Biotechnology and Bioengineering最新文献_第2页

Biomanufacturing Potential of Streamlined Cells. 流线型细胞的生物制造潜力。

IF 3.6 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-25 DOI: 10.1002/bit.70070

Alvaro R Lara, Marie B Andersen, Alexander A V Madsen, Kathrine Gravlund Fønss, Marta Irla, Hilal Taymaz-Nikerel, Luz María Martínez, Max Daniel Dicke, Marcel Mann, Jørgen B Magnus, Guillermo Gosset

{"title":"Biomanufacturing Potential of Streamlined Cells.","authors":"Alvaro R Lara, Marie B Andersen, Alexander A V Madsen, Kathrine Gravlund Fønss, Marta Irla, Hilal Taymaz-Nikerel, Luz María Martínez, Max Daniel Dicke, Marcel Mann, Jørgen B Magnus, Guillermo Gosset","doi":"10.1002/bit.70070","DOIUrl":"https://doi.org/10.1002/bit.70070","url":null,"abstract":"<p><p>A series of Escherichia coli streamlined strains was developed by removing the expression of genes encoding extracellular structures and unessential enzymes. The streamlined strains exhibited improved metabolic performance, including lower overflow metabolism and ATP maintenance coefficient, as well as a higher growth rate, compared to their parental strain. The intracellular levels of ATP were monitored using a genetic sensor, showing the improved resource stewardship of the streamlined cells. The streamlined strains were tested as cell factories to produce plasmid DNA (pDNA) in batch cultures, exhibiting a 23% increase in the specific pDNA production rate, compared to the parental strain. Recombinant protein expression was evaluated in microbioreactors in batch and fed-batch modes. In batch mode, recombinant protein yield from biomass was up to 82% higher in the streamlined strains than in the parental strain. Furthermore, in fed-batch mode, the recombinant protein yield was 79% greater in the streamlined cells compared to the parental strain. Our results show the benefits of reducing cellular complexity on the biomanufacturing of pDNA and recombinant proteins in culture schemes typical of industrial settings.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145136581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Investigation of Oxidative Damage Mechanism of Chlorella sorokiniana by Gas-Liquid Phase Plasma. 气液相等离子体对小球藻氧化损伤机制的研究。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-24 DOI: 10.1002/bit.70071

Dong-Ao Li,Guang-Hui Xu,Jie Shen,Hong Zhang

{"title":"Investigation of Oxidative Damage Mechanism of Chlorella sorokiniana by Gas-Liquid Phase Plasma.","authors":"Dong-Ao Li,Guang-Hui Xu,Jie Shen,Hong Zhang","doi":"10.1002/bit.70071","DOIUrl":"https://doi.org/10.1002/bit.70071","url":null,"abstract":"This study explores the mechanisms of oxidative damage in Chlorella sorokiniana under continuous gas-liquid phase plasma exposure. The oxidative damage mechanism of C. sorokiniana was revealed through multiple analytical techniques, including scanning electron microscopy (SEM), flow cytometry (FCM), and fluorescence excitation-emission matrix (EEM). Plasma-induced damage to algal cell membranes, causing seepage of photosynthetic pigments and dissolved organic matter; chlorophyll fluorescence changes confirmed damage to the photosynthetic system. Alterations in MDA, intracellular ROS/NO, and antioxidant enzyme activities reflect lipid peroxidation and oxidative stress levels. These results show that plasma induces oxidative damage to algal membranes, intracellular components, and photosystems via reactive oxygen and nitrogen species (RONS). C. sorokiniana activates its antioxidant defense system under stress. This gradual, cumulative damage exerts distinct biological effects at different times. The findings support the precise control of low-temperature plasma for beneficial microorganisms and provide a basis for advancing microalgae transformation technologies.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"35 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145127024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Succinic Acid Production by Engineered Mannheimia succiniciproducens and Its Use in Chemoenzymatic Poly(Butylene Succinate) Synthesis. 工程琥珀酸Mannheimia产琥珀酸及其在化学酶合成聚琥珀酸丁二烯中的应用。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-24 DOI: 10.1002/bit.70072

