Biotechnology Progress最新文献_第6页

Deep dive into drug substance HCP patterns for comprehensive safety assessment in recombinant mAb formats. 深入了解原料药HCP模式，以进行重组单抗格式的综合安全性评估。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-07-07 DOI: 10.1002/btpr.88540

Daniel M Waldera-Lupa, Anne Gehrke-Oltmanns, Alireza Dehghani, Melissa Morgenroth, Antje-Alke Betz, Heiner Falkenberg, Thomas Flad, Ulli Backofen, Thomas Waerner

{"title":"Deep dive into drug substance HCP patterns for comprehensive safety assessment in recombinant mAb formats.","authors":"Daniel M Waldera-Lupa, Anne Gehrke-Oltmanns, Alireza Dehghani, Melissa Morgenroth, Antje-Alke Betz, Heiner Falkenberg, Thomas Flad, Ulli Backofen, Thomas Waerner","doi":"10.1002/btpr.88540","DOIUrl":"https://doi.org/10.1002/btpr.88540","url":null,"abstract":"The presence of host cell proteins (HCPs) as process-related impurities in monoclonal antibody (mAb) therapeutics presents risks, including immunogenicity, product instability, and reduced therapeutic efficacy. Therefore, HCP levels are seen as a critical quality attribute which requires monitoring. Characterization of residual HCPs is essential during process development and after process changes in late project phases including post-submission changes. Mass spectrometry-based analysis of HCPs is increasingly employed to overcome the limitations of ELISAs, particularly in identifying individual low-abundance HCPs. In this study, we systematically compared three enrichment strategies: hexapeptide enrichment, immunoaffinity chromatography (IAC), and native digest (ND), to enhance LC-MS/MS-based detection of low-abundance HCPs across five recombinant products expressed in CHO cells. Our results show that both ND and IAC substantially expand HCP detection beyond hexapeptide enrichment and in-solution digest in total number of HCP identification. While IAC supported the detection of several potentially critical HCPs, ND uniquely identified additional low-abundance HCPs not captured by antibody-based methods. Furthermore, pull-down assays revealed product-specific HCPs likely interacting with the active pharmaceutical ingredient, suggesting that standard purification steps may be insufficient for their removal. Together, our findings emphasize the complementary nature of ND and IAC for in-depth HCP profiling. We propose the combined application of both methods to ensure comprehensive detection of potentially critical HCPs in biopharmaceutical products, particularly when facing issues with low product or polysorbate stability caused by HCPs with enzymatic activities. This integrative approach supports enhanced product safety and control of consistent HCP depletion in the process.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88540"},"PeriodicalIF":2.8,"publicationDate":"2026-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148403422","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

CRISPR-Cas9 lipid nanoparticles for targeting cyclin-dependent kinases in the tumor microenvironment 靶向肿瘤微环境中周期蛋白依赖性激酶的CRISPR-Cas9脂质纳米颗粒

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2026-01-28 DOI: 10.1002/btpr.70105

Mohammad Mashreghi, Mahere Rezazade Bazaz, Mahmoud Reza Jaafari

{"title":"CRISPR-Cas9 lipid nanoparticles for targeting cyclin-dependent kinases in the tumor microenvironment","authors":"Mohammad Mashreghi, Mahere Rezazade Bazaz, Mahmoud Reza Jaafari","doi":"10.1002/btpr.70105","DOIUrl":"10.1002/btpr.70105","url":null,"abstract":"Due to the significant roles of cyclin-dependent kinases 4 and 6 (CDK4/6) in cancer progression, this study aimed to introduce clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) plasmid DNA (pDNA) encapsulated in lipid nanoparticles (LNPs) as a novel CDK4/6 inhibitor using a gene knock-out strategy for treating cancer. pDNA-LNP was prepared and characterized using a microfluidic system. The results indicated the hydrodynamic diameter of the pDNA-LNP was 90.0 ± 0.1 nm with the PDI of 0.1 and a negative zeta-potential. The cytotoxicity results demonstrated statistically significant differences at doses of 0.250, 500, and 1 μg of pDNA with the capabilities of pDNA-LNP in the induction of apoptosis, as depicted by the Annexin-V-FITC-PI method. Real-time quantitative PCR (qPCR) also indicated a significant reduction in the expression levels of both CDK4 and 6 in the cells that were treated with pDNA-LNP. The in vivo anti-tumor activities of pDNA-LNP have demonstrated that the formulation has the potential to decrease tumor size and improve survival parameters, including median survival time (MST), which was increased from 31 days for the PBS group to 51 days for the pDNA-LNP group at 0.5 μg. On the other hand, the dose of 1 μg had shown signs of toxicity, indicating the need to optimize dosing in future studies. In summary, these findings indicate that CRISPR-Cas9 encapsulated in the LNP can suppress tumor growth and offer a promising strategy for future cancer treatment approaches.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148249074","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

