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1.
Biotechnol Bioeng ; 2024 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-38993032

RESUMO

Scale-down models (SDM) are pivotal tools for process understanding and improvement to accelerate the development of vaccines from laboratory research to global commercialization. In this study, a 3 L SDM representing a 50 L scale Vero cell culture process of a live-attenuated virus vaccine using microcarriers was developed and qualified based on the constant impeller power per volume principle. Both multivariate data analysis (MVDA) and the traditional univariate data analysis showed comparable and equivalent cell growth, metabolic activity, and product quality results across scales. Computational fluid dynamics simulation further confirmed similar hydrodynamic stress between the two scales.

2.
Data Brief ; 54: 110532, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38868389

RESUMO

Gas chromatography ion mobility spectrometry (GC-IMS) is a robust and sensitive benchtop technique commonly used for non-target screening of volatile organic compounds. It has been applied to authenticity analysis by generating characteristic "fingerprints" of food samples, well suited for chemometric data analysis. This dataset contains headspace GC-IMS spectra from 50 monofloral honey samples from three different botanical origins, 18 acacia honeys (Robinia pseudoacacia), 19 canola honeys (Brassica napus) and 18 honeydew honeys (forest flowers). Honeys were sourced from the beekeepers directly or obtained from governmental food inspectors from Baden-Wuerttemberg, Germany. Authenticity was confirmed by pollen analysis in the framework of the official control of foodstuffs. The data was acquired using a setup based on an Agilent 6890N gas chromatograph (Agilent Technologies, Palo Alto, CA) and an OEM Standalone IMS cell from G.A.S Sensorsysteme m. b. H. (Dortmund, Germany). All samples were recorded in duplicates and spectra are presented as raw data in the .mea file format. The dataset is available on Mendeley Data: https://data.mendeley.com/datasets/jxj2r45t2x.

3.
Foods ; 13(12)2024 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-38928847

RESUMO

Quinoa is an Andean crop that stands out as a high-quality protein-rich and gluten-free food. However, its increasing popularity exposes quinoa products to the potential risk of adulteration with cheaper cereals. Consequently, there is a need for novel methodologies to accurately characterize the composition of quinoa, which is influenced not only by the variety type but also by the farming and processing conditions. In this study, we present a rapid and straightforward method based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to generate global fingerprints of quinoa proteins from white quinoa varieties, which were cultivated under conventional and organic farming and processed through boiling and extrusion. The mass spectra of the different protein extracts were processed using the MALDIquant software (version 1.19.3), detecting 49 proteins (with 31 tentatively identified). Intensity values from these proteins were then considered protein fingerprints for multivariate data analysis. Our results revealed reliable partial least squares-discriminant analysis (PLS-DA) classification models for distinguishing between farming and processing conditions, and the detected proteins that were critical for differentiation. They confirm the effectiveness of tracing the agricultural origins and technological treatments of quinoa grains through protein fingerprinting by MALDI-TOF-MS and chemometrics. This untargeted approach offers promising applications in food control and the food-processing industry.

4.
Spectrochim Acta A Mol Biomol Spectrosc ; 320: 124579, 2024 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-38850824

RESUMO

Among the severe foodborne illnesses, listeriosis resulting from the pathogen Listeria monocytogenes exhibits one of the highest fatality rates. This study investigated the application of near infrared hyperspectral imaging (NIR-HSI) for the classification of three L. monocytogenes serotypes namely serotype 4b, 1/2a and 1/2c. The bacteria were cultured on Brain Heart Infusion agar, and NIR hyperspectral images were captured in the spectral range 900-2500 nm. Different pre-processing methods were applied to the raw spectra and principal component analysis was used for data exploration. Classification was achieved with partial least squares discriminant analysis (PLS-DA). The PLS-DA results revealed classification accuracies exceeding 80 % for all the bacterial serotypes for both training and test set data. Based on validation data, sensitivity values for L. monocytogenes serotype 4b, 1/2a and 1/2c were 0.69, 0.80 and 0.98, respectively when using full wavelength data. The reduced wavelength model had sensitivity values of 0.65, 0.85 and 0.98 for serotype 4b, 1/2a and 1/2c, respectively. The most relevant bands for serotype discrimination were identified to be around 1490 nm and 1580-1690 nm based on both principal component loadings and variable importance in projection scores. The outcomes of this study demonstrate the feasibility of utilizing NIR-HSI for detecting and classifying L. monocytogenes serotypes on growth media.