Ji Yeon Kim,Jong An Lee,Gi Bae Kim,Youngjoon Lee,Sang Yup Lee

{"title":"Succinic Acid Production by Engineered Mannheimia succiniciproducens and Its Use in Chemoenzymatic Poly(Butylene Succinate) Synthesis.","authors":"Ji Yeon Kim,Jong An Lee,Gi Bae Kim,Youngjoon Lee,Sang Yup Lee","doi":"10.1002/bit.70072","DOIUrl":"https://doi.org/10.1002/bit.70072","url":null,"abstract":"Amid growing concerns about climate change, environmental pollution, and fossil resource depletion, bio-based chemicals and materials have garnered significant attention. Succinic acid (SA), a four-carbon dicarboxylic acid, is a key platform chemical for producing industrially important compounds such as poly(butylene succinate) (PBS), a biodegradable polymer. In this paper, we demonstrate the enhanced production of SA using metabolically engineered Mannheimia succiniciproducens PALK (pMS3-gltA) strain, which achieves high SA yields through enhanced biomass production and optimized metabolic pathways. High-purity SA, purified from the fermentation broth, was used in an environmentally friendly two-step polymerization process under mild conditions. In the first step, diethyl succinate was synthesized from SA and ethanol using Amberlyst as a catalyst. This was followed by polymerization with 1,4-butanediol, catalyzed by Candida antarctica lipase B. The resulting PBS exhibited comparable thermal stability and molecular weight distribution to its petrochemical counterpart, with enhanced biodegradability. By integrating metabolic engineering with sustainable polymer synthesis, this study underscores the potential for an eco-friendly approach to developing bio-based polymers, offering a promising solution to the environmental challenges posed by conventional plastics.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"58 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145127165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Metabolic Engineering of Streptomyces leeuwenhoekii C34T to Increase Chaxamycin Production Based on the iVR1007 Genome-Scale Model. 基于iVR1007基因组尺度模型的列文赫氏链霉菌C34T代谢工程提高沙霉素产量

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-16 DOI: 10.1002/bit.70067

Anett Rubio,Valeria Razmilic,Stephanie Brain-Isasi,Barbara Andrews,Juan A Asenjo

{"title":"Metabolic Engineering of Streptomyces leeuwenhoekii C34T to Increase Chaxamycin Production Based on the iVR1007 Genome-Scale Model.","authors":"Anett Rubio,Valeria Razmilic,Stephanie Brain-Isasi,Barbara Andrews,Juan A Asenjo","doi":"10.1002/bit.70067","DOIUrl":"https://doi.org/10.1002/bit.70067","url":null,"abstract":"Streptomyces leeuwenhoekii C34T produces the polyketides chaxamycins A-D and chaxalactins A-C. The former metabolites exhibit antibiotic activity and antitumoral properties. In this study, we selected a subset of overexpression targets, predicted using the FSEOF algorithm, to increase chaxamycin production based on the iVR1007 genome-scale model (GEM). Eight genes were individually cloned under strong constitutive promoters and introduced into the genome of S. leeuwenhoekii C34T, including ugp, tkt, tal, and the acc complex, giving rise to strains S1-S8. The production of chaxamycins by these engineered strains was compared to the wild-type and to a strain overexpressing the pcc complex (S9) (not predicted as target by the model). Chaxamycins were detected in all the samples, with higher amounts observed in strains S1 through S8. In contrast, strain S9 did not show improved chaxamycin A-C production. The best results were obtained with strain tkt (S2), tal (S3), and mcm/mce (S8), which demonstrated 7.41-, 3.67-, and 3.33-fold increases in total chaxamycin production, respectively. These strains overexpress genes associated with AHBA synthesis, pentose phosphate pathway or (2S)-methyl-malonyl-CoA biosynthesis. Extracts from higher producers also showed higher bioactivity against Micrococcus luteus. Thus, the predictions made using the iVR1007 model, were successfully applied to increase chaxamycin production, further validating the utility of the FSEOF algorithm for predicting metabolic engineering targets.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"53 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145068226","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

A Biotechnological Platform Based on the Newly Isolated Streptococcus thermophilus D4 to Obtain Viable Biomass and Exopolysaccharides for Enterocytes Wound Healing. 基于新分离的嗜热链球菌D4获得肠细胞伤口愈合活性生物量和胞外多糖的生物技术平台。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-15 DOI: 10.1002/bit.70063

