Publications

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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
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
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
optimization of parameters affecting on cho cell culture producing recombinant erythropoietin.several factors may affect erythropoietin (epo) sugar structures including designing cell culture procedure, ph, concentration of additives, dissolved oxygen, and other physicochemical parameters. in this study, we investigated the influence of changes in effective parameters and compounds on the growth rate of chinese hamster ovary cell (cho) cells producing recombinant epo. cell culture was performed at different temperature, buffering conditions, and varied concentrations of additives such as ...202032336186
naturally occurring variants in the transmembrane and cytoplasmic domains of the human coxsackie- and adenovirus receptor have no impact on virus internalisation.the coxsackie- and adenovirus receptor (car) mediates homophilic cell-cell contacts and susceptibility to both human pathogenic viruses through its membrane-distal immunoglobulin domain. in the present study, we screened five missense variants of the human car gene for their influence on adenovector or coxsackievirus entry into chinese hamster ovary cells. the car variants facilitated virus internalisation to a similar extent as wild type car. this underlines car's presumed invariance and essent ...202032334832
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
biological filtering and substrate promiscuity prediction for annotating untargeted metabolomics.mass spectrometry coupled with chromatography separation techniques provides a powerful platform for untargeted metabolomics. determining the chemical identities of detected compounds however remains a major challenge. here, we present a novel computational workflow, termed extended metabolic model filtering (emmf), that aims to engineer a candidate set, a listing of putative chemical identities to be used during annotation, through an extended metabolic model (emm). an emm includes not only can ...202032326153
genotoxicity of micro- and nano-particles of kaolin in human primary dermal keratinocytes and fibroblasts.kaolin is a clay mineral with the chemical composition al2si2o5(oh)4. it is an important industrial material, and is also used as a white cosmetic pigment. we previously reported that fine particles of kaolin have genotoxic potency to chinese hamster ovary cho aa8 cells, and to the lungs of c57bl/6 j and icr mice. in the present study, we evaluated the genotoxicity of different particle sizes of kaolin using primary normal human diploid epidermal keratinocytes and primary normal human diploid de ...202032322315
dna nanovaccines prepared using lema antigen protect golden syrian hamsters against leptospira lethal infection.nanoparticles (nps) are viable candidates as carriers of exogenous materials into cells via transfection and can be used in the dna vaccination strategy against leptospirosis.202032321154
multiplex secretome engineering enhances recombinant protein production and purity.host cell proteins (hcps) are process-related impurities generated during biotherapeutic protein production. hcps can be problematic if they pose a significant metabolic demand, degrade product quality, or contaminate the final product. here, we present an effort to create a "clean" chinese hamster ovary (cho) cell by disrupting multiple genes to eliminate hcps. using a model of cho cell protein secretion, we predict that the elimination of unnecessary hcps could have a non-negligible impact on ...202032313013
a high-resolution measurement of nucleotide sugars by using ion-pair reverse chromatography and tandem columns.n-linked glycosylation is a cellular process transferring sugars from glycosyl donors to proteins or lipids. biopharmaceutical products widely produced by culturing mammalian cells such as chinese hamster ovary (cho) cells are typically glycosylated during biosynthesis. for some biologics, the n-linked glycan is a critical quality attribute of the drugs. nucleotide sugars are the glycan donors and impact the intracellular glycosylation process. in current analytical methods, robust separation of ...202032300845
development of a scale-up strategy for chinese hamster ovary cell culture processes using the kl a ratio as a direct indicator of gas stripping conditions.herein, we described a scale-up strategy focused on the dissolved carbon dioxide concentration (dco2 ) during fed-batch cultivation of chinese hamster ovary cells. a fed-batch culture process for a 2000-l scale stainless steel (ss) bioreactor was scaled-up from similarly shaped 200-l scale bioreactors based on power input per unit volume (p/v). however, during the 2000-l fed-batch culture, the dco2 was higher compared with the 200-l scale bioreactor. therefore, we developed an alternative approa ...202032298540
combining lipoic acid to methylene blue reduces the warburg effect in cho cells: from tca cycle activation to enhancing monoclonal antibody production.the warburg effect, a hallmark of cancer, has recently been identified as a metabolic limitation of chinese hamster ovary (cho) cells, the primary platform for the production of monoclonal antibodies (mab). metabolic engineering approaches, including genetic modifications and feeding strategies, have been attempted to impose the metabolic prevalence of respiration over aerobic glycolysis. their main objective lies in decreasing lactate production while improving energy efficiency. although yield ...202032298377
electroceutical silk-silver gel to eradicate bacterial infection.with more than 50% of bacteria resistant to standard antibiotics, new strategies to treat bacterial infection and colonization are needed. based on the concept of targeting the bacteria synergistically on various fronts, it is hypothesized that an electrical insult associated with antibacterial materials may be a highly effective means of killing bacteria. in this work, an injectable conductive gel based on silk fibroin (sf) and silver nanoparticles (ag-nps) is synthesized, capable of coating a ...202032293155
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
lipid catabolism in starved yak is inhibited by intravenous infusion of β-hydroxybutyrate.lipid is the chief energy source for starved animals. β-hydroxybutyrate (bhba) is the main ketone body produced by lipid decomposition. in chinese hamster ovary (cho) cell experiment, it was found that bhba could be used not only as an energy substance, but also as a ligand of gpr109a for regulating lipid metabolism. however, whether bhba can regulate lipid metabolism of yaks, and its effective concentration and signal pathway are not clear. this study investigated the effects and mechanism of s ...202031952136
electron scattering in conventional cell flask experiments and dose distribution dependency.electron beam therapy (ebt) is commonly used for treating superficial and subdermal tumors. previous cellular radiosensitivity research using ebt may be underestimating the contribution from flask wall scattering and the corresponding dose distribution. single cell suspensions of chinese hamster ovary (cho) cells were plated on flasks and irradiated with 3, 4, 7, 9, and 18 mev energy electron beams from two different institutions, and the spatial locations of surviving colonies were recorded. ga ...202031949176
