Publications

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application of a curated genome-scale metabolic model of cho dg44 to an industrial fed-batch process.cho cells have become the favorite expression system for large scale production of complex biopharmaceuticals. however, industrial strategies for upstream process development are based on empirical results, due to a lack of fundamental understanding of intracellular activities. genome scale models of cho cells have been reconstructed to provide an economical way of analyzing and interpreting large-omics datasets, since they add cellular context to the data. here the most recently available cho-d ...201930227251
systematic development of temperature shift strategies for chinese hamster ovary cells based on short duration cultures and kinetic modeling.temperature shift (ts) to a hypothermic condition has been widely used during protein production processes that use chinese hamster ovary (cho) cells. the effect of temperature on cell growth, metabolites, protein titer and quality depends on cell line, product, and other bioreactor conditions. due to the large numbers of experiments, which typically last 2-3 weeks each, limited systematic ts studies have been reported with multiple shift temperatures and steps at different times. here, we syste ...201930230966
the cytosolic peroxisome-targeting signal (pts)-receptors, pex7p and pex5pl, are sufficient to transport pts2 proteins to peroxisomes.proteins harboring peroxisome-targeting signal type-2 (pts2) are recognized in the cytosol by mobile pts2 receptor pex7p and associate with a longer isoform pex5pl of the pts1 receptor. trimeric pts2 protein-pex7p-pex5pl complexes are translocated to peroxisomes in mammalian cells. however, it remains unclear whether pex5pl and pex7p are sufficient cytosolic components in transporting of pts2 proteins to peroxisomes. here, we construct a semi-intact cell import system to define the cytosolic com ...201930296498
a highly efficient and cost-effective recombinant production of a bacterial photolyase from the antarctic isolate hymenobacter sp. uv11.photolyases are dna-repairing flavoproteins that are represented in most phylogenetic taxa with the exception of placental mammals. these enzymes reduce the ultraviolet-induced dna damage; thus, they have features that make them very attractive for dermatological or other medical uses, such as the prevention of human skin cancer and actinic keratosis. in this work, we identified a 50.8 kda photolyase from the uvc-resistant antarctic bacterium hymenobacter sp. uv11. the enzyme was produced by rec ...201930267301
genotoxicity and embryotoxicity study of bicyclol methyl ether, main impurity in bicyclol.to assess the genotoxicity and embryotoxicity of bicyclol methyl ether (bme), the main impurity in bicyclol.201930242590
transient cho expression platform for robust antibody production and its enhanced n-glycan sialylation on therapeutic glycoproteins.large-scale transient expression in mammalian cells is a rapid protein production technology often used to shorten overall timelines for biotherapeutics drug discovery. in this study we demonstrate transient expression in a chinese hamster ovary (cho) host (expicho-s™) cell line capable of achieving high recombinant antibody expression titers, comparable to levels obtained using human embryonic kidney (hek) 293 cells. for some antibodies, expicho-s™ cells generated protein materials with better ...201930299005
helvamide, a new inhibitor of sterol o-acyltransferase produced by the fungus aspergillus nidulans bf-0142.a new piperazine derivative designated helvamide was isolated as a pair of rotamers (1 and 2) from the culture broth of the fungus aspergillus nidulans bf-0142 along with a known helvafuranone (3). the structures of 1 and 2 were elucidated based on spectroscopic analyses by the interpretation of one-dimensional and two-dimensional nuclear magnetic resonance data, roesy (rotational overhauser effect spectroscopy) correlations, and a chemical method. helvamide existed as a rotameric mixture (1 and ...201930305686
thermal expansion of substrate may affect adhesion of chinese hamster fibroblasts to surfaces during freezing.despite success in cryopreservation of cells in suspension, cryopreservation of cells in monolayers is still challenging. one of the major problems is detachment of the cells from the substrate which occurs during cryopreservation. we hypothesized that this detachment may be due to a mismatch in the coefficient of linear thermal expansion αl between glass and the frozen cell layer which manifests as residual stress and stress relaxation. this mismatch results in a difference between the thermal ...201930312591
biomedical potential of chitosan-silver nanoparticles with special reference to antioxidant, antibacterial, hemolytic and in vivo cutaneous wound healing effects.the aim of the present study was to prepare chitosan-pva-silver nanoparticles (cs-agnps) through green method. chitosan and pva polymers acted as stabilizing agents. dls and tem analyses showed that cs-agnps were homogeneously dispersed in matrix with an average size of 190-200 nm. the cs-agnps were tested for their antioxidant and antibacterial properties and the results revealed that they exhibited higher antioxidant activity than cs powder. moreover, cs-agnps were characterized by a low cytot ...201930339915
a recurrent missense variant in slc9a7 causes nonsyndromic x-linked intellectual disability with alteration of golgi acidification and aberrant glycosylation.we report two unrelated families with multigenerational nonsyndromic intellectual disability (id) segregating with a recurrent de novo missense variant (c.1543c>t:p.leu515phe) in the alkali cation/proton exchanger gene slc9a7 (also commonly referred to as nhe7). slc9a7 is located on human x chromosome at xp11.3 and has not yet been associated with a human phenotype. the gene is widely transcribed, but especially abundant in brain, skeletal muscle and various secretory tissues. within cells, slc9 ...201930335141
nickel and cobalt affect galactosylation of recombinant igg expressed in cho cells.glycosylation is an important product quality attribute of antibody biopharmaceuticals. it involves enzymatic addition of oligosaccharides on proteins by sequential action of glycosyltransferases and glycosidases in the endoplasmic reticulum and golgi. some of these enzymes like galactosyltransferase and n-acetylglucosaminyltransferase-i require trace metal cofactors. variations in trace metal availability during production can thus affect glycosylation of recombinant glycoproteins such as monoc ...201930327978
cathepsin l causes proteolytic cleavage of chinese-hamster-ovary cell expressed proteins during processing and storage: identification, characterization, and mitigation.a stochastic approach of copurification of the protease cathepsin l that results in product fragmentation during purification processing and storage is presented. cathepsin l was identified using mass spectroscopy, characterization of proteolytic activity, and comparison with fragmentation patterns observed using recombinant cathepsin l. cathepsin l existed in chinese hamster ovary cell culture fluids obtained from cell lines expressing different products and cleaved a variety of recombinant pro ...201930320962