Alberto Alfano,Darshankumar Parecha,Sergio D'ambrosio,Maria d'Agostino,Donatella Cimini,Chiara Schiraldi

{"title":"A Biotechnological Platform Based on the Newly Isolated Streptococcus thermophilus D4 to Obtain Viable Biomass and Exopolysaccharides for Enterocytes Wound Healing.","authors":"Alberto Alfano,Darshankumar Parecha,Sergio D'ambrosio,Maria d'Agostino,Donatella Cimini,Chiara Schiraldi","doi":"10.1002/bit.70063","DOIUrl":"https://doi.org/10.1002/bit.70063","url":null,"abstract":"There is growing interest in newly isolated lactic acid bacteria from natural sources and traditional food manufacturing and transformation. Streptococcus thermophilus is able to improve the safety of dairy products, showing antioxidant and antimicrobial properties and also to produce bioactive molecules, such as exopolysaccharides (EPSs), usable in foods applications. An integrated bioprocess development was performed using S.thermophilus D4, for the production of viable biomass and EPSs using a media optimized through design of experiments. Plackett-Burman screening method was used twice to optimize biomass and exopolysaccharide production. Different fermentation strategies (batch and fed-batch) were performed to increase yield and productivity. Fed-batch processes increased biomass production approximately 2.5- and 7.5-fold than batch processes with optimized and M17 media, respectively. Furthermore, a significant increase of 1.7- and 6-fold EPS biosynthesis was observed. Finally downstream processes based on membranes were conducted to purify bioactive molecules (EPSs) used on Caco-2 cells showing a significant effect in accelerating wound healing. Finally, purification and recovery of EPSs tested on Caco-2 cells. The results obtained confirm the potential of newly defined medium to replace M17, which has animal-derived nitrogen sources, for probiotic biomass and EPSs production with many applications in different fields.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"169 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145059012","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

M/G-Ratio-Tuned Calcium Alginate Hydrogel Microcarrier for Controlled Delivery of Adeno-Associated Virus for Gene Therapy. 调节M/ g比的海藻酸钙水凝胶微载体用于控制腺相关病毒的基因治疗。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-14 DOI: 10.1002/bit.70066

Aiki Hioki,Shuhei Takatsuka,Yuta Kurashina,Hiroaki Onoe

{"title":"M/G-Ratio-Tuned Calcium Alginate Hydrogel Microcarrier for Controlled Delivery of Adeno-Associated Virus for Gene Therapy.","authors":"Aiki Hioki,Shuhei Takatsuka,Yuta Kurashina,Hiroaki Onoe","doi":"10.1002/bit.70066","DOIUrl":"https://doi.org/10.1002/bit.70066","url":null,"abstract":"This paper proposes the controlled release of adeno-associated virus (AAV) by polymer mesh structures of alginate hydrogel microbeads (gel-beads). The polymer mesh structure of alginate hydrogel can be controlled by adjusting the number of cross-linking points depending on the molecular structure (M/G ratio) of the alginate hydrogel. Scanning electron microscopic analysis on the mesh structures of alginate hydrogels with M/G ratios of 0.96 (MG-0.96) and 0.42 (MG-0.42) indicates that the mesh size distribution of MG-0.42 hydrogel was smaller than that of MG-0.96 hydrogel. Using these characteristics, controlled release of AAVs (AAV-1 or AAV-5) from M/G-ratio-tuned gel-beads (MG-0.96 or MG-0.42), Janus-shaped gel-beads consisting of hemispheres of the MG-0.96 and MG-0.42, and MG-0.64 gel-beads consisting of a mixture of MG-0.96 and MG-0.42 were examined with ELISA for AAV capsids and the GFP fluorescent signals from the transfected cells. The precise control of AAV release by the mesh structure of the alginate hydrogels could be an effective method for creating AAV micro-carriers for gene therapy.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"52 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145058961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Gold Nanoparticles Coated With Hydrophobin-ProteinA Fusion Protein: Development of a Versatile Immunosensing Platform. 包被疏水蛋白融合蛋白的金纳米颗粒：一种多功能免疫传感平台的开发。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-13 DOI: 10.1002/bit.70069

Paola Cicatiello,Bartolomeo Della Ventura,Giulia Fichera,Raffaele Velotta,Paola Giardina,Alessandra Piscitelli