correction of the aprt gene using repair-polypurine reverse hoogsteen hairpins in mammalian cells.in this study, we describe the correction of single-point mutations in mammalian cells by repair-polypurine reverse hoogsteen hairpins (repair-pprhs). these molecules consist of (1) a pprh hairpin core that binds to a polypyrimidine target sequence in the double-stranded dna (dsdna), producing a triplex structure, and (2) an extension sequence homologous to the dna sequence to be repaired but containing the wild-type nucleotide instead of the mutation and acting as a donor dna to correct the mut ...202031945727
photoacoustic effect applied on model membranes and living cells: direct observation with multiphoton excitation microscopy and long-term viability analysis.the photoacoustic effect is generated when a variable light interacts with a strongly light-absorbing material. in water, it may produce hot bubbles and shock waves that could affect the integrity of nearby cellular membranes, opening transient pores (photoporation). in this study, we have evaluated the effect of pulsed laser-irradiated carbon nanoparticles (cnp) on model membranes and on chinese hamster ovary (cho) cells. fluorescence lifetime measurements of calcein-loaded liposomes support th ...202031941922
effective temperature shift strategy development and scale confirmation for simultaneous optimization of protein productivity and quality in chinese hamster ovary cells.temperature shifts to lower culture temperatures are frequently employed in the manufacturing of protein therapeutics in mammalian cells to improve productivity, viability, or quality attributes. the direction and extent to which a temperature shift affects productivity and quality may vary depending on the expression host and characteristics of the expressed protein. we demonstrated here that two chinese hamster ovary (cho) clones expressing different human monoclonal antibodies responded diffe ...202031930722
attenuating apoptosis in chinese hamster ovary cells for improved biopharmaceutical production.chinese hamster ovary (cho) cells are the predominant host cell line for the production of biopharmaceuticals, a growing industry currently worth more than $188 billion usd in global sales. cho cells undergo programmed cell death (apoptosis) following different stresses encountered in cell culture, such as substrate limitation, accumulation of toxic by-products, and mechanical shear, hindering production. genetic engineering strategies to reduce apoptosis in cho cells have been investigated with ...202031930480
kinetic modeling of chinese hamster ovary cell culture: factors and principles.as a host for therapeutic protein expression, chinese hamster ovary (cho) cells are widely utilized in the mainstream biopharmaceutical industry. cell culture process development plays an important role in transitioning laboratory research to manufacturing. among different mathematic tools, kinetic modeling is commonly achieved through analyzing cell culture data to design process parameters, optimize media, and scale up bioreactors. in this review, we examine key factors for upstream process de ...202031928250
targeted optical imaging of the glucagonlike peptide 1 receptor using exendin-4-irdye 800cw.the treatment of choice for insulinomas and focal lesions in congenital hyperinsulinism (chi) is surgery. however, intraoperative detection can be challenging. this challenge could be overcome with intraoperative fluorescence imaging, which provides real-time lesion detection with a high spatial resolution. here, a novel method for targeted near-infrared (nir) fluorescence imaging of glucagonlike peptide 1 receptor (glp-1r)-positive lesions, using the glp-1 agonist exendin-4 labeled with irdye 8 ...202031924726
different localization of lysosomal-associated membrane protein 1 (lamp1) in mammalian cultured cell lines.lysosomal-associated membrane protein 1 (lamp1) mainly localizes to lysosomes and late endosomes. we herein investigated the intracellular localization of lysosomal membrane proteins in five mammalian cultured cell lines. rat lamp1 fused to enhanced green fluorescent protein (egfp) mostly accumulated at a particular cytoplasmic area and barely co-localized with lysotracker® red dnd-99 in golden hamster kidney bhk-21 cells and chinese hamster ovary cho-k1 cells. golden hamster, chinese hamster, a ...202031907597
choice of host cell line is essential for the functional glycosylation of the fc region of human igg1 inhibitors of influenza b viruses.abs are glycoproteins that carry a conserved n-linked carbohydrate attached to the fc whose presence and fine structure profoundly impacts on their in vivo immunogenicity, pharmacokinetics, and functional attributes. the host cell line used to produce igg plays a major role in this glycosylation, as different systems express different glycosylation enzymes and transporters that contribute to the specificity and heterogeneity of the final igg-fc glycosylation profile. in this study, we compare tw ...202031907284
direct optical detection of cell density and viability of mammalian cells by means of uv/vis spectroscopy.the critical process parameters cell density and viability during mammalian cell cultivation are assessed by uv/vis spectroscopy in combination with multivariate data analytical methods. this direct optical detection technique uses a commercial optical probe to acquire spectra in a label-free way without signal enhancement. for the cultivation, an inverse cultivation protocol is applied, which simulates the exponential growth phase by exponentially replacing cells and metabolites of a growing ch ...202031897554
genome-scale reconstructions of the mammalian secretory pathway predict metabolic costs and limitations of protein secretion.in mammalian cells, >25% of synthesized proteins are exported through the secretory pathway. the pathway complexity, however, obfuscates its impact on the secretion of different proteins. unraveling its impact on diverse proteins is particularly important for biopharmaceutical production. here we delineate the core secretory pathway functions and integrate them with genome-scale metabolic reconstructions of human, mouse, and chinese hamster ovary cells. the resulting reconstructions enable the c ...202031896772
summation of disinfection by-product cho cell relative toxicity indices: sampling bias, uncertainty, and a path forward.the cyto- and genotoxic potencies of disinfection by-products (dbps) have been evaluated in published literature by measuring the response of exposed chinese hamster ovary cells. in recent publications, dbp concentrations divided by their individual toxicity indices are summed to predict the relative toxicity of a water sample. we hypothesized that the omission or inclusion of certain dbps over others is equivalent to statistical sampling bias and may result in biased conclusions. to test this h ...202031894210
