Journal of biotechnology最新文献_第2页

Genome-scale metabolic analysis reveals enhanced metabolism and antioxidative stress response in perfusion cell culture under constant hyperosmotic stimulation 基因组尺度的代谢分析揭示了持续高渗刺激下灌注细胞培养中代谢和抗氧化应激反应的增强。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-22 DOI: 10.1016/j.jbiotec.2025.09.010

Qingyuan Ran , Xinran Zhang , Chen Wang , Weijian Zhang , Liang Zhao , Wen-Song Tan , Qian Ye

{"title":"Genome-scale metabolic analysis reveals enhanced metabolism and antioxidative stress response in perfusion cell culture under constant hyperosmotic stimulation","authors":"Qingyuan Ran , Xinran Zhang , Chen Wang , Weijian Zhang , Liang Zhao , Wen-Song Tan , Qian Ye","doi":"10.1016/j.jbiotec.2025.09.010","DOIUrl":"10.1016/j.jbiotec.2025.09.010","url":null,"abstract":"<div><div>Hyperosmotic stimulation is a prevalent strategy to enhance cell culture productivity in fed-batch cultures. Maintaining stable hyperosmotic conditions during perfusion cultures presents a promising strategy, but research on its application in perfusion processes remains limited. In this study, we investigated cellular responses under constant hyperosmolality in Chinese hamster ovary (CHO) cell perfusion cultures using Raman spectroscopy to maintain a constant hyperosmotic environment. Integrated genome-scale metabolic model (GEM) with real-time monitoring of the oxygen uptake rate (OUR) was employed to systematically analyze the metabolic alterations induced by constant hyperosmotic stimulation. Our findings showed that the CHO cells exhibited time-dependent metabolic responses, with rapid changes in nutrient uptake, glycolysis, and TCA cycle activity, while lactate metabolism responded more slowly. The specific productivity (<em>q</em><sub><em>mAb</em></sub>) displayed the slowest changes, stabilizing only after 6–9 days upon the simulation, resulting in a maximum increase up to 168.5 %. Notably, we found shifts in the intracellular redox environment, and the cells enhanced their antioxidative capacity at low dissolved oxygen (DO) levels under hyperosmotic conditions. Even when DO dropped to 10 %, the cells subjected to hyperosmotic stimulation maintained relatively low levels of reactive oxygen species (ROS) while preserving high <em>q</em><sub><em>mAb</em></sub>. Overall, this study provides new insights into cellular responses under constant hyperosmotic condition and provides insights for the application of hyperosmotic strategies in perfusion processes.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 181-191"},"PeriodicalIF":3.9,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145137658","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

Cost-effective melanin production using engineered Escherichia coli with autonomous tyrosine biosynthesis 利用具有自主酪氨酸生物合成的工程大肠杆菌高效生产黑色素

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-19 DOI: 10.1016/j.jbiotec.2025.09.009

Ling Zou , Jian Liu , Yidan Hu , Yan Zhao , Xiaobo Liu

引用次数: 0

Reduction of high mannose glycoforms from monoclonal antibodies by affinity chromatography using a recombinant prokaryotic lectin 利用重组原核凝集素亲和层析法从单克隆抗体中还原高甘露糖糖型。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-18 DOI: 10.1016/j.jbiotec.2025.09.008

