Biotechnology Journal最新文献_第10页

Real-Time Adaptive Inline Acidification Enhances Continuous pH Control for Viral Inactivation 实时自适应内联酸化增强了病毒灭活的连续 pH 值控制。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-11-08 DOI: 10.1002/biot.202400456

Jia Sheng Zach Lee, Tan Dai Nguyen, Zi Ying Zheng, Wei Zhang, Dan Liu

{"title":"Real-Time Adaptive Inline Acidification Enhances Continuous pH Control for Viral Inactivation","authors":"Jia Sheng Zach Lee, Tan Dai Nguyen, Zi Ying Zheng, Wei Zhang, Dan Liu","doi":"10.1002/biot.202400456","DOIUrl":"10.1002/biot.202400456","url":null,"abstract":"<div>\u0000 \u0000 Existing low pH viral inactivation methods for continuous downstream processing of biologics typically rely on predictive models to estimate the necessary pH adjustments. However, these methods are of limited use during the process development stage due to the dynamic nature of capture chromatography, where batch variations can alter the eluted protein titer. This study introduces an inline viral inactivation system (IVIS) that utilizes real-time adaptive control and inline sensor readings to precisely regulate the pH manipulation for inline acidification and continuous viral inactivation. The IVIS, which includes a coiled flow inversion reactor (CFIR), is integrated with a multicolumn capture chromatography system to demonstrate a fully continuous process from protein capture chromatography to inline pH manipulation. The system achieved precise inline pH manipulation within ±0.15 and a narrow residence time distribution of 13.5 min with a relative width of 0.7. The introduction of real-time inline pH manipulation with the IVIS signifies a notable advancement in managing critical process parameters (CPPs) and ensuring consistent product quality across varied production environments for continuous downstream bioprocessing.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 11","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602158","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

Chromatin Accessibility Plays an Important Epigenetic Role on Antibody Expression From CMV Promoter and DNA Elements Flanking the CHO TI Host Landing-Pad CMV启动子和CHO TI宿主着陆垫侧翼DNA元件的染色质可及性对抗体表达起着重要的表观遗传作用

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-29 DOI: 10.1002/biot.202400487

Kavya Ganapathy, Andrew McKay, Steffen Durinck, Minyi Shi, Kristel Dorighi, Cynthia Lam, Yuxin Liang, Amy Shen, Gavin Barnard, Shahram Misaghi

{"title":"Chromatin Accessibility Plays an Important Epigenetic Role on Antibody Expression From CMV Promoter and DNA Elements Flanking the CHO TI Host Landing-Pad","authors":"Kavya Ganapathy, Andrew McKay, Steffen Durinck, Minyi Shi, Kristel Dorighi, Cynthia Lam, Yuxin Liang, Amy Shen, Gavin Barnard, Shahram Misaghi","doi":"10.1002/biot.202400487","DOIUrl":"10.1002/biot.202400487","url":null,"abstract":"<div>\u0000 \u0000 Targeted integration (TI) Chinese hamster ovary (CHO) platforms are commonly used for protein expression. However, the impact of epigenetic modifications on protein expression in TI cell lines remains elusive since almost all the epigenetic studies focus on random integration (RI) of the gene of interest and only within the promoter region. To address the impact of epigenetic modifications on TI CHO cells, we utilized a standard mAb-1 to identify and characterize TI clones with the same transgene copy numbers but different levels of transgene transcription and titer. Surprisingly, while CMV promoters were not methylated and histone acetylation/methylation was present, these epigenetic markers did not trend with mRNA transcription and protein expression in our TI model. Instead, ATAC-seq data analysis revealed that differences in chromatin accessibility within the TI site could be a major factor impacting these observed differences. However, neither chromatin accessibility nor histone acetylation/methylation profiles in early cultures were predictive of high-expressing clones early during the CLD process. Finally, modulation of the histone profiles (H3K27ac and H3K4me3) at the CMV promoters within the TI integration site using dCas9 fusion proteins was not effective in further increasing mAb titers which could have been likely due to interference of the dCas9 fusion proteins with transcription from the CMV promoters. Overall, our data suggests increasing chromatin accessibility at the TI site is the most effective way to increase mRNA transcription and hence, productivity in TI cell lines.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 10","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542311","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