robust analytical methods for the accurate quantification of the total biomass composition of mammalian cells.biomass composition is an important input for genome-scale metabolic models and has a big impact on their predictive capabilities. however, researchers often rely on generic data for biomass composition, e.g. collected from similar organisms. this leads to inaccurate predictions, because biomass composition varies between different cell lines, conditions, and growth phases. in this chapter we present protocols for the determination of the biomass composition of chinese hamster ovary (cho) cells. ...202031893373
small-scale bioreactor supports high density hek293 cell perfusion culture for the production of recombinant erythropoietin.process intensification in mammalian cell culture-based recombinant protein production has been achieved by high cell density perfusion exceeding 108 cells/ml in the recent years. as the majority of therapeutic proteins are produced in chinese hamster ovary (cho) cells, intensified perfusion processes have been mainly developed for this type of host cell line. however, the use of cho cells can result in non-human posttranslational modifications of the protein of interest, which may be disadvanta ...202031891733
cytotoxicity and genotoxicity of dmabee, a co-photoinitiator of resin polymerization, on cho-k1 cells: role of redox and carboxylesterase.the 4-dimethylaminobenzoic acid ethyl ester (dmabee) is an important co-initiator for resin polymerization in dental resinous materials. as a radical forming chemical with high lipophilicity, the genotoxicity and cytotoxicity of dmabee deserve prudent investigation. in this study, we found that dmabee reduced the viability and proliferation of chinese hamster ovary (cho-k1) cells in a dose-dependent manner, and altered cell morphology at higher concentrations. g0/g1 cell cycle arrest was induced ...202031880385
inhibition of endogenous mir-23a/mir-377 in cho cells enhances difficult-to-express recombinant lysosomal sulfatase activity.difficult-to-express (dte) recombinant proteins such as multi-specific proteins, dte monoclonal antibodies, and lysosomal enzymes have seen difficulties in manufacturability using chinese hamster ovary (cho) cells or other mammalian cells as production platforms. cho cells are preferably used for recombinant protein production for their ability to secrete human-like recombinant proteins with posttranslational modification, resistance to viral infection, and familiarity with drug regulators. howe ...202031990124
cytotoxic and genotoxic assessments of 2,4-dichlorophenoxyacetic acid (2,4-d) in in vitro mammalian cells.a combined approach employing alkaline single cell gel electrophoresis (scge) and cytokinesis-blocked micronucleus (mns) cytome bioassays was adopted to assess the deleterious properties of the auxinic 2,4-dichlorophenoxyacetic acid (2,4-d) and its microparticulated low volatility product dedalo elite (30% a.i.) on chinese hamster ovary (cho-k1) cells. cytotoxicity was estimated by neutral red uptake (nru), succinic dehydrogenase activity (mtt) and apoptosis assessment. both compounds were assay ...202031987841
recombinant β-glucocerebrosidase specific immunoaffinity ligands selected from phage-displayed combinatorial scfv libraries.antibodies specific to β-glucocerebrosidase were selected from phage displayed naïve scfv libraries. biopannings were performed against recombinant human protein β-glucocerebrosidase immobilized on polystyrene surface, specific phages were eluted with 50% ethylene glycol in citrate buffer (ph 6.0). several specific binders were discovered and converted to full-size higg1 antibodies leading to highly stable binders with dissociation constants (kd) in the range 10-40 nm. the antibodies were used f ...202031981620
an efficient exogenous gene insertion site in cho cells with high transcription level to enhance aid-induced mutation.antibodies have been extensively used for the purpose of scientific research, clinical diagnosis, and therapy. combination of in vitro somatic hypermutation and mammalian cell surface display has been an efficient technology for antibody or other proteins optimization, in which the efficiency of activation-induced cytidine deaminase (aid) mutations in genes is one of the most important key factors. gene transcriptional level has been found to be positively proportional to aid-induced mutation fr ...202031975519
maximizing antibody production in a targeted integration host by optimization of subunit gene dosage and position.historically, therapeutic protein production in chinese hamster ovary (cho) cells has been accomplished by random integration (ri) of expression plasmids into the host cell genome. more recently, the development of targeted integration (ti) host cells has allowed for recombination of plasmid dna into a predetermined genomic locus, eliminating one contributor to clone-to-clone variability. in this study, a ti host capable of simultaneously integrating two plasmids at the same genomic site was use ...202031965756
macrophages show higher levels of engulfment after disruption of cis interactions between cd47 and the checkpoint receptor sirpα.the macrophage checkpoint receptor sirpα signals against phagocytosis by binding cd47 expressed on all cells - including macrophages. here, we found that inhibiting cis interactions between sirpα and cd47 on the same macrophage increased engulfment ('eating') by approximately the same level as inhibiting trans interactions. antibody blockade of cd47, as pursued in clinical trials against cancer, was applied separately to human-derived macrophages and to red blood cell (rbc) targets for phagocyto ...202031964705
cell electropermeabilisation enhancement by non-thermal-plasma-treated pbs.the effectiveness of electrochemotherapy (ect) in local eradication of tumours in human and veterinary medicine has been proven. ect consists of increasing the uptake of cytotoxic drugs by means of pulsed electric fields (pefs) that transiently permeabilise the cell membrane. still, this tumour treatment includes some drawbacks that are linked to the characteristics of the intense electric pulses (eps) used. meanwhile, the emerging field of cancer therapies that are based on the application of n ...202031963132
defining lncrnas correlated with cho cell growth and igg productivity by rna-seq.how the long non-coding rna (lncrna) genome in recombinant protein producing chinese hamster ovary (cho) cell lines relates to phenotype is not well described. we therefore defined the cho cell lncrna transcriptome from cells grown in controlled miniature bioreactors under fed-batch conditions using rna-seq to identify lncrnas and how the expression of these changes throughout growth and between igg producers. we identify lncrnas including adapt15, linked to er stress, gas5, linked to mtor signa ...202031962234
heterozygous variant fibrinogen γa289v (kanazawa iii) was confirmed as hypodysfibrinogenemia by plasma and recombinant fibrinogens.congenital fibrinogen disorders are classified as afibrinogenemia, hypofibrinogenemia, dysfibrinogenemia, and hypodysfibrinogenemia. however, difficulties are associated with discriminating between dysfibrinogenemia, hypofibrinogenemia, and hypodysfibrinogenemia using routine analyses. we previously reported a heterozygous variant fibrinogen (γa289v; kanazawa iii) as hypodysfibrinogenemia; however, the same variant had previously been described as hypofibrinogenemia. to clarify the production of ...202031957968