{"title":"Gold Nanoparticles Coated With Hydrophobin-ProteinA Fusion Protein: Development of a Versatile Immunosensing Platform.","authors":"Paola Cicatiello,Bartolomeo Della Ventura,Giulia Fichera,Raffaele Velotta,Paola Giardina,Alessandra Piscitelli","doi":"10.1002/bit.70069","DOIUrl":"https://doi.org/10.1002/bit.70069","url":null,"abstract":"Advancing immunosensing technologies hinges on the development of next-generation surface functionalization methods, as the precise anchoring of antibodies on transducer interface is essential for achieving high sensitivity and selectivity. Among the diverse methodologies explored, bioengineered materials have shown significant potential to improve antibody orientation, stability, and functional performance. In this study, we present a chimeric protein created by fusing the adhesive Class I hydrophobin Vmh2 from Pleurotus ostreatus, with the Fc-binding region of protein A from Staphylococcus aureus (SpA). This fusion protein spontaneously adheres on gold nanoparticles (AuNPs) without requiring chemical modification, forming a robust bio-interactive layer for antibody attachment. The platform's adaptability and effectiveness were assessed using an immunoglobulin specific to a fungal laccase to establish the performance of the system, and antibodies against two clinically significant targets- mesothelin, a tumor-associated glycoprotein, and the SARS-CoV-2 spike protein- to showcase the diagnostic potential of the system. A two-step method based on the induced aggregation of the AuNps not bound to the analyte allows underscoring the platform's promise in biosensing applications. Overall, this approach represents a sustainable, versatile, and low-cost route for fabricating biologically active surfaces, with wide-ranging relevance in medical diagnostics, environmental analysis, and biotechnological innovation.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"21 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145043477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Evaluation of Solid-State Fermentation Potential of Sweet Sorghum Stalk by Heyndrickxia coagulans (Bacillus coagulans) L-2 Based on Complete Genome and Phenotypic Analysis. 基于全基因组和表型分析的凝固芽孢杆菌L-2对甜高粱秸秆固态发酵潜力的评价

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-11 DOI: 10.1002/bit.70065

Li Wei,Qian Lu,Hongshen Li,Dong Wei,Xinxin Zhang,Jia Ouyang,Chunying Li,Shizhong Li

{"title":"Evaluation of Solid-State Fermentation Potential of Sweet Sorghum Stalk by Heyndrickxia coagulans (Bacillus coagulans) L-2 Based on Complete Genome and Phenotypic Analysis.","authors":"Li Wei,Qian Lu,Hongshen Li,Dong Wei,Xinxin Zhang,Jia Ouyang,Chunying Li,Shizhong Li","doi":"10.1002/bit.70065","DOIUrl":"https://doi.org/10.1002/bit.70065","url":null,"abstract":"Heyndrickxia coagulans is the dominant strain in L-lactic acid fermentation. To promote the development of green and low-cost L-lactic acid fermentation processes, it is crucial to understand their solid-state fermentation (SSF) performance in lignocellulose raw materials. This study presents the first whole genome analysis of H. coagulans L-2 (single chromosome, 3.58 Mb, 46.28% GC content) to evaluate its SSF potential using sweet sorghum stalk. COG and KEGG annotations revealed that H. coagulans L-2 metabolizes diverse hexoses and pentoses for L-lactic acid production, and the key enzymatic sequences in carbohydrate metabolism exhibited high similarity with other Heyndrickxia strains. Comparative genomics with ATCC7050 demonstrated 70% genome similarity (2148 collinear CDSs), while L-2 has a more comprehensive carbohydrate metabolism network, especially in pentose metabolism pathways, such as arabinose, mannose, and rhamnose. Through the result of batch solid fermentation, H. coagulans L-2 could promote the re-fermentation of L-lactic acid in silage and reduce heterotypic lactic acid fermentation (lactic acid/ethanol ratio increased from 3.890 to 8.103), which indicated that the strain L-2 played a key role in the SSF of sweet sorghum silage and promoted the production of bio-based materials precursors of sweet sorghum. This study provides a new strategy for the high-value utilization of lignocellulosic biomass.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"38 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145035699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Pseudoxanthomonas taiwanensis and Pseudomonas thermotolerans: Novel Thermophilic Amylase Producers in Solid-State Fermentation. 台湾假黄单胞菌和耐热假单胞菌：固态发酵中新的嗜热淀粉酶产生菌。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-10 DOI: 10.1002/bit.70058