a new highly thyrotropin receptor-selective small-molecule antagonist with potential for the treatment of graves' orbitopathy.the thyrotropin receptor (tshr) is the target for autoimmune thyroid stimulating antibodies (tsab) triggering hyperthyroidism. whereas elevated thyroid hormone synthesis by the thyroid in graves' disease can be treated by antithyroid agents, for the pathogenic activation of tshr in retro-orbital fibroblasts of the eye, leading to graves' orbitopathy (go), no causal tshr directed therapy is available.201930351237
whole synthetic pathway engineering of recombinant protein production.the output from protein biomanufacturing systems is a function of total host cell biomass synthetic capacity and recombinant protein production per unit cell biomass. in this study, we describe how these two properties can be simultaneously optimized via design of a product-specific combination of synthetic dna parts to maximize flux through the protein synthetic pathway and the use of a host cell chassis with an increased capability to synthesize both cell and product biomass. using secreted al ...201930345502
structure-activity relationship studies of antiplasmodial cyclometallated ruthenium(ii), rhodium(iii) and iridium(iii) complexes of 2-phenylbenzimidazoles.benzimidazoles, such as albendazole, thiabendazole and omeprazole have antiplasmodial activity against plasmodium falciparum and are widely used as scaffolds for metal-based drug research. incorporating substituents with various lipophilic and electronic properties can influence trans-membrane interactions and concomitantly improve the biological activity. to study structure-activity relationships, a series of 2-phenylbenzimidazoles and their corresponding ru(ii), ir(iii) and rh(iii) cyclometall ...201930342422
label-free independent quantitation of viable and non-viable cells using a multivariable multi-resonant sensor.biological cells are utilized for diverse biotechnological and bioengineering purposes ranging from the production of biopharmaceuticals, to cell therapy, "human-on-a-chip" drug and toxicology assays, and drug-resistance tests. in these and other applications, it is critical to quantify the levels of not only viable but also non-viable cells. while traditional off-line cell-staining methods are available for counting of non-viable cells, many applications cannot periodically remove cells for the ...201930342232
tetrandrine, a novel inhibitor of ether-à-go-go-1 (eag1), targeted to cervical cancer development.mortality-to-incidence ratios in patients with cancer are extremely high, positioning cancer as a major cause of death worldwide. ether-à-go-go-1 (eag1) is an ion channel that plays important roles in tumour proliferation, malignant transformation, invasion, metastasis, recurrence, and prognosis. therefore, identifying potent and specific eag1 channel inhibitors is crucial. in this study, we identified the first natural inhibitor of eag1, the traditional chinese medicine agent tetrandrine, and e ...201930362536
at-line raman spectroscopy and design of experiments for robust monitoring and control of miniature bioreactor cultures.the biopharmaceutical industry is moving toward a more quality by design (qbd) approach that seeks to increase product and process understanding and process control. miniature bioreactor systems offer a high-throughput method enabling the assessment of numerous process variables in a controlled environment. however, the number of off/at-line samples that can be taken is restricted due to the small working volume of each vessel. this limitation may be resolved through the use of raman spectroscop ...201930378770
an alternative exon of caps2 influences catecholamine loading into ldcvs of chromaffin cells.the calcium-dependent activator proteins for secretion (caps) are priming factors for synaptic and large dense-core vesicles (ldcvs), promoting their entry into and stabilizing the release-ready state. a modulatory role of caps in catecholamine loading of vesicles has been suggested. although an influence of caps on monoamine transporter function and on vesicle acidification has been reported, a role of caps in vesicle loading is disputed. using expression of naturally occurring splice variants ...201930389842
mutagenic activity of n-nitrosodiethylamine in cell lines expressing human cyp2e1-adequacy of dimethylsulfoxide as solvent.human cyp2e1 metabolizes many xenobiotics of low-molecular weight, thereby activating various promutagens/procarcinogens. in toxicological studies in vitro, dimethylsulfoxide (dmso) is a common vehicle for organic compounds. however, it was observed to potently inhibit cyp2e1 activity. we were interested in whether it affects cyp2e1-dependent mutagenic responses. in this study, n-nitrosodiethylamine (ndea), which is soluble in both water and dmso, was used as a model promutagen. it induced hprt ...201930387191
efficient development of stable recombinant chinese hamster ovary (rcho) cell lines to produce antibodies by using dimethyl sulfoxide (dmso) in electroporation.development of stable rcho cell lines is still time consuming and labor intensive, although it is a critical step in the commercial development of recombinant antibodies. the current work demonstrates, for the first time, that electroporation of cho cells with dmso can enhance stable expression of recombinant antibodies in rcho cells. electroporation with dmso resulted in an average 3.7-fold and 2.8-fold increases in expression levels of aflibercept and pembrolizumab, respectively, in pools of s ...201930394043
process development for an inducible rituximab-expressing chinese hamster ovary cell line.inducible mammalian expression systems are becoming increasingly available and are not only useful for the production of cytotoxic/cytostatic products, but also confer the unique ability to uncouple the growth and production phases. in this work, we have specifically investigated how the cell culture state at the time of induction influences the cumate-inducible expression of recombinant rituximab by a gs-cho cell line. to this end, cells grown in batch and fed-batch cultures were induced at inc ...201930414355
efficient interleukin-21 production by optimization of codon and signal peptide in chinese hamster ovarian cells.interleukin-21 is a common γ-chain cytokine that controls the immune responses of b cells, t cells, and natural killer cells. targeting il-21 to strengthen the immune system is promising for the development of vaccines as well as anti-infection and anti-tumor therapies. however, the practical application of il-21 is limited by the high production cost. in this study, we improved il-21 production by codon optimization and selection of appropriate signal peptide in cho-k1 cells. codon-optimized or ...201930544288