influence of cytoskeleton organization on recombinant protein expression by cho cells.in this study, we assessed the importance of cytoskeleton organization in the mammalian cells used to produce therapeutic proteins. two cytoskeletal genes, actin alpha cardiac muscle 1 (actc1) and a guanosine triphosphate gtpase-activating protein (tagap), were found to be upregulated in highly productive therapeutic protein-expressing chinese hamster ovary (cho) cells selected by the deprivation of vitamin b5. we report here that the overexpression of the actc1 protein was able to improve signi ...202031956990
overexpression of transcription factor foxa1 and target genes remediate therapeutic protein production bottlenecks in chinese hamster ovary cells.despite extensive research conducted to increase protein production from chinese hamster ovary (cho) cells, cellular bottlenecks often remain, hindering high yields. in this study, a transcriptomic analysis led to the identification of 32 genes that are consistently upregulated in high producer clones and thus might mediate high productivity. candidate genes were associated with functions such as signaling, protein folding, cytoskeleton organization, and cell survival. we focused on two engineer ...202031956982
understanding the effect of high gas entrance velocity on chinese hamster ovary (cho) cell culture performance and its implications on bioreactor scale-up and sparger design.there are three main potential sources for cell shear damage existing in stirred tank bioreactors. one is the potential high energy dissipation in the immediate impeller zones; another from small gas bubble burst; and third is from high gas entrance velocity (gev) emitting from the sparger. while the first two have been thoroughly addressed for the scale-up of chinese hamster ovary (cho) cell culture knowing that a wide tolerable agitation range with non-damaging energy dissipation exists and th ...202032086806
optimized crispr/cas9 strategy for homology-directed multiple targeted integration of transgenes in cho cells.site-specific integration has emerged as a promising strategy for precise chinese hamster ovary (cho) cell line engineering and predictable cell line development (cld). crispr/cas9 with the homology-directed repair (hdr) pathway enables precise integration of transgenes into target genomic sites. however, inherent recalcitrance to hdr-mediated targeted integration (ti) of transgenes results in low targeting efficiency, thus requiring a selection process to find a targeted integrant in cho cells. ...202032086804
identification and functional characterization of a novel splicing variant in the f8 coagulation gene causing severe hemophilia a.we have identified a synonymous f8 variation in a severe hemophilia a (ha) patient who developed inhibitors following factor viii (fviii) prophylaxis. the unreported c.6273 g > a variant targets the consensus splicing site of exon 21.202032078252
altered gene expression in cho cells following polyamine starvation.to investigate the impact of polyamine deprivation on the transcriptome of cho cells results: polyamines play a central but poorly-understood role in cell proliferation. most studies to date have utilised chemical inhibitors to probe polyamine function. here we exploit the fact that cho cells grown in serum-free medium have an absolute requirement for putrescine supplementation due to their deficiency in activity of the enzyme arginase. a gene expression microarray (affymetrix) analysis of cho-k ...202032078082
improvement of intracellular traffic system by overexpression of kdel receptor 1 in antibody-producing cho cells.the localization of soluble endoplasmic reticulum (er) chaperones in the cell organelle is mediated by the c-terminal kdel (lysine, aspartic acid, glutamic acid and leucine) motif. this motif is recognized by the kdel receptor, a seven-transmembrane protein that cycles between the er and cis-golgi to capture missorted kdel chaperones from post-er compartments in a ph-dependent manner. the kdel receptor's target chaperones have a substantial role in protein folding and assembly. in this study, th ...202032073237
[eukaryotic expression, protein purification and biological effects research of human cs1-fc fusion protein].signaling lymphocyte activation family 7 (slamf7/cs1) is a cell surface glycoprotein that is highly expressed in multiple myeloma cells. cs1 is a sensitive and specific biomarker for multiple myeloma. car-t cell immunotherapy is a new method for the treatment of multiple myeloma. cs1 car-t cell immunotherapy has good effect on relapsed refractory multiple myeloma. to detect the expression efficiency of cs1 car on cs1 car-t cells and to find an auxiliary means to car-t cell immunotherapy, we prep ...202032072787
data for engineering lipid metabolism of chinese hamster ovary (cho) cells for enhanced recombinant protein production.the data presented in this article relates to the manuscript entitled 'engineering of chinese hamster ovary cell lipid metabolism results in an expanded er and enhanced recombinant biotherapeutic protein production', published in the journal metabolic engineering [1]. in the article here, we present data examining the overexpression of the lipid metabolism modifying genes scd1 and srebf1 in cho cells by densitometry of western blots and by using mass spectrometry to investigate the impact on spe ...202032071989
the use of catechins in chinese hamster ovary cell media for the improvement of monoclonal antibody yields and a reduction of acidic species.catechin compounds have potential benefits for recombinant monoclonal antibody (mab) production as chemical additives in cell culture media. in this study, four catechin compounds catechin (cat), epicatechin (ec), gallocatechin-gallate (gcg), and epigallocatechin-gallate (egcg) were added to cell culture media (at 50 μm) and their effects on the recombinant chinese hamster ovary (cho) cell culture, specific productivity, and mab quality were assessed. the results indicate that the improvement of ...202032067358
neutralizing antibody against osteopontin attenuates non-alcoholic steatohepatitis in mice.previously, we reported that an extracellular matrix protein, osteopontin (opn), is involved in various autoimmune diseases using a neutralizing polyclonal antibody against opn generated in rabbits. however, the antibody cannot be used for long-term mouse models of chronic inflammatory disease because of the induction of antibodies against anti-opn rabbit igg. in this study, we generated a new antibody, anti-mouse opn mouse igg (35b6). 35b6 inhibited the cell adhesion of mouse and human opn to c ...202032062834
