Biotechnology Progress最新文献_第6页

Continuous glucose feedback control using Raman spectroscopy and deep learning models for biopharmaceutical processes 使用拉曼光谱和生物制药过程的深度学习模型的连续葡萄糖反馈控制。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-04-02 DOI: 10.1002/btpr.70020

Mohammad Rashedi, Matthew Demers, Hamid Khodabandehlou, Tony Wang, Christopher Garvin, Steve Rianna

{"title":"Continuous glucose feedback control using Raman spectroscopy and deep learning models for biopharmaceutical processes","authors":"Mohammad Rashedi, Matthew Demers, Hamid Khodabandehlou, Tony Wang, Christopher Garvin, Steve Rianna","doi":"10.1002/btpr.70020","DOIUrl":"10.1002/btpr.70020","url":null,"abstract":"This study explores the implementation of continuous glucose control strategies in high-consumption, high-complexity cell culture processes using Raman spectroscopy and advanced deep learning models, including convolutional neural networks and variational autoencoder just-in-time learning. By leveraging deep learning-derived process monitoring, the study enhances glucose measurement accuracy and stability, enabling precise control across different glucose set points. This approach allows for a systematic evaluation of glycosylation effects and other critical quality attributes, addressing the impact of glucose variability on product consistency. Continuous glucose control is compared against traditional bolus feeding, demonstrating improved set-point maintenance, reduced high mannose (HM) levels, and enhanced overall titer productivity. To extend these benefits to manufacturing environments where Raman spectroscopy may not be feasible, a continuous glucose calculator (CGC) is developed as a scalable alternative. Experimental validation across multiple cell lines confirmed that both Raman-based and CGC-driven strategies minimized glucose fluctuations, reduced undesirable byproducts, and optimized process yields. These findings highlight the potential of continuous glucose control, combined with deep learning models, to improve bioprocess efficiency and product quality while addressing the challenges of dynamic, high-consumption bioreactor systems.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143762630","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

Cryopreservation practices in clinical and preclinical iPSC-based cell therapies: Current challenges and future directions 低温保存在临床和临床前基于ipsc细胞治疗的实践：当前的挑战和未来的方向。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-04-02 DOI: 10.1002/btpr.70031

Michael Dobruskin, Geoffrey Toner, Ronald Kander

引用次数: 0

Enhancement of D-mannitol production by fine-tuned expression of mannitol-1-phosphate dehydrogenase in Synechocystis sp. PCC6803 聚囊藻PCC6803中甘露醇-1-磷酸脱氢酶的微调表达提高d -甘露醇产量。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-04-02 DOI: 10.1002/btpr.70027

Wenyang Wu, Wei Du, Klaas J. Hellingwerf, Filipe dos Branco dos Santos

{"title":"Enhancement of D-mannitol production by fine-tuned expression of mannitol-1-phosphate dehydrogenase in Synechocystis sp. PCC6803","authors":"Wenyang Wu, Wei Du, Klaas J. Hellingwerf, Filipe dos Branco dos Santos","doi":"10.1002/btpr.70027","DOIUrl":"10.1002/btpr.70027","url":null,"abstract":"D-Mannitol production was achieved in freshwater Synechocystis sp. PCC6803 via the heterologous expression of mannitol-1-phosphate dehydrogenase (mtlD) and mannitol-1-phosphatase (m1p) under control of the strong promoter Ptrc1. However, only 5.54 mg L−1 of mannitol was found extracellularly after 7 days of cultivation, likely due to insufficient expression of a mutated mtlD lacking a methionine at position 332. This study compared mannitol levels using different promoters (Ptrc1, PpsbA2 and PnrsB) to control the expression of (un)mutated versions of mtlD in Synechocystis with co-expression of m1p. Our data suggest that even without the inducer, the weakest promoter, PnrsB, can support the expression of an unmutated mtlD in Synechocystis, which leads to 18.2 mg L−1 of mannitol in 7 days without induction. Such titer is already much higher than the first engineered mannitol-producing Synechocystis. When 5 μM nickel sulfate was added to the medium as an inducer, mannitol production could significantly be increased further, up to 92.9 mg L−1 after 7 days of induction, but it partially inhibited growth. Attempts with the other increasingly stronger promoters always failed to express the unmutated mtlD, probably due to the toxicity caused by the accumulation of the intermediate product, mannitol-1-phosphate. These results clearly suggest that the expression level of mtlD is the bottleneck in achieving a high yield of mannitol in Synechocystis, and consequently, that mannitol production can be enhanced by fine-tuning its expression. Future research is needed to identify bottlenecks that hinder mannitol productivity and long-term stability, facilitating the engineering of more efficient mannitol-producing cyanobacterial strains.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://aiche.onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70027","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143762638","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

