BioTechPub Date : 2025-03-12DOI: 10.3390/biotech14010018
Yao Lu, Qiang Jiang, Yamin Dong, Runzhen Ji, Yiwen Xiao, Du Zhu, Boliang Gao
{"title":"Characterization of a GH43 Bifunctional Glycosidase from Endophytic <i>Chaetomium globosum</i> and Its Potential Application in the Biotransformation of Ginsenosides.","authors":"Yao Lu, Qiang Jiang, Yamin Dong, Runzhen Ji, Yiwen Xiao, Du Zhu, Boliang Gao","doi":"10.3390/biotech14010018","DOIUrl":"https://doi.org/10.3390/biotech14010018","url":null,"abstract":"<p><p>The GH43 family of glycosidases represents an important class of industrial enzymes that are widely utilized across the food, pharmaceutical, and various other sectors. In this study, we identified a GH43 family glycoside hydrolytic enzyme, <i>Xyaf313</i>, derived from the plant endophytic fungus <i>Chaetomium globosum</i> DX-THS3, which is capable of transforming several common ginsenosides. The enzyme function analysis reveals that Xyaf313 exhibits dual functionality, displaying both α-L-arabinofuranosidase and β-D-xylosidase activity. When acting as an α-L-arabinofuranosidase, Xyaf313 achieves optimal enzyme activity of 23.96 U/mg at a temperature of 50 °C and a pH of 7. In contrast, its β-D-xylosidase activity results in a slight reduction in enzyme activity to 23.24 U/mg, with similar optimal temperature and pH conditions to those observed for the α-L-arabinofuranosidase activity. Furthermore, Xyaf313 demonstrates considerable resistance to most metal ions and common chemical reagents. Notably, while the maximum enzyme activity of Xyaf313 occurs at 50 °C, it maintains high activity at room temperature (30 °C), with relative enzyme activity exceeding 90%. Measurements of ginsenoside transformation show that Xyaf313 can convert common ginsenosides Rc, Rb<sub>1</sub>, Rb<sub>2</sub>, and Rb<sub>3</sub> into Rd, underscoring its potential for pharmaceutical applications. Overall, our findings contribute to the identification of a new class of bifunctional GH43 glycoside hydrolases, highlight the significance of plant endophytic fungi as a promising resource for the screening of carbohydrate-decomposing enzymes, and present new candidate enzymes for the biotransformation of ginsenosides.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11940195/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144027439","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A Coupled Model of the Cardiovascular and Immune Systems to Analyze the Effects of COVID-19 Infection.","authors":"Camila Ribeiro Passos, Alexandre Altamir Moreira, Ruy Freitas Reis, Rodrigo Weber Dos Santos, Marcelo Lobosco, Bernardo Martins Rocha","doi":"10.3390/biotech14010019","DOIUrl":"https://doi.org/10.3390/biotech14010019","url":null,"abstract":"<p><p>The COVID-19 pandemic has underscored the importance of understanding the interplay between the cardiovascular and immune systems during viral infections. SARS-CoV-2 enters human cells via the ACE-2 enzyme, initiating a cascade of immune responses. This study presents a coupled mathematical model that integrates the cardiovascular system (CVS) and immune system (IS), capturing their complex interactions during infection. The CVS model, based on ordinary differential equations, describes heart dynamics and pulmonary and systemic circulation, while the IS model simulates immune responses to SARS-CoV-2, including immune cell interactions and cytokine production. A coupling strategy transfers information from the IS to the CVS at specific intervals, enabling the exploration of immune-driven cardiovascular effects. Numerical simulations examined how these interactions influence infection severity and recovery. The coupled model accurately replicated the evolution of cardiac function in survivors and non-survivors of COVID-19. Survivors exhibited a left ventricular ejection fraction (LVEF) reduction of up to 25% while remaining within normal limits, whereas non-survivors showed a severe 4-fold decline, indicative of myocardial dysfunction. Similarly, the right ventricular ejection fraction (RV EF) decreased by approximately 50% in survivors but underwent a drastic 5-fold reduction in non-survivors. These findings highlight the model's capacity to distinguish differential cardiac dysfunction across clinical outcomes and its potential to enhance our understanding of COVID-19 pathophysiology.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11940274/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144017217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-03-08DOI: 10.3390/biotech14010017