cytotoxic and genotoxic effects of e-liquids and their potential associations with nicotine, menthol and phthalate esters.in this study, we determined dna damage and chromosome breakage (indicators of genotoxicity) and cell viability (an indicator of cytotoxicity) in human lymphoblastoid tk6 and chinese hamster ovary (cho) cells treated with 33 e-liquids using in vitro single cell gel (comet), micronucleus (mn), and trypan blue assays, respectively. we also measured the contents of nicotine, five phthalate esters, and dl-menthol in the e-liquids to examine their effects on dna damage, chromosome breakage, and cell ...202032058129
codon and amino acid content are associated with mrna stability in mammalian cells.messenger rna (mrna) degradation plays a critical role in regulating transcript levels in the cell and is a major control point for modulating gene expression. in yeast and other model organisms, codon identity is a powerful determinant of transcript stability, contributing broadly to impact half-lives. general principles governing mrna stability are poorly understood in mammalian systems. importantly, however, the degradation machinery is highly conserved, thus it seems logical that mammalian t ...202032053646
single-step synthesis and in vitro anti-mycobacterial activity of novel nitrofurantoin analogues.the emergence of drug-resistant tuberculosis (dr-tb) as well as the requirement for long, expensive and toxic drug regimens impede efforts to control and eliminate tb. therefore, there's a need for effective and affordable anti-mycobacterial agents which can shorten the duration of therapy and are active against mycobacterium tuberculosis (mtb) in both active and latent phases. nitrofurantoin (nft) is a hypoxic agent with activity against a myriad of anaerobic pathogens and, like the first-line ...202032044516
profiles of plant core-fucosylated n-glycans of acid alpha-glucosidases produced in transgenic rice cell suspension cultures treated with eight different conditions.recombinant human acid alpha-glucosidase (rhgaa) from chinese hamster ovary cells is the only approved treatment for patients with pompe disease. in this study, rhgaas were produced in transgenic rice cell suspension cultures under eight different conditions; untreated, 5 μm of 2-fluoro-l-fucose (2-ff), 50 μm of 2-ff, 100 μm of 2-ff, 100 μm of 2-ff + 0.5% pluronic f-68 (pf-68), 100 μm of 2-ff + 0.05% tween 20 (tw 20), 0.5% pf-68, and 0.05% tw 20. the n-glycans of eight rhgaas were analyzed using ...202032044029
c1q/tnf-related protein 6 is a pattern recognition molecule that recruits collectin-11 from the complement system to ligands.c1q/tnf-related protein (ctrp) 6 is a member of the ctrp protein family associated with the regulation of cellular and endocrine processes. ctrp6 contains collagen and globular structures, resembling the pattern recognition molecules (prms) of the classical and lectin complement pathways. we expressed human ctrp6 in chinese hamster ovary cells and investigated the binding to different putative ligands (acetylated bsa [acbsa], zymosan, mannan, and lps from escherichia coli and salmonella as well ...202032041782
engineering mammalian cells to produce plant-specific n-glycosylation on proteins.protein n-glycosylation is an essential and highly conserved posttranslational modification found in all eukaryotic cells. yeast, plants and mammalian cells, however, produce n-glycans with distinct structural features. these species-specific features not only pose challenges in selecting host cells for production of recombinant therapeutics for human medical use but also provide opportunities to explore and utilize species-specific glycosylation in design of vaccines. here, we used reverse cros ...202032039452
the epilepsy of infancy with migrating focal seizures: identification of de novo mutations of the kcnt2 gene that exert inhibitory effects on the corresponding heteromeric kna1.1/kna1.2 potassium channel.the epilepsy of infancy with migrating focal seizures (eimfs; previously called malignant migrating partial seizures of infancy) are early-onset epileptic encephalopathies (eoee) that associate multifocal ictal discharges and profound psychomotor retardation. eimfs have a genetic origin and are mostly caused by de novo mutations in the kcnt1 gene, and much more rarely in the kcnt2 gene. kcnt1 and kcnt2 respectively encode the kna1.1 (slack) and kna1.2 (slick) subunits of the sodium-dependent vol ...202032038177
inhibition of cholesteryl ester synthesis by polyacetylenes from atractylodes rhizome.using activity guided purification, four known compounds, sesquiterpene atractylenolide iii (1), and the polyacetylenes 14-acetoxy-12-senecioyloxytetradeca-2e,8e,10e-trien-4,6-diyn-1-ol (2), 14-acetoxy-12-α-methylbutyl-2e,8e,10e-trien-4,6-diyn-1-ol (3), and 14-acetoxy-12-β -methylbutyl-2e,8e,10e-trien-4,6-diyn-1-ol (4), were isolated from a traditional herbal medicine, atractylodes rhizome. structurally similar 3 and 4 (3/4 mixture) were obtained as a mixture. in intact chinese hamster ovary (ch ...202032035699
protective effects of compounds from cimicifuga dahurica against amyloid beta production in vitro and scopolamine-induced memory impairment in vivo.cimicifuga dahurica has traditionally been used as an antipyretic, analgesic, and anti-inflammatory agent and as a treatment for uterine and anal prolapse. this study has investigated the potential beneficial effects of this medicinal plant and its components on alzheimer's disease (ad) with a focus on amyloid beta (aβ) production and scopolamine-induced memory impairment in mice. an ethanol extract from c. dahurica roots decreased aβ production in app-cho cells [chinese hamster ovarian (cho) ce ...202032031796
glucose monitoring in cell culture with online ultrasound-assisted near-infrared spectroscopy.near-infrared (nir) spectroscopic analysis for suspensions often requires a pretreatment by dilution or filtering of suspended solids to maintain the sensitivity of the measurements. an online ultrasound-assisted spectroscopy (uas) unit enabling pretreatment-free and noncontact analysis for bioprocessing is proposed and evaluated with a model suspension containing 3-μm-diameter polystyrene latex particles (psls) with the density of 5.1 × 108 counts/ml and chinese hamster ovary (cho) cell culture ...202032023047
differential effects of 2-hydroxypropyl-cyclodextrins on lipid accumulation in npc1-null cells.niemann-pick disease type c (npc) is an autosomal recessive disorder characterized by abnormal accumulation of free cholesterol and sphingolipids in lysosomes. the iminosugar miglustat, which inhibits hexosylceramide synthesis, is used for npc treatment, and 2-hydroxypropyl-β-cyclodextrin (hp-β-cd), a cyclic oligosaccharide derivative, is being developed to treat npc. moreover, therapeutic potential of 2-hydroxypropyl-γ-cyclodextrin (hp-γ-cd) was shown in npc models, although its mechanism of ac ...202032019132
3-quinuclidinyl-α-methoxydiphenylacetate: a multi-targeted ligand with antimuscarinic and antinicotinic effects designed for the treatment of anticholinesterase poisoning.racemic 3-quinuclidinyl-α-methoxydiphenylacetate (mb266) was synthesised. its activity at muscarinic acetylcholine receptors (machrs), and muscle and neuronal nicotinic acetylcholine receptors (nachrs), was compared to that of atropine and racemic 3-quinucidinyl benzilate (qnb) using a functional assay based on agonist-induced elevation of intracellular calcium ion concentration in cn21, chinese hamster ovary (cho) and shsy5y human cell lines. mb266 acted as an antagonist at acetylcholine recept ...202032017982