Triton CG-110 as an alternative for cell lysis in manufacturing of adeno-associated virus-based gene therapy Triton CG-110在制造腺相关病毒基因治疗中作为细胞裂解的替代品。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-26 DOI: 10.1002/btpr.70025

Yixuan Ming, Tianyi Zhou, Bin Lu, Yemaiza Ojeda-Lassalle, Pasquale Valerio, Lu Wang, Mi Jin

引用次数: 0

Effect of nucleophilic additives on phosphoric acid pretreatment of lignocelluloses 亲核添加剂对木质纤维素磷酸预处理的影响。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-25 DOI: 10.1002/btpr.70026

Xin Tan, Xuan Wu, Wei Wang, Jiale An, Qin Zhang, Song Tang, Bangxiang He, Chenhuan Lai, Yequan Sheng

{"title":"Effect of nucleophilic additives on phosphoric acid pretreatment of lignocelluloses","authors":"Xin Tan, Xuan Wu, Wei Wang, Jiale An, Qin Zhang, Song Tang, Bangxiang He, Chenhuan Lai, Yequan Sheng","doi":"10.1002/btpr.70026","DOIUrl":"10.1002/btpr.70026","url":null,"abstract":"The inhibition of lignin condensation during biomass pretreatment is crucial for enhancing enzymatic hydrolysis efficiency, since the formation of rigid cross-linked lignin networks hinders cellulose accessibility and enzyme activity. This study investigates the effects of nucleophilic additives, including ascorbic acid (AsA), 2-naphthol (2N), 3-hydroxy-2-naphthoic acid (3H2NA), and 2-naphthol-7-sulfonate (7S2NA), as potential agents to suppress lignin condensation on the phosphoric acid pretreatment of poplar. The phosphoric acid pretreatment demonstrated a remarkable efficacy in the removal of xylan (100%) and lignin (18.06%–31.35%) from poplar, both with and without the inclusion of nucleophilic additives. An enzymatic hydrolysis yield ranging from 71.41% to 100% was achieved with the incorporation of AsA, 2N, 3H2NA, and 7S2NA, compared to a yield of 66.15% for substrates pretreated solely with phosphoric acid. The enhancement in enzymatic hydrolysis yield upon the addition of nucleophilic additives was probably due to the improved cellulose accessibility and the enhanced proportion of cellulose II in the pretreated substrates. The analysis of total phenolic content in the prehydrolysates revealed that 3H2NA and 7S2NA, characterized by their strong hydrophilic groups within their chemical structures, significantly facilitated lignin fractionation during phosphoric acid pretreatment.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699201","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

Iterative hybrid model based optimization of rAAV production 基于迭代混合模型的rAAV生产优化。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-24 DOI: 10.1002/btpr.70006

Claudio Müller, Gerald Siegwart, Susanne Heider, Michael Sokolov, Angela Botros, Alexandra Umprecht, Moritz von Stosch, Mariano Nicolas Cruz Bournazou

{"title":"Iterative hybrid model based optimization of rAAV production","authors":"Claudio Müller, Gerald Siegwart, Susanne Heider, Michael Sokolov, Angela Botros, Alexandra Umprecht, Moritz von Stosch, Mariano Nicolas Cruz Bournazou","doi":"10.1002/btpr.70006","DOIUrl":"10.1002/btpr.70006","url":null,"abstract":"Changes in serotype or genetic payload of recombinant adeno associated virus (rAAVs) gene therapies require adapting the transfection conditions of the upstream HEK293 cultivations. This study adopts an iterative model-based experiment design approach, where increasing data availability is leveraged to evolve models of different complexity. Initial models based on data from shaker flask runs guided the design of the first round at Ambr250 scale. With Ambr250 data becoming available, hybrid models capturing process state evolutions and historical models incorporating these evolutions to predict rAAV titer, were developed. These models were then combined into a full model approach, which was utilized within a Bayesian Optimization framework for the design of a second round of Ambr250 scale runs. The iterative approach was tested across different projects applying transfer learning to enhance the predictive power and improve the subsequent optimization. The approach was benchmarked against a statistical Design of Experiment method. The results show that the model-based experiment design consistently (and across projects) produces higher rAAV titer values than the benchmark approach (Project C: 4.4% or 7.0% increases in titer values relative to the response surface modeling approach for ELISA and ddPCR, respectively; Project D: 32.4% or 10.9% increases in titer values relative to the standard DoE-screening pick for ELISA and ddPCR, respectively), effectively optimizing the transfection mixture composition. The combination of propagation and historical models, augmented by transfer learning and an ever-increasing amount of data, enhanced the process design workflow, contributing to improved rAAV production through efficient transfection strategies.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://aiche.onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70006","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699206","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

