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1.
Int J Mol Sci ; 25(17)2024 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-39273253

RESUMO

Remote ischemic preconditioning (RIPC) reduces ischemia-reperfusion injury in aortocoronary bypass surgery, potentially via extracellular vesicles (EVs) and their micro-RNA content. Clinical data implicate that propofol might inhibit the cardioprotective RIPC effect. This prospective, randomized study investigated the influence of different anesthetic regimes on RIPC efficacy and EV micro-RNA signatures. We also assessed the impact of propofol on cell protection after hypoxic conditioning and EV-mediated RIPC in vitro. H9c2 rat cardiomyoblasts were subjected to hypoxia, with or without propofol, and subsequent simulated ischemia-reperfusion injury. Apoptosis was measured by flow cytometry. Blood samples of 64 patients receiving anesthetic maintenance with propofol or isoflurane, along with RIPC or sham procedures, were analyzed, and EVs were enriched using a polymer-based method. Propofol administration corresponded with increased Troponin T levels (4669 ± 435.6 pg/mL), suggesting an inhibition of the cardioprotective RIPC effect. RIPC leads to a notable rise in miR-21 concentrations in the group receiving propofol anesthesia (fold change 7.22 ± 6.6). In vitro experiments showed that apoptosis reduction was compromised with propofol and only occurred in an EV-enriched preconditioning medium, not in an EV-depleted medium. Our study could clinically and experimentally confirm propofol inhibition of RIPC protection. Increased miR-21 expression could provide evidence for a possible inhibitory mechanism.


Assuntos
Apoptose , Doença da Artéria Coronariana , Vesículas Extracelulares , Propofol , Vesículas Extracelulares/metabolismo , Animais , Propofol/farmacologia , Ratos , Humanos , Doença da Artéria Coronariana/metabolismo , Masculino , Apoptose/efeitos dos fármacos , Precondicionamento Isquêmico/métodos , Feminino , Pessoa de Meia-Idade , MicroRNAs/genética , MicroRNAs/metabolismo , Precondicionamento Isquêmico Miocárdico/métodos , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Traumatismo por Reperfusão Miocárdica/metabolismo , Idoso , Anestésicos/farmacologia , Estudos Prospectivos , Linhagem Celular
2.
Biotechnol Prog ; : e3507, 2024 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-39329353

RESUMO

Technology scale-up and transfer are a fundamental and critical part of process development in biomanufacturing. Important bioreactor hydrodynamic characteristics such as working volume, overhead gas flow rate, volumetric power input (P/V), impeller type, agitation regimen, sparging aeration strategy, sparger type, and kLa must be selected based on key performance indicators (KPI) to ensure a smooth and seamless process scale-up and transfer. Finding suitable operational setpoints and developing an efficient feeding regimen to ensure process efficacy and consistency are instrumental. In this investigation, process development of a cumate inducible Chinese hamster ovary (CHO) stable pool expressing trimeric SARS-CoV-2 spike protein in 1.8 L benchtop stirred-tank bioreactors is detailed. Various dissolved oxygen levels and aeration air caps were studied to determine their impact on cell growth and metabolism, culture longevity, and endpoint product titers. Once hydrodynamic conditions were tuned to an optimal zone, various feeding strategies were explored to increase culture performance. Dynamic feedings such as feeding based on current culture volume, viable cell density (VCD), oxygen uptake rate (OUR), and bio-capacitance signals were tested and compared to standard bolus addition. Increases in integral of viable cell concentration (IVCC) (1.25-fold) and protein yield (2.52-fold), as well as greater culture longevity (extension of 5 days) were observed in dynamic feeding strategies when compared to periodic bolus feeding. Our study emphasizes the benefits of designing feeding strategies around metabolically relevant signals such as OUR and bio-capacitance signals.

