Biotechnology Progress最新文献_第3页

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":"https://doi.org/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":" ","pages":"e70014"},"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":"https://doi.org/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":" ","pages":"e70009"},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","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":"","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":"https://doi.org/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":" ","pages":"e70011"},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","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":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Improving outcomes in intensified processing via optimization of the cell line development workflow. 通过优化细胞系开发工作流程，改善强化处理的结果。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70003

Vincent Balassi, Mary Otto, Corey Kretzmer, Amber Petersen, Channing McLaurin, Jana Mahadevan, Jason Gustin, Trissa Borgschulte, David Razafsky

{"title":"Improving outcomes in intensified processing via optimization of the cell line development workflow.","authors":"Vincent Balassi, Mary Otto, Corey Kretzmer, Amber Petersen, Channing McLaurin, Jana Mahadevan, Jason Gustin, Trissa Borgschulte, David Razafsky","doi":"10.1002/btpr.70003","DOIUrl":"https://doi.org/10.1002/btpr.70003","url":null,"abstract":"As the industry continues to explore the benefits of continuous and intensified manufacturing, it is important to assure that the cell line development (CLD) workflows in practice today are well suited to generate clones that meet the unique challenges associated with these processes. Most cell lines used in intensified processes are currently developed using traditional fed-batch CLD workflows followed by adaptation of these cell lines to perfusion processes. This method maybe suboptimal as fed-batch CLD workflows select clones which produce high volumetric titers irrespective of cell growth rate and specific productivity (qP). Although sufficient for fed-batch processes, performance of cells derived from this traditional CLD workflow may not be maintained in perfusion processes, where an intricate balance of performance parameters is needed. Until now, a thorough investigation into the effect of the CLD workflow on top clone performance in perfusion processes has not been conducted. Here, we show how the CLD workflow impacts cell performance in both fed-batch and perfusion processes, emphasizing the advantages of adopting a perfusion-specific CLD workflow which includes the use of medium specially designed for expansion and production in a perfusion setting, scale-down models which more accurately simulate perfusion process, and the adoption of perfusion-specific cell line selection criteria. Together, this results in the development of more efficient cell lines, fit for continuous and intensified processing.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70003"},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448015","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 real-time cell culture process monitoring through the integration of advanced machine learning techniques: A comparative analysis of Raman and capacitance spectroscopies. 通过集成先进的机器学习技术增强实时细胞培养过程监测：拉曼光谱和电容光谱的比较分析。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70013

Feng Xu, Nuno Pinto, George Zhou, Sanjeev Ahuja

{"title":"Enhancing real-time cell culture process monitoring through the integration of advanced machine learning techniques: A comparative analysis of Raman and capacitance spectroscopies.","authors":"Feng Xu, Nuno Pinto, George Zhou, Sanjeev Ahuja","doi":"10.1002/btpr.70013","DOIUrl":"https://doi.org/10.1002/btpr.70013","url":null,"abstract":"Machine learning (ML) techniques have emerged as an important tool improving the capabilities of online process monitoring and control in cell culture process for biopharmaceutical manufacturing. A variety of advanced ML algorithms have been evaluated in this study for cell growth monitoring using spectroscopic tools, including Raman and capacitance spectroscopies. While viable cell density can be monitored real-time in the cell culture process, online monitoring of cell viability has not been well established. A thorough comparison between the advanced ML techniques and traditional linear regression method (e.g., Partial Least Square regression) reveals a significant improvement in accuracy with the leading ML algorithms (e.g., 31.7% with Random Forest regressor), addressing the unmet need of continuous monitoring viability in a real time fashion. Both Raman and capacitance spectroscopies have demonstrated success in viability monitoring, with Raman exhibiting superior accuracy compared to capacitance. In addition, the developed methods have shown better accuracy in a relatively higher viability range (>90%), suggesting a great potential for early fault detection during cell culture manufacturing. Further study using ML techniques for VCD monitoring also showed an increased accuracy (27.3% with Raman spectroscopy) compared to traditional linear modeling. The successful integration of ML techniques not only amplifies the potential of process monitoring but also makes possible the development of advanced process control strategies for optimized operations and maximized efficiency.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70013"},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448013","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

The production of succinate with more CO₂ fixation reactions facilitated by RuBisCO-based engineered Escherichia coli. 基于rubisco的工程大肠杆菌促进了更多CO2固定反应的琥珀酸盐生产。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-19 DOI: 10.1002/btpr.70015