Synthetic modeling: A cell-free approach for faster implementation of Raman spectroscopy in cell culture 合成建模：在细胞培养中更快实现拉曼光谱的无细胞方法。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-24 DOI: 10.1002/btpr.70018

Célia Sanchez, Hadi El Radi, Nathan Gay, Johan Cailletaud, Kévin Grollier, Fabrice Thomas, Thierry Gonthiez

{"title":"Synthetic modeling: A cell-free approach for faster implementation of Raman spectroscopy in cell culture","authors":"Célia Sanchez, Hadi El Radi, Nathan Gay, Johan Cailletaud, Kévin Grollier, Fabrice Thomas, Thierry Gonthiez","doi":"10.1002/btpr.70018","DOIUrl":"10.1002/btpr.70018","url":null,"abstract":"Monitoring cell culture is crucial for gaining a deeper understanding of processes and ensuring the production of safe and high-quality products. The capability to measure in real time several parameters of interest can be achieved with Raman spectroscopy. However, before using Raman spectroscopy to monitor a specific process, a calibration phase is required to develop chemometric models that correlate Raman spectra with the target parameters. It is mandatory to conduct this phase with multiple batches to build robust models that account for biological variability. This model building phase can be time-consuming and require a lot of resources. The industry is actively seeking solutions to simplify and expedite this step without compromising accuracy. Moreover, the current approach has limitations regarding changing cell culture media, celllines, or process scale. The novel synthetic model approach provides a significant gain of time and resources for the calibration phase, which is reduced to just a few days. The methodology involves using cell-free samples of cell culture media that are spiked with various concentrations of target compounds. The results indicate that the innovative approach enables accurate measurement for glucose and lactate parameters in real process conditions comparable to a standard modeling methodology.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://aiche.onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143690733","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

5-Aza combined with VPA reprograms human T lineage acute leukemia Jurkat cells into B-cell-like cells by epigenetic activation of PAX5 5-Aza联合VPA通过PAX5的表观遗传激活将人T系急性白血病Jurkat细胞重编程为b细胞样细胞。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-17 DOI: 10.1002/btpr.70023

Wenjin Xi, Guoxu Zheng, Xu Chen, Baile Zuo, Wei Wang, Yufang Li, Chunmei Zhang, Jie Chu, Xiuli Mu, Weihong Wen, Tao Wang, An-Gang Yang

{"title":"5-Aza combined with VPA reprograms human T lineage acute leukemia Jurkat cells into B-cell-like cells by epigenetic activation of PAX5","authors":"Wenjin Xi, Guoxu Zheng, Xu Chen, Baile Zuo, Wei Wang, Yufang Li, Chunmei Zhang, Jie Chu, Xiuli Mu, Weihong Wen, Tao Wang, An-Gang Yang","doi":"10.1002/btpr.70023","DOIUrl":"10.1002/btpr.70023","url":null,"abstract":"Epigenetic regulation plays an important role in cell fate reprogramming. Here, we found that inhibitors of epigenetic modifiers, including VPA, TSA, and 5-Aza-2'-deoxycytidine, can induce phenotypic transformation from Jurkat cells into B-cell-like cells. When Jurkat cells were treated with 5-Aza combined with VPA, B cell and stem cell marker expression was observed. These gene expression pattern changes were most remarkable in the optimized B cell induction conditions provided by the cocultured and genetically modified murine bone marrow OP9 cells. In such conditions, Jurkat cells were endowed with the ability to secrete B cell cytokines, and B lymphocyte-related genes and pathways were activated. In studying the mechanism underlying Jurkat cell reprogramming by 5-Aza and VPA, we found that PAX5, the key transcription factor regulating B cell development, was significantly upregulated. Treatment with 5-Aza and VPA inhibited the methylation of CpG islands and upregulated the acetylated H3K9 modification in the PAX5 promoter region, respectively, thus epigenetically activating the expression of PAX5 and promoting the reprogramming of Jurkat cells. Similar reprogramming results were also observed in primary CD4+T cells following treatment with 5-Aza and VPA. Our results provide a de novo paradigm for the reprogramming of T cells through epigenetic modifications.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://aiche.onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70023","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143647073","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