combined effects of glycosylation precursors and lactate on the glycoprofile of igg produced by cho cells.the glycosylation profile of therapeutic monoclonal antibodies (mabs) is a crucial quality parameter for industrial immunoglobulin g (igg) production. several alternative carbon sources, which function as glycosylation precursors, have been reported to impact the glycosylation pattern. since the cells give priority to glucose uptake, the presence of this substrate can lower the effects of alternative sugars on the glycosylation. in order to get a better understanding of the influence of alternat ...201930423367
effective bioreactor ph control using only sparging gases.ph control is critical in bioreactor operations, typically realized through a two-sided control loop, where co2 sparging and base addition are used in bicarbonate-buffered media. though a common approach, base addition could compromise culture performance due to the potential impact from ph excursions and osmolality increase in large-scale bioreactors. in this study, the feasibility of utilizing control of sparge gas composition as part of the ph control loop was assessed in chinese hamster ovar ...201930421525
two human mars effectively increase transgene expression in transfected cho cells.matrix attachment regions (mars) can enhance the expression level of transgene in chinese hamster ovaries (cho) cell expression system. however, improvements in function and analyses of the mechanism remains unclear. in this study, we screened two new and more functional mar elements from the human genome dna. the human mar-3 and mar-7 element were cloned and inserted downstream of the polya site in a eukaryotic vector. the constructs were transfected into cho cells, and screened under g418 to p ...201930450759
characterizing and enhancing virus removal by protein a chromatography.protein a chromatography is an effective capture step to separate fc-containing biopharmaceuticals from cell culture impurities but is generally not effective for virus removal, which tends to vary among different products. previous findings have pointed to the differences in feedstocks to protein a, composed of the products and other cell culture-related impurities. to separate the effect of the feedstock components on virus removal, and understand why certain monoclonal antibody (mab) products ...201930450554
improving recombinant bone morphogenetic protein-4 (bmp-4) production by autoregulatory feedback loop removal using bmp receptor-knockout cho cell lines.a chinese hamster ovary (cho) cell line producing recombinant human bone morphogenetic protein-4 (rhbmp-4) (cho-bmp-4), which expresses essential components of bmp signal transduction, underwent autocrine bmp-4 signaling. rna seq analysis on cho host cells (dg44) treated with rhbmp-4 (20 µg/ml) suggested that rhbmp-4 induced signaling in cho cells could be a critical factor in limiting rhbmp-4 production and should be removed to improve rhbmp-4 production in recombinant cho (rcho) cells. the inh ...201930447330
study of the cytotoxic and genotoxic potential of the carbonyl ruthenium(ii) compound, ct-[rucl(co)(dppb)(bipy)]pf6 [dppb = 1,4-bis(diphenylphosphino)butane and bipy = 2,2'-bipyridine], by in vitro and in vivo assays.considering the promising previous results of ct-[rucl(co)(dppb)(bipy)]pf6 (where dppb = 1,4-bis(diphenylphosphino)butane and bipy = 2,2'-bipyridine) as an antitumor agent, novel biological assays evaluating its toxicogenic potential were performed. the genotoxicity of the compound was evaluated by the in vitro micronucleus test (v79, chinese hamster lung fibroblasts; hepg2, hepatocellular carcinoma cells), in vivo bone marrow micronucleus test and comet assay in hepatocytes (swiss mice). the an ...201930460706
epigenetic regulation of gene expression in chinese hamster ovary cells in response to the changing environment of a batch culture.the existence of dynamic cellular phenotypes in changing environmental conditions is of major interest for cell biologists who aim to understand the mechanism and sequence of regulation of gene expression. in the context of therapeutic protein production by chinese hamster ovary (cho) cells, a detailed temporal understanding of cell-line behavior and control is necessary to achieve a more predictable and reliable process performance. of particular interest are data on dynamic, temporally resolve ...201930512195
production of a recombinant monoclonal antibody to herpes simplex virus glycoprotein d for immunoaffinity purification of tagged proteins.we have developed a stable chinese hamster ovary (cho) cell line for the production of a recombinant monoclonal antibody (mab) to a short protein sequence derived from the n-terminus of human herpes simplex virus type 1 glycoprotein d (hsv-1 gd). the antibody (designated r34.1) provides a useful tool for the immunoaffinity purification of hsv-1 gd tagged proteins, and provides a generic purification system by which various proteins and peptides can be purified. recombinant 34.1 was assembled usi ...201930502324
azure b affects amyloid precursor protein metabolism in ps70 cells.alzheimer's disease (ad), the most common form of dementia, is characterized by abundant deposition of amyloid-β (aβ) peptide that is the result of sequential cleavage of amyloid precursor protein (app) by β-secretase and γ-secretase. several studies have documented that inhibition of aβ peptide synthesis or facilitating its degradation is one of the attractive therapeutic strategies in ad. methylene blue (methb), which has recently been investigated in phase ii clinical trials, is a prominent i ...201930500345
a novel muc1 aptamer-modified plga-epirubicin-pβae-antimir-21 nanocomplex platform for targeted co-delivery of anticancer agents in vitro and in vivo.conventional chemotherapy suffers from several drawbacks, including toxic side effects together with the development of resistance to the chemical agents. therefore, exploring alternative therapeutic approaches as well as developing targeted delivery systems are in demand. oligonucleotide-based therapy has emerged as a promising and alternative procedure for treating malignancies involving gene-related diseases. in the current study, a targeted delivery system was designed to target cancer cells ...201930537619
amber suppression coupled with inducible surface display identifies cells with high recombinant protein productivity.cell line development (cld) for biotherapeutics is a time- and resource-intensive process requiring the isolation and screening of large numbers of clones to identify high producers. novel methods aimed at enhancing cell line screening efficiency using markers predictive of productivity early in the cld process are needed to reliably generate high-yielding cell lines. to enable efficient and selective isolation of antibody expressing chinese hamster ovary cells by fluorescence-activated cell sor ...201930536645