Xiuyuan Zhou, Linqing Li, Shengjie Sun, Peng Xiong, Xiutao Liu

{"title":"The production of succinate with more CO2 fixation reactions facilitated by RuBisCO-based engineered Escherichia coli.","authors":"Xiuyuan Zhou, Linqing Li, Shengjie Sun, Peng Xiong, Xiutao Liu","doi":"10.1002/btpr.70015","DOIUrl":"https://doi.org/10.1002/btpr.70015","url":null,"abstract":"Redesigning metabolic pathways to enhance the efficiency of carbon fixation during chemical biosynthesis is a promising approach for achieving cleaner and greener production of multi-carbon compounds. In this study, we established a model of cell growth in Escherichia coli that is dependent on the RuBisCO-Prk pathway by regulating its central metabolism. This rewiring ensures that growth depends on RuBisCO's carboxylation, allowing heterotrophic growth to rely on carbon fixation. This model was verified by detecting the growth curve, and it was used to screen four RuBisCO genes, of which the gene from Rhodospirillum rubrum ATCC 11170 serves as a growth advantage for E.coli. In addition, this model was applied to construct an efficient succinate biosynthetic pathway that can produce two moles of succinate from one mole of xylose and three moles of CO2. Compared to conventional succinate biosynthesis, this strategy has a CO2 fixation capacity that is 1.5 times greater. Furthermore, to optimize succinate production, various approaches were employed, including the optimization of key enzymes, substrate transport, and the supply of inorganic carbon. The resulting strain was capable of producing succinate at a level of 2.09 ± 0.14 g/L, which is nearly 22.4 times that of the original strain. In conclusion, this study was developed for the production of two moles of succinate by implementing three moles of carbon fixation reactions and demonstrated the feasibility of various optimization strategies in biological carbon fixation.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70015"},"PeriodicalIF":2.5,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143447639","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

pH and conductivity transients during elution of IgG from protein A columns. 从A蛋白柱中洗脱IgG时的pH值和电导率变化。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-18 DOI: 10.1002/btpr.3534

Rainer Hahn, Lukas Berger, Jürgen Beck, Giorgio Carta

{"title":"pH and conductivity transients during elution of IgG from protein A columns.","authors":"Rainer Hahn, Lukas Berger, Jürgen Beck, Giorgio Carta","doi":"10.1002/btpr.3534","DOIUrl":"https://doi.org/10.1002/btpr.3534","url":null,"abstract":"The evolution of pH and conductivity waves during elution of IgG from protein A columns is studied for the resin MabSelect PrismA as well as other commercial resins using glycine and acetate buffers as eluents. The effects of buffer composition, IgG load, and residence time are explored. For glycine buffers, conductivity and pH waves travel through the column at different speeds, with the conductivity wave emerging in the column void volume and the pH waves emerging in 1 to 6 column volumes (CV) dependent on the buffer composition. The pH effluent temporarily overshoots the feed value, followed by a sharp drop as the pH approaches the eluent pH. For these conditions, elution of IgG is delayed and, at high loads, most of the IgG loaded elutes from the column at high pH values. Complex pH profiles, involving overshoots and delays between conductivity and pH waves are also observed when eluting with dilute sodium acetate (50 mM) or with acetic acid, but to a lesser extent. No overshoots or delays are observed for more concentrated sodium acetate (100 mM). A mechanistic model is developed to predict elution with glycine buffers. Model predictions agree with the experimental trends. In particular, the model shows that when eluting a partially loaded column, IgG desorbed near the column entrance is re-adsorbed downstream of the pH front eventually emerging at high concentration and high pH. The model can be used to design optimized buffers and predict the pH of the IgG elution pool.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3534"},"PeriodicalIF":2.5,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439855","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

The combination of oleic acid, linoleic acid, palmitoleic acid, and α-linolenic acid promoted the expansion of NK-92 cells in vitro. 油酸、亚油酸、棕榈油酸和α-亚麻酸联合作用可促进NK-92细胞体外扩增。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-18 DOI: 10.1002/btpr.70000

Shumin Zhang, Huimin Huang, Jingwei Zhang, Yuanyuan Zhao, Wen-Song Tan, Haibo Cai

