In the realm of clinical surgery, the unceasing pursuit of hemostatic materials with rapid hemostasis, excellent biocompatibility, degradability, wound-healing promotion, and economic viability is of utmost importance. This study, inspired by the preparation and hemostatic mechanism of Surgicel®, a prevalent surgical hemostatic agent, low-cost silk fibroin (SF) was modified. By adding carboxyl groups while maintaining the SF micron-fiber structure, the modified SF showed enhanced water and blood absorption, as well as a significantly improved Ca²⁺ transport capacity compared to unmodified SF. In vitro coagulation tests revealed that the modified SF had excellent hemostatic efficacy, with the quickest time at 76.75 ± 2.17 s. In the animal bleeding model, its fastest hemostatic time was 93 ± 9.63 s, with a mean blood loss of 0.28 ± 0.05 g, similar to Surgicel® (100.64 ± 2.87 s, 0.24 ± 0.05 g). Moreover, the modified SF hemostatic yarns exhibited favorable blood, cell, and biocompatibility, making them a promising option for surgical wound hemostasis, especially in organ surgery.
{"title":"Silk Fibroin With Enhanced Hemostatic Property by the Introduction of Carboxyl Moieties and Preservation of Micrometer Scale Fiber Architecture.","authors":"Mingyu Jiang, Fuping Wang, Jun Zhang, Changfu Hu, Guobao Chen, Zhongmin Chen","doi":"10.1002/bit.70148","DOIUrl":"https://doi.org/10.1002/bit.70148","url":null,"abstract":"<p><p>In the realm of clinical surgery, the unceasing pursuit of hemostatic materials with rapid hemostasis, excellent biocompatibility, degradability, wound-healing promotion, and economic viability is of utmost importance. This study, inspired by the preparation and hemostatic mechanism of Surgicel®, a prevalent surgical hemostatic agent, low-cost silk fibroin (SF) was modified. By adding carboxyl groups while maintaining the SF micron-fiber structure, the modified SF showed enhanced water and blood absorption, as well as a significantly improved Ca²⁺ transport capacity compared to unmodified SF. In vitro coagulation tests revealed that the modified SF had excellent hemostatic efficacy, with the quickest time at 76.75 ± 2.17 s. In the animal bleeding model, its fastest hemostatic time was 93 ± 9.63 s, with a mean blood loss of 0.28 ± 0.05 g, similar to Surgicel® (100.64 ± 2.87 s, 0.24 ± 0.05 g). Moreover, the modified SF hemostatic yarns exhibited favorable blood, cell, and biocompatibility, making them a promising option for surgical wound hemostasis, especially in organ surgery.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145862061","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sophie Weiser, Sidney Jung, Bettina Bardl, Johann E Kufs, Slavica Janevska, Vito Valiante, Dirk Hoffmeister, Lars Regestein
Psilocybin, an indole alkaloid of psychedelic mushrooms, has the potential to sustainably improve the treatment of several psychiatric diseases. So far, the psilocybin demand for clinical trials has been met by chemical synthesis. In this study, we pursued the biotechnological approach to develop a psilocybin production process utilizing an overproduction strain of Aspergillus nidulans. The developed shake flask cultivation regime was characterized rheologically and was evaluated concerning the sensitivity to changes in oxygen availability and power input. Due to the strong impact of power input on viscosity and thus, (oxygen) mass transfer and mixing of the filamentous culture broth, the bioprocess was scaled up from shake flask to 7 L stirred tank reactor according to the specific power input. Utilizing a pressure reactor, the oxygen supply of the viscous culture broth was enhanced. Subsequently, the nitrogen limitation was addressed by supplementing the cultivation medium with additional ammonium sulfate to provide sufficient building blocks for protein biosynthesis. By producing 542 mg L-1 psilocybin within 68 h from glucose, a robust and efficient batch bioprocess for psilocybin production was developed to potentially contribute to the future supply of psilocybin for pharmaceutical purposes. Moreover, we demonstrated the suitability of pressurized bioprocesses to counteract oxygen limitations for shear-sensitive, filamentous organisms.