Alessandro A Dos Santos, Camila Nader, Mateus B de Freitas, César F Ribeiro, Geovanna de Oliveira Costa, Louis P Sandjo, Alex S Poltronieri, Roberto B Derner, Marciel J Stadnik
{"title":"Chemical Profiling and Bioactivity of Microalgae Extracts for Enhancing Growth and Anthracnose Resistance in the Common Bean (<i>Phaseolus vulgaris</i> L.).","authors":"Alessandro A Dos Santos, Camila Nader, Mateus B de Freitas, César F Ribeiro, Geovanna de Oliveira Costa, Louis P Sandjo, Alex S Poltronieri, Roberto B Derner, Marciel J Stadnik","doi":"10.3390/biotech14010017","DOIUrl":"https://doi.org/10.3390/biotech14010017","url":null,"abstract":"<p><p>The present study aimed to chemically profile the hydroalcoholic extracts from the microalgae (MEs) <i>Nannochloropsis oculata</i>, <i>Phaeodactylum tricornutum</i>, <i>Tetradesmus obliquus</i>, and <i>Tetraselmis tetrathele</i> and evaluate their effects on the development of <i>Colletotrichum lindemuthianum</i> and anthracnose symptoms, as well as on the initial growth of bean plants. For this, MEs were analyzed using UPLC coupled with a mass spectrometer, allowing the identification of peaks and annotation of potential metabolites. Fungal mycelial growth was assessed seven days after inoculation, and conidial germination was measured 72 h after incubation, using ME concentrations of 0, 0.1, 0.5, and 1.0 mg·mL<sup>-1</sup>. Bean seeds of the IPR Uirapuru cultivar were sown and treated with 3 mL of extracts at four time points: at sowing and 72 h after each previous treatment. After 11 days of cultivation in a growth chamber, the plants were divided into two groups: one for anthracnose control assessment and the other for evaluating growth promotion by MEs. Plant length as well as fresh and dry weights of shoots and roots were determined, leaf pigments were quantified, and anthracnose severity was assessed using a diagrammatic scale. The UPLC analysis identified 32 compounds in the extracts of the four microalgae, belonging to different chemical and functional groups, with lipids being the most significant fraction. The extracts exhibited variability and diversity in chemical composition depending on the microalgal species. MEs did not affect mycelial growth yet increased the germination of <i>C. lindemuthianum</i> conidia, regardless of the dose or species used. Anthracnose severity was not affected by the microalgae extracts. Regarding growth promotion, the extracts showed varying effects but generally increased shoot and root length, fresh biomass, and leaf pigment content.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11940543/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144022314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-03-07DOI: 10.3390/biotech14010016
William Y Oyadomari, Elizangela A Carvalho, Gabriel E Machado, Ana Júlia O Machado, Gabriel S Santos, Marcelo Marcondes, Vitor Oliveira
{"title":"Towards a Survival-Based Cellular Assay for the Selection of Protease Inhibitors in <i>Escherichia coli</i>.","authors":"William Y Oyadomari, Elizangela A Carvalho, Gabriel E Machado, Ana Júlia O Machado, Gabriel S Santos, Marcelo Marcondes, Vitor Oliveira","doi":"10.3390/biotech14010016","DOIUrl":"https://doi.org/10.3390/biotech14010016","url":null,"abstract":"<p><p>We describe a method tailored to the in-cell selection of protease inhibitors. In this method, a target protease is co-expressed with a selective substrate, the product of which kills host cells. Therefore, the method can be applied to identify potential inhibitors based on cell host survival when inhibition of the target protease occurs. The TEV protease was chosen for this proof-of-concept experiment. The genetically encoded selective substrate is a single polypeptide chain composed of three parts: (1) a ccdB protein, which can cause host cell death when it accumulates inside the cell; (2) a protease cleavage sequence that can be changed according to the target protease, in this case the TEV substrate ENLYFQ↓G (↓-predicted cleavage site); and (3) the ssrA sequence (AANDENYALAA), which drives the polypeptide to degradation by the ClpX/ClpP complex inside host <i>E. coli</i> cells. In our experiment, co-expression of the active TEV protease and this selective substrate (ccdB-ENLYFQG-ssrA) caused the death of a significant host cell population, while control assays with an inactive mutant TEV Asp81Asn did not. Details of the methodology used are given, providing the basis for the application of similar systems for other proteases of interest.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11940102/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144027444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-03-06DOI: 10.3390/biotech14010015