4.
Nanotechnology ; 35(43)2024 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-39133056

RESUMO

This study describes the fabrication of composite electrodes comprising TiO2and reduced graphene oxide layers using a moderate-temperature hydrothermal method. The morphology, crystalline structure, chemical composition, and optical features of the prepared composites were analyzed by FE-SEM, x-ray diffraction, FTIR, and UV-visible spectroscopy. The cyclic voltammetry (CV) and Nyquist plots were used to assess the electrochemical and impedance responses of the composite electrodes, respectively. The analysis revealed that the incorporation of RGO reduced the TiO2bandgap to 3.87 eV 3.02 eV and improved the specific capacitance, enhancing the TiO2-RGO electrode's supercapacitive performance. CV studies highlight that the TiO2-RGO composite has a high specific capacitance of 152 F g-1at a substantially faster scan rate of 25 mV s-1in a 1.0 M-KOH dilute electrolyte. These findings confirmed the applicability of the fabricated electrodes as prospective supercapacitor electrodes.

5.
Environ Res ; 261: 119759, 2024 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-39122163

RESUMO

Fabrication of ternary composited photocatalytic nanomaterials with strong interaction is vital to deriving the fast charge separation for efficient photodegradation of organic contaminants in wastewater under visible light. In this work, novel ternary 2D/3D/2D MoS2-In2O3-WS2 multi-nanostructures were synthesized using facile hydrothermal processes. XRD, FTIR, and XPS results confirmed the phase, functional groups, and element composition of pure MoS2, MoS2-In2O3, and MoS2-In2O3-WS2 hybrids. UV-DRS spectra of the MoS2-In2O3-WS2 ternary hybrid indicate maximum absorption in the visible light range with a band-gap energy value of 2.4 eV. The surface of the 2D WS2 nanosheet structure tightly blends and densely disperses 2D MoS2 nanosheets and 3D In2O3 nanocubes. This confirmed the formation of the MoS2-In2O3-WS2 ternary hybrid in the form of 2D/3D/2D multi-nanostructures, which is also indicated from SEM and HR-TEM images. The synthesized MoS2-In2O3-WS2 ternary hybrid showed maximum photocatalytic activity under visible-light for antimicrobial agents such as triclosan (TCS) and trichlorocarban (TCC). The photocatalytic activity of TCS was revealed to be 95% at 90 min, while that of TCC was 93% at 100 min. The reusability and stability tests of the prepared MoS2-In2O3-WS2 ternary hybrid after four consecutive photocatalytic cycles were analyzed by FTIR and SEM, which indicated that the prepared ternary hybrid was very stable. Overall results suggested that the developed MoS2-In2O3-WS2 (2D/3D/2D) multi-nanostructures are environmentally friendly and low-cost nanocomposites as a potential photocatalyst for the removal of antimicrobial agents from wastewater.


Assuntos
Dissulfetos , Luz , Molibdênio , Nanocompostos , Fotólise , Molibdênio/química , Nanocompostos/química , Dissulfetos/química , Catálise , Anti-Infecciosos/química , Sulfetos/química , Poluentes Químicos da Água/química , Águas Residuárias/química
7.
RSC Adv ; 14(31): 22690-22700, 2024 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-39027038

RESUMO

This study reports a facile approach for examining surface morphology transitions in semiconductor nanoparticles (NPs), with a focus on pristine and magnesium-doped cadmium oxide NPs. Mg-doped CdO NPs are synthesized via co-precipitation, and their composition, structure, and elemental distribution are analyzed through X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), Raman spectra, and X-ray photoelectron spectroscopy (XPS), along with optical characterization and impedance analysis. Doping with Mg2+ changes the morphology from rod-like to quasi-spherical, reduces the crystallite size, and impacts their structural and functional properties. Optical transmittance analysis revealed that Mg2+ doping resulted in a reduction of the band gap energy. Impedance spectroscopy demonstrates improved dielectric constant and electrical conductivity for Mg-doped CdO NPs. The Nyquist plots show grain effects and the equivalent circuit analysis corresponds to a R(CR)(CR) circuit. These advancements point to the potential of spherical Mg-doped CdO NPs in semiconductor applications due to their superior structural and functional characteristics.