evaluation of pta-derived ruthenium(ii) and iridium(iii) quinoline complexes against chloroquine-sensitive and resistant strains of the plasmodium falciparum malaria parasite.cationic 1,3,5‑triaza‑phosphaadamantane (pta) quinoline ruthenium(ii) and iridium(iii) complexes were successfully synthesized and characterized using standard spectroscopic and analytical techniques. the complexes were evaluated for their in vitro antiplasmodial activities against the chloroquine-sensitive (cqs) nf54 and chloroquine-resistant (cqr) k1 strains of the plasmodium falciparum species of the malaria parasite and were found to exhibit good activities in the sensitive strain but modera ...201930529881
multistage antiplasmodium activity of astemizole analogues and inhibition of hemozoin formation as a contributor to their mode of action.a drug repositioning approach was leveraged to derivatize astemizole (ast), an antihistamine drug whose antimalarial activity was previously identified in a high-throughput screen. the multistage activity potential against the plasmodium parasite's life cycle of the subsequent analogues was examined by evaluating against the parasite asexual blood, liver, and sexual gametocytic stages. in addition, the previously reported contribution of heme detoxification to the compound's mode of action was i ...201930525439
the mouse-canine chimeric anti-dog podoplanin antibody p38b exerts antitumor activity in mouse xenograft models.podoplanin (pdpn) is a type i transmembrane heavily glycosylated sialoglycoprotein that is expressed in normal tissues such as pulmonary type i alveolar cells, renal podocytes, and lymphatic endothelial cells. pdpn overexpression in cancerous tissue is associated with hematogenous metastasis through interactions with the c-type lectin-like receptor 2 (clec-2). previously, we have reported the development of a mouse monoclonal antibody (mab), pmab-38 (igg1, kappa) against dog pdpn (dpdpn). pmab-3 ...201930519645
multiple modes of action of hydrophobic amines and their guanidine analogues on asic1a.hydrophobic monoamines containing only a hydrophobic/aromatic moiety and protonated amino group are a recently described class of acid-sensing ion channel (asic) modulators. intensive studies have revealed a number of active compounds including endogenous amines and pharmacological agents and shown that these compounds potentiate and inhibit asics depending on their specific structure and on subunit composition of the target channel. the action of monoamines also depends on the application proto ...201930557561
muscarinic agonist, (±)-quinuclidin-3-yl-(4-fluorophenethyl)(phenyl)carbamate: high affinity, but low subtype selectivity for human m1 - m5 muscarinic acetylcholine receptors.novel quinuclidinyl n-phenylcarbamate analogs were synthesized, and binding affinities at m1-m5 muscarinic acetylcholine receptor (machr) subtypes were determined using chinese hamster ovary (cho) cell membranes stably expressing one specific subtype of human machr. although not subtype selective, the lead analog (±)-quinuclidin-3-yl-(4-fluorophenethyl)(phenyl)carbamate (3c) exhibited the highest affinity (ki = 2.0, 13, 2.6, 2.2, 1.8 nm) at each of the m1-m5 machrs, respectively. based on result ...201930554957
the effect of hyperosmolality application time on production, quality, and biopotency of monoclonal antibodies produced in cho cell fed-batch and perfusion cultures.hyperosmolality has been commonly investigated due to its effects on the production and quality characteristics of monoclonal antibodies (mabs) produced in cho cell fed-batch cultures. however, the application of hyperosmolality at different times and its effect on biopotency have seldom been researched, especially in perfusion culture. in our study, different degrees of hyperosmolality induced by sodium chloride were investigated in anti-ige rcho cell fed-batch cultures and anti-cd52 rcho cell ...201930554388
impact of cell culture media additives on igg glycosylation produced in chinese hamster ovary cells.glycosylation is a key critical quality attribute for monoclonal antibodies and other recombinant proteins because of its impact on effector mechanisms and half-life. in this study, a variety of compounds were evaluated for their ability to modulate glycosylation profiles of recombinant monoclonal antibodies produced in chinese hamster ovary cells. compounds were supplemented into the cell culture feed of fed-batch experiments performed with a cho k1 and a cho dg44 cell line expressing a recombi ...201930552760
cxcl10 production in equine monocytes is stimulated by interferon-gamma.c-x-c motif ligand 10 (cxcl10) is a pro-inflammatory chemokine and has been extensively evaluated in people and mice. in horses, cxcl10 and its involvement in host immunity has rarely been analyzed due to the lack of specific antibodies. we generated a mab specific for the equine chemokine cxcl10 using hybridoma technology. antibody specificity was confirmed by intracellular staining and flow cytometric analysis of chinese hamster ovary (cho) cells expressing equine rcxcl10, while cho cells expr ...201930593347
utilizing a regulated targeted integration cell line development approach to systematically investigate what makes an antibody difficult to express.chinese hamster ovary (cho) cells are conventionally used to generate therapeutic cell lines via random integration (ri), where desired transgenes are stably integrated into the genome. targeted integration (ti) approaches, which involve integration of a transgene into a specific locus in the genome, are increasingly utilized for cho cell line development (cld) in recent years. none of these cld approaches, however, are suitable for expression of toxic or difficult-to-express molecules, or for d ...201930592567
secretory leakage of igg1 aggregates from recombinant chinese hamster ovary cells.aggregation of therapeutic antibodies is one of the most important issues to be resolved in manufacturing processes because of reduced efficacy and immunogenicity. despite aggregation studies in vitro, little is known about the aggregation mechanism in cell culture processes. in this study, we investigated the process of aggregate formation of igg1 antibodies during the culture of chinese hamster ovary (cho) cells to determine how aggregation occurs. a recombinant cho cell line was cultivated in ...201930580968
investigating the effects of stove emissions on ocular and cancer cells.more than a third of the world's population relies on solid fuels for cooking and heating, with major health consequences. although solid fuel combustion emissions are known to increase the prevalence of illnesses such as chronic obstructive pulmonary disease and lung cancer, however, their effect on the eyes is underexplored. this study assesses the acute toxicity of solid fuel combustion emissions on healthy ocular cells and a cancer cell line. three healthy ocular cell lines (corneal, lens, a ...201930755694
chloramination of iodide-containing waters: formation of iodinated disinfection byproducts and toxicity correlation with total organic halides of treated waters.the formation of iodinated disinfection byproducts (i-dbps) in drinking waters is of a concern due to their higher cyto- and genotoxicity than their chlorinated and brominated analogues. this study investigated the formation of i-dbps under chloramination conditions using preformed chloramine and associated cyto- and geno-toxicities obtained with chinese hamster ovary (cho) cell assay. cyto- and geno-toxicity of the samples were also calculated using dbp toxicity index values and correlated with ...201931484087
