Biotechnology Progress最新文献_第4页

Addressing challenging impurities in fusion proteins: A comprehensive review and cost-effective strategy for HCP and low molecular weight species control 解决融合蛋白中具有挑战性的杂质：HCP和低分子量物种控制的综合综述和成本效益策略。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-04-23 DOI: 10.1002/btpr.88512

Anindita Das, Madhava Ram Paranandi, Karthikeyan Subbiramani, Ranganadha Reddy

{"title":"Addressing challenging impurities in fusion proteins: A comprehensive review and cost-effective strategy for HCP and low molecular weight species control","authors":"Anindita Das, Madhava Ram Paranandi, Karthikeyan Subbiramani, Ranganadha Reddy","doi":"10.1002/btpr.88512","DOIUrl":"10.1002/btpr.88512","url":null,"abstract":"Fusion proteins represent a rapidly expanding class of biotherapeutics engineered by combining functional domains from different proteins to enhance therapeutic efficacy, stability, and pharmacokinetics. Their unique design enables extended half-life, reduced dosing frequency, and multi-targeting capabilities, making them highly versatile for diverse therapeutic indications. However, the manufacturing of fusion proteins presents significant challenges, particularly in managing complex product- and process-related impurities such as host cell proteins (HCPs), fragments, and low-molecular-weight (LMW) species. These impurities are difficult to eliminate due to their structural and physicochemical similarity to the target protein, complicating purification and scale-up. Part I of this paper provides a comprehensive review of published studies addressing impurity characterization and control strategies in fusion protein manufacturing. The review highlights critical impurity types, analytical tools for detection and quantification, and process strategies for effective removal while maintaining product integrity and regulatory compliance. Part II presents a case study from Kemwell Biopharma, illustrating a cost-effective platform purification strategy for complex fusion proteins. The approach is applicable to non-tagged, Fc-based, and multispecific fusion protein formats and demonstrates efficient removal of HCPs and LMW impurities while achieving high yield and purity. Together, the review and case study provide both a broad scientific understanding and a practical demonstration of scalable impurity control approaches, offering valuable insights for the development of robust and economically viable manufacturing processes for next-generation fusion protein therapeutics.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147760810","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

Disulfide bond reduction of antibodies during clarification: A bench-scale assay and a mitigation tool 澄清过程中抗体的二硫键减少：一种实验规模的测定和缓解工具。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-04-03 DOI: 10.1002/btpr.88507

Hendri Tjandra, Deepa Surendar, Isu Yoon, Florence Rusly, Yan Su, Vu Thai, Yi Liu, Ashley Hesslein

{"title":"Disulfide bond reduction of antibodies during clarification: A bench-scale assay and a mitigation tool","authors":"Hendri Tjandra, Deepa Surendar, Isu Yoon, Florence Rusly, Yan Su, Vu Thai, Yi Liu, Ashley Hesslein","doi":"10.1002/btpr.88507","DOIUrl":"10.1002/btpr.88507","url":null,"abstract":"We investigated side-by-side clarification with a two-stage depth filter system against a single-stage charge chromatographic clarifier on a number of Chinese hamster ovary (CHO) cell cultures. We found that the one-stage fibrous anion exchange (AEX) clarification met or exceeded the performance of the conventional two-stage depth filtration systems in terms of product recovery, removal of process-related impurities, and throughput. The one-stage anion clarification approach had a smaller footprint, a shorter set-up time, an easier setup, consumed less buffer flush volume, and operated at a lower differential pressure relative to those of conventional systems. In a comparability study at 200 L scale, antibodies in cell culture fluid (CCF) clarified by depth filtration system exhibited product quality degradation due to disulfide bond reduction (DSBR), whereas antibodies clarified by AEX fiber method were not. We surmised that the lower operating differential pressure and electrostatic adsorption mechanism of the anion clarification system minimized the potential of product to undergo DSBR. We developed a bench-scale process model that allowed us to assay DSBR and predict the process results. We then utilized it to demonstrate that the antibody was reduced when clarified with a two-stage 3 M™ Zeta Plus™ Depth Filter system but not with the anionic clarification system. This bench-scale DSBR assay allowed us to rank the relative disulfide bond reduction potentials of various monoclonal antibodies (mAbs) and demonstrate the reduction-mitigating impacts of the fibrous clarification approach on a reduction-prone antibody culture at 2000 L processing scale.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13441350/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147607475","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