Elizabeth Matthews , Neha Tushar Dalvi , Michael Butler

{"title":"Reduction of high mannose glycoforms from monoclonal antibodies by affinity chromatography using a recombinant prokaryotic lectin","authors":"Elizabeth Matthews , Neha Tushar Dalvi , Michael Butler","doi":"10.1016/j.jbiotec.2025.09.008","DOIUrl":"10.1016/j.jbiotec.2025.09.008","url":null,"abstract":"<div><div>Glycan profiles of monoclonal antibodies are critical quality attributes for their use as therapeutic agents. However, the control of glycosylation can prove difficult during large-scale manufacture. An on-going problem is the increase in high-mannose glycans (HM) that can occur unexpectedly under cell culture conditions that disrupt the glycosylation pathway. We have developed a method for removal of high-mannose glycans from a heterogeneous mixture of mAb glycoforms purified from a bioprocess. This involves the use of a prokaryotic lectin (RPL-Man2) bound to agarose beads to selectively bind reversibly to mAbs carrying HM-glycans. The unbound flow-through from this step contains the remainder of the mAb preparation with a reduced HM-glycan content. In this paper we demonstrate the application of this method using two distinct antibodies derived from Chinese hamster ovary (CHO) cells purposely grown under conditions that enhanced the content of HM-glycans. The presence of an elevated level of mannose in the culture of the producer cells as well as supplementation with a specific mannosidase inhibitor resulted in purified mAbs containing significantly higher levels of HM-glycans than would be expected normally under standard conditions. We showed that HM-glycans in the purified mAbs were reduced significantly by the application of lectin chromatography. Analysis by both UPLC using fluorescence detection as well as by mass spectrometry showed that the removal of the HM-glycoforms was selective and did not affect the profile of the remaining glycoforms. This method has potential for use in large-scale biomanufacturing for the reduction or elimination of HM-glycoforms of mAbs.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 158-167"},"PeriodicalIF":3.9,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145102709","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

Preparation of enzymatic microchannel reactor with immobilized urease-crosslinked enzyme aggregates and its performance 固定化脲酶-交联酶聚集体酶微通道反应器的制备及性能研究。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-17 DOI: 10.1016/j.jbiotec.2025.09.007

Siqi Ma, Chang Liu, Chenfeng Ma, Shuguang Wang

{"title":"Preparation of enzymatic microchannel reactor with immobilized urease-crosslinked enzyme aggregates and its performance","authors":"Siqi Ma, Chang Liu, Chenfeng Ma, Shuguang Wang","doi":"10.1016/j.jbiotec.2025.09.007","DOIUrl":"10.1016/j.jbiotec.2025.09.007","url":null,"abstract":"<div><div>Urine recovery has been achieved through physical-chemical technologies on space station presently, but it requires harsh operation conditions and careful equipment maintenance. Here, a novel microreactor that combined microchannel and urease-crosslinked enzyme aggregates (urease-CLEAs) was prepared to mildly reclaim water and nitrogen nutrient from urine. First, the material that was suitable to make microchannel reactor and immobilize urease-CLEAs was selected from five organic materials, and poly(methyl methacrylate) was the best one. Second, the immobilization conditions for urease-CLEAs were optimized. Microchannel reactor with immobilized urease-CLEAs (CLEAs-microreactor) showed the highest activity of 0.54 U/cm<sup>2</sup> under the optimal conditions of precipitant 20 % acetone, crosslinker 2.0 % glutaraldehyde, and 2.0-hour precipitation/crosslinking. CLEAs-microreactor showed good stability of pH, thermal, and storage compared with the microchannel reactor with immobilized free-urease (Free-microreactor). The efficiency of CLEAs-microreactor reached the highest of 101.8 μmol/h, which was 2.24-fold of Free-microreactor. This study provides an alternative technology for the recovery of water and nitrogen nutrient from urine on space station.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 142-150"},"PeriodicalIF":3.9,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091458","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

Synergistic strategy for high-yield 2,3-butanediol and acetoin production in Bacillus licheniformis MW03 based on metabolic engineering 基于代谢工程的地衣芽孢杆菌MW03高产2,3-丁二醇和乙酰素协同策略

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-16 DOI: 10.1016/j.jbiotec.2025.09.006

Yinhao Gao , Yazi Zhou , Liqian Wang , Na Zhang , Weishuai Qin , Wu Meng , Cuixia Zhou