peptide-driven targeted drug-delivery system comprising turn-on near-infrared fluorescent xanthene-cyanine reporter for real-time monitoring of drug release.targeted drug delivery (tdd) is an efficient strategy for cancer treatment. however, the real-time monitoring of drug delivery is still challenging because of a pronounced lack of tdd systems capable of providing a near-infrared (nir) fluorescence signal for the detection of drug-release events. herein, a new tdd system, comprising a turn-on nir fluorescent reporter attached to an anticancer drug and targeting peptide, is reported. this system provides both tdd and nir fluorescence monitoring of ...201931403246
consequences of trace metal variability and supplementation on chinese hamster ovary (cho) cell culture performance: a review of key mechanisms and considerations.trace metals are supplied to chemically-defined media (cdm) for optimal chinese hamster ovary (cho) cell culture performance during the production of monoclonal antibodies and other therapeutic proteins. however, lot-to-lot and vendor-to-vendor variability in raw materials consequently leads to an imbalance of trace metals that are supplied to cdm. this imbalance can yield detrimental effects rooted in several primary mechanisms and pathways including oxidative stress, apoptosis, lactate accumul ...201931403183
zinc supplementation increases protein titer of recombinant cho cells.in order to study the impact of zinc and copper on the titer levels of mab and recombinant protein in cho cells, the igg-expressing (dp12) and epo-expressing (sk15) cell lines were cultured in chemically defined media with increasing concentrations of either metal. supplementation with 25 mg/l in cdm media resulted in a significant increase in epo (1.7-fold) and igg (2.6-fold) titers compared to control (no added zinc). titers at this zn concentration in cdm containing the insulin replacing agen ...201931396753
interphase cytogenetic analysis of micronucleated and multinucleated cells supports the premature chromosome condensation hypothesis as the mechanistic origin of chromothripsis.the discovery of chromothripsis in cancer genomes challenges the long-standing concept of carcinogenesis as the result of progressive genetic events. despite recent advances in describing chromothripsis, its mechanistic origin remains elusive. the prevailing conception is that it arises from a massive accumulation of fragmented dna inside micronuclei (mn), whose defective nuclear envelope ruptures or leads to aberrant dna replication, before main nuclei enter mitosis. an alternative hypothesis i ...201931390832
recombinant hemagglutinin produced from chinese hamster ovary (cho) stable cell clones and a pelc/cpg combination adjuvant for h7n9 subunit vaccine development.the novel h7n9 avian influenza a virus has caused human infections in china since 2013; some isolates from the fifth wave of infections have emerged as highly pathogenic avian influenza viruses. recombinant hemagglutinin proteins of h7n9 viruses can be rapidly and efficiently produced with low-level biocontainment facilities. in this study, recombinant h7 antigen was obtained from engineered stable clones of chinese hamster ovary (cho) cells for subsequent large-scale production. the stable cho ...201931383491
treatment with 3-aminobenzamide negates the radiofrequency-induced adaptive response in two cell models.in previous investigations, we demonstrated that pre-exposure of different cell cultures to radiofrequency fields can reduce the damage induced by genotoxic agents, an effect resembling the so-called adaptive response. in this study, we pre-exposed human peripheral blood lymphocytes and chinese hamster lung fibroblast cell line to 1950 mhz, umts (universal mobile telecommunication system) signal, for 20 h, and then treated cultures with mitomycin-c. after confirming the induction of an adaptive ...201931382475
four ways to fit an ion channel model.mathematical models of ionic currents are used to study the electrophysiology of the heart, brain, gut, and several other organs. increasingly, these models are being used predictively in the clinic, for example, to predict the risks and results of genetic mutations, pharmacological treatments, or surgical procedures. these safety-critical applications depend on accurate characterization of the underlying ionic currents. four different methods can be found in the literature to fit voltage-sensit ...201931493859
dynamics of mechanosensitive nascent adhesion formation.cellular migration is a tightly regulated process that involves actin cytoskeleton, adaptor proteins, and integrin receptors. forces are transmitted extracellularly through protein complexes of these molecules, called adhesions. adhesions anchor the cell to its substrate, allowing it to migrate. in chinese hamster ovary cells, three classes of adhesion can be identified: nascent adhesions (nas), focal complexes, and focal adhesions, ranked here ascendingly based on size and stability. to underst ...201931493858
impacts of intentional mycoplasma contamination on cho cell bioreactor cultures.mycoplasma contamination events in biomanufacturing facilities can result in loss of production and costly cleanups. mycoplasma may survive in mammalian cell cultures with only subtle changes to the culture and may penetrate the 0.2 µm filters often used in the primary clarification of harvested cell culture fluid. culture cell-based and indicator cell-based assays that are used to detect mycoplasma are highly sensitive but can take up to 28 days to complete and cannot be used for real-time deci ...201931478189
at1aa (angiotensin ii type-1 receptor autoantibodies): cause or consequence of human primary aldosteronism?at1aa (angiotensin ii type-1 receptor autoantibodies) were first detected in patients with primary aldosteronism (pa) because of aldosterone-producing adenoma (apa) with an in-house developed assay, but it remained unclear if they can be ascertained also with commercially available assays and if they have a functional role. aims of our study were to investigate if (1) commercially available kits allow detection of raised at1aa titer in apa; (2) this titer is normalized by adrenalectomy; and (3) ...201931476908
a role for alternative end-joining factors in homologous recombination and genome editing in chinese hamster ovary cells.crispr technologies greatly foster genome editing in mammalian cells through site-directed dna double strand breaks (dsbs). however, precise editing outcomes, as mediated by homologous recombination (hr) repair, are typically infrequent and outnumbered by undesired genome alterations. by using knockdown and overexpression studies in chinese hamster ovary (cho) cells as well as characterizing repaired dna junctions, we found that efficient hr-mediated genome editing depends on alternative end-joi ...201931476574