Multivariate phase-dependent optimization of bioprocesses boosts performance and quality—Why timing (of exposure) matters 生物过程的多变量相位依赖优化提高了性能和质量——为什么（暴露）时间很重要。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-05-17 DOI: 10.1002/btpr.88508

Samuel Kienzle, Lisa Junghans, Anja Wittmann, Stefan Wieschalka, Ralf Takors, Nicole Erika Radde, Beate Presser, Verena Nold

{"title":"Multivariate phase-dependent optimization of bioprocesses boosts performance and quality—Why timing (of exposure) matters","authors":"Samuel Kienzle, Lisa Junghans, Anja Wittmann, Stefan Wieschalka, Ralf Takors, Nicole Erika Radde, Beate Presser, Verena Nold","doi":"10.1002/btpr.88508","DOIUrl":"10.1002/btpr.88508","url":null,"abstract":"Applying a single parameter set to describe complex mammalian kinetics often is too simplistic, as it fails to capture sensitive cell-to-environment interactions that may be exploited to optimize production performance. To resolve this time dependency, intra-experimental parameter shifts as part of design of dynamic experiments (DoDE) can be performed to study mammalian growth and production kinetics in fed-batch processes. This enables growth phase-dependent optimization, aligned with cellular requirements. Here, we provide a comprehensive, head-to-head comparison of our phase-dependent optimization approach with intra-experimental shifts of process parameters to a static optimization that retains parameter settings through the entire bioprocess. Showcasing a monoclonal antibody production process development scenario, the study examines growth phase-dependent effects of temperature (T) and dissolved oxygen (DO) together with time-invariant parameters for feed and seeding cell density. While the static optimization suggests settings near the center of the design space, phase-dependent optimization finds an optimum by shifting T and DO between the exponential growth, transition, and production phases. Overall, the phase-wise optimized process gives an experimentally validated ~30% increase in product titer while maintaining comparable product quality. Furthermore, the approach breaks the correlation between product titer and acidic charged variants: both depend on T but at different timeframes. Additionally, DoDE uncovers a crucial interaction between T and DO, with low T and high DO during the exponential growth phase, leading to strong lactate accumulation. The data demonstrate the advantages of phase-dependent optimization enabled by DoDE. The results may serve as a good practice example for follow-up research.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13441397/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147961963","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

Machine learning-assisted clone selection for intensified cell culture processes 强化细胞培养过程的机器学习辅助克隆选择。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-05-21 DOI: 10.1002/btpr.88519

Nicolas Wolnick, John Schmitt, Christian Siltanen, Julia Armstrong, Shannon Hendrick, Ariel Atchley, Brandon Downey