Preparation of Thermosensitive Lignocellulose Hollow Fiber Membrane Grafted With PNIPAAm and Its Application as a Cell Culture Carrier in a RSOC Dynamic Culture 接枝 PNIPAAm 的热敏木质纤维素中空纤维膜的制备及其在 RSOC 动态培养中作为细胞培养载体的应用

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-29 DOI: 10.1002/biot.202400444

Jiaqi Liu, Hao Wang, Zijiao Feng, Hailin Ma, Yuen Yee Cheng, Jie Xu, Yanchun Guan, Shuo Wu, Kedong Song

{"title":"Preparation of Thermosensitive Lignocellulose Hollow Fiber Membrane Grafted With PNIPAAm and Its Application as a Cell Culture Carrier in a RSOC Dynamic Culture","authors":"Jiaqi Liu, Hao Wang, Zijiao Feng, Hailin Ma, Yuen Yee Cheng, Jie Xu, Yanchun Guan, Shuo Wu, Kedong Song","doi":"10.1002/biot.202400444","DOIUrl":"10.1002/biot.202400444","url":null,"abstract":"Currently, the cells, which are urgently required for large-scale application in biomedical-related fields, harvested by traditional trypsin digestion are usually subject to repeated digestion, leading to a reduction of cell activity. In this study, poly (N-isopropylacrylamide) (PNIPAAm) was grafted onto the lignocellulose hollow fiber membranes (HFMs) with cerium ammonium nitrate (CAN) as the initiator to prepare thermosensitive HFMs, which was combined with a rotation system of culture (RSOC) to achieve dynamic culture and non-destructive harvesting of cells from the HFMs. The results of ATR-FTIR, elemental analysis, and SEM confirmed the successful preparation of PNIPAAm-grafted-HFMs, which also showed good biocompatibility to apply for cell culture carriers. In cooling detachment, the HFMs-0.01 group could completely detach the cells within 1 h with a cell separation efficiency of more than 90%. The laminin (LN) and fibronectin (FN) harvested by cooling detachment of P8 generation PC12 cells reached 0.0531 ± 0.0032 and 2.5045 ± 0.0001 pg/cell, respectively, which were significantly higher than that by trypsin digestion. In addition, the cells on the thermosensitive HFMs proliferated fastest in RSOC at 30 rpm with higher glucose consumption and lactate metabolism than in static conditions. Moreover, the cells that had dynamic detachment at 20 rpm had the highest cell density and activity. Therefore, the thermosensitive HFMs could be applied as cell culture carriers in RSOC for cell culturing at 30 rpm and harvesting at 20 rpm, which would provide considerable potential for large-scale cell culture in vitro.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 10","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542313","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

Enhanced Fecal Norovirus Detection Using Magneto-Nanocatalys–Based Immunoassay 利用基于磁性纳米催化的免疫测定法加强粪便诺罗病毒检测

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-29 DOI: 10.1002/biot.202400447

Indra Memdi Khoris, Kenta Tsuruga, Jirayu Boonyakida, Enoch Y. Park

{"title":"Enhanced Fecal Norovirus Detection Using Magneto-Nanocatalys–Based Immunoassay","authors":"Indra Memdi Khoris, Kenta Tsuruga, Jirayu Boonyakida, Enoch Y. Park","doi":"10.1002/biot.202400447","DOIUrl":"10.1002/biot.202400447","url":null,"abstract":"<div>\u0000 \u0000 A new method has been developed to improve the detection of norovirus (NoV) in complex fecal samples using nanocatalyst-based immunoassays. The method involves using multifunctional trimetallic nanoparticles, known as Ag@Fe3O4@Au NPs. These nanoparticles consist of a core of silver (Ag) and a shell of iron oxide (Fe3O4) decorated with isolated gold nanoparticles (Au NPs). The nanoparticles have enhanced catalytic activity, making them an ideal nanocatalyst for reducing 4-nitrophenol (4-NP, yellow) to 4-aminophenol (4-AP, colorless). The developed Ag@Fe3O4@Au NPs-based immunoassay achieved a limit of detection (LOD) of 1.9 pg/mL for norovirus-like particles (NoV-LP) and 6.97 RNA copy number/mL for fecal NoV. In fecal sample analysis for NoV, a heat treatment at 65°C was necessary to prevent degradation of the target protein, ensuring sensitive detection. This work successfully combined multifunctional nanocatalysts for advanced immunoassays, which could contribute to developing nano-biosensing platforms.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 10","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542312","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