8.
Nanoscale Horiz ; 9(8): 1230-1249, 2024 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-38958571

RESUMO

The growth of graphene on a metal substrate using chemical vapor deposition (CVD), assisted by hydrocarbons such as CH4, C3H8, C2H6, etc. leads to the formation of carbon clusters, amorphous carbon, or any other structure. These carbon species are considered as unwanted impurities; thus a conventional etching step is used simultaneously with CVD graphene growth to remove them using an etching agent. Meanwhile, art etching is a specific method of producing controlled non-Euclidean and Euclidean geometries by employing intricate and precise etching parameters or integrated growth/etching modes. Agents such as H2, O2, CH4, Ar, and others are applied as art etching agents to support the art etching technology. This technique can generate nanopores and customize the properties of graphene, facilitating specific applications such as nanodevices, nanosensors, nanofilters, etc. This comprehensive review investigates how precursor gases concurrently induce graphene growth and art etching during a chemical vapor deposition process, resulting in beautifully etched patterns. Furthermore, it discusses the techniques leading to the creation of these patterns. Finally, the challenges, uses, and perspectives of these non-Euclidean and Euclidean-shaped art etched graphene geometries are discussed.

9.
Data Brief ; 54: 110285, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38962208

RESUMO

The bacterium Paenibacillus elgii YSY-1.2 was recently isolated from soil collected from Yok Don National Park in Vietnam. Previous experiments showed this bacterium possesses high chitin-degrading activity, plant-growth promotion, and biocontrol capacity. Here, we report the draft genome sequence of strain YSY-1.2 for further characterizations related to crop production. The genome sequencing was performed using the DNBSeq-G99 with the Illumina platform. The draft genome of P. elgii YSY-1.2 has 8,240,519 bp in length and comprises 135 contigs. It has an N50 of 315,408 bp and a GC% of 52.8%. The genome contains 7498 protein-coding genes, 87 tRNA genes, and 1 rRNA gene. Among the protein-coding sequences, 6610 were assigned by COG, while 3230 were assigned by KEGG. The genome possesses at least 61 genes involved in environmental adaptation and plant growth promotion. Additionally; there are 258 carbohydrate-active enzymes deduced from the genome; among them, at least 14 may contribute to the biocontrol capacity. The chitin-degrading system of strain YSY-1.2 contains 16 chitinolytic enzymes, comprising 10 chitinases, 4 ß-N-acetylhexosaminidases, and 2 auxiliary activities. Furthermore, 32 gene clusters encoding antimicrobial metabolites were identified from the genome, with 17 showing no sequence similarities to reported clusters. Data provide an insight into the genomic information of strain YSY-1.2 and could lead to valuable further explorations and applications in crop production. This is the first report describing the genome sequence of P. elgii isolated from Vietnam.

10.
Nanotechnology ; 35(38)2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-38958589

RESUMO

Green energy technology is generally becoming one of hot issues that need to be solved due to the adverse effects on the environment of fossil fuels. One of the strategies being studied and developed by theorists and experimentalists is the use of photoelectrochemical (PEC) cells, which are emerging as a candidate to produce hydrogen from water splitting. However, creating photoelectrodes that meet the requirements for PEC water splitting has emerged as the primary obstacle in bringing this technology to commercial fruition. Here, we construct a heterostructure, which consists of MoS2/TiO2/Au nanoparticles (NPs) to overcome the drawbacks of the photoanode. Owing to the dependence on charge transfer, the bandgap of MoS2/TiO2and the utilization the Au NPs as a stimulant for charges separation of TiO2by localized surface plasmon resonances effect as well as the increase of hot electron injection to cathode, leading to photocatalytic activities are improved. The results have recorded a significant increase in the photocurrent density from 2.3µAcm-2of TiO2to approximately 16.3µAcm-2of MoS2/TiO2/Au NPs. This work unveils a promising route to enhance the visible light adsorption and charge transfer in photo-electrode of the PEC cells by combining two-dimensional materials with metal NPs.