Sandeep Nair, Gerald H Lushington, Mohan Purushothaman, Bernard Rubin, Eldon Jupe, Santosh Gattam
{"title":"Prediction of Lupus Classification Criteria via Generative AI Medical Record Profiling.","authors":"Sandeep Nair, Gerald H Lushington, Mohan Purushothaman, Bernard Rubin, Eldon Jupe, Santosh Gattam","doi":"10.3390/biotech14010015","DOIUrl":"https://doi.org/10.3390/biotech14010015","url":null,"abstract":"<p><p>Systemic lupus erythematosus (SLE) is a complex autoimmune disease that poses serious long-term patient burdens. <b>(1)</b> Background: SLE patient classification and care are often complicated by case heterogeneity (diverse variations in symptoms and severity). Large language models (LLMs) and generative artificial intelligence (genAI) may mitigate this challenge by profiling medical records to assess key medical criteria. <b>(2)</b> Methods: To demonstrate genAI-based profiling, ACR (American College of Rheumatology) 1997 SLE classification criteria were used to define medically relevant LLM prompts. Records from 78 previously studied patients (45 classified as having SLE; 33 indeterminate or negative) were computationally profiled, via five genAI replicate runs. <b>(3)</b> Results: GenAI determinations of the \"Discoid Rash\" and \"Pleuritis or Pericarditis\" classification criteria yielded perfect concurrence with clinical classification, while some factors such as \"Immunologic Disorder\" (56% accuracy) were statistically unreliable. Compared to clinical classification, our genAI approach achieved a 72% predictive success rate. <b>(4)</b> Conclusions: GenAI classifications may prove sufficiently predictive to aid medical professionals in evaluating SLE patients and structuring care strategies. For individual criteria, accuracy seems to correlate inversely with complexities in clinical determination, implying that improvements in AI patient profiling tools may emerge from continued advances in clinical classification efficacy.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11940096/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144030765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-02-27DOI: 10.3390/biotech14010014
Anna Panozzo, Pranay Kumar Bolla, Giuseppe Barion, Alessandro Botton, Teofilo Vamerali
{"title":"Phytohormonal Regulation of Abiotic Stress Tolerance, Leaf Senescence and Yield Response in Field Crops: A Comprehensive Review.","authors":"Anna Panozzo, Pranay Kumar Bolla, Giuseppe Barion, Alessandro Botton, Teofilo Vamerali","doi":"10.3390/biotech14010014","DOIUrl":"https://doi.org/10.3390/biotech14010014","url":null,"abstract":"<p><p>Field crops are expected to be increasingly threatened by climate change, which will negatively impact plant development, growth and yield. Phytohormones play a crucial role in regulating specific signalling pathways to induce rapid adaptive responses to environmental stresses. Exogenous phytohormone application alters hormonal balance, thereby enhancing plant adaptation to adverse conditions. While several studies have advanced our understanding of the use of phytohormones in field crops, yield responses and species-specific application strategies remain inconsistent and rarely assessed under field conditions. The application of cytokinins (CKs), abscisic acid (ABA), and gibberellic acid (GA) has been shown to maintain prolonged photosynthetic activity, stabilize plasma membrane, and reduce lipid peroxidation and ion accumulation under salinity stress in wheat. Additionally, inhibitors of ethylene synthesis and receptors can mitigate stress symptoms under drought and heat stress, which typically accelerates senescence and shortens the grain-filling period in cereal crops. In this way, exogenous application of CKs, GA, and ethylene inhibitors can delay senescence by sustaining leaf photosynthetic activity and postponing nutrient remobilization. However, these benefits may not consistently translate into improvements in grain yield and quality. This review explores the molecular mechanisms of phytohormones in abiotic stress tolerance, delineates their specific functions and evaluates experimental findings from field applications. It also summarizes the potential of phytohormone applications in field crops, emphasizing the need for species-specific investigations on application timing and dosages under open-field conditions to optimize their agronomic potential.