability of mycoplasma arginini to persist in a single-use, perfusion rocking bioreactor system containing a chinese hamster ovary (cho) dg4 ...202032436993
overexpression of the mitochondrial pyruvate carrier reduces lactate production and increases recombinant protein productivity in cho cells.chinese hamster ovary (cho) cells are characterized by a low glucose catabolic efficiency, resulting in undesirable lactate production. here, it is hypothesized that such low efficiency is determined by the transport of pyruvate into the mitochondria. the mitochondrial pyruvate carrier (mpc), responsible for introducing pyruvate into the mitochondria, is formed by two subunits, mpc1 and mpc2. stable cho cell lines, overexpressing the genes of both subunits, were constructed to facilitate the ent ...202032436990
rich production media as a platform for cho cell line development.recent cell culture media for mammalian cells can be abundantly formulated with nutrients supporting production, but such media can be limited to use in host cell culture, transfection, cell cloning, and cell growth under the low cell density conditions. in many cases, appropriate platform media are used for cell line development, and then replaced with rich media for production. in this study, we demonstrate rich chemically defined media for chinese hamster ovary (cho) cells that are suitable a ...202032415509
biomanufacturing evolution from conventional to intensified processes for productivity improvement: a case study.process intensification has shown great potential to increase productivity and reduce costs in biomanufacturing. this case study describes the evolution of a manufacturing process from a conventional processing scheme at 1000-l scale (process a, n = 5) to intensified processing schemes at both 1000-l (process b, n = 8) and 2000-l scales (process c, n = 3) for the production of a monoclonal antibody by a chinese hamster ovary cell line. for the upstream part of the process, we implemented an inte ...202032425110
synergistic and antagonistic interactions among organic and metallic components of the ambient particulate matter (pm) for the cytotoxicity measured by chinese hamster ovary cells.although pm2.5 toxicity is known to be related to its chemical composition, the effect of interactions among various particles' components on the toxicity is not well explored. to understand these interactions, especially metals and organic compounds on pm2.5 cytotoxicity, we chose several redox-active substances known to be present in the ambient particles such as metals (cu, fe, and mn) and quinones [9,10-phenanthraquinone (pq), 1,2-naphthoquinone (1,2-nq), 1,4-naphthoquinone (1,4-nq), and 5-h ...202032474273
development of a proline-based selection system for reliable genetic engineering in chinese hamster ovary cells.chinese hamster ovary (cho) cells are the superior host cell culture models used for the bioproduction of therapeutic proteins. one of the prerequisites for bioproduction using cho cell lines is the need to generate stable cho cell lines with optimal expression output. antibiotic selection is commonly employed to isolate and select cho cell lines with stable expression, despite its potential negative impact on cellular metabolism and expression level. herein, we present a novel proline-based sel ...202032470293
a doubly auxotrophic cho-k1 cell line for the production of recombinant monoclonal antibodies.chinese hamster ovary (cho) cells are the most widely used mammalian hosts for recombinant protein production due to their hardiness, ease of transfection, and production of glycan structures similar to those in natural human monoclonal antibodies. to enhance the usefulness of cho-k1 cells we developed a new selection system based on double auxotrophy. we used crispr-cas9 to knockout the genes that encode the bifunctional enzymes catalyzing the last two steps in the de novo synthesis of pyrimidi ...202032346859
atypical chemokine receptor ackr2-v41a has decreased ccl2 binding, scavenging, and activation, supporting sustained inflammation and increased alzheimer's disease risk.a recent genome-wide association study (gwas) of 59 cerebrospinal fluid (csf) proteins with a connection to alzheimer's disease (ad) demonstrated an association between increased levels of chemokine ligand 2 (ccl2) with an atypical chemokine receptor chemokine-binding protein 2 variant v41a (ackr2-v41a; rs2228467). high levels of ccl2 are associated with increased risk of ad development as well as other inflammatory diseases. in this study we characterized the biological function of the ackr2-v4 ...202032415244
antimicrobial activity, toxicity and accumulated hard-tissue debris (ahtd) removal efficacy of several chelating agents.to evaluate the antimicrobial, toxicity and cleaning effectiveness of ethylenediaminetetraacetic acid (edta) and maleic acid (ma) alone and combined with cetrimide (ctr).202032344451
