Biotechnology Progress最新文献_第2页

5-Aza combined with VPA reprograms human T lineage acute leukemia Jurkat cells into B-cell-like cells by epigenetic activation of PAX5. 5-Aza联合VPA通过PAX5的表观遗传激活将人T系急性白血病Jurkat细胞重编程为b细胞样细胞。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-17 DOI: 10.1002/btpr.70023

Wenjin Xi, Guoxu Zheng, Xu Chen, Baile Zuo, Wei Wang, Yufang Li, Chunmei Zhang, Jie Chu, Xiuli Mu, Weihong Wen, Tao Wang, An-Gang Yang

{"title":"5-Aza combined with VPA reprograms human T lineage acute leukemia Jurkat cells into B-cell-like cells by epigenetic activation of PAX5.","authors":"Wenjin Xi, Guoxu Zheng, Xu Chen, Baile Zuo, Wei Wang, Yufang Li, Chunmei Zhang, Jie Chu, Xiuli Mu, Weihong Wen, Tao Wang, An-Gang Yang","doi":"10.1002/btpr.70023","DOIUrl":"https://doi.org/10.1002/btpr.70023","url":null,"abstract":"Epigenetic regulation plays an important role in cell fate reprogramming. Here, we found that inhibitors of epigenetic modifiers, including VPA, TSA, and 5-Aza-2'-deoxycytidine, can induce phenotypic transformation from Jurkat cells into B-cell-like cells. When Jurkat cells were treated with 5-Aza combined with VPA, B cell and stem cell marker expression was observed. These gene expression pattern changes were most remarkable in the optimized B cell induction conditions provided by the cocultured and genetically modified murine bone marrow OP9 cells. In such conditions, Jurkat cells were endowed with the ability to secrete B cell cytokines, and B lymphocyte-related genes and pathways were activated. In studying the mechanism underlying Jurkat cell reprogramming by 5-Aza and VPA, we found that PAX5, the key transcription factor regulating B cell development, was significantly upregulated. Treatment with 5-Aza and VPA inhibited the methylation of CpG islands and upregulated the acetylated H3K9 modification in the PAX5 promoter region, respectively, thus epigenetically activating the expression of PAX5 and promoting the reprogramming of Jurkat cells. Similar reprogramming results were also observed in primary CD4+T cells following treatment with 5-Aza and VPA. Our results provide a de novo paradigm for the reprogramming of T cells through epigenetic modifications.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70023"},"PeriodicalIF":2.5,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143647073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Predictive mechanistic model for separation of monoclonal antibody, fab fragment, and aggregate species on multimodal chromatography. 单克隆抗体、fab片段和聚集体的多模态色谱分离预测机制模型。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-17 DOI: 10.1002/btpr.70022

Lalita Kanwar Shekhawat, Todd Markle, Jean-Luc Maloisel, Gunnar Malmquist

{"title":"Predictive mechanistic model for separation of monoclonal antibody, fab fragment, and aggregate species on multimodal chromatography.","authors":"Lalita Kanwar Shekhawat, Todd Markle, Jean-Luc Maloisel, Gunnar Malmquist","doi":"10.1002/btpr.70022","DOIUrl":"https://doi.org/10.1002/btpr.70022","url":null,"abstract":"The specific selectivities offered by multimodal ligands drive the increased application of multimodal chromatography in the purification of complex new \"multispecific\" antibodies, which requires improved understanding of the protein-multimodal ligand interaction mechanism. In the present study, a mechanistic model is developed to predict monoclonal antibody (mAb1)-Fab fragment (Fab) and heterogeneous aggregates separation on Capto™ MMC ImpRes multimodal resin based on the general rate model coupled with the proposed preferential interaction (PI) analysis-based Langmuir non-linear binding model. The model input value of binding parameters is obtained from Perkin et al. developed PI model, fit to the characteristic 'U'-shaped curve for isocratic retention factors of mAb1, Fab, and aggregates as a function of NaCl salt concentrations. The model successfully simulates mAb1 and Fab elution peaks, whereas in the absence of deconvoluted peaks of heterogeneous aggregates, aggregates are modeled as a single species, giving satisfactory prediction of elution peak position, describing the average of the multiple (majority as double peaks) aggregate elution peaks. The physical significance of model estimated binding parameters is obtained from model estimated total number of released counter salt ions and water molecules for each species during binding, found to be consistent with their isocratic retention data. The underlying mechanism of double peak elution of aggregates during linear gradient elution was investigated based on mechanistic model estimated equilibrium constant. The proposed predictive mechanistic model was successfully validated by predicting mAb1, Fab, and aggregates elution peaks for the multimodal column operated in hydrophobic interaction mode and can be successfully implemented for process development.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70022"},"PeriodicalIF":2.5,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143647078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Improving the expression yield of recombinant adeno-associated virus serotype 2 using dimethyl sulfoxide DMSO as an additive to the triple transient transfection process of HEK293 cells. 二甲亚砜DMSO在HEK293细胞三重瞬时转染过程中提高重组腺相关病毒血清型2的表达量