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11939854/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144017232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-02-25DOI: 10.3390/biotech14010013
Despoina P Kiouri, Georgios C Batsis, Thomas Mavromoustakos, Alessandro Giuliani, Christos T Chasapis
{"title":"Structure-Based Modeling of the Gut Bacteria-Host Interactome Through Statistical Analysis of Domain-Domain Associations Using Machine Learning.","authors":"Despoina P Kiouri, Georgios C Batsis, Thomas Mavromoustakos, Alessandro Giuliani, Christos T Chasapis","doi":"10.3390/biotech14010013","DOIUrl":"https://doi.org/10.3390/biotech14010013","url":null,"abstract":"<p><p>The gut microbiome, a complex ecosystem of microorganisms, plays a pivotal role in human health and disease. The gut microbiome's influence extends beyond the digestive system to various organs, and its imbalance is linked to a wide range of diseases, including cancer and neurodevelopmental, inflammatory, metabolic, cardiovascular, autoimmune, and psychiatric diseases. Despite its significance, the interactions between gut bacteria and human proteins remain understudied, with less than 20,000 experimentally validated protein interactions between the host and any bacteria species. This study addresses this knowledge gap by predicting a protein-protein interaction network between gut bacterial and human proteins. Using statistical associations between Pfam domains, a comprehensive dataset of over one million experimentally validated pan-bacterial-human protein interactions, as well as inter- and intra-species protein interactions from various organisms, were used for the development of a machine learning-based prediction method to uncover key regulatory molecules in this dynamic system. This study's findings contribute to the understanding of the intricate gut microbiome-host relationship and pave the way for future experimental validation and therapeutic strategies targeting the gut microbiome interplay.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11940256/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144050181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-02-25DOI: 10.3390/biotech14010012
Hwan-Hee Yu, Jong-Suk Park, Sanghyun Lee
{"title":"Enhanced Bioactive Compounds and Antioxidant Activity in Germinated Seeds of the New Peanut Variety.","authors":"Hwan-Hee Yu, Jong-Suk Park, Sanghyun Lee","doi":"10.3390/biotech14010012","DOIUrl":"https://doi.org/10.3390/biotech14010012","url":null,"abstract":"<p><p>The sprout market in Korea is expanding as consumers seek healthier food options and farmers strive to increase added value and competitiveness. This study examined the changes in the phytochemical composition of Sinpalkwang (SPK), a peanut variety developed in Korea, during germination. Four samples (SPK1, SPK2, SPK3, and SPK4) were collected at different growth stages and analyzed for total polyphenol content (TPC), total flavonoid content (TFC), and antioxidant activities using ABTS<sup>+</sup> and DPPH assays. The levels of <i>trans</i>-resveratrol and soyasaponin Bb were quantified using high-performance liquid chromatography (HPLC) with a photo-diode array (PDA). Among the samples, SPK2 exhibited the highest TFC (1.61 mg QE/g ext.) and <i>trans</i>-resveratrol content (0.054 mg/g ext.), while SPK4 showed the highest TPC (29.38 mg TAE/g ext.) and soyasaponin Bb content (6.543 mg/g ext.). In terms of radical scavenging activities, SPK2 and SPK3 performed best in the ABTS<sup>+</sup> and DPPH assays, respectively. Germinated samples demonstrated strong results across all analyses, highlighting the benefits of germination in enhancing phytochemical properties. This study provides foundational information on the phytochemical composition of SPK and the effects of germination. Future research will focus on optimizing germination conditions to further enhance the functionality and value of this Korean-bred variety as a source of high-value bioactive ingredients.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11940248/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144053047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-02-20DOI: 10.3390/biotech14010011