a different perspective: how much innovation is really needed for monoclonal antibody production using mammalian cell technology?as biopharmaceutical companies have optimized cell line and production culture process development, titers of recombinant antibodies have risen steadily to 3-8 g/l for fed-batch mammalian cultures at production scales of 10 kl or larger. most new antibody products are produced from chinese hamster ovary (cho) cell lines, and there are relatively few alternative production hosts under active evaluation. many companies have adopted a strategy of using the same production cell line for early clinic ...201929721583
transcriptome analysis of cho cell size increase during a fed-batch process.in a chinese hamster ovary (cho) cell fed-batch process, arrest of cell proliferation and an almost threefold increase in cell size occurred, which is associated with an increase in cell-specific productivity. in this study, transcriptome analysis is performed to identify the molecular mechanisms associated with this. cell cycle analysis reveals that the cells are arrested mainly in the g0 /g1 phase. the cell cycle arrest is associated with significant up-regulation of cyclin-dependent kinases i ...201930024106
brassica rapa hairy root based expression system leads to the production of highly homogenous and reproducible profiles of recombinant human alpha-l-iduronidase.the brassica rapa hairy root based expression platform, a turnip hairy root based expression system, is able to produce human complex glycoproteins such as the alpha-l-iduronidase (idua) with an activity similar to the one produced by chinese hamster ovary (cho) cells. in this article, a particular attention has been paid to the n- and o-glycosylation that characterize the alpha-l-iduronidase produced using this hairy root based system. this analysis showed that the recombinant protein is charac ...201930058762
leaky expression of the tet-on system hinders control of endogenous mirna abundance.with the ability to affect multiple genes and fundamental pathways simultaneously, mirna engineering of chinese hamster ovary (cho) cells has significant advantages over single gene expression or repression. tight control of these molecular triggers is desirable as it could in theory allow on/off or even tunable regulation of desirable cellular phenotypes. the present study investigated the potential of employing a tetracycline inducible (tet-on) system for conditional knockdown of specific mirn ...201929989353
crispr/cas9-mediated knockout of six glycosyltransferase genes in nicotiana benthamiana for the production of recombinant proteins lacking β-1,2-xylose and core α-1,3-fucose.plants offer fast, flexible and easily scalable alternative platforms for the production of pharmaceutical proteins, but differences between plant and mammalian n-linked glycans, including the presence of β-1,2-xylose and core α-1,3-fucose residues in plants, can affect the activity, potency and immunogenicity of plant-derived proteins. nicotiana benthamiana is widely used for the transient expression of recombinant proteins so it is desirable to modify the endogenous n-glycosylation machinery t ...201929969180
analysis of intracellular igg secretion in chinese hamster ovary cells to improve igg production.the production of biopharmaceutical immunoglobulin g (igg) using cultured mammalian cells, especially chinese hamster ovary (cho) cells is well established and has been markedly improved through the modification of cells and cell culture engineering technologies. the establishment of high-production cell lines remains a challenge. the intracellular secretion of igg has been investigated to identify and solve the rate-limiting steps in antibody production. however, strategies that regulate the ex ...201930017708
lion/web: a web-based ontology enrichment tool for lipidomic data analysis.a major challenge for lipidomic analyses is the handling of the large amounts of data and the translation of results to interpret the involvement of lipids in biological systems.201931141612
establishment of a monoclonal antibody pmab-231 for tiger podoplanin.podoplanin (pdpn), also known as t1alpha, has been used as a lung type i alveolar cell marker in the pathophysiological condition. although we have established several monoclonal antibodies (mabs) against mammalian pdpns, mabs against tiger pdpn (tigpdpn), which are useful for immunohistochemical analysis, remain to be developed. in this study, we immunized mice with tigpdpn-overexpressing chinese hamster ovary (cho)-k1 cells (cho/tigpdpn) and screened hybridomas producing mabs against tigpdpn u ...201931009336
development of an anti-bear podoplanin monoclonal antibody pmab-247 for immunohistochemical analysis.sensitive and specific monoclonal antibodies (mabs) targeting podoplanin (pdpn) are needed for immunohistochemical analyses using formalin-fixed paraffin-embedded tissues because pdpn is known as a lymphatic endothelial cell maker in pathology. recently, we established anti-pdpn mabs against many species, such as human, mouse, rat, rabbit, dog, cat, bovine, pig, horse, goat, tiger, alpaca, and tasmanian devil. however, anti-bear pdpn (bpdpn) has not been established yet. in this study, we immuni ...201931061899
establishment of a monoclonal antibody pmab-233 for immunohistochemical analysis against tasmanian devil podoplanin.monoclonal antibodies (mabs) against not only human, mouse, and rat but also rabbit, dog, cat, bovine, pig, and horse podoplanins (pdpns) have been established in our previous studies. pdpn is used as a lymphatic endothelial cell marker in pathological diagnoses. however, mabs against tasmanian devil pdpn (taspdpn), which are useful for immunohistochemical analysis, remain to be developed. herein, mice were immunized with taspdpn-overexpressing chinese hamster ovary (cho)-k1 (cho/taspdpn) cells, ...201930984883
intact bioactivities and improved pharmacokinetic of the sl335-ifn-β-1a fusion protein that created by genetic fusion of sl335, a human anti-serum albumin fab, and human interferon-β.recombinant human interferon beta (rifn-β) has long been used as a first-line treatment for multiple sclerosis (ms), and any attempt to develop a long-acting rifn-β is desirable since only one pegylated version of long-acting rifn-β-1a (plegridy) is currently available in clinics. previously, we reported that sl335, a human fab molecule specific to serum albumin, exhibits an extended serum half-life via utilizing the fcrn recycling mechanism. with the ultimate goal of developing a long-acting ri ...201930684504
construction of a mammalian ires-based expression vector to amplify a bispecific antibody; blinatumomab.blinatumomab, the bispecific t cell engager antibody (bsab), has been demonstrated as the most successful bsab to date. throughout the past decade, vector design has great importance for the expression of monoclonal antibody in chinese hamster ovary (cho) cells. it has been indicated that expression vectors based on the elongation factor-1 alpha (ef-1 alpha) gene and dhfr selection marker can be highly effective to produce populations of stably transfected cells in the selection medium. moreover ...201932184875