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70017

Alexander Burns, Daniel Ramos-Sono, Saurav Datta

{"title":"Improving the expression yield of recombinant adeno-associated virus serotype 2 using dimethyl sulfoxide DMSO as an additive to the triple transient transfection process of HEK293 cells.","authors":"Alexander Burns, Daniel Ramos-Sono, Saurav Datta","doi":"10.1002/btpr.70017","DOIUrl":"https://doi.org/10.1002/btpr.70017","url":null,"abstract":"One of the widely used techniques for producing recombinant adeno-associated virus serotype 2 (rAAV2) particles, as viral vectors for gene therapy applications, is the triple transient (TT) transfection of human embryonic kidney 293 (HEK293) cells. It is desirable to optimize this transfection process for more efficient manufacturing of rAAV viral vectors for gene therapy purposes. We examined the application of dimethyl sulfoxide (DMSO) as an additive to this transfection technique to improve the expression yield of rAAV2 particles with HEK293 cells in adherent and suspension cell culture modalities. This assistance by DMSO should increase the trafficking of plasmid DNA (pDNA) through the cell membrane, and thus, increase the viral titer of rAAV2 full capsids at the time of harvesting the cell culture. The study demonstrated that DMSO as an additive for the TT transfection process led to an 8.2-fold increase in the expression yield of full AAV2 capsids using HEK293 cells in adherent cell culture modality, and also led to a 4.0-fold increase in the expression yield of full AAV2 capsids using HEK293 cells in suspension cell culture modality. There are no reported studies on the application of DMSO as an additive to the TT transfection process of HEK293 cells for the production of AAV particles. This is a novel, simple, and inexpensive method to improve the yield of rAAV2 full capsids with the TT transfection process of HEK293 cells, using a well-known cryoprotectant agent (CPA), as an additive to this transfection process.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70017"},"PeriodicalIF":2.5,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhancing cryopreservation of human induced pluripotent stem cells: Bottom-up versus conventional freezing geometry. 强化人诱导多能干细胞的低温保存：自下而上与传统冷冻几何的对比。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70019

Fernando Teodoro, Soukaina El-Guendouz, Rafaela Neves, Andreia Duarte, Miguel A Rodrigues, Eduardo P Melo

