Biotechnology Progress最新文献_第7页

Enhancing cryopreservation of human induced pluripotent stem cells: Bottom-up versus conventional freezing geometry 强化人诱导多能干细胞的低温保存：自下而上与传统冷冻几何的对比。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70019

Fernando Teodoro, Soukaina El-Guendouz, Rafaela Neves, Andreia Duarte, Miguel A. Rodrigues, Eduardo P. Melo

{"title":"Enhancing cryopreservation of human induced pluripotent stem cells: Bottom-up versus conventional freezing geometry","authors":"Fernando Teodoro, Soukaina El-Guendouz, Rafaela Neves, Andreia Duarte, Miguel A. Rodrigues, Eduardo P. Melo","doi":"10.1002/btpr.70019","DOIUrl":"10.1002/btpr.70019","url":null,"abstract":"Induced pluripotent stem cells (iPSCs) hold large potential in regenerative medicine due to their pluripotency and unlimited self-renewal capacity without the ethical issues of embryonic stem cells. To provide quality-controlled iPSCs for clinical therapies, it is essential to develop safe cryopreservation protocols for long-term storage, preferably amenable to scale-up and automation. We have compared the impact of two different freezing geometries (bottom-up and conventional radial freezing) on the viability and differentiation potential of human iPSCs. Our results demonstrate that bottom-up freezing under optimized conditions significantly increases iPSC viability, up to 9% for cell membrane integrity and up to 21% for cell metabolic state, compared to conventional freezing. The improvement achieved for bottom-up versus conventional freezing was maintained after scale-up from cryogenic vials to 30 mL bags, highlighting its potential for clinical applications. These findings show that bottom-up freezing can offer a more controlled and scalable cryopreservation strategy for iPSCs, promoting their application in regenerative medicine.","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":"143603834","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

Process optimization mitigated the retention loss of an Fc-fusion protein during ultrafiltration/diafiltration 工艺优化减轻了超滤/滤除过程中fc融合蛋白的保留损失。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70021

Hao Yu, Li Fei

{"title":"Process optimization mitigated the retention loss of an Fc-fusion protein during ultrafiltration/diafiltration","authors":"Hao Yu, Li Fei","doi":"10.1002/btpr.70021","DOIUrl":"10.1002/btpr.70021","url":null,"abstract":"In the downstream processing of antibody-based therapeutics, ultrafiltration/diafiltration (UF/DF) is commonly applied for concentration and buffer exchange in the final formulation. For a given molecule, various factors such as membrane type, feed flux, and transmembrane pressure (TMP) can significantly influence the performance of UF/DF, impacting yield, buffer exchange efficiency, and product quality. Conventional membrane pore size selection is based on product molecular weight to ensure high retention. While working on an Fc-fusion protein, we found that the pH of load material had a critical effect on the retention of the molecule due to conformational changes at different pH values, as evidenced by the size-exclusion chromatography (SEC). Meanwhile, optimization of the UF/DF process underscored the importance of concentration polarization to protein retention. Approaches to reduce concentration polarization, such as increasing feed flux and lowering TMP, resulted in less protein loss in the permeate stream. High retention of this Fc-fusion protein during the UF/DF step can be achieved not only by utilizing a 5 kDa membrane but also by employing a 10 kDa membrane with optimized process parameters such as load conditions, feed flux, and TMP. These observations provide important insights on the factors impacting protein retention beyond the molecular weight cutoff (MWCO) of UF/DF membrane.","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":"143603837","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

Enhancing the performance of an in vitro RNA biosensor through iterative design of experiments 通过实验迭代设计提高体外RNA生物传感器的性能。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70005

Rochelle Aw, Karen Polizzi

{"title":"Enhancing the performance of an in vitro RNA biosensor through iterative design of experiments","authors":"Rochelle Aw, Karen Polizzi","doi":"10.1002/btpr.70005","DOIUrl":"10.1002/btpr.70005","url":null,"abstract":"The quality control of RNA has become increasingly crucial with the rise of mRNA-based vaccines and therapeutics. However, conventional methods such as LC–MS often require specialized equipment and expertise, limiting their applicability to high throughput experiments. Here, we optimize a previously characterized RNA integrity biosensor, that provides a simple colorimetric output, using Design of Experiments (DoE). Through iterative rounds of a Definitive Screening Design (DSD) and experimental validation, we systematically explored different assay conditions to enhance the biosensor's performance. Optimization led to a 4.1-fold increase in dynamic range and reduced RNA concentration requirements by one-third, significantly improving usability. Notable modifications included reducing the concentrations of reporter protein and poly-dT oligonucleotide and increasing DTT concentration, suggesting a reducing environment for optimal functionality. Importantly, the optimized biosensor retained its ability to discriminate between capped and uncapped RNA even at lower RNA concentrations. Overall, our improved biosensor offers enhanced performance and reduced sample requirements, paving the way for rapid, cost-effective RNA quality control in diverse settings, including resource-limited environments.","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":"https://aiche.onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603835","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