cellular reference materials for dna damage using electrochemical oxidation.reference materials are needed to quantify the level of dna damage in cells, to assess sources of measurement variability and to compare results from different laboratories. the comet assay (single cell gel electrophoresis) is a widely used method to determine dna damage in the form of strand breaks. here we examine the use of electrochemical oxidation to produce dna damage in cultured mammalian cells and quantify its percentage using the comet assay. chinese hamster ovary (cho) cells were grown ...202032411438
proteoglycan-dependent endo-lysosomal fusion affects intracellular survival of salmonella typhimurium in epithelial cells.proteoglycans (pgs) are glycoconjugates which are predominately expressed on cell surfaces and consist of glycosaminoglycans (gags) linked to a core protein. an initial step of gags assembly is governed by the β-d-xylosyltransferase enzymes encoded in mammals by the xylt1/xylt2 genes. pgs are essential for the interaction of a cell with other cells as well as with the extracellular matrix. a number of studies highlighted a role of pgs in bacterial adhesion, invasion, and immune response. in this ...202032411142
chromosome-scale scaffolds for the chinese hamster reference genome assembly to facilitate the study of the cho epigenome.the chinese hamster genome serves as a reference genome for the study of chinese hamster ovary (cho) cells, the preferred host system for biopharmaceutical production. recent re-sequencing of the chinese hamster genome resulted in the refseq picr meta-assembly, a set of highly accurate scaffolds that filled over 95% of the gaps in previous assembly versions. however, these scaffolds did not reach chromosome-scale due to the absence of long-range scaffolding information during the meta-assembly p ...202032410221
the relation of bleomycin delivery efficiency to microbubble sonodestruction and cavitation spectral characteristics.the concurrent assessment of principal sonoporation factors has been accomplished in a single systemic study. microbubble sonodestruction dynamics and cavitation spectral characteristics, ultrasound scattering and attenuation, were examined in relation to the intracellular delivery of anticancer drug, bleomycin. experiments were conducted on chinese hamster ovary cells coadministered with sonovue microbubbles. detailed analysis of the scattering and attenuation temporal functions culminated in q ...202032385397
receptor-targeted photodynamic therapy of glucagon-like peptide 1 receptor-positive lesions.treatment of hyperinsulinemic hypoglycemia is challenging. surgical treatment of insulinomas and focal lesions in congenital hyperinsulinism is invasive and carries major risks of morbidity. medication to treat nesidioblastosis and diffuse congenital hyperinsulinism has varying efficacy and causes significant side effects. here, we describe a novel method for therapy of hyperinsulinemic hyperglycemia, highly selectively killing β-cells by receptor-targeted photodynamic therapy (rtpdt) with exend ...202032385165
detrimental effect of zwitterionic buffers on lysosomal homeostasis in cell lines and ipsc-derived neurons.good's buffers are commonly used for cell culture and, although developed to have minimal to no biological impact, they cause alterations in cellular processes such as autophagy and lysosomal enzyme activity. using chinese hamster ovary cells and induced pluripotent stem cell-derived neurons, this study explores the effect of zwitterionic buffers, specifically hepes, on lysosomal volume and ca2+ levels. certain zwitterionic buffers lead to lysosomal expansion and reduced lysosomal ca2+. care sho ...202032490352
harnessing the potential of artificial neural networks for predicting protein glycosylation.kinetic models offer incomparable insight on cellular mechanisms controlling protein glycosylation. however, their ability to reproduce site-specific glycoform distributions depends on accurate estimation of a large number of protein-specific kinetic parameters and prior knowledge of enzyme and transport protein levels in the golgi membrane. herein we propose an artificial neural network (ann) for protein glycosylation and apply this to four recombinant glycoproteins produced in chinese hamster ...202032489858
excito-repellency and biological safety of β-caryophyllene oxide against aedes albopictus and anopheles dirus (diptera: culicidae).the activity of β-caryophyllene oxide as either a contact or noncontact repellent was evaluated against two laboratory strains (aedes albopictus and anopheles dirus) using an excito-repellency test system. n, n-diethyl-3-methylbenzamide (deet) was used as a standard reference baseline for comparative purposes. β-caryophyllene oxide and deet were tested at concentrations of 0.1, 0.25, 0.5 and 1.0% (v/v). in addition, the phototoxic and genotoxic effects of β-caryophyllene oxide were investigated ...202032485168