analysis of chemometric models applied to raman spectroscopy for monitoring key metabolites of cell culture.the food and drug administration (fda) initiative of process analytical technology (pat) encourages the monitoring of biopharmaceutical manufacturing processes by innovative solutions. raman spectroscopy and the chemometric modeling tool partial least squares (pls) have been applied to this aim for monitoring cell culture process variables. this study compares the chemometric modeling methods of support vector machine radial (svmr), random forests (rf), and cubist to the commonly used linear pls ...202032012476
feeding tricarboxylic acid cycle intermediates improves lactate consumption and antibody production in chinese hamster ovary cell cultures.media components play an important role in modulating cell metabolism and improving product titer in mammalian cell cultures. to sustain cell productivity, highly active oxidative metabolism is desired. here we explored the effect of tricarboxylic acid (tca) cycle intermediates supplementation on lactate metabolism and productivity in chinese hamster ovary fed-batch cultures. direct addition of 5 mm alpha-ketoglutarate (α-kg), malic acid, or succinic acid in the basal medium did not have any sig ...202032012447
structure-function relationships of the soluble form of the antiaging protein klotho have therapeutic implications for managing kidney disease.the fortuitously discovered antiaging membrane protein αklotho (klotho) is highly expressed in the kidney, and deletion of the klotho gene in mice causes a phenotype strikingly similar to that of chronic kidney disease (ckd). klotho functions as a co-receptor for fibroblast growth factor 23 (fgf23) signaling, whereas its shed extracellular domain, soluble klotho (sklotho), carrying glycosidase activity, is a humoral factor that regulates renal health. low sklotho in ckd is associated with diseas ...202032005658
anti-d monoclonal antibodies from 23 human and rodent cell lines display diverse igg fc-glycosylation profiles that determine their clinical efficacy.anti-d immunoglobulin (anti-d ig) prophylaxis prevents haemolytic disease of the fetus and newborn. monoclonal igg anti-ds (mab-ds) would enable unlimited supplies but have differed in efficacy in fcγriiia-mediated adcc assays and clinical trials. structural variations of the oligosaccharide chains of mab-ds are hypothesised to be responsible. quantitative data on 12 fc-glycosylation features of 23 mab-ds (12 clones, 5 produced from multiple cell lines) and one blood donor-derived anti-d ig were ...202032001734
antibody-drug conjugates using mouse-canine chimeric anti-dog podoplanin antibody exerts antitumor activity in a mouse xenograft model.antibody-drug conjugates (adcs), which consist of a monoclonal antibody (mab), a linker, and a payload, can deliver a drug to cancer tissues. we previously produced an anti-dog podoplanin (dpdpn) mab, pmab-38, which reacts with dpdpn-expressing canine melanomas and squamous cell carcinomas (sccs), but not with dpdpn-expressing canine type i alveolar cells or lymphatic endothelial cells, indicating that pmab-38 possesses cancer specificity. in this study, we developed an adc, p38b-dm1, using the ...202032182186
cell function profiling to assess clone stability.in cell line development the identification of stable chinese hamster ovary cells for production is a critical but onerous task. the stability trial focus upon high-level attributes can mask profound underlying cellular changes, leading to unstable clones mistakenly being chosen for production. the challenge is to assay underlying cell pathways and subsystems without pushing up cell line development costs. chemstress® cell function profiling is a simple, multiwell plate-based assay that uses a p ...202032181887
hydrocyclones as cell retention device for cho perfusion processes in single-use bioreactors.in this study, a hydrocyclone (hc) especially designed for mammalian cell separation was applied for the separation of chinese hamster ovary cells. the effect of key features on the separation efficiency, such as type of pumphead in the peristaltic feed pump, use of an auxiliary pump to control the perfusate flow rate, and tubing size in the recirculation loop were evaluated in batch separation tests. based on these preliminary batch tests, the hc was then integrated to 50-l disposable bioreacto ...202032181883
slowing of the time course of acidification decreases the acid-sensing ion channel 1a current amplitude and modulates action potential firing in neurons.acid-sensing ion channels (asics) are h+-activated neuronal na+ channels. they are involved in fear behavior, learning, neurodegeneration after ischemic stroke and in pain sensation. asic activation has so far been studied only with fast ph changes, although the ph changes associated with many roles of asics are slow. it is currently not known whether slow ph changes can open asics at all. here, we investigated to which extent slow ph changes can activate asic1a channels and induce action potent ...202032180707
multi-omics reveals impact of cysteine feed concentration and resulting redox imbalance on cellular energy metabolism and specific productivity in cho cell bioprocessing.chinese hamster ovary (cho) cells are currently the primary host cell lines used in biotherapeutic manufacturing of monoclonal antibodies (mabs) and other biopharmaceuticals. cellular energy metabolism and endoplasmic reticulum (er) stress are known to greatly impact cell growth, viability, and specific productivity of a biotherapeutic; but the molecular mechanisms are not fully understood. the authors previously employed multi-omics profiling to investigate the impact of a reduction in cysteine ...202032170810
evaluation of glycoprotein e subunit and live attenuated varicella-zoster virus vaccines formulated with a single-strand rna-based adjuvant.varicella-zoster virus (vzv), a human alphaherpesvirus 3, elicits both chickenpox and shingles and/or postherpetic neuralgia. a live attenuated vaccine (lav) and glycoprotein e (ge) subunit vaccine were developed to prevent vzv-induced diseases. we recently reported that single-strand rna (ssrna) based on the intergenic region of the internal ribosome entry site of cricket paralysis virus (crpv) is an effective adjuvant for protein-based and virus-like particle-based vaccines. here, chinese hams ...202032167678
review of the current methods of chinese hamster ovary (cho) cells cultivation for production of therapeutic protein.most of clinical approved protein-based drugs or under in clinical trial have a profound impact in the treatment of critical diseases. the mammalian eukaryotic cells culture approaches, particularly the cho (chinese hamster ovary) cells are mainly used in the biopharmaceutical industry for the mass-production of therapeutic protein. recent advances in cho cell bioprocessing to yield recombinant proteins and monoclonal antibodies have enabled the expression of quality protein. the developments of ...202032164511