Silvestre Ruano-Rodríguez, Mar Navarro-Alonso, Benito Domínguez-Velasco, Manuel Álvarez-Dolado, Francisco J Esteban
{"title":"STXBP1 Syndrome: Biotechnological Advances, Challenges, and Perspectives in Gene Therapy, Experimental Models, and Translational Research.","authors":"Silvestre Ruano-Rodríguez, Mar Navarro-Alonso, Benito Domínguez-Velasco, Manuel Álvarez-Dolado, Francisco J Esteban","doi":"10.3390/biotech14010011","DOIUrl":"https://doi.org/10.3390/biotech14010011","url":null,"abstract":"<p><p>STXBP1 syndrome is a severe early-onset epileptic encephalopathy characterized by developmental delay and intellectual disability. This review addresses key challenges in STXBP1 syndrome research, focusing on advanced therapeutic approaches and experimental models. We explore gene therapy strategies, including CRISPR-Cas9, adeno-associated viral (AAV) vectors, and RNA therapies such as antisense oligonucleotides (ASOs), aimed at correcting STXBP1 genetic dysfunctions. This review presents in vivo and in vitro models, highlighting their contributions to understanding disease mechanisms. Additionally, we provide a proposal for a detailed bioinformatic analysis of a Spanish cohort of 41 individuals with STXBP1-related disorders, offering insights into specific mutations and their biological implications. Clinical and translational perspectives are discussed, emphasizing the potential of personalized medicine approaches. Future research directions and key challenges are outlined, including the identification of STXBP1 interactors, unexplored molecular pathways, and the need for clinically useful biomarkers. This comprehensive review underscores the complexity of STXBP1-related infantile epileptic encephalopathy and opens new avenues for advancing the understanding and treatment of this heterogeneous disease.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11939967/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144064389","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioTechPub Date : 2025-02-14DOI: 10.3390/biotech14010010
Jorge L Arias-Arias, Laura Monturiol-Gross, Eugenia Corrales-Aguilar
{"title":"A Live-Cell Imaging-Based Fluorescent SARS-CoV-2 Neutralization Assay by Antibody-Mediated Blockage of Receptor Binding Domain-ACE2 Interaction.","authors":"Jorge L Arias-Arias, Laura Monturiol-Gross, Eugenia Corrales-Aguilar","doi":"10.3390/biotech14010010","DOIUrl":"10.3390/biotech14010010","url":null,"abstract":"<p><p>Neutralization assays have become an important tool since the beginning of the COVID-19 pandemic for testing vaccine responses and therapeutic antibodies as well as for monitoring humoral immunity to SARS-CoV-2 in epidemiological studies. The spike glycoprotein (S) present on the viral surface contains a receptor binding domain (RBD) that recognizes the angiotensin-converting enzyme 2 receptor (ACE2) in host cells, allowing virus entry. The gold standard for determining SARS-CoV-2 neutralizing antibodies is the plaque reduction neutralization test (PRNT), which relies on live-virus replication performed exclusively in biosafety level 3 (BSL-3) laboratories. Here, we report the development of a surrogate live-cell imaging-based fluorescent SARS-CoV-2 neutralization assay, applicable to BSL-1 or BSL-2 laboratories, by antibody-mediated blockage of the interaction between recombinant RBD with overexpressed ACE2 receptor in a genetically modified HEK 293T stable cell line. Our approach was able to detect neutralizing antibodies both in COVID-19-positive human serum samples and polyclonal equine formulations against SARS-CoV-2. This new cell-based surrogate neutralization assay represents a virus-free fluorescence imaging alternative to the reported approaches, which can be used to detect antibody-neutralizing capabilities toward SARS-CoV-2. This assay could also be extrapolated in the future to other established and emergent viral agents.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843899/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143469467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}