one-step enrichment of intact glycopeptides from glycoengineered chinese hamster ovary cells.recently, the glycoproteomic analysis of intact glycopeptides has emerged as an effective approach to decipher the glycan modifications of glycoproteins at the site-specific level. a rapid method to enrich intact glycopeptides is essential for the analysis of glycoproteins, especially for biopharmaceutical proteins. in this study, we established a one-step method for the rapid capture of intact glycopeptides for analysis by mass spectrometry. compared to the conventional sequential enrichment me ...202032363175
gene editing in cho cells to prevent proteolysis and enhance glycosylation: production of hiv envelope proteins as vaccine immunogens.several candidate hiv subunit vaccines based on recombinant envelope (env) glycoproteins have been advanced into human clinical trials. to facilitate biopharmaceutical production, it is necessary to produce these in cho (chinese hamster ovary) cells, the cellular substrate used for the manufacturing of most recombinant protein therapeutics. however, previous studies have shown that when recombinant env proteins from clade b viruses, the major subtype represented in north america, europe, and oth ...202032470085
voluhemins, new inhibitors of sterol o-acyltransferase, produced by volutella citrinella bf-0440.new compounds, designated voluhemins a (1) and b (2), are isolated from the culture broth of the fungal strain volutella citrinella bf-0440 along with structurally related known nk12838 (3). spectroscopic data, including 1d and 2d nmr, elucidated their structures. compounds 1-3 have a common indoline-diterpene core and two additional isoprenyl moieties. compounds 1 and 3 contain a hemiaminal unit, while 2 is o-methylated 1. their inhibitory activities toward sterol o-acyltransferase (soat) 1 and ...202032467602
the use of site-specific recombination and cassette exchange technologies for monoclonal antibody production in chinese hamster ovary cells: retrospective analysis and future directions.chinese hamster ovary (cho) cell-based platforms are the most widely used for the biomanufacturing of complex therapeutic proteins, such as monoclonal antibodies (mabs). the development of high-producing clones that are stable and amenable to large-scale cultures is essential to advance a molecule toward clinical evaluation. nevertheless, the generation of such clones generally relies on random integration of an expression plasmid encoding the therapeutic protein gene into the host genome. the e ...202032456474
a general evidence-based sequence variant control limit for recombinant therapeutic protein development.sequence variants (svs) resulting from unintended amino acid substitutions in recombinant therapeutic proteins have increasingly gained attention from both regulatory agencies and the biopharmaceutical industry given their potential impact on efficacy and safety. with well-optimized production systems, such sequence variants usually exist at very low levels in the final protein products due to the high fidelity of dna replication and protein biosynthesis process in mammalian expression systems s ...202032744138
roles of lys191 and lys179 in regulating thermodynamic binding forces of ligands to determine their binding affinity for human histamine h1 receptors.docking simulations based on the crystal structure of human histamine h1 receptors have predicted crucial roles of lys1915.39 and lys179ecl2, which exist at the entrance of the ligand-binding pocket, in increasing the h1-receptor selectivity for carboxylated second-generation antihistamines via electrostatic interaction. in this study, we evaluated the roles of lys1915.39 and lys179ecl2 in regulating the thermodynamic binding forces of non-carboxylated and carboxylated antihistamines that determ ...202032738199
expanding the activity profile of pyrido[1,2-a]benzimidazoles: synthesis and evaluation of novel n1-1-phenylethanamine derivatives against schistosoma mansoni.praziquantel is the only widely available drug to treat schistosomiasis. with very few candidates currently in the drug development pipeline, there is an urgent need to discover and develop novel antischistosomal drugs. in this regard, the pyrido[1,2-a]benzimidazole (pbi) scaffold has emerged as a promising chemotype in hit-to-lead efforts. here, we report a novel series of antischistosomal pbis with potent in vitro activity (ic50 values of 0.08-1.43 μm) against schistosoma mansoni newly transfo ...202032786285