{"title":"Machine learning-assisted clone selection for intensified cell culture processes","authors":"Nicolas Wolnick, John Schmitt, Christian Siltanen, Julia Armstrong, Shannon Hendrick, Ariel Atchley, Brandon Downey","doi":"10.1002/btpr.88519","DOIUrl":"10.1002/btpr.88519","url":null,"abstract":"Intensified fed-batch processes are becoming increasingly prevalent among biomanufacturers due to their superior space–time yields relative to traditional, non-intensified fed-batch processes. However, the shift towards intensified manufacturing has unexpectedly made optimal clone selection more challenging. Clone selection, traditionally an empirical screening of candidates on the basis of productivity, has been complicated by the observation that clonal productivity rankings can vary substantially between intensified and non-intensified processes. In the absence of a small-scale intensified empirical screen, clones destined for intensified manufacturing are selected based on their performance in the non-intensified process. This mismatch risks selection of suboptimal clones, representing a missed opportunity to generate additional product at no extra cost. This study presents a machine learning approach to improve clone selection for intensified processes by predicting and ranking clonal productivity. Models were trained on non-intensified clone performance data routinely collected during cell line development to facilitate seamless integration into existing capabilities. To evaluate generalizability and simulate real-world application, independent clone panels expressing two distinct monoclonal antibodies were withheld from model training and used in comparative testing against the legacy clone selection method. The ranking model identified the most productive clone of those expressing the first mAb, representing a 68.5% higher titer than the clone selected by the legacy method. For the second mAb, the ranking model identified the second most productive clone, resulting in a marginally lower titer. These results demonstrate the potential of machine learning models as practical tools for improving the selection of high productivity clones in intensified bioprocessing.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13441383/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147986427","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

Regenerative perfusion bioreactor for biologics production with lower COGs. 用于低COGs生物制剂生产的再生灌注生物反应器。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 DOI: 10.1002/btpr.88550

Eric Wynne, Dohyun Park, Hourieh Ahi, Jaeweon Lee, Seongkyu Yoon, Jongyoon Han

{"title":"Regenerative perfusion bioreactor for biologics production with lower COGs.","authors":"Eric Wynne, Dohyun Park, Hourieh Ahi, Jaeweon Lee, Seongkyu Yoon, Jongyoon Han","doi":"10.1002/btpr.88550","DOIUrl":"https://doi.org/10.1002/btpr.88550","url":null,"abstract":"Perfusion bioreactor systems enable intensified bioprocessing with high cell densities and volumetric productivities in monoclonal antibody (mAb) manufacturing. However, their widespread adoption remains constrained by the high consumption of chemically defined cell culture media. In this study, we introduce a multi-stage electrokinetic waste separation process that regenerates spent media, recovering up to 87.5% of the spent-medium volume for reuse. Using a mock-perfusion model, the regeneration-recycle processes were shown to sustain high-density Chinese hamster ovary cell growth, compared with spent media recycling without regeneration. Integration into a self-recycling perfusion bioreactor (350 mL working volume) operating with 75% regenerated media sustained the target viable cell density and antibody titer, with limited reductions to cell-specific growth rate (~31%) and cell-specific productivity (~9%), supporting the feasibility of high-ratio media recycling. Techno-economic modeling of large-scale scenarios revealed that media regeneration could offer a path to $15/g COGs, thereby increasing the accessibility of mAbs. Techno-economic modeling of hypothetical large-scale manufacturing scenarios indicated that the experimentally demonstrated process could reduce cost of goods (COG) by approximately 9%, and identified a theoretical pathway toward ~$15/g COG contingent upon multi-cycle regeneration processes that could be developed in the future. These results support regenerative perfusion as a promising process-intensification strategy that, with further development, may be adaptable to other mammalian cell culture platforms where media cost and sustainability are critical bottlenecks.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88550"},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148676967","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

Strategies for removing two “problematic” host cell proteins, using an “end-to-end” approach 使用“端到端”方法去除两种“有问题”的宿主细胞蛋白的策略。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-04-23 DOI: 10.1002/btpr.88514

Greg Evangelist, Chris Kwiatkowski, Krishnakumar Malu, Suli Liu, Linda Yi, Sarwat Khattak, Brad Stanley