11.
Chemosphere ; 362: 142724, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38950748

RESUMO

A large number of studies on organophosphate esters (tri-OPEs) in marine organisms have not assessed the simultaneous occurrence of tri-OPEs and their metabolites (di-OPEs) in these species. This research investigated the concentration and geographical distribution of 15 tri-OPEs and 7 di-OPEs in 172 samples of Pampus argenteus that were collected annually from 2021 to 2023 at three distinct locations along the Vietnamese coast. As a result, tri-OPEs and di-OPEs were detected in numerous fish samples, indicating their widespread spatial and temporal occurrence in marine fish and pointing out the importance of monitoring their levels. The tri-OPEs and di-OPEs ranged within 2.1-38.9 ng g-1 dry weight (dw) and 3.2-263.4 ng g-1 dw, respectively. The mean concentrations of tri-OPEs ranged from 0.4 (TIPrP) to 5.4 ng g-1 dw (TBOEP), with TBOEP and TEHP having the highest mean values. In addition, the profiles of tri-OPEs in fish exhibited a descending order: Σalkyl OPEs > ΣCl-alkyl OPEs > Σaryl OPEs. The di-OPEs, namely BEHP and DMP, had the highest mean levels, measuring 33.4 ng g-1 dw and 23.8 ng g-1 dw, respectively. Furthermore, there have been significant findings of strong positive correlations between di-OPEs and tri-OPE pairs (p < 0.05). It is worth noting that there is a noticeable difference in the composition of tri-OPEs between the North and other regions. Despite these findings, the presence of OPE-contaminated fish did not pose any health risks to Vietnam's coastal population.


Assuntos
Monitoramento Ambiental , Ésteres , Organofosfatos , Perciformes , Poluentes Químicos da Água , Animais , Vietnã , Organofosfatos/análise , Organofosfatos/metabolismo , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/metabolismo , Ésteres/análise , Ésteres/metabolismo , Perciformes/metabolismo , Análise Espaço-Temporal , Peixes/metabolismo , População do Sudeste Asiático
12.
Biomolecules ; 14(6)2024 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-38927132

RESUMO

Lung cancer is the leading cause of cancer deaths globally, necessitating effective early detection methods. Traditional diagnostics like low-dose computed tomography (LDCT) often yield high false positive rates. SHOX2 gene methylation has emerged as a promising biomarker. This study aimed to develop and validate a novel semi-nested real-time PCR assay enhancing sensitivity and specificity for detecting SHOX2 methylation using extendable blocking probes (ExBPs). The assay integrates a semi-nested PCR approach with ExBPs, enhancing the detection of low-abundance methylated SHOX2 DNA amidst unmethylated sequences. It was tested on spiked samples with varied methylation levels and on clinical samples from lung cancer patients and individuals with benign lung conditions. The assay detected methylated SHOX2 DNA down to 0.01%. Clinical evaluations confirmed its ability to effectively differentiate between lung cancer patients and those with benign conditions, demonstrating enhanced sensitivity and specificity. The use of ExBPs minimized non-target sequence amplification, crucial for reducing false positives. The novel semi-nested real-time PCR assay offers a cost-effective, highly sensitive, and specific method for detecting SHOX2 methylation, enhancing early lung cancer detection and monitoring, particularly valuable in resource-limited settings.


Assuntos
Metilação de DNA , Proteínas de Homeodomínio , Neoplasias Pulmonares , Reação em Cadeia da Polimerase em Tempo Real , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/metabolismo , Metilação de DNA/genética , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/métodos , Biomarcadores Tumorais/genética , Sensibilidade e Especificidade
13.
IUCrdata ; 9(Pt 4): x240299, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38720996

RESUMO

In the title compound, C22H11N3O3S, dihedral angle between the phenyl rings on the periphery of the molecule is 8.05 (18)°. In the crystal, aromatic π-π stacking distance and short C-H⋯O contacts are observed. The maximum absorption occurs at 688 nm.