{"title":"Psilocybin Production With Genetically Modified Aspergillus nidulans Under Pressurized Conditions.","authors":"Sophie Weiser, Sidney Jung, Bettina Bardl, Johann E Kufs, Slavica Janevska, Vito Valiante, Dirk Hoffmeister, Lars Regestein","doi":"10.1002/bit.70137","DOIUrl":"10.1002/bit.70137","url":null,"abstract":"<p><p>Psilocybin, an indole alkaloid of psychedelic mushrooms, has the potential to sustainably improve the treatment of several psychiatric diseases. So far, the psilocybin demand for clinical trials has been met by chemical synthesis. In this study, we pursued the biotechnological approach to develop a psilocybin production process utilizing an overproduction strain of Aspergillus nidulans. The developed shake flask cultivation regime was characterized rheologically and was evaluated concerning the sensitivity to changes in oxygen availability and power input. Due to the strong impact of power input on viscosity and thus, (oxygen) mass transfer and mixing of the filamentous culture broth, the bioprocess was scaled up from shake flask to 7 L stirred tank reactor according to the specific power input. Utilizing a pressure reactor, the oxygen supply of the viscous culture broth was enhanced. Subsequently, the nitrogen limitation was addressed by supplementing the cultivation medium with additional ammonium sulfate to provide sufficient building blocks for protein biosynthesis. By producing 542 mg L<sup>-1</sup> psilocybin within 68 h from glucose, a robust and efficient batch bioprocess for psilocybin production was developed to potentially contribute to the future supply of psilocybin for pharmaceutical purposes. Moreover, we demonstrated the suitability of pressurized bioprocesses to counteract oxygen limitations for shear-sensitive, filamentous organisms.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145854370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Patients around the world, especially in low‐ or medium‐income countries (LMICs), are unable to afford the prohibitively high cost of monoclonal antibodies (mAbs). One promising approach to this issue of accessibility and affordability is intensifying the manufacturing process by employing continuous processing technology, which improves sustainability, efficiency, and process compactness, while reducings the operational expenses without sacrificing product safety or quality. Even though the biomanufacturing sector has been discussing continuous processing for more than 15 years, its adoption has not been as rapid as expected, with most companies still relying on traditional batch‐based production systems. This brings up important inquiries: is it necessary to have a completely end‐to‐end continuous processing, or are hybrid and selectively intensified methods enough? In this article, we offer a perspective on the current status of continuous biomanufacturing, challenges associated with it and the production costs of five different intensification scenarios using a process simulation tool, incorporating both traditional batch processing and fully integrated continuous processing, and identifying the manufacturing hot‐spots that result in the significant cost savings. Our findings suggest that instead of completely replacing the batch‐process equipment, mAb manufacturing should strategically engage continuous technologies when they deliver clear value. Sustainable and efficient biopharmaceutical production that enables broad access and affordabilty is achievable through an evolutionary strategy based on planned intensification and risk‐managed implementation. So, the question is not whether end‐to‐end continuous processing is required, but rather how to maximize its advantages while efficiently handling its complexity.
{"title":"Do We Really Need End‐To‐End Continuous Processing for Biomanufacturing of Monoclonal Antibodies?","authors":"Anurag S. Rathore, Subhankar Metya, Nitika Nitika","doi":"10.1002/bit.70147","DOIUrl":"https://doi.org/10.1002/bit.70147","url":null,"abstract":"Patients around the world, especially in low‐ or medium‐income countries (LMICs), are unable to afford the prohibitively high cost of monoclonal antibodies (mAbs). One promising approach to this issue of accessibility and affordability is intensifying the manufacturing process by employing continuous processing technology, which improves sustainability, efficiency, and process compactness, while reducings the operational expenses without sacrificing product safety or quality. Even though the biomanufacturing sector has been discussing continuous processing for more than 15 years, its adoption has not been as rapid as expected, with most companies still relying on traditional batch‐based production systems. This brings up important inquiries: is it necessary to have a completely end‐to‐end continuous processing, or are hybrid and selectively intensified methods enough? In this article, we offer a perspective on the current status of continuous biomanufacturing, challenges associated with it and the production costs of five different intensification scenarios using a process simulation tool, incorporating both traditional batch processing and fully integrated continuous processing, and identifying the manufacturing hot‐spots that result in the significant cost savings. Our findings suggest that instead of completely replacing the batch‐process equipment, mAb manufacturing should strategically engage continuous technologies when they deliver clear value. Sustainable and efficient biopharmaceutical production that enables broad access and affordabilty is achievable through an evolutionary strategy based on planned intensification and risk‐managed implementation. So, the question is not whether end‐to‐end continuous processing is required, but rather how to maximize its advantages while efficiently handling its complexity.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"51 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145847108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Constraint-based reconstruction and analysis (COBRA) is a powerful systems biology approach for computational bioengineering. Synechococcus elongatus PCC 11801 and PCC 11802 are fast-growing, stress-tolerant cyanobacteria that are promising platforms for photosynthetic biomanufacturing. Here, we present constraint-based models (CBMs) iLV1052 and iLV1087 of PCC 11801 and PCC 11802, respectively, to facilitate and streamline strain engineering efforts. Following draft reconstruction using a template model, the models underwent extensive manual curation to reduce redundancy, and verification using BiGG, KEGG, and BRENDA databases. We added 281 and 69 new reactions for PCC 11801 and PCC 11802, respectively, associated with stress tolerance, growth stability, antioxidant defense, energy regulation, and sulfur acquisition. The models were refined through iterative debugging and validation using flux balance analysis, flux variability analysis, and single gene/reaction deletion analysis. Gene essentiality predictions gave 69% accuracy for PCC 11801 and 83% for PCC 11802. The flux maps captured key features of cyanobacterial metabolism, including an incomplete TCA cycle. The final PCC 11801 CBM contained 1130 reactions, 1052 genes, and 930 metabolites, while the PCC 11802 CBM included 1199 reactions, 1087 genes, and 951 metabolites. The simulations predicted that succinic acid exhibited the highest theoretical yield among the tested target products in both strains. Using the Optknock framework, phosphoenolpyruvate carboxylase was identified as a metabolic hotspot for future bioengineering efforts aimed at the production of valuable products like ethanol, butanol, and butanediol.