evaluation of antibacterial and cytotoxic properties of a fluorinated diamond-like carbon coating for the development of antibacterial medical implants.peri-implant infection is a serious complication in surgical procedures involving implants. we conducted an in vitro study to determine whether the use of a fluorinated diamond-like carbon (f-dlc) coating on a titanium alloy surface can prevent peri-implant infection. after applying the f-dlc, we evaluated its antibacterial and cytotoxic properties. the coating groups, containing controlled fluorine concentrations of 5.44%, 17.43%, 24.09%, and 30%, were examined for the presence of staphylococcu ...202032784861
a direct rt qpcr method for quantification of retrovirus-like particles in biopharmaceutical production with cho cells.chinese hamster ovary (cho) cells are the host cell of choice for manufacturing biologic drugs, like monoclonal antibody, in the biopharmaceutical industry. retrovirus-like particles (rvlps) are made during the manufacturing process with cho cells and it is incumbent upon the manufacturer to perform risk assessment based on levels of rvlp in unprocessed bulk. quantification of rvlp using electron microscopy (em) is the standard method. however, reverse transcription based real-time pcr (rt qpcr) ...202032693202
dynamism of pi4-phosphate during interactions with human erythrocytes in entamoeba histolytica.phosphatidylinositol phosphates (pips) are involved in many cellular events as important secondary messengers. in entamoeba histolytica, a human intestinal protozoan parasite, virulence-associated mechanisms such as cell motility, vesicular traffic, trogo- and phagocytosis are regulated by pips. it has been well established that pi3p, pi4p, and pi(3,4,5)p3 play specific roles during amoebic trogo- and phagocytosis. in the present study, we demonstrated the nuclear localization of pi4p in e. hist ...202032679800
sub-microsecond electrotransfection using new modality of high frequency electroporation.micro-millisecond range electric field pulses have been used for decades to facilitate dna transfer into cells and tissues, while the growing number of clinical trials underline the strong potential of dna electroporation. in this work, we present new sub-microsecond range protocols and methodology enabling successful electrotransfection in the sub-microsecond range. to facilitate dna transfer, a 3 kv/60 a and high frequency (1 mhz) sub-microsecond range square wave generator was applied in the ...202032679337
toxicity of chlorinated algal-impacted waters: formation of disinfection byproducts vs. reduction of cyanotoxins.seasonal algal blooms in surface waters can impact water quality through an input of algal organic matter (aom) to the pool of dissolved organic matter as well as the release of cyanotoxins. the formation and speciation of disinfection byproducts (dbps) during chlorination of algal-impacted waters, collected from growth of microcystis aeruginosa were studied. second-order rate constants for the reactions of microcystins (mcs) with chlorine and bromine were determined. finally, the toxicity of ch ...202032771689
cisplatin decreases enac activity contributing to renal salt wasting syndrome.cisplatin (cddp) is an important anticancer drug. a common side effect of cddp is renal salt and water-wasting syndrome (rsws). the origin of rsws is obscure. emerging evidence, though, suggests that broad inhibition of sodium transport proteins by cddp may result in decreases in tubular reabsorption, causing increases in sodium and water excretion. in this sense, cddp would be acting like a diuretic. the effect of cddp on the epithelial na+ channel (enac), which is the final arbiter fine-tuning ...202032752278
establishment of an anti-cd20 monoclonal antibody (c20mab-60) for immunohistochemical analyses.sensitive and specific monoclonal antibodies (mabs) are needed for detecting cd20. this antigen, one of several b lymphocyte antigens, is a target for every application used in the diagnosis of b cell lymphoma. many anti-cd20 mabs have been established, although applications of these antibodies are limited. this study aims to establish sensitive and specific anti-cd20 mabs suitable for broad application, such as flow cytometry, western blotting, and immunohistochemical analyses. using the cell-b ...202032667234
a novel dextran-grafted tetrapeptide resin for antibody purification.short peptide biomimetic affinity chromatography as a novel antibody separation chromatography is a potential alternative to protein a chromatography. however, if directly attaching ligand to matrix, the adsorption capacity and mass transfer rate would be affected by pore blockage and steric effect. grafting resin is an effective method to solve this problem by using polymer as a bridge between matrix and ligand. in this work, a novel resin was prepared by grafting a tetrapeptide to the dextran- ...202032729191