eco-friendly synthesis of glutathione-capped cdte/cdse/znse core/double shell quantum dots: their cytotoxicity and genotoxicity effects on chinese hamster ovary cells.in this work, we report green one-pot synthesis, cytotoxicity and genotoxicity of glutathione-capped cdte/cdse/znse heterostructured quantum dots (qds) using a label-free xcelligence rtca system as well as the cytokinesis blocked micronucleus assay. the as-synthesised nanocrystals displayed good optical properties and were spherical in shape with an average particle diameter of 5.9 ± 1.13 nm. the intracellular uptake study showed that most of the as-synthesised glutathione stabilized qds penetra ...201932055393
discovery of the arp2 protein as a determining molecule in tumor cell death.cancer is a multifactorial disease that constitutes a serious public health problem worldwide. prostate cancer advanced stages are associated with the development of androgen-independent tumors and an apoptosis-resistant phenotype that progresses to metastasis. by studying androgen-independent lymphoid nodule carcinoma of the prostate (lncap) cells induced to apoptosis by serum elimination, we identified the activation of a non-selective cationic channel of 23ps conductance that promotes incomin ...201932091029
the gβ1 and gβ3 subunits differentially regulate rat vascular kv7 channels.within the vasculature kv7 channels are key regulators of basal tone and contribute to a variety of receptor mediated vasorelaxants. the kv7.4 isoform, abundant within the vasculature, is key to these processes and was recently shown to have an obligatory requirement of g-protein βγ subunits for its voltage dependent activity. there is an increasing appreciation that with 5 gβ subunits and 12 gγ subunits described in mammalian cells that different gβ x γ x combinations can confer selectivity in ...201931992990
gene disruption of ribosomal protein l5 (rpl5) decreased the sensitivity of cho-k1 cells to uncoupler carbonylcyanide-3-chlorophenylhydrazone.protonophoric uncoupler carbonylcyanide-3-chlorophenylhydrazone (cccp) decreases the proton motive force (δp) of the mitochondrial inner membrane and results in inhibition of oxidative phosphorylation. in this study, a cccp-resistant clone was isolated from a random gene trap insertional mutant library of chinese hamster ovary (cho)-k1 cells which was constructed by infecting a retrovirus vector, rosaβgeo. although we expected the isolation of the mutants defective in nuclear genes responsible f ...201931956228
an in vitro study on the cytotoxicity and genotoxicity of silver sulfide quantum dots coated with meso-2,3-dimercaptosuccinic acid.silver sulfide (ag2s) quantum dots (qds) are highly promising nanomaterials in bioimaging systems due to their high activities for both imaging and drug/gene delivery. there is insufficient research on the toxicity of ag2s qds coated with meso-2,3-dimercaptosuccinic acid (dmsa). in this study, we aimed to determine the cytotoxicity of ag2s qds coated with dmsa in chinese hamster lung fibroblast (v79) cells over a wide range of concentrations (5-2000 μg/ml).201932454726
increased mab production in amplified cho cell lines is associated with increased interaction of creb1 with transgene promoter.most therapeutic monoclonal antibodies in biopharmaceutical processes are produced in chinese hamster ovary (cho) cells. technological advances have rendered the selection procedure for higher producers a robust protocol. however, information on molecular mechanisms that impart the property of hyper-productivity in the final selected clones is currently lacking. in this study, an igg-producing industrial cell line and its methotrexate (mtx)-amplified progeny cell line were analyzed using transcr ...201932577618
retrovectors packaged in cho cells to generate glp-1-fc stable expression cho cell lines.chinese hamster ovary (cho) cells are the most dependable mammalian cells for the production of recombinant proteins. replication-incompetent retroviral vector (retrovector) is an efficient tool to generate stable cell lines. multiple copies of integrated genes by retrovector transduction results in improved recombinant protein yield. hek-293 and their genetic derivatives are principal cells for retrovector production. retrovectors packaged in hek-293 cells pose a risk of infectious agent transm ...201932288149
toxicity studies of lactobacillus plantarum ps128tm isolated from spontaneously fermented mustard greens.: probiotics are extensively available to consumers; however, the use of probiotics may not always be safe, and there are few reports on their side effects, including those of lactobacillus. lactobacillus plantarum strain ps128tm isolated from spontaneously fermented mustard greens in taiwan was recently reported to exhibit probiotic properties. in this study, we aimed to assess the safety of strain ps128tm for use in humans via examining genotoxic and oral toxic effects using in vitro and in vi ...201931835837
assessment of dna vaccine encoding toxoplasma gondii microneme complete gene and il-12 as adjuvant in balb/c mice.toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis in humans and animals. micronemes (mics) are effective candidates for dna vaccine.201931579446
functional analysis of ldlr (low-density lipoprotein receptor) variants in patient lymphocytes to assess the effect of evinacumab in homozygous familial hypercholesterolemia patients with a spectrum of ldlr activity.homozygous familial hypercholesterolemia is a rare disease usually caused by ldlr (low-density lipoprotein receptor) mutations. homozygous familial hypercholesterolemia is characterized by markedly elevated ldl-c (low-density lipoprotein cholesterol) levels and an extremely high risk of premature atherosclerotic cardiovascular disease. a phase 2, proof-of-concept study (nct02265952) demonstrated that evinacumab, a fully human monoclonal antibody to angptl3 (angiopoietin-like 3 protein), reduced ...201931578082
the comet assay: assessment of in vitro and in vivo dna damage.anthropogenic activities, indiscriminate and rapid industrialization as well as pursuance of a better life has led to an increase in the concentration of chemicals, like pesticides, automobile exhausts, and new chemical entities, in the environment, which have an adverse effect on all living organisms including humans. sensitive and robust test systems are thus required for accurate hazard identification and risk assessment. the comet assay has been used widely as a simple, rapid, and sensitive ...201931473963
identification of lark as a novel and conserved g-quadruplex binding protein in invertebrates and vertebrates.double-stranded dnas are usually present in the form of linear b-form double-helix with the base pairs of adenine (a) and thymine (t) or cytosine (c) and guanine (g), but g-rich dna can form four-stranded g-quadruplex (g4) structures, which plays important roles in transcription, replication, translation and protection of telomeres. in this study, a rna recognition motif (rrm)-containing protein, bmlark, was identified and demonstrated to bind g4 structures in the promoters of a transcription fa ...201931165881