{"title":"Enhancing cryopreservation of human induced pluripotent stem cells: Bottom-up versus conventional freezing geometry.","authors":"Fernando Teodoro, Soukaina El-Guendouz, Rafaela Neves, Andreia Duarte, Miguel A Rodrigues, Eduardo P Melo","doi":"10.1002/btpr.70019","DOIUrl":"https://doi.org/10.1002/btpr.70019","url":null,"abstract":"Induced pluripotent stem cells (iPSCs) hold large potential in regenerative medicine due to their pluripotency and unlimited self-renewal capacity without the ethical issues of embryonic stem cells. To provide quality-controlled iPSCs for clinical therapies, it is essential to develop safe cryopreservation protocols for long-term storage, preferably amenable to scale-up and automation. We have compared the impact of two different freezing geometries (bottom-up and conventional radial freezing) on the viability and differentiation potential of human iPSCs. Our results demonstrate that bottom-up freezing under optimized conditions significantly increases iPSC viability, up to 9% for cell membrane integrity and up to 21% for cell metabolic state, compared to conventional freezing. The improvement achieved for bottom-up versus conventional freezing was maintained after scale-up from cryogenic vials to 30 mL bags, highlighting its potential for clinical applications. These findings show that bottom-up freezing can offer a more controlled and scalable cryopreservation strategy for iPSCs, promoting their application in regenerative medicine.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70019"},"PeriodicalIF":2.5,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Process optimization mitigated the retention loss of an Fc-fusion protein during ultrafiltration/diafiltration. 工艺优化减轻了超滤/滤除过程中fc融合蛋白的保留损失。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70021

Hao Yu, Li Fei

{"title":"Process optimization mitigated the retention loss of an Fc-fusion protein during ultrafiltration/diafiltration.","authors":"Hao Yu, Li Fei","doi":"10.1002/btpr.70021","DOIUrl":"https://doi.org/10.1002/btpr.70021","url":null,"abstract":"In the downstream processing of antibody-based therapeutics, ultrafiltration/diafiltration (UF/DF) is commonly applied for concentration and buffer exchange in the final formulation. For a given molecule, various factors such as membrane type, feed flux, and transmembrane pressure (TMP) can significantly influence the performance of UF/DF, impacting yield, buffer exchange efficiency, and product quality. Conventional membrane pore size selection is based on product molecular weight to ensure high retention. While working on an Fc-fusion protein, we found that the pH of load material had a critical effect on the retention of the molecule due to conformational changes at different pH values, as evidenced by the size-exclusion chromatography (SEC). Meanwhile, optimization of the UF/DF process underscored the importance of concentration polarization to protein retention. Approaches to reduce concentration polarization, such as increasing feed flux and lowering TMP, resulted in less protein loss in the permeate stream. High retention of this Fc-fusion protein during the UF/DF step can be achieved not only by utilizing a 5 kDa membrane but also by employing a 10 kDa membrane with optimized process parameters such as load conditions, feed flux, and TMP. These observations provide important insights on the factors impacting protein retention beyond the molecular weight cutoff (MWCO) of UF/DF membrane.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70021"},"PeriodicalIF":2.5,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603837","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhancing the performance of an in vitro RNA biosensor through iterative design of experiments. 通过实验迭代设计提高体外RNA生物传感器的性能。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70005

Rochelle Aw, Karen Polizzi

{"title":"Enhancing the performance of an in vitro RNA biosensor through iterative design of experiments.","authors":"Rochelle Aw, Karen Polizzi","doi":"10.1002/btpr.70005","DOIUrl":"https://doi.org/10.1002/btpr.70005","url":null,"abstract":"The quality control of RNA has become increasingly crucial with the rise of mRNA-based vaccines and therapeutics. However, conventional methods such as LC-MS often require specialized equipment and expertise, limiting their applicability to high throughput experiments. Here, we optimize a previously characterized RNA integrity biosensor, that provides a simple colorimetric output, using Design of Experiments (DoE). Through iterative rounds of a Definitive Screening Design (DSD) and experimental validation, we systematically explored different assay conditions to enhance the biosensor's performance. Optimization led to a 4.1-fold increase in dynamic range and reduced RNA concentration requirements by one-third, significantly improving usability. Notable modifications included reducing the concentrations of reporter protein and poly-dT oligonucleotide and increasing DTT concentration, suggesting a reducing environment for optimal functionality. Importantly, the optimized biosensor retained its ability to discriminate between capped and uncapped RNA even at lower RNA concentrations. Overall, our improved biosensor offers enhanced performance and reduced sample requirements, paving the way for rapid, cost-effective RNA quality control in diverse settings, including resource-limited environments.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70005"},"PeriodicalIF":2.5,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of multiparametric bioprinting method for generation of 3D printed cell-laden structures. 3D打印细胞负载结构的多参数生物打印方法的发展。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70016