Assuntos
Imageamento Hiperespectral , Listeria monocytogenes , Análise de Componente Principal , Sorogrupo , Espectroscopia de Luz Próxima ao Infravermelho , Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/classificação , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Imageamento Hiperespectral/métodos , Análise Discriminante , Análise dos Mínimos Quadrados
5.
J Agric Food Chem ; 72(26): 15040-15052, 2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-38906536

RESUMO

Wheat species with various ploidy levels may be different regarding their immunoreactive potential in celiac disease (CD), but a comprehensive comparison of peptide sequences with known epitopes is missing. Thus, we used an untargeted liquid chromatography tandem mass spectrometry method to analyze the content of peptides with CD-active epitope in the five wheat species common wheat, spelt, durum wheat, emmer, and einkorn. In total, 494 peptides with CD-active epitope were identified. Considering the average of the eight cultivars of each species, spelt contained the highest number of different peptides with CD-active epitope (193 ± 12, mean ± SD). Einkorn showed the smallest variability of peptides (63 ± 4) but higher amounts of certain peptides compared to the other species. The wheat species differ in the presence and distribution of CD-active epitopes; hence, the entirety of peptides with CD-active epitope is crucial for the assessment of their immunoreactive potential.


Assuntos
Doença Celíaca , Epitopos , Proteínas de Plantas , Proteômica , Triticum , Doença Celíaca/imunologia , Triticum/química , Triticum/imunologia , Epitopos/imunologia , Epitopos/química , Proteínas de Plantas/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Humanos , Espectrometria de Massas em Tandem , Peptídeos/imunologia , Peptídeos/química
6.
Eur J Pharm Sci ; 200: 106836, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38901784

RESUMO

Principal component analysis (PCA) and partial least squares regression (PLS) were combined in this study to identify key material descriptors determining tabletability in direct compression and roller compaction. An extensive material library including 119 material descriptors and tablet tensile strengths of 44 powders and roller compacted materials with varying drug loads was generated to systematically elucidate the impact of different material descriptors, raw API and filler properties as well as process route on tabletability. A PCA model was created which highlighted correlations between different powder descriptors and respective characterization methods and, thus, can enable reduction of analyses to save resources to a certain extent. Subsequently, PLS models were established to identify key material attributes for tabletability such as density and particle size but also surface energy, work of cohesion and wall friction, which were for the first time demonstrated by PLS as highly relevant for tabletability in roller compaction and direct compression. Further, PLS based on extensive material characterization enabled the prediction of tabletability of materials unknown to the model. Thus, this study highlighted how PCA and PLS are useful tools to elucidate the correlations between powder and tabletability, which will enable more robust prediction of manufacturability in formulation development.

7.
Food Chem ; 456: 139982, 2024 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-38876062

RESUMO

Fermentation stage is a crucial factor for flavor profiles formation of hawthon wine. Thus, comprehensive knowledge of dynamic relationship between nonvolatile (NVOCs) and volatile aroma compounds (VOCs) from hawthorn wine at different fermentation stages was investigated by GC-MS and HPLC coupled with multivariate analysis. The increase of alcohols/esters/acids but decrease of terpenes/aldehydes/ketones was observed as fermentation extension. Specifically, OAV of ethyl acetate, ethyl caprylate, and ethyl caprate was > 50 from the 3rd day to 10th day, giving more fruity properties. Multivariate analysis showed that 1-hexanol, ethyl myristate, isobutyric acid, et al., were linked to the sensory evaluation of "sweet", "floral" and "fruity", and fructose, glucose and bitter amino acids were responsible for reduction of "bitterness" and "astringency". Additionally, VOCs were positively correlated with organic acids while negative to amino acids/soluble sugars, probably due to metabolization as precursors, providing references for aroma enhancement by regulating NVOCs precursors.