the identification of nuclear αvβ3 integrin in ovarian cancer: non-paradigmal localization with cancer promoting actions.nuclear translocation of transmembrane proteins was reported in high-grade serous ovarian cancer (hgsoc), a highly aggressive gynecological malignancy. although the membrane receptor αvβ3 integrin is amply expressed in hgsoc and involved in disease progression, its nuclear localization was never demonstrated. nuclear αvβ3 was explored in hgsoc cells (ovcar3, kuramochi, and jhos4), nuclear localization signal (nls) modified β3 ovcar3, chinese hamster ovaries (cho-k1) and human embryonic kidney (h ...202032728020
analysis of damage-associated molecular pattern molecules due to electroporation of cells in vitro.background tumor cells can die via immunogenic cell death pathway, in which damage-associated molecular pattern molecules (damps) are released from the cells. these molecules activate cells involved in the immune response. both innate and adaptive immune response can be activated, causing a destruction of the remaining infected cells. activation of immune response is also an important component of tumor treatment with electrochemotherapy (ect) and irreversible electroporation (ire). we thus expl ...202032726295
generation of fx-/- and gmds-/- chozn host cell lines for the production of afucosylated therapeutic antibodies.antibody-dependent cellular cytotoxicity (adcc) is the primary mechanism of actions for several marketed therapeutic antibodies (mabs) and for many more in clinical trials. the adcc efficacy is highly dependent on the ability of therapeutic mabs to recruit effector cells such as natural killer cells, which induce the apoptosis of targeted cells. the recruitment of effector cells by mabs is negatively affected by fucose modification of n-glycans on the fc; thus, utilization of afucosylated mabs h ...202032748555
sensitivity of a per.c6® cell line to bis(2,4-di-tert-butylphenyl)phosphate and evaluation of a new, biocompatible single-use film.single-use technologies have brought numerous advantages to the biopharmaceutical industry. in particular, single-use bags made from multi-layered polymeric films have been adopted for cell culture and liquid handling operations in place of traditional stainless-steel systems. despite the advantages, leachable compounds originating from the film's materials of construction present a new challenge. in 2013, bis(2,4-di-tert-butylphenyl)phosphate (bdtbpp) was identified as a common leachable from s ...202032748552
toxicity variability of urban road stormwater during storage processes in shenzhen, china: identification of primary toxicity contributors and implications for reuse safety.urban road stormwater reuse is one of the most important ways to mitigate water resource shortage. generally, stormwater is stored prior to reuse or further treatment. this study explored the stormwater toxicity variability during two types of storages, closed and open storages using chinese hamster ovary cells (cho), which are mammalian cells. the toxicity test by cho cells can indirectly represent the risk related to human health. both rainfall (without reaching ground surfaces) and urban road ...202032717603
novel peptide ligands for antibody purification provide superior clearance of host cell protein impurities.the quest for ligands alternative to protein a for the purification of monoclonal antibodies (mabs) has been pursued for almost three decades. yet, the igg-binding peptides known to date still fall short of the host cell protein (hcp) logarithmic removal value (lrv) set by protein a media (2.5-3.1). in this study, we present an integrated computational-experimental approach leading to the discovery of peptide ligands that provide hcp lrvs on par with protein a. first, the screening of 60,000 pep ...202032709313
chromatographic clarification overcomes chromatin-mediated hitch-hiking interactions on protein a capture column.protein a capture chromatography is a critical unit operation in the clearance of host cell protein (hcp) impurities in monoclonal antibody (mab) purification processes. though one of the most effective purification steps, variable levels of protein impurities are often observed in the eluate. coelution of hcp impurities is suggested to be strongly affected by the presence of chromatin complexes (gagnon et al., 2014; koehler et al., 2019). we investigated the effect of removal of dna complex and ...202032706389
characterization of glutathione proteome in cho cells and its relationship with productivity and cholesterol synthesis.glutathione (gsh) plays a central role in the redox balance maintenance in mammalian cells. previous studies of industrial chinese hamster ovary cell lines have demonstrated a relationship between gsh metabolism and clone productivity. however, a thorough investigation is required to understand this relationship and potentially highlight new targets for cell engineering. in this study, we have modulated the gsh intracellular content of an industrial cell line under bioprocess conditions to furth ...202032662871