Sophie Lipshutz, Yoontae Kim, Micaila Curtis, Leanne Friedrich, Stella Alimperti

{"title":"Development of multiparametric bioprinting method for generation of 3D printed cell-laden structures.","authors":"Sophie Lipshutz, Yoontae Kim, Micaila Curtis, Leanne Friedrich, Stella Alimperti","doi":"10.1002/btpr.70016","DOIUrl":"https://doi.org/10.1002/btpr.70016","url":null,"abstract":"The organ transplantation field requires new approaches for replacing and regenerating tissues due to the lack of adequate transplant methods. Three-dimensional (3D) extrusion-based bioprinting is a rapid prototyping approach that can engineer 3D scaffolds for tissue regeneration applications. In this process, 3D printed cell-based constructs, consisting of biomaterials, growth factors, and cells, are formed by the extrusion of bioinks from nozzles. However, extrusion applies shear stresses to cells, often leading to cellular damage or membrane rupture. To address this limitation, herein, we developed and optimized a 3D bioprinting approach by evaluating the effect of key extrusion-based 3D bioprinting parameters-bioink viscosity, nozzle size, shape, and printing speed-on cell viability. Our results revealed that cells printed in higher-viscosity bioinks, with smaller, cylindrical nozzles, exhibited lower viability due to their exposure to high shear stresses. Translational flow speed had a cell-dependent impact, as different cell types have different sensitivities to the magnitude and duration of shear stress inside the nozzle. Overall, evaluating these parameters could facilitate the development of 3D high-resolution bioprinted constructs for tissue regeneration applications, offering a more efficient alternative to traditional fabrication methods, which are often labor intensive, expensive, and repetitive.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70016"},"PeriodicalIF":2.5,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Assessing the probability of clonality achieved by single-cell cloning of CHO cells through cell deposition combined with imaging using distinguishable cells. 通过细胞沉积结合可区分细胞成像，评估CHO细胞单细胞克隆获得克隆性的可能性。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-04 DOI: 10.1002/btpr.70012

Genevieve H Nonet, Elena Scut, Raymond Ogawa, Milan T Tomic

{"title":"Assessing the probability of clonality achieved by single-cell cloning of CHO cells through cell deposition combined with imaging using distinguishable cells.","authors":"Genevieve H Nonet, Elena Scut, Raymond Ogawa, Milan T Tomic","doi":"10.1002/btpr.70012","DOIUrl":"https://doi.org/10.1002/btpr.70012","url":null,"abstract":"Mammalian cell lines used for clinical studies and post-approval production of recombinant DNA-derived biotherapeutics are expected to be derived from a single cell, and regulatory submissions are expected to provide robust evidence of monoclonality. Imaged single-cell deposition followed by whole-well imaging using specialized instruments has, in many cell line development labs, replaced the \"gold standard\" of two rounds of limiting dilution due to its increased speed and the assurance of clonality provided by orthogonal images. However, there is still a lack of information on how the procedures used to define these clonal cell lines perform. Here we use a mixture of two distinguishable Chinese hamster ovary (CHO) cells to document that a greater than 99% probability of clonality can be obtained from our single-cell cloning method that uses our preparation procedures, the VIPS® single-cell deposition instrument, the Cell Metric® whole-well imager, and a comprehensive visual review. Together with the assurance of cell/well images, the determination of the probability of clonality of our VIPS+Cell Metric method provides a strong package of evidence of single-cell derivation of a recombinant CHO cell line.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70012"},"PeriodicalIF":2.5,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143540029","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Efficient expression of recombinant proteins in Bacillus subtilis using a rewired gene circuit of quorum sensing. 重组蛋白在枯草芽孢杆菌群体感应基因回路中的高效表达。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70007

Wenliang Hao, Shihao Yang, Yuou Sheng, Chengfeng Ye, Laichuang Han, Zhemin Zhou, Wenjing Cui