Development of multiparametric bioprinting method for generation of 3D printed cell-laden structures 3D打印细胞负载结构的多参数生物打印方法的发展。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-12 DOI: 10.1002/btpr.70016

Sophie Lipshutz, Yoontae Kim, Micaila Curtis, Leanne Friedrich, Stella Alimperti

{"title":"Development of multiparametric bioprinting method for generation of 3D printed cell-laden structures","authors":"Sophie Lipshutz, Yoontae Kim, Micaila Curtis, Leanne Friedrich, Stella Alimperti","doi":"10.1002/btpr.70016","DOIUrl":"10.1002/btpr.70016","url":null,"abstract":"The organ transplantation field requires new approaches for replacing and regenerating tissues due to the lack of adequate transplant methods. Three-dimensional (3D) extrusion-based bioprinting is a rapid prototyping approach that can engineer 3D scaffolds for tissue regeneration applications. In this process, 3D printed cell-based constructs, consisting of biomaterials, growth factors, and cells, are formed by the extrusion of bioinks from nozzles. However, extrusion applies shear stresses to cells, often leading to cellular damage or membrane rupture. To address this limitation, herein, we developed and optimized a 3D bioprinting approach by evaluating the effect of key extrusion-based 3D bioprinting parameters—bioink viscosity, nozzle size, shape, and printing speed—on cell viability. Our results revealed that cells printed in higher-viscosity bioinks, with smaller, cylindrical nozzles, exhibited lower viability due to their exposure to high shear stresses. Translational flow speed had a cell-dependent impact, as different cell types have different sensitivities to the magnitude and duration of shear stress inside the nozzle. Overall, evaluating these parameters could facilitate the development of 3D high-resolution bioprinted constructs for tissue regeneration applications, offering a more efficient alternative to traditional fabrication methods, which are often labor intensive, expensive, and repetitive.","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":"https://aiche.onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603833","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

Assessing the probability of clonality achieved by single-cell cloning of CHO cells through cell deposition combined with imaging using distinguishable cells 通过细胞沉积结合可区分细胞成像，评估CHO细胞单细胞克隆获得克隆性的可能性。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-03-04 DOI: 10.1002/btpr.70012

Genevieve H. Nonet, Elena Scut, Raymond Ogawa, Milan T. Tomic

{"title":"Assessing the probability of clonality achieved by single-cell cloning of CHO cells through cell deposition combined with imaging using distinguishable cells","authors":"Genevieve H. Nonet, Elena Scut, Raymond Ogawa, Milan T. Tomic","doi":"10.1002/btpr.70012","DOIUrl":"10.1002/btpr.70012","url":null,"abstract":"Mammalian cell lines used for clinical studies and post-approval production of recombinant DNA-derived biotherapeutics are expected to be derived from a single cell, and regulatory submissions are expected to provide robust evidence of monoclonality. Imaged single-cell deposition followed by whole-well imaging using specialized instruments has, in many cell line development labs, replaced the “gold standard” of two rounds of limiting dilution due to its increased speed and the assurance of clonality provided by orthogonal images. However, there is still a lack of information on how the procedures used to define these clonal cell lines perform. Here we use a mixture of two distinguishable Chinese hamster ovary (CHO) cells to document that a greater than 99% probability of clonality can be obtained from our single-cell cloning method that uses our preparation procedures, the VIPS® single-cell deposition instrument, the Cell Metric® whole-well imager, and a comprehensive visual review. Together with the assurance of cell/well images, the determination of the probability of clonality of our VIPS+Cell Metric method provides a strong package of evidence of single-cell derivation of a recombinant CHO cell line.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143540029","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

Efficient expression of recombinant proteins in Bacillus subtilis using a rewired gene circuit of quorum sensing 重组蛋白在枯草芽孢杆菌群体感应基因回路中的高效表达。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70007

Wenliang Hao, Shihao Yang, Yuou Sheng, Chengfeng Ye, Laichuang Han, Zhemin Zhou, Wenjing Cui