Transcriptional Downregulation of Methanol Metabolism Key Genes During Yeast Death in Engineered Pichia pastoris 酵母死亡过程中甲醇代谢关键基因的转录下调

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-15 DOI: 10.1002/biot.202400328

Chenbo Wang, Wei Jiang, Chang Yu, Jianye Xia

{"title":"Transcriptional Downregulation of Methanol Metabolism Key Genes During Yeast Death in Engineered Pichia pastoris","authors":"Chenbo Wang, Wei Jiang, Chang Yu, Jianye Xia","doi":"10.1002/biot.202400328","DOIUrl":"https://doi.org/10.1002/biot.202400328","url":null,"abstract":"<div>\u0000 \u0000 Pichia pastoris possesses the unique ability to utilize methanol as its sole carbon source, which makes it a proper host for producing various high-value-added products via metabolic engineering. Nevertheless, cell death has been observed during the fermentation of modified P. pastoris, with limited literature elucidating the underlying causes and mechanisms. Understanding the death mechanisms during methanol-based fermentation is crucial for optimizing fermentation strategies, enhancing the accumulation of target products, and reducing production costs. Here, we first sought to eliminate the potential causes of cell death during fermentation, such as inadequate inorganic salts and toxic by-product accumulation. The elimination of these potential causes was achieved efficiently utilizing the high-throughput fermentation equipment. Subsequently, we established a correlation between yeast cell death and the duration of the methanol metabolism period by monitoring the growth of the yeast at different fermentation stages. A critical revelation from this work came from analyzing the yeast's transcriptomic data at various stages of methanol metabolism. It was observed that a significant characteristic of yeast cell death during fermentation was the marked down-regulation of transcript levels of key enzymes involved in the methanol assimilation pathway and genes related to their biosynthesis process. The findings of this work are crucial for better understanding the causes and mechanisms of cell death for engineered P. pastoris during methanol-utilized fermentation.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 10","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142443539","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

Thermoresponsive and Supramolecular Polymers: Interesting Biomaterials for Drug Delivery 热致伸缩性和超分子聚合物：有趣的药物输送生物材料。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-09 DOI: 10.1002/biot.202400379

Ahmad Darvishi, Mojtaba Ansari

引用次数: 0

A Novel and Simplified Anion Exchange Flow-Through Polishing Approach for the Separation of Full From Empty Adeno-Associated Virus Capsids 一种新颖、简化的阴离子交换流式抛光方法，用于从空腺病毒囊壳中分离完整的腺病毒囊壳。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-09 DOI: 10.1002/biot.202400430

Frederik Meierrieks, Alisa Weltken, Karl Pflanz, Andreas Pickl, Benjamin Graf, Michael W. Wolff

{"title":"A Novel and Simplified Anion Exchange Flow-Through Polishing Approach for the Separation of Full From Empty Adeno-Associated Virus Capsids","authors":"Frederik Meierrieks, Alisa Weltken, Karl Pflanz, Andreas Pickl, Benjamin Graf, Michael W. Wolff","doi":"10.1002/biot.202400430","DOIUrl":"10.1002/biot.202400430","url":null,"abstract":"Adeno-associated viruses (AAV) are widely used viral vectors for in vivo gene therapy. The purification of AAV, particularly the separation of genome-containing from empty AAV capsids, is usually time-consuming and requires expensive equipment. In this study, we present a novel laboratory scale anion exchange flow-through polishing method designed to separate full and empty AAV. Once the appropriate conditions are defined, this method eliminates the need for a chromatography system. Determination of optimal polishing conditions using a chromatography system revealed that the divalent salt MgCl2 resulted in better separation of full and empty AAV than the monovalent salt NaCl. The efficacy of the method was demonstrated for three distinct AAV serotypes (AAV8, AAV5, and AAV2) on two different stationary phases: a membrane adsorber and a monolith, resulting in a 4- to 7.5-fold enrichment of full AAV particles. Moreover, the method was shown to preserve the AAV capsids’ functional potency and structural integrity. Following the successful establishment of the flow-through polishing approach, it was adapted to a manual syringe-based system. Manual flow-through polishing using the monolith or membrane adsorber achieved 3.6- and 5.4-fold enrichment of full AAV, respectively. This study demonstrates the feasibility of separating full and empty AAV without complex linear or step gradient elution and the necessity of specialized equipment. Flow-through polishing provides a rapid and easy-to-perform platform for polishing multiple vector preparations, addressing a critical aspect in the research and development of novel gene therapies.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 10","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.202400430","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142386595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Electrochemical Glucose Sensors: Classification, Catalyst Innovation, and Sampling Mode Evolution 电化学葡萄糖传感器：分类、催化剂创新和采样模式演变。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-09 DOI: 10.1002/biot.202400349