{"title":"Efficient expression of recombinant proteins in Bacillus subtilis using a rewired gene circuit of quorum sensing.","authors":"Wenliang Hao, Shihao Yang, Yuou Sheng, Chengfeng Ye, Laichuang Han, Zhemin Zhou, Wenjing Cui","doi":"10.1002/btpr.70007","DOIUrl":"https://doi.org/10.1002/btpr.70007","url":null,"abstract":"Bacillus subtilis is a favored chassis for high productivity of several high value-added product in synthetic biology. Efficient production of recombinant proteins is critical but challenging using this chassis because these expression systems in use, such as constitutive and inducible expression systems, demand for coordination of cell growth with production and addition of chemical inducers. These systems compete for intracellular resources with the host, eventually resulting in dysfunction of cell survival. To overcome the problem, in this study, LuxRI quorum sensing (QS) system from Aliivibrio fischeri was functionally reconstituted in B. subtilis for achieving coordinated protein overproduction with cell growth in a cell-density-dependent manner. Furthermore, the output-controlling promoter, PluxI, was engineered through two rounds of evolution, by which we identified four mutants, P22, P47, P56, and P58 that exhibited elevated activity compared to the original PluxI. By incorporating a strong terminator (TB5) downstream of the target gene further enhanced expression level. The expression level of this system surpasses commonly used promoter-based systems in B. subtilis like P43 and PylbP. The LuxRI QS system proves to be a potent platform for recombinant protein overproduction in B. subtilis.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70007"},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448011","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Microcontact printing of lectin self-assembled monolayers for arbovirus detection. 用于虫媒病毒检测的凝集素自组装单层微接触印刷。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70008

Raiza P S Lucena, Alberto G Silva-Junior, Isaac A M Frías, Laura H V Gil, Marli T Cordeiro, Abdelhamid E El Salhi, Cesar A S Andrade, Maria D L Oliveira

{"title":"Microcontact printing of lectin self-assembled monolayers for arbovirus detection.","authors":"Raiza P S Lucena, Alberto G Silva-Junior, Isaac A M Frías, Laura H V Gil, Marli T Cordeiro, Abdelhamid E El Salhi, Cesar A S Andrade, Maria D L Oliveira","doi":"10.1002/btpr.70008","DOIUrl":"https://doi.org/10.1002/btpr.70008","url":null,"abstract":"Arboviruses significantly burden public health in Brazil, constituting a constant challenge for health authorities. The diagnosis and, consequently, clinical management and the reporting of arbovirus infections in regions where multiple arboviruses coexist are complex processes. Herein, we report the development of a new electrochemical biosensor based on Concanavalin A (ConA) to identify carbohydrate patterns in the viral structure of Dengue 3 (DENV-3), Zika (ZIKV) and Chikungunya (CHIKV) viruses. The biorecognition of arboviruses was carried out through functionalization with 4-aminophenylacetic acid (CMA) on poly (ethylene terephthalate) (PET) substrate coated with a gold layer combining microcontact printing (μCP). Bovine serum albumin (BSA) was used after ConA immobilization to block binding to nonspecific sites. Subsequently, the interaction between ConA and arbovirus was characterized by standard atomic force microscopy (AFM), fluorescence microscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). Fluorescent imaging was conducted to confirm the occurrence of the DENV-3, ZIKV, and CHIKV detection processes. The obtained results demonstrated the success of the biosensor (CMA-ConA-BSA) manufactured on a PET substrate using μCP for detecting medically significant arboviruses. RCT values showed an increase in impedimetric response total of the system after exposition to DENV-3 (RCT = 68.82 kΩ) and a lower recognition to CHIKV (RCT = 44.44 kΩ). The present biosensor platform reveals the applicability of the ConA lectin in the viral biorecognition process based on flexible biosensors for differential detection of DENV-3, ZIKV, and CHIKV. ConA-based electrochemical biosensor provide high selectivity, real-time detection, and low volumes of analytes.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70008"},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0