Scalable desolvation synthesis, storage stability, and comprehensive biophysical characterization of human hemoglobin nanoparticles as a potential hemoglobin-based oxygen carrier 人血红蛋白纳米颗粒作为潜在的血红蛋白基氧载体的可扩展溶解合成、储存稳定性和综合生物物理特性。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2026-02-13 DOI: 10.1002/btpr.70114

Alisyn Greenfield, Andre F. Palmer

{"title":"Scalable desolvation synthesis, storage stability, and comprehensive biophysical characterization of human hemoglobin nanoparticles as a potential hemoglobin-based oxygen carrier","authors":"Alisyn Greenfield, Andre F. Palmer","doi":"10.1002/btpr.70114","DOIUrl":"10.1002/btpr.70114","url":null,"abstract":"Hemoglobin (Hb)-based oxygen carriers (HBOCs) are materials that exploit Hb's native oxygen transport while avoiding the adverse side-effects associated with the exposure of the cell-free, unmodified Hb tetramer (i.e. vasoconstriction, systemic hypertension, and oxidative tissue injury) in circulation. There are many synthetic routes to generate HBOCs including chemical polymerization, polymer surface conjugation, and protein encapsulation that meet the material and O2 demands of different biomedical applications (e.g. emergency transfusion medicine, ex vivo organ perfusion, tumor oxygenation, etc.) especially when red blood cells may not be immediately available or desirable. There is currently no FDA approved HBOC in the US, therefore leaving room in the field for further HBOC design and optimization. An ideal HBOC should effectively bind and release O2, have a scalable synthesis to meet material demand, and limit batch-to-batch variability to yield consistent, uniform material. This work demonstrates the successful synthesis of human Hb nanoparticles using a desolvation technique (hHb-dNPs) as a new HBOC candidate. hHb-dNPs demonstrated a monodisperse size distribution between 130 and 150 nm with spherical morphology, a negative surface charge, and hemocompatibility. hHb-dNPs exhibited high O2 affinity with decreased cooperativity similar to that of relaxed-state polymerized Hb. The methodology demonstrated scalability from 5 to 500 mL starting volume, producing products with similar biophysical properties regardless of the scale. hHb-dNPs demonstrated structural and biophysical stability for a month at 4°C and −80°C with various cryoprotectants, with the optimal combination being hHb-dNPs co-stored at −80°C with HSA. Considering these results, hHb-dNPs are a promising potential next generation HBOC.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70114","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146193804","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

Utilizing Cole–Cole parameters for in-line feedback: Cell culture process adjustments based on cell health 利用Cole-Cole参数进行在线反馈：基于细胞健康的细胞培养过程调整。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2025-12-28 DOI: 10.1002/btpr.70096

Nithya Krishnan, Tingting Jiang, Jacob Crowe, Stefano Gotta, Theresa Wittmer, Jennifer Staffenhagen, Barbara Alvarado, Pooja Jambunathan, Shahid Rameez

{"title":"Utilizing Cole–Cole parameters for in-line feedback: Cell culture process adjustments based on cell health","authors":"Nithya Krishnan, Tingting Jiang, Jacob Crowe, Stefano Gotta, Theresa Wittmer, Jennifer Staffenhagen, Barbara Alvarado, Pooja Jambunathan, Shahid Rameez","doi":"10.1002/btpr.70096","DOIUrl":"10.1002/btpr.70096","url":null,"abstract":"In the biopharmaceutical industry, effective process control strategies are essential for enhancing drug substance quality and yield. This study presents dielectric spectroscopy as a novel approach for in-line monitoring of cell apoptosis, enabling earlier detection of apoptotic events and related cellular changes. Utilizing permittivity measurements, we assessed Cole-Cole parameters, specifically critical frequency (fc) and delta epsilon (Δε), as key performance indicators (KPIs) for real-time monitoring of cell health in CHO cell cultures during batch and perfusion processes. Our findings demonstrate that variations in these parameters correlate with cellular stress responses, such as nutrient limitation and high shear conditions, providing timely signals for process monitoring and control. By integrating these in-line measurements, we can enhance feeding strategies, ultimately improving cell viability and productivity. This approach not only streamlines the monitoring process but also offers a robust framework for proactive adjustments in bioprocessing, thereby optimizing overall performance and resource utilization.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145848651","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