8.
Foods ; 13(11)2024 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-38890893

RESUMO

In the last decade, "expressions" of grape marc spirits aged in wooden barrels of characteristic amber color and complex sensory attributes have been introduced. Yet studies on constituents migrating from the barrel to the beverage are scarce, and their metabolic profile remains unexplored. Furthermore, the literature on the assessment of their antioxidant activity is limited. NMR metabolomics and spectrophotometry have been implemented in 38 samples to elucidate the impact of the aging procedure on the metabolites' composition and establish whether these beverages exhibit antioxidant activity. Provenance was related to fusel alcohols, esters, acetaldehyde, methanol, saccharides, and 2-phenylethanol, while ethyl acetate and ethyl lactate contributed to discriminating samples of the same winery. Identified metabolites such as vanillin, syringaldehyde, and sinapaldehyde were related to the aging procedure. The maturation in the barrel was also associated with an increase in xylose, glucose, fructose, and arabinose. The antioxidant potential of the aged Greek grape marc spirits resulting from their maturation in oak barrels was highlighted. The metabolic profiling and antioxidant potential of aged Greek grape marc spirits were assessed for the first time. Finally, the enrichment of the aromatic region was noted with the presence of metabolites with a furanic and phenolic ring derived, respectively, from the polysaccharides' degradation or the thermal decomposition of lignin.

9.
Phytochem Anal ; 2024 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-38768954

RESUMO

INTRODUCTION: The Olive (Olea europaea L.) is one of the most popular edible oil-producing fruits, consumed worldwide for its myriad nutritional and health benefits. Olive oil production generates huge quantities of by-products from the fruit, which are considered environmental hazards. Recently, more and more efforts have been made to valorize olive by-products as a source of low-cost, value-added food applications. OBJECTIVE: The main objective of this study was to globally assess the metabolome of olive fruit by-products, including olive mill wastewater, olive pomace, and olive seeds from fruits from two areas, Siwa and Anshas, Egypt. METHODS: Gas chromatography-mass spectrometry (GC-MS) and ultra-high-performance liquid chromatography with mass spectrometry (UPLC-MS) were used for profiling primary and secondary metabolites in olive by-products. Also, multivariate data analyses were used to assess variations between olive by-product samples. RESULTS: A total of 103 primary metabolites and 105 secondary metabolites were identified by GC-MS and UPLC-MS, respectively. Fatty acids amounted to a major class in the olive by-products at 53-91%, with oleic acid dominating, especially in the pomace of Siwa. Mill wastewater was discriminated from other by-products by the presence of phenolics mainly tyrosol, hydroxyl tyrosol, and α-tocopherol as analyzed by UPLC-MS indicating their potential antioxidant activity. Pomace and seeds were rich in fatty acids/esters and hydroxy fatty acids and not readily distinguishable from each other. CONCLUSION: The current work discusses the metabolome profile of olive waste products for valorization purposes. Pomace and seeds were enriched in fatty acids/esters, though not readily distinguishable from each other.