{"title":"Synergistic strategy for high-yield 2,3-butanediol and acetoin production in Bacillus licheniformis MW03 based on metabolic engineering","authors":"Yinhao Gao , Yazi Zhou , Liqian Wang , Na Zhang , Weishuai Qin , Wu Meng , Cuixia Zhou","doi":"10.1016/j.jbiotec.2025.09.006","DOIUrl":"10.1016/j.jbiotec.2025.09.006","url":null,"abstract":"<div><div><em>Bacillus licheniformis</em> is an efficient platform for 2,3-butanediol (2,3-BD) and acetoin production due to its rapid glucose utilization rate and adaptability to industrial fermentation conditions. Here, we isolated the <em>B. licheniformis</em> strain MW03 with high yield of acetoin and 2,3-BD, which carried genetic mutations in <em>acoR</em> and <em>budC</em>, respectively encoding an acetoin dehydrogenase regulator and meso-2,3-BD dehydrogenase. To further confirm the physiological effects on acetoin and 2,3 BD biosynthesis, gene editing was performed using the CRISPR-Cas9 system, followed by phenotypic screening and genotype validation. The knockout of <em>acoR</em> and <em>budC</em> increased the acetoin maximum titer by 21.2 % and 49.2 %, respectively. Moreover, the optical purity of D-(-)-2,3-BD reached 92.7 % following the knockout of <em>budC</em>. Heterologous expression of <em>acoR</em> from <em>B. licheniformis</em> 2709 in both the wild type and <em>acoR</em> knockout mutant strongly inhibited acetoin accumulation compared to native <em>acoR</em>, which emphasized the regulatory role of AcoR in acetoin accumulation. Conversely, complementation of <em>budC</em> restored the synthesis of meso-2,3-BD synthesis, emphasizing its importance in this process. Overexpression of <em>alsD</em> in the <em>acoR</em> mutant increased the 2,3-BD titer by 61.9 % to 121.97 g/L, while the productivity reached 2.03 g/L·h. Finally, co-expression of <em>bdhA</em> and <em>gldA</em> increased 2,3-BD production by 25.6 %. This study elucidated the dual regulatory roles of <em>acoR</em> and <em>budC</em> in acetoin and 2,3-BD metabolism, establishing a \"knockout-overexpression\" synergic strategy, which offers theoretical support and practical guidance for further strain optimization.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 232-243"},"PeriodicalIF":3.9,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145080838","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

Enhanced volatile fatty acids production from Chinese cabbage waste by pretreatment 预处理提高大白菜废弃物挥发性脂肪酸产量

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-12 DOI: 10.1016/j.jbiotec.2025.09.005

Yingmeng Shen, Longlong Liu, Sen Yang, Xiaofeng Ji, Zhengang Chen, Jiying Zhu

{"title":"Enhanced volatile fatty acids production from Chinese cabbage waste by pretreatment","authors":"Yingmeng Shen, Longlong Liu, Sen Yang, Xiaofeng Ji, Zhengang Chen, Jiying Zhu","doi":"10.1016/j.jbiotec.2025.09.005","DOIUrl":"10.1016/j.jbiotec.2025.09.005","url":null,"abstract":"<div><div>Effects of cellulase pretreatment, thermal pretreatment, and a combined cellulase–thermal pretreatment on the anaerobic fermentation performance of Chinese cabbage waste (CCW) were investigated. The combined pretreatment enhanced the structure loosenness and porosity of CCW, resulting in a 10.2 % increase in the hydrolysis rate and a 7.4 % increase in reducing sugar content. In this group, acid productivity and caproic acid yields were 0.57 g VFA/g VS and 7351 mg COD/L, respectively, which were 18.1 % and 30 % higher than those in the control group. Cellulase pretreatment significantly enhanced the butyrate production, which was consistent with the higher <em>Clostridium</em> spp. abundance observed in this group. Although thermal pretreatment significantly enhanced the release of soluble chemical oxygen demand (SCOD), resulting in substantial increases of 38.7 % in acetic acid and 73.8 % in propionic acid production, respectively, it did not improve the total volatile fatty acids (VFAs) yield. <em>Lactobacillus</em> and <em>Weissella</em> were selectively enriched in this group, leading to lactic acid accumulation during the initial fermentation stage and a high yield of propionic acid in the later stages. In the cellulase-thermal pretreatment group, the relative abundance of <em>Prevotella</em> increased to 20.70 %. This major rumen bacterium can degrade polysaccharides in plant cell walls, thereby facilitating carboxylic acid production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 91-100"},"PeriodicalIF":3.9,"publicationDate":"2025-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145057170","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 universal single-plasmid, single-promoter system for efficient protein display and RNA encapsulation in MS2 virus-like particles 在MS2病毒样颗粒中高效蛋白展示和RNA封装的通用单质粒、单启动子系统。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-10 DOI: 10.1016/j.jbiotec.2025.09.004