effects of titanium dioxide nanoparticles on the hprt gene mutations in v79 hamster cells.the genotoxicity of anatase/rutile tio2 nanoparticles (tio2 nps, nm105 at 3, 15 and 75 µg/cm2) was assessed with the mammalian in-vitro hypoxanthine guanine phosphoribosyl transferase (hprt) gene mutation test in chinese hamster lung (v79) fibroblasts after 24 h exposure. two dispersion procedures giving different size distribution and dispersion stability were used to investigate whether the effects of tio2 nps depend on the state of agglomeration. tio2 nps were fully characterised in the previ ...202032150818
comparison of the pharmacological profiles of arginine vasopressin and oxytocin analogs at marmoset, macaque, and human vasopressin 1a receptor.arginine vasopressin (avp) and oxytocin (ot) are nonapeptides that bind to g-protein coupled receptors and influence social behaviors. consensus mammalian avp and ot (leu8-ot) sequences are highly conserved. in marmosets, an amino acid change in the 8th position of the peptide (pro8-ot) exhibits unique structural and functional properties. there is ∼85 % structural homology between the ot receptor (otr) and vasopressin 1a receptor (v1ar) resulting in significant cross-reactivity between the liga ...202032145592
influence of polymer charge on the localization and dark- and photo-induced toxicity of a potential type i photosensitizer in cancer cell models.a current trend within photo-dynamic therapy (pdt) is the development of molecular systems targeting hypoxic tumors. thus, type i pdt sensitizers could here overcome traditional type ii molecular systems that rely on the photo-initiated production of toxic singlet oxygen. here, we investigate the cell localization properties and toxicity of two polymeric anthracene-based fluorescent probes (neutral ant-phea and cationic ant-pim). the cell death and dna damage of chinese hamster ovary cancer cell ...202032138280
enhancement of sialylation in rigg in glyco-engineered chinese hamster ovary cells.since about 70% of commercial biopharmaceutical products have been produced in chinese hamster ovary (cho) cells, this cell line is undeniably a workhorse for biopharmaceuticals production. meanwhile, sialic acid terminals were reported to affect anti-inflammatory activity, antibody-dependent cellular cytotoxicity efficacy of igg antibodies. taking these findings together, we aimed to establish cho cell lines that highly produce sialic acid terminals by overexpressing two n-acetylneuraminic acid ...202032125558
ascorbic acid 2-glucoside pretreatment protects cells from ionizing radiation, uvc, and short wavelength of uvb.ascorbic acid 2-glucoside (aa2g), glucosylated ascorbic acid (aa), has superior properties for bioavailability and stability compared to aa. although aa2g has shown radioprotective properties similar to aa, effects for uv light, especially uvc and uvb, are not studied. aa2g was tested for cytotoxicity and protective effects against ionizing radiation, uvc, and broadband and narrowband uvb in chinese hamster ovary (cho) cells and compared to aa and dimethyl sulfoxide (dmso). pretreatment with dms ...202032106443
circulating ficolin-2 and ficolin-3 form heterocomplexes.the complement system constitutes an important part of the innate immune system. the collectins and the ficolins are soluble pattern recognition molecules that contribute to complement activation via the lectin pathway. during previous experiments with ficolin-2 and ficolin-3, we have observed that the molecules may interact. we therefore hypothesized the existence of stable ficolin-2/-3 heterocomplexes. we could demonstrate ficolin-2/-3 heterocomplexes in normal human serum and plasma by elisa ...202032094208
mrna transfection into cho-cells reveals production bottlenecks.obtaining highly productive chinese hamster ovary (cho)-cell clones for the production of therapeutic proteins relies on multiple time-consuming selection steps. several cho-cell strains with high degrees of genomic and epigenetic variation are available. each harbor potential advantages and disadvantages for any given product, particularly those considered difficult to express. a simple test system to quickly assess compatibility of cell line and product may therefore prove useful. transient pl ...202031475780
hplc-esi-qqq based standardization, mutagenic and genotoxic potential of methanol extract of ziziphus mauritiana lam leaves.the leaves of ziziphus mauritiana lam have been an integral part of the traditional system of medicine for the treatment of inflammation, wounds, fever, asthma and liver disorders. the leaves are utilised as an edible vegetable in rural parts of india and indonesia. despite its pharmacological significance, ziziphus mauritiana lam lacks scientific evidence on its mutagenic and genotoxic potential.202031493461
role of human n-acetyltransferase 2 genetic polymorphism on aromatic amine carcinogen-induced dna damage and mutagenicity in a chinese hamster ovary cell mutation assay.carcinogenic aromatic amines such as 4-aminobiphenyl (abp) and 2-aminofluorene (af) require metabolic activation to form electrophilic intermediates that mutate dna leading to carcinogenesis. bioactivation of these carcinogens includes n-hydroxylation catalyzed by cyp1a2 followed by o-acetylation catalyzed by arylamine n-acetyltransferase 2 (nat2). to better understand the role of nat2 genetic polymorphism in abp- and af-induced mutagenesis and dna damage, nucleotide excision repair-deficient (u ...202031490564
process intensification in fed-batch production bioreactors using non-perfusion seed cultures.although process intensification by continuous operation has been successfully applied in the chemical industry, the biopharmaceutical industry primarily uses fed-batch, rather than continuous or perfusion methods, to produce stable monoclonal antibodies (mabs) from chinese hamster ovary (cho) cells. conventional fed-batch bioreactors may start with an inoculation viable cell density (vcd) of ~0.5 × 106 cells/ml. increasing the inoculation vcd in the fed-batch production bioreactor (referred to ...202031379298
intracerebroventricular enzyme replacement therapy with β-galactosidase reverses brain pathologies due to gm1 gangliosidosis in mice.autosomal recessive mutations in the galactosidase β1 (glb1) gene cause lysosomal β-gal deficiency, resulting in accumulation of galactose-containing substrates and onset of the progressive and fatal neurodegenerative lysosomal storage disease, gm1 gangliosidosis. here, an enzyme replacement therapy (ert) approach in fibroblasts from gm1 gangliosidosis patients with recombinant human β-gal (rhβ-gal) produced in chinese hamster ovary cells enabled direct and precise rhβ-gal delivery to acidified ...202031481471
phosphorylation of unique c-terminal sites of the mu-opioid receptor variants 1b2 and 1c1 influences their gs association following chronic morphine.we recently demonstrated in rat spinal cord that a regimen of escalating doses of systemic morphine, analogous to regimens used clinically for chronic pain management, selectively up-regulates the mu-opioid receptor (mor) splice variants mor-1b2 and mor-1c1 mrna and functional protein. this study investigated the potential relevance of up-regulating mor-1b2 and mor-1c1 to the ability of chronic morphine to shift mor signaling from predominantly gi /go inhibitory to gs stimulatory. specifically, ...202031479519