10.
Plants (Basel) ; 13(9)2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38732385

RESUMO

The Italian garlic ecotype "Vessalico" possesses distinct characteristics compared to its French parent cultivars Messidor and Messidrôme, used for sowing, as well as other ecotypes in neighboring regions. However, due to the lack of a standardized seed supply method and cultivation protocol among farmers in the Vessalico area, a need to identify garlic products that align with the Vessalico ecotype arises. In this study, an NMR-based approach followed by multivariate analysis to analyze the chemical composition of Vessalico garlic sourced from 17 different farms, along with its two French parent cultivars, was employed. Self-organizing maps allowed to identify a homogeneous subset of representative samples of the Vessalico ecotype. Through the OPLS-DA model, the most discriminant metabolites based on values of VIP (Variable Influence on Projections) were selected. Among them, S-allylcysteine emerged as a potential marker for distinguishing the Vessalico garlic from the French parent cultivars by NMR screening. Additionally, to promote sustainable agricultural practices, the potential of Vessalico garlic extracts and its main components as agrochemicals against Xanthomonas campestris pv. campestris, responsible for black rot disease, was explored. The crude extract exhibited a MIC of 125 µg/mL, and allicin demonstrated the highest activity among the tested compounds (MIC value of 31.25 µg/mL).

11.
Food Res Int ; 186: 114346, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38729720

RESUMO

Specialty coffee beans are those produced, processed, and characterized following the highest quality standards, toward delivering a superior final product. Environmental, climatic, genetic, and processing factors greatly influence the green beans' chemical profile, which reflects on the quality and pricing. The present study focuses on the assessment of eight major health-beneficial bioactive compounds in green coffee beans aiming to underscore the influence of the geographical origin and post-harvesting processing on the quality of the final beverage. For that, we examined the non-volatile chemical profile of specialty Coffea arabica beans from Minas Gerais state, Brazil. It included samples from Cerrado (Savannah), and Matas de Minas and Sul de Minas (Atlantic Forest) regions, produced by two post-harvesting processing practices. Trigonelline, theobromine, theophylline, chlorogenic acid derivatives, caffeine, caffeic acid, ferulic acid, and p-coumaric acid were quantified in the green beans by high-performance liquid chromatography with diode array detection. Additionally, all samples were roasted and subjected to sensory analysis for coffee grading. Principal component analysis suggested that Cerrado samples tended to set apart from the other geographical locations. Those samples also exhibited higher levels of trigonelline as confirmed by two-way ANOVA analysis. Samples subjected to de-pulping processing showed improved chemical composition and sensory score. Those pulped coffees displayed 5.8% more chlorogenic acid derivatives, with an enhancement of 1.5% in the sensory score compared to unprocessed counterparts. Multivariate logistic regression analysis pointed out altitude, ferulic acid, p-coumaric acid, sweetness, and acidity as predictors distinguishing specialty coffee beans obtained by the two post-harvest processing. These findings demonstrate the influence of regional growth conditions and post-harvest treatments on the chemical and sensory quality of coffee. In summary, the present study underscores the value of integrating target metabolite analysis with statistical tools to augment the characterization of specialty coffee beans, offering novel insights for quality assessment with a focus on their bioactive compounds.


Assuntos
Coffea , Café , Manipulação de Alimentos , Sementes , Brasil , Coffea/química , Sementes/química , Manipulação de Alimentos/métodos , Café/química , Alcaloides/análise , Cromatografia Líquida de Alta Pressão , Humanos , Paladar , Análise de Componente Principal
12.
Stud Health Technol Inform ; 314: 178-182, 2024 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-38785027

RESUMO

The characterization of local improved varieties as well as the reduction of synthetic chemical fertilizers are sustainable approaches in the vision of a new precision Farming. Aim of our study was to improve the geographical characterization of local ecotypes and to identify peculiar features of new crops in terms of bioactive compounds. NMR and LC-MS metabolite profiling approaches followed by multivariate data analysis were applied to characterize local rosemary and garlic ecotypes. With the aim of applying for a protected designation of origin, orthogonal partial least squares discriminant analysis (OPLS-DA) was used to identify representative sensory quality indicators for Vessalico garlic and rosemary "Eretto Liguria" local ecotypes, Variable Influence on Projections (VIP) values of OPLS-DA indicated six metabolites as quality indicators for Vessalico garlic and sixteen metabolites as quality indicators for rosemary "Eretto Liguria". Finally, to discover and utilize new ecotypes in a sustainable way, Vessalico garlic extracts antiviral activity, previously evaluated against Tomato brown rugose fruit virus (ToBRFV), a Tobamovirus affecting tomato crops, was extended to Pepino mosaic virus (PepMV) with positive results.