Ping Chen , Hao-Min Li , Chu-Yu Huang , Jie-Jing Liu , Lin-Yi Luo , Jing Luo , Xing-Jun Huo , Mei Huang , Ying-Lin Yin , Lei Mou , Yong-Chang Ouyang

{"title":"A universal single-plasmid, single-promoter system for efficient protein display and RNA encapsulation in MS2 virus-like particles","authors":"Ping Chen , Hao-Min Li , Chu-Yu Huang , Jie-Jing Liu , Lin-Yi Luo , Jing Luo , Xing-Jun Huo , Mei Huang , Ying-Lin Yin , Lei Mou , Yong-Chang Ouyang","doi":"10.1016/j.jbiotec.2025.09.004","DOIUrl":"10.1016/j.jbiotec.2025.09.004","url":null,"abstract":"<div><div>Bacteriophage MS2 virus-like particles (VLPs) represent a promising platform for biomedical applications. However, challenges remain in integrating protein display, RNA encapsulation, and convenient purification methods. Here, we developed a universal single-plasmid single-promoter system (USS) that enables modular assembly of multifunctional VLPs through designated <em>Pst</em>I (protein insertion) and <em>Kpn</em>I (RNA encapsulation) sites. The USS system achieved: 1) Efficient display of large protein molecules (e.g., EGFP, Nanobody N16) on each VLP surface while encapsulating target RNA; 2) Rapid one-step purification via His-tag affinity chromatography, bypassing ultracentrifugation; 3) The resulting VLPs exhibit thermal stability (60°C for 15 min) and nuclease resistance. Functional validation demonstrated: 1) Diagnostics: <em>CoviN</em>-encapsulated VLPs served as RT-qPCR controls for COVID-19 detection (linear range: 10<sup>2</sup>–10<sup>8</sup> copies/μL); 2) Dual functionality for co-delivery: N16-VLPs with <em>shPCSK9</em> suppressed protein PCSK9 expression by 68 % (<em>p</em> < 0.01) in Hepa1–6 cells and bound PD-L1 with nanomolar affinity (EC50 = 1.6 μg/mL). Crucially, the simplicity of USS eliminated the need for multi-plasmid constructs, providing a streamlined approach to VLPs production that is both efficient and scalable for applications from molecular diagnostics to combination therapies.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 132-141"},"PeriodicalIF":3.9,"publicationDate":"2025-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145053486","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

The impact of charged particles on beta-glucosidase and its molecular dynamics mechanism: A deep insight into the effects of charged peptides on enzyme activity and conformation 带电粒子对β -葡萄糖苷酶的影响及其分子动力学机制：深入了解带电肽对酶活性和构象的影响。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-09 DOI: 10.1016/j.jbiotec.2025.08.019

Qi Yang , Xiaoyu Yang , Lulu Wang , Jiayi Ji , Xinyue Jiang , Liang Dong , Chunying Liu , Shaohua Dou

{"title":"The impact of charged particles on beta-glucosidase and its molecular dynamics mechanism: A deep insight into the effects of charged peptides on enzyme activity and conformation","authors":"Qi Yang , Xiaoyu Yang , Lulu Wang , Jiayi Ji , Xinyue Jiang , Liang Dong , Chunying Liu , Shaohua Dou","doi":"10.1016/j.jbiotec.2025.08.019","DOIUrl":"10.1016/j.jbiotec.2025.08.019","url":null,"abstract":"<div><div>Beta-Glucosidase(BGL) is an enzyme present in all organisms and plays a key role in a variety of biological processes. In this study, three short peptides Z1, O1, and F1 with positive, neutral, and negative charges, respectively, were designed and synthesised, which were added to the BGL system of action, and the changes in BGL activity and conformation induced by the charged short peptides were investigated by measuring changes in enzyme activities and zeta potentials, as well as by applying spectroscopy and computer simulation. The results showed that Z1 increased BGL activity and zeta potential by 54.6 %, on the contrary, the negatively charged short peptide F1 inhibited its enzyme activity and decreased the potential by 38.3 %, and O1 had less effect on BGL. The results of spectroscopy and computer simulation showed that the addition of the charged short peptide led to significant changes in the apparent morphology and structure of BGL, but it did not bind to BGL to form a complex, but rather affected its surface charge distribution and altered the enzymatic activity and structure of BGL.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 112-120"},"PeriodicalIF":3.9,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145040234","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