construction of an expression vector mediated by the dual promoter for prokaryotic and mammalian cell expression system.the aim of this study was to construct an expression vector mediated by the dual promoter that can simultaneously drive the recombinant protein production in eukaryotic and prokaryotic cells. the prokaryotic t7 promoter and ribosome binding site (rbs) was cloned downstream of cmv promoter in the eukaryotic expression vector pires-neo, and t7 termination sequence was inserted upstream of neomycin phosphotransferase gene to generate the dual promoter vector. the enhanced green fluorescent protein ...202032564228
investigation into the impact of tyrosine on the product formation and quality attributes of mabs in rcho cell cultures.tyrosine (tyr) is crucial to the maintenance of the monoclonal antibody (mab) titers and quality attributes in fed-batch cultures of recombinant chinese hamster ovary (rcho) cells. however, the relation between tyrosine and these aspects is not yet fully defined. in order to further elucidate such a relation, two groups of fed-batch experiments with high tyrosine (h-t) or low tyrosine (l-t) additions producing an igg1 monoclonal antibody against cd20 were implemented to investigate the intracell ...202032577803
immunoglobulin fragment f(ab')2 against rbd potently neutralizes sars-cov-2 in vitro.covid-19, which is caused by the emerging human coronavirus sars-cov-2, has become a global pandemic that poses a serious threat to human health. to date, no vaccines or specific antiviral drugs have been approved for the treatment of this disease in clinic. herein, therapeutic antibodies for sars-cov-2 were obtained from hyperimmune equine plasma. first, a recombinant sars-cov-2 spike protein receptor-binding domain (rbd) was obtained in gram-level quantities through high-cell density fermentat ...202032659292
rational optimization of a monoclonal antibody improves the aggregation propensity and enhances the cmc properties along the entire pharmaceutical process chain.the discovery of therapeutic monoclonal antibodies (mabs) primarily focuses on their biological activity favoring the selection of highly potent drug candidates. these candidates, however, may have physical or chemical attributes that lead to unfavorable chemistry, manufacturing, and control (cmc) properties, such as low product titers, conformational and colloidal instabilities, or poor solubility, which can hamper or even prevent development and manufacturing. hence, there is an urgent need to ...202032658605
enabling speed to clinic for monoclonal antibody programs using a pool of clones for ind-enabling toxicity studies.the importance of speed to clinic for medicines that may address unmet medical needs puts pressure on product development timelines. historically, both toxicology and first-in-human clinical materials are generated using the same clonal-derived cells to ensure safety and minimize any development risks. however, cell line development with single cell cloning is time consuming, and aggravated by the time needed to screen for a lead clone based on cell line stability and manufacturability. in order ...202032449878
an efficient screening system in yeast to select a hyperactive piggybac transposase for mammalian applications.as non-viral transgenic vectors, the piggybac transposon system represents an attractive tool for gene delivery to achieve a long-term gene expression in immunotherapy applications due to its large cargo capacity, its lack of a trace of transposon and of genotoxic potential, and its highly engineered structure. however, further improvements in transpose activity are required for industrialization and clinical applications. herein, we established a one-plasmid effective screening system and a two ...202032357554
kcnq1 antibodies for immunotherapy of long qt syndrome type 2.patients with long qt syndrome (lqts) are predisposed to life-threatening arrhythmias. a delay in cardiac repolarization is characteristic of the disease. pharmacotherapy, implantable cardioverter-defibrillators, and left cardiac sympathetic denervation are part of the current treatment options, but no targeted therapy for lqts exists to date. previous studies indicate that induced autoimmunity against the voltage-gated kcnq1 k+ channels accelerates cardiac repolarization.202032354382