14.
IUCrdata ; 9(Pt 4): x240357, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38721004

RESUMO

The title compound, C22H12N2S2, crystallizes in space group P21/c with four mol-ecules in the asymmetric unit. The heterocyclic mol-ecule is quasi-planar with a dihedral angle between the phenyl rings on the periphery of the mol-ecule of 1.73 (19)°. Short H⋯S (2.92 Å) and C-H⋯π [2.836 (3) Å] contacts are observed in the crystal with shorted π-π stacking distances of 3.438 (3) Šalong the b axis. Surprisingly, and unlike a closely related material, this mol-ecule readily forms large crystals by sublimation and by slow evaporation from di-chloro-methane. The maximum absorbance in the UV-Vis spectrum is at 533 nm. Emission was measured upon excitation at 533 nm with a fluorescence λmax of 658 nm and cutoff of 900 nm.

15.
Beilstein J Org Chem ; 20: 931-939, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38711584

RESUMO

The remediation of the carbohydrate data of the Protein Data Bank (PDB) has brought numerous enhancements to the findability and interpretability of deposited glycan structures, yet crucial quality indicators are either missing or hard to find on the PDB pages. Without a way to access wider glycochemical context, problematic structures may be taken as fact by keen but inexperienced scientists. The Privateer software is a validation and analysis tool that provides access to a number of metrics and links to external experimental resources, allowing users to evaluate structures using carbohydrate-specific methods. Here, we present the Privateer database, a free resource that aims to complement the growing glycan content of the PDB.

16.
ACS Nano ; 18(23): 14841-14876, 2024 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-38810109

RESUMO

Atomically thin 2D films and their van der Waals heterostructures have demonstrated immense potential for breakthroughs and innovations in science and technology. Integrating 2D films into electronics and optoelectronics devices and their applications in electronics and optoelectronics can lead to improve device efficiencies and tunability. Consequently, there has been steady progress in large-area 2D films for both front- and back-end technologies, with a keen interest in optimizing different growth and synthetic techniques. Parallelly, a significant amount of attention has been directed toward efficient transfer techniques of 2D films on different substrates. Current methods for synthesizing 2D films often involve high-temperature synthesis, precursors, and growth stimulants with highly chemical reactivity. This limitation hinders the widespread applications of 2D films. As a result, reports concerning transfer strategies of 2D films from bare substrates to target substrates have proliferated, showcasing varying degrees of cleanliness, surface damage, and material uniformity. This review aims to evaluate, discuss, and provide an overview of the most advanced transfer methods to date, encompassing wet, dry, and quasi-dry transfer methods. The processes, mechanisms, and pros and cons of each transfer method are critically summarized. Furthermore, we discuss the feasibility of these 2D film transfer methods, concerning their applications in devices and various technology platforms.

17.
Nucleic Acids Res ; 52(11): 6392-6405, 2024 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-38676944

RESUMO

We report that the Escherichia coli chromosome includes novel GC-rich genomic structural elements that trigger formation of post-replication gaps upon replisome passage. The two nearly perfect 222 bp repeats, designated Replication Risk Sequences or RRS, are each 650 kb from the terminus sequence dif and flank the Ter macrodomain. RRS sequence and positioning is highly conserved in enterobacteria. At least one RRS appears to be essential unless a 200 kb region encompassing one of them is amplified. The RRS contain a G-quadruplex on the lagging strand which impedes DNA polymerase extension producing lagging strand ssDNA gaps, $ \le$2000 bp long, upon replisome passage. Deletion of both RRS elements has substantial effects on global genome structure and topology. We hypothesize that RRS elements serve as topological relief valves during chromosome replication and segregation. There have been no screens for genomic sequences that trigger transient gap formation. Functional analogs of RRS could be widespread, possibly including some enigmatic G-quadruplexes in eukaryotes.


Assuntos
Replicação do DNA , Escherichia coli , Quadruplex G , Genoma Bacteriano , Replicação do DNA/genética , Escherichia coli/genética , Escherichia coli/metabolismo , DNA Bacteriano/metabolismo , DNA Bacteriano/genética , Cromossomos Bacterianos/genética , Cromossomos Bacterianos/metabolismo , DNA de Cadeia Simples/metabolismo , DNA de Cadeia Simples/genética , DNA Polimerase Dirigida por DNA/metabolismo , DNA Polimerase Dirigida por DNA/genética , Sequências Repetitivas de Ácido Nucleico/genética
18.
Chem Soc Rev ; 53(10): 5190-5226, 2024 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-38586901