{"title":"The combination of oleic acid, linoleic acid, palmitoleic acid, and α-linolenic acid promoted the expansion of NK-92 cells in vitro.","authors":"Shumin Zhang, Huimin Huang, Jingwei Zhang, Yuanyuan Zhao, Wen-Song Tan, Haibo Cai","doi":"10.1002/btpr.70000","DOIUrl":"https://doi.org/10.1002/btpr.70000","url":null,"abstract":"Cell culture medium is an important factor affecting the expansion of NK cells in vitro. As an important component of cell culture medium, lipids participate in various complex physiological activities of cells and significantly affect the expansion of cells. Using NK-92 cells as a model, the lipid metabolism of NK cells in vitro was analyzed, and combined with the kinetic relationship between lipid metabolism and NK cell expansion. Four fatty acids, oleic acid, linoleic acid, palmitoleic acid, and α-linolenic acid, were preliminatively identified as the key lipid combinations. The combination was preliminarily verified on the self-developed serum-free medium. It was found that when the key lipid combination was added according to the concentration in the serum, NK-92 cells expansion reached 188.03 ± 33.34-folds, which was significantly higher than 105.28 ± 13.23-folds in the basic medium. Additionally, NK-92 cells expanded by adding key lipid combinations could maintain cell killing function. Overall, this research provides technical support for the development of NK cell serum-free medium.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70000"},"PeriodicalIF":2.5,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439857","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

Mechanistic insights into hypoxia-induced metabolic reprogramming in colorectal cancer through genome-scale modeling. 通过基因组尺度模型对低氧诱导的结直肠癌代谢重编程的机制见解。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-18 DOI: 10.1002/btpr.70002

Sonal Omer, Subasree Sridhar, D Karunagaran, G K Suraishkumar

{"title":"Mechanistic insights into hypoxia-induced metabolic reprogramming in colorectal cancer through genome-scale modeling.","authors":"Sonal Omer, Subasree Sridhar, D Karunagaran, G K Suraishkumar","doi":"10.1002/btpr.70002","DOIUrl":"https://doi.org/10.1002/btpr.70002","url":null,"abstract":"The hypoxic colorectal cancer (CRC) microenvironment is a complex niche. Hence, in vivo, the metabolism occurring in the cancer cell is not fully known due to difficulties in estimating metabolic fluxes and metabolite exchanges. Genome-scale metabolic modeling helps estimate such metabolic fluxes to gain insights into the metabolic behavior of individual cancer cell types under various tumor microenvironments (TME). We developed a simplified approach to apply proteomics data-based enzyme usage constraints and integrated reactive species (RS) reactions in a context-specific genome-scale metabolic model (GSMM) of HCT116, a CRC cell line. The combined modeling approach reproduced several phenotypes of HCT116 under hypoxia such as the Warburg effect. The integration of the RS module with the hypoxic HCT116 context-specific GSMM highlighted the hypoxia-mediated dysregulation occurring in important metabolic pathways such as hyaluronan metabolism in which 80% of the reactions from the total reactions corresponding to this metabolic pathway were dysregulated. Similarly, 23% of reactions in the urea cycle, 26% of reactions in eicosanoid metabolism and 38% of reactions in glyoxylate and dicarboxylate metabolism were dysregulated.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70002"},"PeriodicalIF":2.5,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439853","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

Flocculation-based clarification for production of protein therapeutics in Pichia pastoris: Recombinant human serum albumin as a case study. 基于絮凝澄清的毕赤酵母蛋白治疗剂生产：重组人血清白蛋白为例研究。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2025-02-18 DOI: 10.1002/btpr.70001

Vishwanath Hebbi, Jashwant Kumar, Anurag S Rathore

{"title":"Flocculation-based clarification for production of protein therapeutics in Pichia pastoris: Recombinant human serum albumin as a case study.","authors":"Vishwanath Hebbi, Jashwant Kumar, Anurag S Rathore","doi":"10.1002/btpr.70001","DOIUrl":"https://doi.org/10.1002/btpr.70001","url":null,"abstract":"Pichia pastoris has been used as an expression system for multiple biotherapeutic products due to the unique advantages it offers with respect to cell density, protein titer, extracellular expression, and other such advantages. However, clarification of cell broth presents a significant challenge, primarily due to the high cell density (up to 50% W/V). Additionally, the abundance of host cell proteins complicates secondary clarification, impacting subsequent chromatographic, and filtration steps. In this study, a flocculation-based cell clarification method has been developed for the primary recovery of protein therapeutic products from Pichia broth. Human serum albumin (HSA) has been used as a case study. Unlike polymer-based flocculants, which introduce challenges in process clearance, the proposed method employs process-compatible salts. The approach has been designed and optimized using Quality by Design (QbD) principles, achieving a clarification efficiency with up to 90% recovery and a reduction of host cell proteins by up to 30%. The proposed methodology would be applicable to other biotherapeutic applications involving protein production in P. pastoris.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70001"},"PeriodicalIF":2.5,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439850","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