A practical framework for clone selection, media-feed screening, and upstream process parameter optimization for a mAb CHO cell culture process 一个实用的框架克隆选择，培养基筛选，和上游工艺参数优化的单克隆CHO细胞培养过程。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2026-03-11 DOI: 10.1002/btpr.88503

Shreya Shah, Dhananajay Kumar, Aadesh Dhamdhere, Sonali Devi, Swapnali Patekar, Velu Mahalingam, Laxmi Adhikary, Viki Chopda

{"title":"A practical framework for clone selection, media-feed screening, and upstream process parameter optimization for a mAb CHO cell culture process","authors":"Shreya Shah, Dhananajay Kumar, Aadesh Dhamdhere, Sonali Devi, Swapnali Patekar, Velu Mahalingam, Laxmi Adhikary, Viki Chopda","doi":"10.1002/btpr.88503","DOIUrl":"10.1002/btpr.88503","url":null,"abstract":"This study presents a data-driven workflow for upstream process development in biologics manufacturing, aimed at improving consistency, efficiency, and decision-making using established statistical and machine learning tools. Rather than relying on subjective interpretation, the framework integrates multiple analytical methods in a structured manner to support key process development decisions. For clone selection, K-means clustering was applied and benchmarked with hierarchical clustering to identify top-performing candidates from a large clone pool. By incorporating multiple product quality attributes into the evaluation, the approach improves selection consistency and reduces subjectivity. In the media and feed screening stage, principal component analysis (PCA) was used to explore how specific combinations influence glycosylation patterns, enabling rapid identification of promising conditions for further optimization while reducing experimental burden. For upstream process parameter refinement, a sequential learning strategy based on multi-objective Bayesian optimization (MOBO) was applied to adaptively explore trade-offs among competing quality attributes and titer. This approach enabled more efficient identification of improved operating conditions compared with static experimental designs while minimizing experimental runs. While the methods used are well-established, their structured integration into a cohesive workflow demonstrates practical utility for industrial applications. Aligned with Bioprocessing 4.0 principles, this approach supports improved process understanding and informed decision-making in upstream bioprocess development.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147430494","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

Rapid development of a transferable Raman model using high-throughput cell culture for monitoring monoclonal antibody titer 利用高通量细胞培养监测单克隆抗体滴度的可转移拉曼模型的快速发展。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2026-04-05 DOI: 10.1002/btpr.88506

Alexandra Umprecht, Nicholas Uth, Haenah Kim, Oliver Spadiut, Yang Yang

{"title":"Rapid development of a transferable Raman model using high-throughput cell culture for monitoring monoclonal antibody titer","authors":"Alexandra Umprecht, Nicholas Uth, Haenah Kim, Oliver Spadiut, Yang Yang","doi":"10.1002/btpr.88506","DOIUrl":"10.1002/btpr.88506","url":null,"abstract":"Monoclonal antibody (mAb) titer monitoring is a key capability during process development and optimization, enabling timely decision making and increasing the speed of development. Raman spectroscopy is a prominent process analytical technology (PAT), but resource-efficient calibration strategies for the development of transferable models are limited. This work demonstrates the development and successful transfer of a calibration model for monoclonal antibody concentration between two different cell lines with varied metabolic profiles expressing different antibodies. The root mean square error of prediction (RMSEP) for titer in the source cell line (0.266 g L−1) was comparable to that of the target cell line (0.325 g L−1). The transferable model was achieved by conducting a spiking study in a high-throughput parallel bioreactor system. Different experimental approaches with models trained on spiked versus native samples were compared. This analysis revealed that model transferability was influenced by the degree of correlation of lactate with antibody titer in the source process, emphasizing the importance of process knowledge in the development of Raman calibration models. Overall, the study presents evidence of the feasibility of transferable titer models, marking a significant advancement in process monitoring capabilities for high-throughput cell culture as well as introducing a generic methodology for calibration of transferable models in the context of upstream bioprocessing.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.88506","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147621729","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