Assuntos
Ecótipo , Extratos Vegetais/uso terapêutico , Alho/química , Rosmarinus/química , Agroquímicos
13.
Heliyon ; 10(10): e30498, 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38803972

RESUMO

The composition of honey is mostly determined by the species-specific characteristics of flowering plants, which is reflected in the significant deviations in composition of honey varieties. The high-quality acacia honey is assessed based on both physical-chemical parameters and melissopalynology. The appearance of rape pollen in acacia honey makes the acacia honey be sorted into the multifloral honey category. Over carrying out melissopalynology, the149 samples of various honeys (acacia, rape and multifloral) have also been analysed by using physical-chemical and elemental analysis. Multivariate data analysis revealed that multifloral honey is much closer to acacia honey than to rape honey, as it can be observed from the examined unique parameters. By the PCA (Principal Component Analysis) analysis based on united set of physico-chemical and melissopalynology results the acacia and rape honey samples are entirely separated for each other, while multifloral honey samples are very close to acacia honey group and partially overlap with it. On ignoring the pollen analysis and based on the rest of the results, the multifloral honey category is almost indistinguishable from the declared and verified acacia honey category.

14.
J Agric Food Chem ; 2024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38803291

RESUMO

Cereal grains play an important role in human health as a source of macro- and micronutrients, besides phytochemicals. The metabolite diversity was investigated in cereal crops and their milling fractions by untargeted metabolomics ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) of 69 samples: 7 species (barley, oat, pearl millet, rye, sorghum, triticale, and wheat), 23 genotypes, and 4 milling fractions (husk, bran, flour, and wholegrain). Samples were also analyzed by in vitro antioxidant activity. UHPLC-MS/MS signals were processed using XCMS, and metabolite annotation was based on SIRIUS and GNPS libraries. Bran and husk showed the highest antioxidant capacity and phenolic content/diversity. The major metabolite classes were phenolic acids, flavonoids, fatty acyls, and organic acids. Sorghum, millet, barley, and oats showed distinct metabolite profiles, especially related to the bran fraction. Molecular networking and chemometrics provided a comprehensive insight into the metabolic profiling of cereal crops, unveiling the potential of coproducts and super cereals such as sorghum and millet as sources of polyphenols.

15.
Spectrochim Acta A Mol Biomol Spectrosc ; 315: 124261, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38608560

RESUMO

Food safety is always of paramount importance globally due to the devasting social and economic effects of foodborne disease outbreaks. There is a high consumption rate of meat worldwide, making it an essential protein source in the human diet, hence its microbial safety is of great importance. The food industry stakeholders are always in search of methods that ensure safe food whilst maintaining food quality and excellent sensory attributes. Currently, there are several methods used in microbial food analysis, however, these methods are often time-consuming and do not allow real-time analysis. Considering the recent technological breakthroughs in artificial intelligence and machine learning, it raises the question of whether these advancements could be leveraged within the meat industry to improve turnaround time for microbial assessments. Hyperspectral imaging (HSI) is a highly prospective technology worth exploring for microbial analysis. The rapid, non-destructive method has the potential to be integrated into food production systems and allows foodborne pathogen detection in food samples, thus saving time. Although there has been a substantial increase in research on the utilisation of HSI in food applications over the past years, its use in the microbial assessment of meat is not yet optimal. This review aims to provide a basic understanding of the visible-near infrared HSI system, recent applications in the microbial assessment of meat products, challenges, and possible future applications.