{"title":"Efficient expression of recombinant proteins in Bacillus subtilis using a rewired gene circuit of quorum sensing","authors":"Wenliang Hao, Shihao Yang, Yuou Sheng, Chengfeng Ye, Laichuang Han, Zhemin Zhou, Wenjing Cui","doi":"10.1002/btpr.70007","DOIUrl":"10.1002/btpr.70007","url":null,"abstract":"Bacillus subtilis is a favored chassis for high productivity of several high value-added product in synthetic biology. Efficient production of recombinant proteins is critical but challenging using this chassis because these expression systems in use, such as constitutive and inducible expression systems, demand for coordination of cell growth with production and addition of chemical inducers. These systems compete for intracellular resources with the host, eventually resulting in dysfunction of cell survival. To overcome the problem, in this study, LuxRI quorum sensing (QS) system from Aliivibrio fischeri was functionally reconstituted in B. subtilis for achieving coordinated protein overproduction with cell growth in a cell-density-dependent manner. Furthermore, the output-controlling promoter, PluxI, was engineered through two rounds of evolution, by which we identified four mutants, P22, P47, P56, and P58 that exhibited elevated activity compared to the original PluxI. By incorporating a strong terminator (TB5) downstream of the target gene further enhanced expression level. The expression level of this system surpasses commonly used promoter-based systems in B. subtilis like P43 and PylbP. The LuxRI QS system proves to be a potent platform for recombinant protein overproduction in B. subtilis.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448011","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

Microcontact printing of lectin self-assembled monolayers for arbovirus detection 用于虫媒病毒检测的凝集素自组装单层微接触印刷。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70008

Raiza P. S. Lucena, Alberto G. Silva-Junior, Isaac A. M. Frías, Laura H. V. Gil, Marli T. Cordeiro, Abdelhamid E. El Salhi, Cesar A. S. Andrade, Maria D. L. Oliveira

{"title":"Microcontact printing of lectin self-assembled monolayers for arbovirus detection","authors":"Raiza P. S. Lucena, Alberto G. Silva-Junior, Isaac A. M. Frías, Laura H. V. Gil, Marli T. Cordeiro, Abdelhamid E. El Salhi, Cesar A. S. Andrade, Maria D. L. Oliveira","doi":"10.1002/btpr.70008","DOIUrl":"10.1002/btpr.70008","url":null,"abstract":"Arboviruses significantly burden public health in Brazil, constituting a constant challenge for health authorities. The diagnosis and, consequently, clinical management and the reporting of arbovirus infections in regions where multiple arboviruses coexist are complex processes. Herein, we report the development of a new electrochemical biosensor based on Concanavalin A (ConA) to identify carbohydrate patterns in the viral structure of Dengue 3 (DENV-3), Zika (ZIKV) and Chikungunya (CHIKV) viruses. The biorecognition of arboviruses was carried out through functionalization with 4-aminophenylacetic acid (CMA) on poly (ethylene terephthalate) (PET) substrate coated with a gold layer combining microcontact printing (μCP). Bovine serum albumin (BSA) was used after ConA immobilization to block binding to nonspecific sites. Subsequently, the interaction between ConA and arbovirus was characterized by standard atomic force microscopy (AFM), fluorescence microscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). Fluorescent imaging was conducted to confirm the occurrence of the DENV-3, ZIKV, and CHIKV detection processes. The obtained results demonstrated the success of the biosensor (CMA-ConA-BSA) manufactured on a PET substrate using μCP for detecting medically significant arboviruses. RCT values showed an increase in impedimetric response total of the system after exposition to DENV-3 (RCT = 68.82 kΩ) and a lower recognition to CHIKV (RCT = 44.44 kΩ). The present biosensor platform reveals the applicability of the ConA lectin in the viral biorecognition process based on flexible biosensors for differential detection of DENV-3, ZIKV, and CHIKV. ConA-based electrochemical biosensor provide high selectivity, real-time detection, and low volumes of analytes.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448017","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

Recent trends in biotechnological production, engineering, and applications of lysophospholipases 溶血磷脂酶的生物技术生产、工程和应用的最新趋势。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70014