dynamic modeling of cho cell metabolism using the hybrid cybernetic approach with a novel elementary mode analysis strategy.chinese hamster ovary (cho) cell culture has a major importance on the production of biopharmaceuticals, including recombinant therapeutic proteins such as monoclonal antibodies (mab). mathematical modeling of biological systems can successfully assess metabolism complexity while providing logical and systematic methods for relevant genetic target and culture parameter identification toward cell growth and productivity improvements. most modeling approaches on cho cells have been performed under ...202032351947
subphysiological temperature induces pervasive alternative splicing in chinese hamster ovary cells.rna sequencing (rnaseq) has been widely used to associate alterations in chinese hamster ovary (cho) cell gene expression with bioprocess phenotypes; however, alternative messenger rna (mrna) splicing, has thus far, received little attention. in this study, we utilized rnaseq for transcriptomic analysis of a monoclonal antibody (mab) producing cho k1 cell line subjected to a temperature shift. more than 2,465 instances of differential splicing were observed 24 hr after the reduction of cell cult ...202032346860
effects of gentiana delavayi flower extract on app processing in app/ps1 cho cells.gentiana delavayi franch. (gentianaceae) as an ethnomedicinal plant contains a variety of effective active ingredients and exhibits diverse pharmacological actions, such as hepatoprotective, anti-inflammatory and central nervous system effects. in this study we investigated the influence of g. delavayi flower extract on amyloid precursor protein (app) processing at molecular and cellular levels. app/ps1 chinese hamster ovary (cho) cells were treated with chloroform extract of g. delavayi flower ...202032378555
expression vector cassette engineering for recombinant therapeutic production in mammalian cell systems.human tissue plasminogen activator was the first recombinant therapy protein that successfully produced in chinese hamster ovary cells in 1986 and approved for clinical use. since then, more and more therapeutic proteins are being manufactured in mammalian cells, and the technologies for recombinant protein production in this expression system have developed rapidly, with the optimization of both upstream and downstream processes. one of the most promising strategies is expression vector cassett ...202032372203
short communication: the essential role of n-glycosylation in the transport activity of bovine peptide transporter 2.bovine peptide transporter 2 (bpept2), which mediates the absorption of di- and tripeptides in the bovine mammary gland, was predicted to contain multiple putative n-glycosylation sites of asparagine residues. n-linked glycosylation is proven to be essential for the folding, stability, localization, and substrate binding of nutrient transporters and could therefore potentially have an essential role in the function of bpept2. this study investigated the effect of mutagenesis of n-glycosylation s ...202032331895
enzyme capacity-based genome scale modelling of cho cells.chinese hamster ovary (cho) cells are most prevalently used for producing recombinant therapeutics in biomanufacturing. recently, more rational and systems approaches have been increasingly exploited to identify key metabolic bottlenecks and engineering targets for cell line engineering and process development based on the cho genome-scale metabolic model which mechanistically characterizes cell culture behaviours. however, it is still challenging to quantify plausible intracellular fluxes and d ...202032330653
immunogenicity risk assessment for an engineered human cytokine analogue expressed in different cell substrates.the purpose of this article is to illustrate how performance of an immunogenicity risk assessment at the earliest stage of product development can be instructive for critical early decision-making such as choice of host system for expression of a recombinant therapeutic protein and determining the extent of analytical characterization and control of heterogeneity in co- and post-translational modifications. application of a risk-based approach for a hypothetical recombinant dna analogue of a hum ...202032291556
patches and blebs: a comparative study of the composition and biophysical properties of two plasma membrane preparations from cho cells.this study was aimed at preparing and characterizing plasma membranes (pm) from chinese hamster ovary (cho) cells. two methods of pm preparation were applied, one based on adhering cells to a poly-lysine-coated surface, followed by hypotonic lysis and removal of intracellular components, so that pm patches remain adhered to each other, and a second one consisting of bleb induction in cells, followed by separation of giant plasma membrane vesicles (gpmv). both methods gave rise to pm in sufficien ...202032290157
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