Assuntos
Microbiologia de Alimentos , Imageamento Hiperespectral , Carne , Imageamento Hiperespectral/métodos , Carne/análise , Carne/microbiologia , Microbiologia de Alimentos/métodos , Animais , Bactérias/isolamento & purificação , Humanos , Espectroscopia de Luz Próxima ao Infravermelho/métodos
16.
Biotechnol Bioeng ; 121(7): 2175-2192, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38613199

RESUMO

In the era of Biopharma 4.0, process digitalization fundamentally requires accurate and timely monitoring of critical process parameters (CPPs) and quality attributes. Bioreactor systems are equipped with a variety of sensors to ensure process robustness and product quality. However, during the biphasic production of viral vectors or replication-competent viruses for gene and cell therapies and vaccination, current monitoring techniques relying on a single working sensor can be affected by the physiological state change of the cells due to infection/transduction/transfection step required to initiate production. To address this limitation, a multisensor (MS) monitoring system, which includes dual-wavelength fluorescence spectroscopy, dielectric signals, and a set of CPPs, such as oxygen uptake rate and pH control outputs, was employed to monitor the upstream process of adenovirus production in HEK293 cells in bioreactor. This system successfully identified characteristic responses to infection by comparing variations in these signals, and the correlation between signals and target critical variables was analyzed mechanistically and statistically. The predictive performance of several target CPPs using different multivariate data analysis (MVDA) methods on data from a single sensor/source or fused from multiple sensors were compared. An MS regression model can accurately predict viable cell density with a relative root mean squared error (rRMSE) as low as 8.3% regardless of the changes occurring over the infection phase. This is a significant improvement over the 12% rRMSE achieved with models based on a single source. The MS models also provide the best predictions for glucose, glutamine, lactate, and ammonium. These results demonstrate the potential of using MVDA on MS systems as a real-time monitoring approach for biphasic bioproduction processes. Yet, models based solely on the multiplicity and timing of infection outperformed both single-sensor and MS models, emphasizing the need for a deeper mechanistic understanding in virus production prediction.


Assuntos
Adenoviridae , Reatores Biológicos , Humanos , Células HEK293 , Reatores Biológicos/virologia , Adenoviridae/genética , Análise Multivariada , Cultura de Vírus/métodos
17.
Foods ; 13(8)2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38672895

RESUMO

Data processing and data extraction are the first, and most often crucial, steps in metabolomics and multivariate data analysis in general. There are several software solutions for these purposes in GC-MS metabolomics. It becomes unclear which platform offers what kind of data and how that information influences the analysis's conclusions. In this study, selected analytical platforms for GC-MS metabolomics profiling, SpectConnect and XCMS as well as MestReNova software, were used to process the results of the HS-SPME/GC-MS aroma analyses of several blackberry varieties. In addition, a detailed analysis of the identification of the individual components of the blackberry aroma club varieties was performed. In total, 72 components were detected in the XCMS platform, 119 in SpectConnect, and 87 and 167 in MestReNova, with automatic integral and manual correction, respectively, as well as 219 aroma components after manual analysis of GC-MS chromatograms. The obtained datasets were fed, for multivariate data analysis, to SIMCA software, and underwent the creation of PCA, OPLS, and OPLS-DA models. The results of the validation tests and VIP-pred. scores were analyzed in detail.

18.
Front Vet Sci ; 11: 1323420, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38596461

RESUMO

Amid the surge in data volume generated across various fields of knowledge, there is an increasing necessity for advanced analytical methodologies to effectively process and utilize this information. Particularly in the field of animal health, this approach is pivotal for enhancing disease understanding, surveillance, and management. The main objective of the study was to conduct a comprehensive livestock and environmental characterization of Colombian municipalities and examine their relationship with the distribution of vesicular stomatitis (VS). Utilizing satellite imagery to delineate climatic and land use profiles, along with data from the Colombian Agricultural Institute (ICA) concerning animal populations and their movements, the research employed Principal Component Analysis (PCA) to explore the correlation between environmental and livestock-related variables. Additionally, municipalities were grouped through a Hierarchical Clustering process. The assessment of risk associated with VS was carried out using a Generalized Linear Model. This process resulted in the formation of four distinct clusters: three primarily characterized by climatic attributes and one predominantly defined by livestock characteristics. Cluster 1, identified as "Andino" due to its climatic and environmental features, exhibited the highest odds ratio for VS occurrence. The adopted methodology not only provides a deeper understanding of the local population and its context, but also offers valuable insights for enhancing disease surveillance and control programs.