Image analysis method for measurement and prediction of intra-matrix IgG diffusion 用于测量和预测基质内IgG扩散的图像分析方法。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2025-11-17 DOI: 10.1002/btpr.70085

Riya Debbarma, Antonio C. F. dos Santos, Michael Ladisch

{"title":"Image analysis method for measurement and prediction of intra-matrix IgG diffusion","authors":"Riya Debbarma, Antonio C. F. dos Santos, Michael Ladisch","doi":"10.1002/btpr.70085","DOIUrl":"10.1002/btpr.70085","url":null,"abstract":"Measurement and imaging of intra-matrix protein therapeutics diffusion is important due to the emergence of injectable biologics currently in various stages of research and clinical testing. These therapeutics are developed for delivery to hyaluronic acid (HA)-rich anatomical sites such as subcutaneous tissue, the vitreous humor, and knee joints, depending on the target tissue. Understanding their diffusion behavior is essential for optimizing drug delivery strategies. Our work presents an image analysis method suited for tracking IgG diffusion in low viscosity HA matrices representative of the vitreous humor, where diffusion occurs more rapidly unlike a previously reported analysis method for higher viscosity matrices where protein diffusion is significantly slower. The current method utilizes scanner images at 6.3 MP resolution, and an algorithm that removes background and calculates protein mass and concentration measured directly within matrices formulated to represent HA in an intravitreal environment. We report and demonstrate a robust method for predicting protein diffusion coefficient from images of label-free protein diffusing in a low viscosity HA matrix.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70085","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145533805","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

Innovative methodology for flexible monitoring of various bioprocesses by using pre-existing Raman data coupled with automated transfer learning technique 通过使用预先存在的拉曼数据与自动迁移学习技术相结合，灵活监测各种生物过程的创新方法。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2026-02-10 DOI: 10.1002/btpr.70111

Adèle Schini, Hadi El Radi, Johan Cailletaud, Célia Sanchez, Nathan Gay, Bruno Ebel, Emmanuel Guedon, Laurent Jourdainne

{"title":"Innovative methodology for flexible monitoring of various bioprocesses by using pre-existing Raman data coupled with automated transfer learning technique","authors":"Adèle Schini, Hadi El Radi, Johan Cailletaud, Célia Sanchez, Nathan Gay, Bruno Ebel, Emmanuel Guedon, Laurent Jourdainne","doi":"10.1002/btpr.70111","DOIUrl":"10.1002/btpr.70111","url":null,"abstract":"This study introduces an innovative approach for the flexible monitoring of bioprocesses using Raman spectroscopy coupled with automated transfer learning. Traditional Raman spectroscopy requires extensive process-specific calibration, limiting its transferability across different conditions. To address this, we developed an automated method that utilizes the dynamic orthogonal projection (DOP) algorithm to use pre-existing chemometric models built from one process (“Input Process”) to monitor a distinct process (“Target Process”) which lacked its own Raman models. These new processes conditions varied in terms of culture mode, cell line, media, analyzer, and acquisition settings. This method used spectral data from Target Process to modify the existing spectral data from Input Process, aligning them with the new conditions. The approach was validated on Chinese Hamster Ovary cell cultures, targeting critical metabolic parameters such as glucose, lactate, glutamine, and viable cell density. The results showed that with only one batch used for the transfer, the average relative errors compared to offline values were around 10% for glucose and lactate but remained high for VCD and glutamine. After a second batch to perform the transfer on two batches, the relative errors were further reduced below 10% for most parameters. By effectively transferring models across different processes, this approach minimizes the need for extensive recalibrations, enhancing the efficiency and applicability of Raman spectroscopy in diverse bioprocess environments.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146155845","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

Design of experiments-guided membrane capture facilitates integration of continuous virus inactivation using Phi6 as a surrogate virus in an intensified downstream process 实验引导的膜捕获设计有助于整合在强化的下游过程中使用Phi6作为替代病毒的连续病毒灭活。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2026-03-25 DOI: 10.1002/btpr.88504