identification of cp77 as the third orthopoxvirus samd9 and samd9l inhibitor with unique specificity for a rodent samd9l.orthopoxviruses (opxvs) have a broad host range in mammalian cells, but chinese hamster ovary (cho) cells are nonpermissive for vaccinia virus (vacv). here, we revealed a species-specific difference in host restriction factor samd9l as the cause for the restriction and identified orthopoxvirus cp77 as a unique inhibitor capable of antagonizing chinese hamster samd9l (chsamd9l). two known vacv inhibitors of samd9 and samd9l (samd9&l), k1 and c7, can bind human and mouse samd9&l, but neither can b ...201930918078
new insight into the classification and evolution of glucose transporters in the metazoa.because glucose is an essential energy source for living organisms, glucose transporters (gluts) are present in all species worldwide. encoded by the solute carrier family 2 gene family, the glut proteins generally have 12 transmembrane helices (tmhs). in total, 14 glut proteins have been identified in humans (hgluts), and they are divided into 3 classes on the basis of their transport characteristics and sequence similarities. herein, we report the use of protein sequence similarity networks (s ...201930888203
simultaneous expression of clophensor and slc26a3 reveals the nature of endogenous oxalate transport in cho cells.clophensor, a fluorescent fusion protein, is a dual function biosensor that has been utilized as a tool for the simultaneous measurement of intracellular chloride and ph in cells. clophensor has traditionally been used in conjunction with fluorescence microscopy for single cell measurements. here, we present a promising multi-well format advancement for the use of clophensor as a potential high-throughput method capable of measuring fluorescence signal intensity across a well of confluent cells ...201930837228
identification of verapamil binding sites within human kv1.5 channel using mutagenesis and docking simulation.the phenylalkylamine class of l-type ca2+ channel antagonist verapamil prolongs the effective refractory period (erp) of human atrium, which appears to contribute to the efficacy of verapamil in preventing reentrant-based atrial arrhythmias including atrial fibrillation. this study was designed to investigate the molecular and electrophysiological mechanism underlying the action of verapamil on human kv1.5 (hkv1.5) channel that determines action potential duration and erp in human atrium.201930816676
incorporation of cd40 ligand or granulocyte-macrophage colony stimulating factor into hantaan virus (htnv) virus-like particles significantly enhances the long-term immunity potency against htnv infection.hantavirus infections cause severe haemorrhagic fever with renal syndrome (hfrs) in humans and are associated with high fatality rates. in 2017, numerous outbreaks were reported in china and germany. this represents a significant public-healthcare issue with no effective hfrs vaccines that offer a long-term immune response. in this study, we investigated the long-term humoral and cellular immune responses and protective immunity of hantaan virus (htnv) granulocyte-macrophage colony stimulating f ...201930657443
kcnq1 rescues tmc1 plasma membrane expression but not mechanosensitive channel activity.transmembrane channel-like protein isoform 1 (tmc1) is essential for the generation of mechano-electrical transducer currents in hair cells of the inner ear. tmc1 disruption causes hair cell degeneration and deafness in mice and humans. although thought to be expressed at the cell surface in vivo, tmc1 remains in the endoplasmic reticulum when heterologously expressed in standard cell lines, precluding determination of its roles in mechanosensing and pore formation. here, we report that the kcnq ...201930613966
glyco-engineered cho cell lines producing alpha-1-antitrypsin and c1 esterase inhibitor with fully humanized n-glycosylation profiles.recombinant chinese hamster ovary (cho) cells are able to provide biopharmaceuticals that are essentially free of human viruses and have n-glycosylation profiles similar, but not identical, to humans. due to differences in n-glycan moieties, two members of the serpin superfamily, alpha-1-antitrypsin (a1at) and plasma protease c1 inhibitor (c1inh), are currently derived from human plasma for treating a1at and c1inh deficiency. deriving therapeutic proteins from human plasma is generally a cost-in ...201930513349
exposure to the fungicide captan induces dna base alterations and replicative stress in mammalian cells.the classification of the fungicide captan (cas number: 133-06-2) as a carcinogen agent is presently under discussion. despite the mutagenic effect detected by the ames test and carcinogenic properties observed in mice, the genotoxicity of this pesticide in humans is still unclear. new information is needed about its mechanism of action in mammalian cells. here, we show that chinese hamster ovary (cho) cells exposed to captan accumulate fpg-sensitive dna base alterations. in cho and hela cells, ...201930471166
na+ /ca2+ exchanger 1 is a key mechanosensitive molecule of the esophageal myenteric neurons.our earlier studies showed that mechanical stretch activates inhibitory motor neurons of the oesophagus; however, the underlying molecular mechanisms are unclear. here, we sought to examine if na+ /ca2+ exchanger 1 (ncx1) is responsible for the mechanosensitivity in the esophageal myenteric neurons (emn) of rats and humans.201930466198
early porcine sapovirus infection disrupts tight junctions and uses occludin as a coreceptor.the genus sapovirus belongs to the family caliciviridae, and its members are common causative agents of severe acute gastroenteritis in both humans and animals. some caliciviruses are known to use either terminal sialic acids or histo-blood group antigens as attachment factors and/or cell surface proteins, such as cd300lf, cd300ld, and junctional adhesion molecule 1 of tight junctions (tjs), as receptors. however, the roles of tjs and their proteins in sapovirus entry have not been examined. in ...201930463963
production of soluble pmhc-i molecules in mammalian cells using the molecular chaperone tapbpr.current approaches for generating major histocompatibility complex (mhc) class-i proteins with desired bound peptides (pmhc-i) for research, diagnostic and therapeutic applications are limited by the inherent instability of empty mhc-i molecules. using the properties of the chaperone tap-binding protein related (tapbpr), we have developed a robust method to produce soluble, peptide-receptive mhc-i molecules in chinese hamster ovary cells at high yield, completely bypassing the requirement for la ...201932725167
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
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
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
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
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