{"title":"Strategies for removing two “problematic” host cell proteins, using an “end-to-end” approach","authors":"Greg Evangelist, Chris Kwiatkowski, Krishnakumar Malu, Suli Liu, Linda Yi, Sarwat Khattak, Brad Stanley","doi":"10.1002/btpr.88514","DOIUrl":"10.1002/btpr.88514","url":null,"abstract":"Host cell proteins (HCPs) are critical process-related impurities of recombinant protein biopharmaceuticals that have the potential to impact product safety and efficacy. In this study, two residual HCPs, heat shock protein 90 beta and perilipin-4-like, produced from a CHO cell line, were identified during the development of a late-stage platform Immunoglobulin G subclass 1 (IgG1) monoclonal antibody process. A risk assessment was performed that included identification of HCPs, homology with their human protein counterparts, and prior non-clinical and clinical experience. The outcome deemed these two species as “problematic” and target levels were established to guide approaches for removal. Given the high productivity of the upstream and downstream platform processes, the goal was to explore conditions that minimize deviations from the platform. An end-to-end approach was performed that evaluated downstream levers, including Protein A washes, polishing chromatography operational parameters, and exploration of depth filter media. Upstream levers were also explored, evaluating effects of temperature shift and modulation of iron and citrate to help control levels of both HCP species. The results presented in this study demonstrated the upstream and downstream conditions achieved effective removal of the two HCP species to meet drug substance targets.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147760794","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

Evaluating the impact of ionic strength on retention of minute virus of mice during small virus-retentive filtration 小病毒保留过滤过程中离子强度对小鼠微小病毒保留的影响。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-04-20 DOI: 10.1002/btpr.88511

Cody Murphy, Ashley Slocum, Matthew Tierson, Benjamin Cacace, Paul Genest, Kathryn Martin Remington, Justin Hale

{"title":"Evaluating the impact of ionic strength on retention of minute virus of mice during small virus-retentive filtration","authors":"Cody Murphy, Ashley Slocum, Matthew Tierson, Benjamin Cacace, Paul Genest, Kathryn Martin Remington, Justin Hale","doi":"10.1002/btpr.88511","DOIUrl":"10.1002/btpr.88511","url":null,"abstract":"Small virus-retentive filtration (VRF) is a critical unit operation utilized in the viral clearance package to ensure the viral safety of biotherapeutics. This step is relied upon to provide effective parvovirus retention of ≥4 log reduction value (LRV) (e.g., minute virus of mice; MVM), and robust retention of larger viruses. Biomolecules, including multispecific antibodies and recombinant proteins, present purification challenges that require multiple chromatography columns to achieve final purity specifications. These additional polishing steps may utilize high ionic strength solution conditions to meet purity targets, and it is critical to assess if those process conditions impact the viral retention performance of the subsequent VRF operations. In one instance, retention performance was lower than expected (≤4 LRV) and was attributed to the use of a high ionic strength buffer. Two possible mechanisms were investigated, that is, reduction in the effective size of the MVM virus (~25 nm) and/or an increase in the membrane filter pore size, as observed with increase in the virus filter permeability. Based on modeling and experimental data presented here, at high ionic strength solution conditions the hydrodynamic size of MVM is reduced while virus-retentive membrane effective pore size is larger, as compared to low ionic strength conditions. Additionally, MVM LRV and membrane permeability of multiple commercial-grade virus filters were impacted by high ionic strength and process interruptions. These results demonstrate that high ionic strength solution conditions could impact parvovirus retention performance, and these findings may guide process development for biotherapeutics operating under similar atypical solution conditions.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147728196","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

Protein hydrolysates in cell culture: Toward multi-omics characterization 细胞培养中的蛋白水解物：迈向多组学表征。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-04-28 DOI: 10.1002/btpr.88505

Michelle Combe, Brandon M. Wrage, Angel Varela-Rohena, Stanislav Sokolenko

引用次数: 0

On-deck pH measurement with integrated feedback control on a micro-scale platform for monoclonal antibody low pH viral inactivation 微尺度单克隆抗体低pH病毒灭活平台上集成反馈控制的甲板pH测量。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-05-05 DOI: 10.1002/btpr.88516

Paras Sharma, Nikolas von den Eichen, Sabine Schweisgut, Lars Robbel, Michael Schmitt, Daniel G. Bracewell