assessment of complex genomic alterations induced by azt, 3tc, and the combination azt +3tc.highly active antiretroviral therapy (haart) regimens are based on the use of nucleoside reverse transcriptase inhibitors (nrtis), which are the main drugs used by patients infected with the human immunodeficiency virus (hiv). the use of nrtis combinations has afforded clear clinical benefits to patients undergoing haart. however, the combination of two nrtis may increase the risk of genomic instability in comparison with the drugs administered individually. we analyzed the ability of zidovudine ...202030208744
the development and study of recombinant immunoglobulin a to hemagglutinins of the influenza virus.we obtained recombinant variants of human antibody fi6 broadly specific to hemagglutinins of the influenza a virus. on the basis of a bi-promoter (cmv, hef1-htlv) vector, we developed genetic constructs for the expression of the heavy and light chains of the immunoglobulins of iga1-, iga2m1-, and igg-isotypes. following transfection and selection, stable chinese hamster ovary (cho) cell lines were produced. the antibodies of iga1-, iga2m1-, and igg-isotypes were purified from culture media. we p ...202030116613
variable readthrough responsiveness of nonsense mutations in hemophilia a.readthrough therapy relies on the use of small molecules that enable premature termination codons in mrna open reading frames to be misinterpreted by the translation machinery, thus allowing the generation of full-length, potentially functional proteins from mrna carrying nonsense mutations. in patients with hemophilia a, nonsense mutations potentially sensitive to readthrough agents represent approximately 16% of the point mutations. the aim of this study was to measure the readthrough effect o ...202031197069
characterization of chinese hamster ovary cells with disparate chromosome numbers: reduction of the amount of mrna relative to total protein.chromosomes in chinese hamster ovary (cho) cells are labile. we have shown that high-chromosome-number cho cells have greater potential to become robust producers of recombinant proteins. one explanation being the increase in transgene integration sites. however, high-chromosome-number cell clones produce more igg3 following culture of single-cell clones, even under conditions that yield the same number of integrations as cells with normal chromosome numbers. here, we characterized high-chromoso ...202031303495
impact of fc n-glycan sialylation on igg structure.human igg antibodies containing terminal alpha 2,6-linked sialic acid on their fc n-glycans have been shown to reduce antibody-dependent cell-mediated cytotoxicity and possess anti-inflammatory properties. although terminal sialylation on complex n-glycans can happen via either an alpha 2,3-linkage or an alpha 2,6-linkage, sialic acids on human serum igg fc are almost exclusively alpha 2,6-linked. recombinant iggs expressed in chinese hamster ovary (cho) cells, however, have sialic acids through ...202031411531
identification of novel phytocannabinoids from ganoderma by label-free dynamic mass redistribution assay.located throughout the body, cannabinoid receptors (cb1 and cb2) are therapeutic targets for obesity/metabolic diseases, neurological/mental disorders, and immune modulation. phytocannabinoids are greatly important for the development of new medicines with high efficacy and/or minor side effects. plants and fungi are used in traditional medicine for beneficial effects to mental and immune system. the current research studied five fungi from the genus ganoderma and five plants: ganoderma hainanen ...202031494202
advancing biologics development programs with legacy cell lines: advantages and limitations of genetic testing for addressing clonality concerns prior to availability of late stage process and product consistency data.the bioprocessing industry uses recombinant mammalian cell lines to generate therapeutic biologic drugs. to ensure consistent product quality of the therapeutic proteins, it is imperative to have a controlled production process. regulatory agencies and the biotechnology industry consider cell line "clonal origin" an important aspect of maintaining process control. demonstration of clonal origin of the cell substrate, or production cell line, has received considerable attention in the past few ye ...202031519780
glycan residues balance analysis - greba: a novel model for the n-linked glycosylation of igg produced by cho cells.the structure of n-linked glycosylation is a very important quality attribute for therapeutic monoclonal antibodies. different carbon sources in cell culture media, such as mannose and galactose, have been reported to have different influences on the glycosylation patterns. accurate prediction and control of the glycosylation profile are important for the process development of mammalian cell cultures. in this study, a mathematical model, that we named glycan residues balance analysis (greba), w ...202031539564
the impact of sialylation linkage-type on the pharmacokinetics of recombinant butyrylcholinesterases.chinese hamster ovary (cho) cells typically produce glycoproteins with n-glycans terminating in α-2,3 sialylation. human cells produce glycoproteins that include α-2,3 and α-2,6 sialic acids. to examine the impact of altering protein sialylation on pharmacokinetic properties, recombinant human butyrylcholinesterase (bche) was produced in cho cells by knocking out the α-2,3 sialyltransferase genes followed by overexpression of the α-2,6 sialyltransferase (26bche) enzyme. the n-glycan composition ...202031544955
impact of micro and macroporous tff membranes on product sieving and chromatography loading for perfusion cell culture.bioprocess intensification can be achieved through high cell density perfusion cell culture with continuous protein capture integration. protein passage and cell retention are commonly accomplished using tangential flow filtration systems consisting of microporous membranes. significant challenges, including low efficiency and decaying product sieving over time, are commonly observed in these cell retention devices. here, we demonstrate that a macroporous membrane overcomes the product sieving c ...202031612989
integration of high-throughput analytics and cell imaging enables direct early productivity and product quality assessment during chinese hamster ovary cell line development for a complex multi-subunit vaccine antigen.mammalian cell line generation typically includes stable pool generation, single cell cloning and several rounds of clone selection based on cell growth, productivity and product quality criteria. individual clone expansion and phenotype-based ranking is performed initially for hundreds or thousands of mini-scale cultures, representing the major operational challenge during cell line development. automated cell culture and analytics systems have been developed to enable high complexity clone sel ...202031568688
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