Anna Taulé,Teresa Gea,Alejandra Cerda,Esther Molina-Peñate

{"title":"Pseudoxanthomonas taiwanensis and Pseudomonas thermotolerans: Novel Thermophilic Amylase Producers in Solid-State Fermentation.","authors":"Anna Taulé,Teresa Gea,Alejandra Cerda,Esther Molina-Peñate","doi":"10.1002/bit.70058","DOIUrl":"https://doi.org/10.1002/bit.70058","url":null,"abstract":"Amylases are key industrial enzymes, and thermostable variants are particularly valuable for robust bioprocessing. This study investigates amylase production via solid-state fermentation (SSF) using non-sterile potato peel as substrate, comparing the performance of the autochthonous microbial population with that of the inoculated fungus Thermomyces lanuginosus. Non-inoculated batch reactors reached maximum productivities of 3920 U g-¹DM day-¹, more than double of the inoculated ones (1823 U g-¹DM day-¹), highlighting the potential of native thermophiles. Sequential batch reactor (SBR) strategies were applied to promote microbial selection and monitor T. lanuginosus persistence over seven cycles. Although the inoculum initially increased overall microbial activity (sOUR), T. lanuginosus was not detected, and no sustained improvement in amylase productivity was observed. Three thermophilic amylase-producing strains were isolated: Bacillus coagulans, Pseudoxanthomonas taiwanensis, and Pseudomonas thermotolerans, the last two being newly reported amylase producers. These findings demonstrate that efficient, scalable amylase production can be achieved through non-sterile SSF relying on native microbial communities, supporting circular bioeconomy strategies and potentially reducing the need for external inoculation. Further work is needed to confirm the generalizability of these results and to better understand the interactions between inoculated and native strains.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"17 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145032177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Elucidating the Interplay Between Resin Media, Feed Constituents, and Process Conditions in AAV Affinity Column Fouling With Confocal Microscopy. 用共聚焦显微镜研究树脂介质、进料成分和工艺条件在AAV亲和柱污染中的相互作用。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-09-10 DOI: 10.1002/bit.70060

Harshal Soni,Sergey Pryshchep,Thomas F Johnson,Steven M Cramer

{"title":"Elucidating the Interplay Between Resin Media, Feed Constituents, and Process Conditions in AAV Affinity Column Fouling With Confocal Microscopy.","authors":"Harshal Soni,Sergey Pryshchep,Thomas F Johnson,Steven M Cramer","doi":"10.1002/bit.70060","DOIUrl":"https://doi.org/10.1002/bit.70060","url":null,"abstract":"In this work, confocal microscopy is employed to study the loading and fouling behavior in AAV affinity resins as well as the implications of resin reuse with several commercial chromatographic materials and feed mixtures. Resin samples are obtained from both batch and column experiments, and confocal microscopy is carried out to examine the adsorption profiles in the beads after loading, wash, elution, and CIP steps. A comparison of PSDVB-based POROS CaptureSelect (PCS) AAV resins with agarose-based AVIPure AAV9 resins revealed distinct differences in both AAV transport and resin fouling. While AAVs were able to fully access the entire PCS AAV resin under loading onto virgin resin materials, they were restricted to the surface region of the AVIPure AAV9 resin. High resolution X-ray CT scans indicate that the large pores of the PCS media are likely responsible for this enhanced transport. However, AAV transport into full PCS AAV resin volume was found to only occur during the first AAV loading experiment and was not observed when clarified lysate was employed or upon resin reuse. Further, confocal microscopy indicated that fouling of the PCS AAV affinity resins occurred due to deposition and carryover of AAV and residual impurities upon column re-use, likely due to incomplete elution and ineffective column CIP. Interestingly, this residual binding in the region near the resin surface resulted in limiting solute transport in subsequent cycles, likely due to pore occlusion. In contrast, AAV bound to the AVIPure resin was readily eluted and removed during CIP. This study elucidates the impact of resin characteristics, feed constituents, and process conditions on the lifetime and fouling of AAV affinity columns.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"48 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145026021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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