RESUMO

Etching technology - one of the representative modern semiconductor device makers - serves as a broad descriptor for the process of removing material from the surfaces of various materials, whether partially or entirely. Meanwhile, thinning technology represents a novel and highly specialized approach within the realm of etching technology. It indicates the importance of achieving an exceptionally sophisticated and precise removal of material, layer-by-layer, at the nanoscale. Notably, thinning technology has gained substantial momentum, particularly in top-down strategies aimed at pushing the frontiers of nano-worlds. This rapid development in thinning technology has generated substantial interest among researchers from diverse backgrounds, including those in the fields of chemistry, physics, and engineering. Precisely and expertly controlling the layer numbers of 2D materials through the thinning procedure has been considered as a crucial step. This is because the thinning processes lead to variations in the electrical and optical characteristics. In this comprehensive review, the strategies for top-down thinning of representative 2D materials (e.g., graphene, black phosphorus, MoS2, h-BN, WS2, MoSe2, and WSe2) based on conventional plasma-assisted thinning, integrated cyclic plasma-assisted thinning, laser-assisted thinning, metal-assisted splitting, and layer-resolved splitting are covered in detail, along with their mechanisms and benefits. Additionally, this review further explores the latest advancements in terms of the potential advantages of semiconductor devices achieved by top-down 2D material thinning procedures.

19.
J Int Med Res ; 52(4): 3000605241240999, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38606734

RESUMO

OBJECTIVE: This study aimed to assess the association of the neutrophil-to-lymphocyte ratio (NLR) with the occurrence of venous thromboembolism (VTE) and arterial thrombosis (AT). METHODS: This was a retrospective cross-sectional study including 585 medical records obtained from all consecutive patients who were suspected of having thrombosis. RESULTS: The AT group had a higher neutrophil count and NLR and a lower lymphocyte count than the non-thrombosis group. Receiver operating characteristic curve analysis showed the ability of the NLR to predict the presence of AT. The cut-off value for the NLR was 4.44. No distinction was found in the NLR between the VTE and non-thrombosis groups. Regression analysis showed that a high NLR was an independent factor related to the presence of AT. Patients with an NLR ≥ 4.44 had a higher risk of AT than those with an NLR < 4.44 (odds ratio = 2.015, 95% confidence interval: 1.180-3.443). CONCLUSION: A high NLR may be considered a predictive factor for the occurrence of AT, but an association with the presence of VTE was not found.


Assuntos
Trombose , Tromboembolia Venosa , Humanos , Neutrófilos , Tromboembolia Venosa/diagnóstico , Estudos Retrospectivos , Estudos Transversais , Linfócitos , Curva ROC , Prognóstico
20.
Biotechnol Prog ; : e3467, 2024 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-38660973

RESUMO

The recent COVID-19 pandemic revealed an urgent need to develop robust cell culture platforms which can react rapidly to respond to this kind of global health issue. Chinese hamster ovary (CHO) stable pools can be a vital alternative to quickly provide gram amounts of recombinant proteins required for early-phase clinical assays. In this study, we analyze early process development data of recombinant trimeric spike protein Cumate-inducible manufacturing platform utilizing CHO stable pool as a preferred production host across three different stirred-tank bioreactor scales (0.75, 1, and 10 L). The impact of cell passage number as an indicator of cell age, methionine sulfoximine (MSX) concentration as a selection pressure, and cell seeding density was investigated using stable pools expressing three variants of concern. Multivariate data analysis with principal component analysis and batch-wise unfolding technique was applied to evaluate the effect of critical process parameters on production variability and a random forest (RF) model was developed to forecast protein production. In order to further improve process understanding, the RF model was analyzed with Shapley value dependency plots so as to determine what ranges of variables were most associated with increased protein production. Increasing longevity, controlling lactate build-up, and altering pH deadband are considered promising approaches to improve overall culture outcomes. The results also demonstrated that these pools are in general stable expressing similar level of spike proteins up to cell passage 11 (~31 cell generations). This enables to expand enough cells required to seed large volume of 200-2000 L bioreactor.

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