19.
J Pharm Sci ; 113(5): 1168-1176, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38447668

RESUMO

In recent years, multivariate data analysis (MVDA) has been widely used for process characterization and fault diagnosis in the biopharmaceutical industry. This study aims to investigate the feasibility of using MVDA for the development and scale-up of a perfusion process for HEK293 cell-based recombinant adenovirus zoster vaccine (Ad-HER) production. The Principal Component Analysis (PCA) results suggested comparable performance among the ATF, PATFP, and BFP perfusion systems in benchtop-scale stirred-tank bioreactor (STR). Then a Batch Evolution Model (BEM) was built using representative data from 10 L STR with a BFP system to assess the Ad-HER perfusion process performance at pilot-scale bioreactor (50 L STR and 50 L wave bioreactor). Furthermore, another BEM model and Batch Level Model (BLM) were built to monitor process parameters over time and predict the final adenovirus titer in 50 L wave bioreactor. The loading plot revealed that lactate dehydrogenase activity, viable cell diameter, and base-added during the virus production phase could be used as preliminary indicators of adenovirus yield. Finally, an adenovirus titer of 2.0±0.3×1010 IFU/mL was achieved in the 50 L wave bioreactor with BFP system, highlighting the robustness of the Ad-HER perfusion process at pilot-scale. Overall, this study emphasizes the effectiveness of MVDA as a tool for advancing the understanding of recombinant adenovirus vaccine perfusion production process development and scale-up.


Assuntos
Vacinas contra Adenovirus , Vacina contra Herpes Zoster , Humanos , Técnicas de Cultura de Células/métodos , Adenoviridae , Células HEK293 , Reatores Biológicos
20.
BMC Bioinformatics ; 25(1): 93, 2024 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-38438871

RESUMO

An organism's observable traits, or phenotype, result from intricate interactions among genes, proteins, metabolites and the environment. External factors, such as associated microorganisms, along with biotic and abiotic stressors, can significantly impact this complex biological system, influencing processes like growth, development and productivity. A comprehensive analysis of the entire biological system and its interactions is thus crucial to identify key components that support adaptation to stressors and to discover biomarkers applicable in breeding programs or disease diagnostics. Since the genomics era, several other 'omics' disciplines have emerged, and recent advances in high-throughput technologies have facilitated the generation of additional omics datasets. While traditionally analyzed individually, the last decade has seen an increase in multi-omics data integration and analysis strategies aimed at achieving a holistic understanding of interactions across different biological layers. Despite these advances, the analysis of multi-omics data is still challenging due to their scale, complexity, high dimensionality and multimodality. To address these challenges, a number of analytical tools and strategies have been developed, including clustering and differential equations, which require advanced knowledge in bioinformatics and statistics. Therefore, this study recognizes the need for user-friendly tools by introducing Holomics, an accessible and easy-to-use R shiny application with multi-omics functions tailored for scientists with limited bioinformatics knowledge. Holomics provides a well-defined workflow, starting with the upload and pre-filtering of single-omics data, which are then further refined by single-omics analysis focusing on key features. Subsequently, these reduced datasets are subjected to multi-omics analyses to unveil correlations between 2-n datasets. This paper concludes with a real-world case study where microbiomics, transcriptomics and metabolomics data from previous studies that elucidate factors associated with improved sugar beet storability are integrated using Holomics. The results are discussed in the context of the biological background, underscoring the importance of multi-omics insights. This example not only highlights the versatility of Holomics in handling different types of omics data, but also validates its consistency by reproducing findings from preceding single-omics studies.


Assuntos
Beta vulgaris , Multiômica , Melhoramento Vegetal , Biologia Computacional , Análise por Conglomerados
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