Chenyang Song, Jian Guo, Yuhan Wang, Hongying Xiang, Yufeng Yang

{"title":"Electrochemical Glucose Sensors: Classification, Catalyst Innovation, and Sampling Mode Evolution","authors":"Chenyang Song, Jian Guo, Yuhan Wang, Hongying Xiang, Yufeng Yang","doi":"10.1002/biot.202400349","DOIUrl":"10.1002/biot.202400349","url":null,"abstract":"<div>\u0000 \u0000 Glucose sensors are essential tools for monitoring blood glucose concentration in diabetic patients. In recent years, with the increasing number of individuals suffering from diabetes, blood glucose monitoring has become extremely necessary, which expedites the iteration and upgrade of glucose sensors greatly. Currently, two main types of glucose sensors are available for blood glucose testing: enzyme-based glucose sensor (EBGS) and enzyme-free glucose sensor (EFGS). For EBGS, several progresses have been made to comprehensively improve detection performance, ranging from enhancing enzyme activity, thermostability, and electron transfer properties, to introducing new materials with superior properties. For EFGS, more and more new metallic materials and their oxides are being applied to further optimize its blood glucose monitoring. Here the latest progress of electrochemical glucose sensors, their manufacturing methods, electrode materials, electrochemical parameters, and applications were summarized, the development glucose sensors with various noninvasive sampling modes were also compared.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 10","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142386598","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

Metabolic Engineering of Saccharomyces cerevisiae for Fermentative Production of Heme 发酵生产血红素的酿酒酵母代谢工程。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-09 DOI: 10.1002/biot.202400351

Hyun-Jae Lee, Dong Joo Shin, Soo Bin Nho, Ki Won Lee, Sun-Ki Kim

{"title":"Metabolic Engineering of Saccharomyces cerevisiae for Fermentative Production of Heme","authors":"Hyun-Jae Lee, Dong Joo Shin, Soo Bin Nho, Ki Won Lee, Sun-Ki Kim","doi":"10.1002/biot.202400351","DOIUrl":"10.1002/biot.202400351","url":null,"abstract":"Heme is a key ingredient required to mimic the color and flavor of meat in plant-based alternatives. This study aimed to develop a yeast-based microbial cell factory for efficient and sustainable production of heme. To this end, first, Hem12p (uroporphyrinogen decarboxylase) was identified as the rate-limiting enzyme in the heme biosynthetic pathway present in Saccharomyces cerevisiae D452-2. Next, we investigated the effects of disruption of the genes involved in the competition for heme biosynthesis precursors, transcriptional repression, and heme degradation (HMX1) on heme production efficiency. Of the knock-out strains constructed in this study, only the HMX1-deficient strain produced heme at a higher concentration than the background strain without gene disruption. In addition, overexpression of PUG1 encoding a plasma membrane transporter involved in protoporphyrin IX (the precursor to heme biosynthesis) uptake led to a significant increase in intracellular heme concentration. As a result, among the various engineered strains constructed in this study, the ΔHMX1/H3&12 + PUG1 strain, the HMX1-deficient strain overexpressing HEM3, HEM12, and PUG1, produced the highest concentration of heme (4.6 mg/L) in batch fermentation, which was 3.9-fold higher than that produced by the wild-type D452-2 strain. In a glucose-limited fed-batch fermentation, the ΔHMX1/H3&12 + PUG1 strain produced 28 mg/L heme in 66 h.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 10","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.202400351","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142386601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Issue Information: Biotechnology Journal 10/2024 发行信息：生物技术期刊 10/2024

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-10-09 DOI: 10.1002/biot.202470101

引用次数: 0