Developing a robust and scalable platform for AAV8 production 为AAV8生产开发一个健壮且可扩展的平台。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-09 DOI: 10.1016/j.jbiotec.2025.09.002

André Nascimento , Tiago Q. Faria , Tiago Nunes , Joana G. Oliveira , José Mendes , António Roldão , Cristina Peixoto

{"title":"Developing a robust and scalable platform for AAV8 production","authors":"André Nascimento , Tiago Q. Faria , Tiago Nunes , Joana G. Oliveira , José Mendes , António Roldão , Cristina Peixoto","doi":"10.1016/j.jbiotec.2025.09.002","DOIUrl":"10.1016/j.jbiotec.2025.09.002","url":null,"abstract":"<div><div>Creating robust and scalable bioprocesses is essential for the production of viral vectors, particularly adeno-associated virus (AAV), which are in growing demand for gene therapy applications. This study presents the design and implementation of a scalable AAV8 production platform, leveraging extensive in-house expertise. Three production campaigns were conducted: two at the 2-liter scale and one at the 50-liter scale. The robustness of the upstream and downstream processes was confirmed in the 2-liter campaigns, yielding consistent titers and recoveries close to 80 %. Scalability was validated, demonstrating successful translation from the 2–50 L scale without compromising titers, recoveries, or product quality with a 3-fold enrichment of full capsids. Key modifications, such as adjustments to inoculation concentration, the choice of nuclease, and the direct loading of clarified material onto affinity chromatography, were evaluated to enhance process economics. These modifications did not adversely impact the production process and resulted in significant cost savings. Noteworthy, this study highlights a three-fold enrichment of full capsids, showcasing the process's ability to deliver a higher quality product. While the process is optimized for AAV8, only the polishing step is serotype-specific. The rest of the operations can be broadly applied to other AAV serotypes with minor adjustments. This flexibility suggests potential for wider applications in gene therapy and other fields.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 72-79"},"PeriodicalIF":3.9,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145040267","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

Improving AAV8 purification with continuous affinity capture: From batch to continuous multicolumn chromatography 用连续亲和捕获改进AAV8的纯化：从批处理到连续多柱色谱。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-09 DOI: 10.1016/j.jbiotec.2025.09.003

Salomé Neto , Greig Rankine , Franziska Bollmann , Manuel J.T. Carrondo , Ricardo J.S. Silva

{"title":"Improving AAV8 purification with continuous affinity capture: From batch to continuous multicolumn chromatography","authors":"Salomé Neto , Greig Rankine , Franziska Bollmann , Manuel J.T. Carrondo , Ricardo J.S. Silva","doi":"10.1016/j.jbiotec.2025.09.003","DOIUrl":"10.1016/j.jbiotec.2025.09.003","url":null,"abstract":"<div><div>The increasing demand for adeno-associated viruses (AAVs) in gene therapy is driving the development of more efficient and scalable purification processes. This study explored different batch and continuous multicolumn chromatography (MCC) approaches for AAV8 affinity resin purification using the Resolute® BioSMB PD platform. Starting with a description of the breakthrough behaviour, we defined a continuous loading strategy by dividing a single chromatography bed into smaller, connected columns. This continuous batch multicolumn chromatography (B-MCC) process was then modified into a version where the columns were overloaded to enhance productivity and resin utilization. The different approaches were compared regarding recovery, productivity, buffer consumption, and resin utilization. The continuous sequential multicolumn chromatography (S-MCC) process developed enables higher AAV recovery in comparison to the B-MCC process, with a recovery of 76 % for the highest load concentration evaluated, with a productivity of 1.8 × 10<sup>17</sup> VP h<sup>−1</sup> mL<sup>−1</sup>, 1.5 times higher than the B-MCC. Additionally, for the same volume of processed feed, the S-MCC halves the buffer consumption compared to the B-MCC. Our results demonstrate that S-MCC significantly improves overall process efficiency, offering higher recovery and productivity while reducing resource consumption. These results strongly support the broader application of continuous processing techniques in the biopharmaceutical industry, particularly for high-demand biologics such as AAV.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 101-111"},"PeriodicalIF":3.9,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145040244","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