Predictive mechanistic model for separation of monoclonal antibody, fab fragment, and aggregate species on multimodal chromatography 单克隆抗体、fab片段和聚集体的多模态色谱分离预测机制模型。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-17 DOI: 10.1002/btpr.70022

Lalita Kanwar Shekhawat, Todd Markle, Jean-Luc Maloisel, Gunnar Malmquist

{"title":"Predictive mechanistic model for separation of monoclonal antibody, fab fragment, and aggregate species on multimodal chromatography","authors":"Lalita Kanwar Shekhawat, Todd Markle, Jean-Luc Maloisel, Gunnar Malmquist","doi":"10.1002/btpr.70022","DOIUrl":"10.1002/btpr.70022","url":null,"abstract":"The specific selectivities offered by multimodal ligands drive the increased application of multimodal chromatography in the purification of complex new “multispecific” antibodies, which requires improved understanding of the protein-multimodal ligand interaction mechanism. In the present study, a mechanistic model is developed to predict monoclonal antibody (mAb1)-Fab fragment (Fab) and heterogeneous aggregates separation on Capto™ MMC ImpRes multimodal resin based on the general rate model coupled with the proposed preferential interaction (PI) analysis-based Langmuir non-linear binding model. The model input value of binding parameters is obtained from Perkin et al. developed PI model, fit to the characteristic ‘U’-shaped curve for isocratic retention factors of mAb1, Fab, and aggregates as a function of NaCl salt concentrations. The model successfully simulates mAb1 and Fab elution peaks, whereas in the absence of deconvoluted peaks of heterogeneous aggregates, aggregates are modeled as a single species, giving satisfactory prediction of elution peak position, describing the average of the multiple (majority as double peaks) aggregate elution peaks. The physical significance of model estimated binding parameters is obtained from model estimated total number of released counter salt ions and water molecules for each species during binding, found to be consistent with their isocratic retention data. The underlying mechanism of double peak elution of aggregates during linear gradient elution was investigated based on mechanistic model estimated equilibrium constant. The proposed predictive mechanistic model was successfully validated by predicting mAb1, Fab, and aggregates elution peaks for the multimodal column operated in hydrophobic interaction mode and can be successfully implemented for process development.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143647078","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

Improving the expression yield of recombinant adeno-associated virus serotype 2 using dimethyl sulfoxide DMSO as an additive to the triple transient transfection process of HEK293 cells 二甲亚砜DMSO在HEK293细胞三重瞬时转染过程中提高重组腺相关病毒血清型2的表达量

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70017

Alexander Burns, Daniel Ramos-Sono, Saurav Datta

{"title":"Improving the expression yield of recombinant adeno-associated virus serotype 2 using dimethyl sulfoxide DMSO as an additive to the triple transient transfection process of HEK293 cells","authors":"Alexander Burns, Daniel Ramos-Sono, Saurav Datta","doi":"10.1002/btpr.70017","DOIUrl":"10.1002/btpr.70017","url":null,"abstract":"One of the widely used techniques for producing recombinant adeno-associated virus serotype 2 (rAAV2) particles, as viral vectors for gene therapy applications, is the triple transient (TT) transfection of human embryonic kidney 293 (HEK293) cells. It is desirable to optimize this transfection process for more efficient manufacturing of rAAV viral vectors for gene therapy purposes. We examined the application of dimethyl sulfoxide (DMSO) as an additive to this transfection technique to improve the expression yield of rAAV2 particles with HEK293 cells in adherent and suspension cell culture modalities. This assistance by DMSO should increase the trafficking of plasmid DNA (pDNA) through the cell membrane, and thus, increase the viral titer of rAAV2 full capsids at the time of harvesting the cell culture. The study demonstrated that DMSO as an additive for the TT transfection process led to an 8.2-fold increase in the expression yield of full AAV2 capsids using HEK293 cells in adherent cell culture modality, and also led to a 4.0-fold increase in the expression yield of full AAV2 capsids using HEK293 cells in suspension cell culture modality. There are no reported studies on the application of DMSO as an additive to the TT transfection process of HEK293 cells for the production of AAV particles. This is a novel, simple, and inexpensive method to improve the yield of rAAV2 full capsids with the TT transfection process of HEK293 cells, using a well-known cryoprotectant agent (CPA), as an additive to this transfection process.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603836","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