{"title":"On-deck pH measurement with integrated feedback control on a micro-scale platform for monoclonal antibody low pH viral inactivation","authors":"Paras Sharma, Nikolas von den Eichen, Sabine Schweisgut, Lars Robbel, Michael Schmitt, Daniel G. Bracewell","doi":"10.1002/btpr.88516","DOIUrl":"10.1002/btpr.88516","url":null,"abstract":"The increasing demand for efficient monoclonal antibody manufacturing has accelerated the adoption of high throughput process development (HTPD) platforms, which enable rapid, automated screening of downstream operations. However, the integration of non-chromatographic steps such as low pH viral inactivation (VI) within automated workflows remains limited, largely due to the absence of a micro-scale method for accurate, on-deck pH measurement and control. This study presents the development and implementation of an automated pH measurement and feedback control system using optical pH sensors immobilized within 96-well microplates. The approach enables non-invasive, real-time monitoring of pH across the acidic range required for VI and is fully compatible with standard liquid-handling platforms. Integration of a feedback control algorithm allowed autonomous acid and base addition to achieve precise target pH values during both acidification and neutralization phases. The method achieved strong agreement between measured and expected pH values following optimization of measurement conditions, including ionic strength adjustment. The system was further integrated with Sartobind® Q and cation exchange chromatography steps to demonstrate an end-to-end automated workflow. Systematic assessment of cation exchange chromatography performance under controlled loading conditions enabled direct visualization of separation behavior and early identification of sub-optimal operating regions, demonstrating the platform's capability to expand experimental space and accelerate mechanistic process understanding. This work establishes a micro-scale, fully automated downstream platform with pH control, bridging a critical technological gap and advancing the vision of an end-to-end HTPD system for biopharmaceutical purification.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13441358/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147833040","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

Data-driven design of deterministic lateral displacement microfluidic devices for CTC separation 确定性横向位移微流控分离装置的数据驱动设计。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-04-29 DOI: 10.1002/btpr.88515

Tanbir Sarowar, Sadia Anjum Mim, Xiaolin Chen

{"title":"Data-driven design of deterministic lateral displacement microfluidic devices for CTC separation","authors":"Tanbir Sarowar, Sadia Anjum Mim, Xiaolin Chen","doi":"10.1002/btpr.88515","DOIUrl":"10.1002/btpr.88515","url":null,"abstract":"Circulating tumor cells (CTCs) are established biomarkers for cancer diagnosis and therapeutic monitoring, yet their extreme rarity in peripheral blood and size overlaps with leukocytes present substantial technical barriers to clinical deployment. Deterministic lateral displacement (DLD) is a promising microfluidic technique for label-free, continuous CTC separation. However, due to the extreme heterogeneity of cancer cells, rational device design requires computationally expensive iterative simulation across multi-dimensional geometric parameter spaces. Here we present a data-driven design framework that integrates validated computational fluid dynamics (CFD) simulations with supervised machine learning to enable rapid prediction of particle separation behavior and critical diameter in DLD arrays. A dataset of approximately 8.5 million trajectory data points was generated across 1160 unique combinations of array period number (N = 3–48) and particle diameter (1–20 μm) using COMSOL-based CFD. Four regression algorithms, Gradient Boosting, Random Forest, k-Nearest Neighbors, and Multi-Layer Perceptron, were trained and evaluated on trajectory prediction and critical diameter estimation. Random Forest achieved the highest accuracy, with a test-set R2 of 0.994 and a mean absolute error of 0.31 μm in critical diameter prediction. A clinical case study targeting colorectal CTC separation from blood demonstrated that the ML-guided design workflow converged on an optimal device configuration (N = 15, Dc = 10–12 μm) in under 2.3 s, representing a speedup exceeding four orders of magnitude compared to equivalent CFD iteration. The framework enables direct translation of CTC size distributions into DLD device configurations and provides a scalable foundation for reproducible microfluidic process development in biomedical applications.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147760831","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