Arshia Nazir, Muhammad Sajjad

{"title":"Recent trends in biotechnological production, engineering, and applications of lysophospholipases","authors":"Arshia Nazir, Muhammad Sajjad","doi":"10.1002/btpr.70014","DOIUrl":"10.1002/btpr.70014","url":null,"abstract":"Oil degumming process involves the removal of gums, which is required to improve the physicochemical and storage properties of the vegetable oils. Degumming of oils can be carried out by using chemicals, membranes (polymeric, inorganic, and ceramic), or enzymes, for example, phospholipases. Phospholipases are enzymes of tremendous significance in the degumming process as they convert gums to fatty acids and lipophilic substances. They provide a cost-effective and safe alternative to other degumming processes without affecting the oil yield. Lysophospholipases (LPLs) are highly valuable tools for degumming vegetable oils. LPLs can hydrolyze fatty acyl ester bonds of phosphatidylcholine at the sn-1 and sn-2 positions of glycerol moiety. In addition, they have the ability to catalyze hydrolysis lysophospholipids' ester bond either at sn-1 or sn-2 position. In this review, biotechnological production and biochemical characteristics of LPLs from three domains of life are highlighted. In comparison to bacterial and eukaryotic LPLs, archaeal LPLs were found to be active at high temperatures. Broad substrate specificity and thermostability of archaeal LPLs make them ideal candidates for the industrial degumming of oils. However, improvement of activity and substrate specificity of archaeal LPLs is required for enhancing their industrial utility. In the current review, various protein-engineering approaches (directed evolution, rational design, site-saturation mutagenesis, and fusion technology) as well as in silico tools have been discussed to increase the commercial significance of LPLs.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143447616","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 synthetic platform for developing recombinant adeno-associated virus type 8 producer cell lines 重组腺相关病毒8型产生细胞系的合成平台。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70009

Yu-Chieh Lin, Han-Jung Kuo, Min Lu, Thomas Mahl, George Aslanidi, Wei-Shou Hu

{"title":"A synthetic platform for developing recombinant adeno-associated virus type 8 producer cell lines","authors":"Yu-Chieh Lin, Han-Jung Kuo, Min Lu, Thomas Mahl, George Aslanidi, Wei-Shou Hu","doi":"10.1002/btpr.70009","DOIUrl":"10.1002/btpr.70009","url":null,"abstract":"Recombinant adeno-associated virus (rAAV) is one of the most widely used viral vectors for gene therapy. It is used in very high doses for the treatment of many diseases, making large-scale production for clinical applications challenging. We have established a synthetic biology-based platform to construct stable production cell lines, which can be induced to produce rAAV2. In this study, we extended our cell line construction pipelines for rAAV2 to rAAV8, a serotype whose tropism makes it attractive for gene delivery in multiple tissues. The Genome Module, encoding the rAAV2 genome, and Replication Modules, containing Rep68, DBP and E4orf6 coding sequences, originally used for rAAV2 were retained, but the Packaging Module was modified to replace the AAV2 intron-less cap gene (VP123) with that of AAV8. These three genetic modules were integrated into HEK293 genome to generate four rAAV8 producer cell lines VH1-4, which all produced rAAV8 upon induction. Their productivity was similar to the initial rAAV2 producer cell lines GX2/6 constructed using the same pipeline, but was much lower than conventional triple plasmid transfection. We identified Cap protein production and capsid formation as a potential limiting factor, just as we observed in GX2/6. By integrating more copies of AAV8 VP123 into VH3 clone, the encapsidated rAAV8 titer increased 20-fold to a level comparable to triple transfection. By tuning induction conditions to modulate capsid production, the full particle content could be elevated. This study demonstrated that our rAAV producer cell line development platform is robust and applicable to different AAV serotypes.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448007","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

Detergent/surfactant retention during ultrafiltration in the formulation of biotherapeutics 生物治疗制剂超滤过程中洗涤剂/表面活性剂的保留。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70011

Liang-Kai Chu, Zhuoshi Du, Matthew Billups, Hee Jeung Oh, Andrew L. Zydney

{"title":"Detergent/surfactant retention during ultrafiltration in the formulation of biotherapeutics","authors":"Liang-Kai Chu, Zhuoshi Du, Matthew Billups, Hee Jeung Oh, Andrew L. Zydney","doi":"10.1002/btpr.70011","DOIUrl":"10.1002/btpr.70011","url":null,"abstract":"Surfactants like polysorbate (Tween®) are commonly used as excipients in the production of monoclonal antibodies and other recombinant proteins. The retention behavior of these excipients in the final ultrafiltration step can be difficult to predict due to the presence of both monomers and micelles. This study examined the retention of polysorbate during ultrafiltration through cellulose and polyethersulfone membranes with nominal molecular weight cutoffs of 10, 30, and 100 kDa. Novel flux stepping experiments were performed to examine the effects of concentration polarization on surfactant transmission. Polysorbate 20 transmission through the 30 kDa membrane was a strong function of the surfactant concentration, decreasing from nearly 100% for a 2.5 mg/L solution to <10% for a 50 mg/L solution due to high retention of the micelles. Polysorbate transmission was lower for the polyethersulfone membrane due to polysorbate adsorption. A simple mathematical model was developed to describe the polysorbate transmission accounting for the effects of concentration polarization as well as the presence of surfactant monomers and micelles. Model calculations were in good agreement with the experimental data, providing a framework for the analysis and design of ultrafiltration/diafiltration processes for biopharmaceutical formulations containing surfactants.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"41 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.70011","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448009","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