Mario Grünberg, Lisa Lipski, Gabriel Fisicaro, Thomas Duvignau, Karolina Meyer-Heinrichs, Diana Carmen Mocsy, Thomas-Josef Filz, Bastian Quaas, Alexandra Stützer

{"title":"Design of experiments-guided membrane capture facilitates integration of continuous virus inactivation using Phi6 as a surrogate virus in an intensified downstream process","authors":"Mario Grünberg, Lisa Lipski, Gabriel Fisicaro, Thomas Duvignau, Karolina Meyer-Heinrichs, Diana Carmen Mocsy, Thomas-Josef Filz, Bastian Quaas, Alexandra Stützer","doi":"10.1002/btpr.88504","DOIUrl":"10.1002/btpr.88504","url":null,"abstract":"The growing demand for cost-efficient and flexible biomanufacturing has increased interest in process-intensified downstream platforms. This study evaluates an intensified monoclonal antibody (mAb) purification sequence, where two unit operations traditionally performed in batch mode—Protein A capture and low pH virus inactivation (VI)—are redesigned to enhance productivity and minimize resource usage. Rapid-cycling Protein A membrane chromatography was optimized using a design-of-experiments approach to address inherent membrane challenges such as buffer consumption. Wash step volumes were systematically reduced without compromising host cell protein or host cell DNA clearance, yielding a 70% reduction in total wash buffer consumption. At manufacturing scale, membrane adsorbers achieved critical quality attributes comparable to a benchmark resin, while increasing productivity ~20-fold and lowering capture-step costs. Protein A eluates were processed in a novel continuous virus inactivation (cVI) system using bacteriophage Phi6 as a surrogate for enveloped viruses. At residence times of 35 and 70 min, the cVI system achieved a ≥5-log reduction, equaling conventional batch performance without compromising mAb quality. The study demonstrates that membrane-based Protein A capture and continuous VI can be seamlessly integrated into an intensified DSP framework. This approach effectively maintains product quality while significantly reducing buffer usage and cost, thus supporting modular intensification strategies for clinical-scale mAb manufacturing.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.88504","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147509230","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

Improved methodology for direct monitoring of large-scale manufacturing cell culture using Raman spectroscopy data from process development 改进的方法，直接监测大规模生产细胞培养使用拉曼光谱数据从工艺发展。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-06-15 Epub Date: 2026-05-13 DOI: 10.1002/btpr.88513

Hadi El Radi, Hui Chen, Xiande Su, Weiwei Hu, Peng Zhang, Linfeng Jin, Xisheng Cao, Hui Wang, Jiansheng Lu

{"title":"Improved methodology for direct monitoring of large-scale manufacturing cell culture using Raman spectroscopy data from process development","authors":"Hadi El Radi, Hui Chen, Xiande Su, Weiwei Hu, Peng Zhang, Linfeng Jin, Xisheng Cao, Hui Wang, Jiansheng Lu","doi":"10.1002/btpr.88513","DOIUrl":"10.1002/btpr.88513","url":null,"abstract":"Process analytical technology (PAT) is a framework that encourages the implementation of analytical tools for real-time monitoring of critical batch components during the production of pharmaceuticals. In mammalian cell cultures, Raman spectroscopy as a PAT tool has demonstrated its value for accurate in-line monitoring of various process parameters as well as quality attributes. However, as process development cultures are transferred to manufacturing scales, the small-scale models are often adjusted either with new calibration data or with transfer learning algorithms. This model adjustment step makes the overall process transfer to manufacturing less straightforward from an analytical standpoint, requiring additional resources and specific data treatment expertise to fine tune the models. In this work, we compare a specific spectra processing approach against existing approaches reported in the literature for processing Raman spectra in the context of scale-up challenges. The proposed novelty is a preprocessing strategy which computes spectra normalization based on the 3100–3700 cm−1 band. Data generated at 3 L scale were used to monitor a 3000 L bioreactor without model recalibration or transfer learning with equivalent accuracy as a same scale 3 L bioreactor monitoring. While broader validation case would be warranted, these results demonstrate a direct scale-up model implementation in large scale manufacturing under the studied conditions.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147925788","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