{"title":"Constraint-Based Metabolic Modeling of Synechococcus elongatus PCC 11801 and PCC 11802 for Photosynthetic Biomanufacturing.","authors":"Lokesh Venkatesh Babu, Pramod P Wangikar","doi":"10.1002/bit.70146","DOIUrl":"https://doi.org/10.1002/bit.70146","url":null,"abstract":"<p><p>Constraint-based reconstruction and analysis (COBRA) is a powerful systems biology approach for computational bioengineering. Synechococcus elongatus PCC 11801 and PCC 11802 are fast-growing, stress-tolerant cyanobacteria that are promising platforms for photosynthetic biomanufacturing. Here, we present constraint-based models (CBMs) iLV1052 and iLV1087 of PCC 11801 and PCC 11802, respectively, to facilitate and streamline strain engineering efforts. Following draft reconstruction using a template model, the models underwent extensive manual curation to reduce redundancy, and verification using BiGG, KEGG, and BRENDA databases. We added 281 and 69 new reactions for PCC 11801 and PCC 11802, respectively, associated with stress tolerance, growth stability, antioxidant defense, energy regulation, and sulfur acquisition. The models were refined through iterative debugging and validation using flux balance analysis, flux variability analysis, and single gene/reaction deletion analysis. Gene essentiality predictions gave 69% accuracy for PCC 11801 and 83% for PCC 11802. The flux maps captured key features of cyanobacterial metabolism, including an incomplete TCA cycle. The final PCC 11801 CBM contained 1130 reactions, 1052 genes, and 930 metabolites, while the PCC 11802 CBM included 1199 reactions, 1087 genes, and 951 metabolites. The simulations predicted that succinic acid exhibited the highest theoretical yield among the tested target products in both strains. Using the Optknock framework, phosphoenolpyruvate carboxylase was identified as a metabolic hotspot for future bioengineering efforts aimed at the production of valuable products like ethanol, butanol, and butanediol.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145854439","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Continuous bioprocessing with Protein A affinity chromatography has demonstrated great potential to increase productivity and reduce the cost of goods in monoclonal antibody (mAb) production. However, maintaining process stability and responding to dynamic changes remains significant challenges, particularly in the real‐time optimization and control of multi‐column periodic counter‐current chromatography (PCC) for Protein A affinity chromatography, due to the computational complexity of rapidly solving mechanistic models. To address this challenge, this study developed distilled physics‐informed neural networks (PINNs) based on the general rate model (GRM) to accelerate and enhance the breakthrough curve fitting and four‐column PCC (4C‐PCC) process optimization. The distilled PINNs achieved a balance between prediction accuracy and computational speed. The 157k‐parameter distilled PINN enabled the breakthrough curve fitting and 4C‐PCC process optimization approximately 10 times faster than numerical methods while improving accuracy by about 40%. A smaller 2k‐parameter model achieved a 22‐fold acceleration with an acceptable trade‐off in accuracy, and the optimization time was reduced to 1.44 s. Explainability analyses confirmed the PINN's capability to capture nonlinear and interactive effects among key process parameters. The PINN‐accelerated GRM was then integrated with real‐time model predictive control (MPC) and applied to a lab‐scale continuous manufacturing process. PINN‐based MPC maintained robust control of binding capacity and yield, achieving a productivity of 35 g/L resin/h and resin capacity utilization of 90%, despite resin capacity decay and upstream variability. This work demonstrates that the PINNs can provide a computationally efficient and physically consistent framework for real‐time optimization and control of continuous processes. Integrating a mechanistic model with neural networks can enhance process understanding and robustness, supporting the implementation of continuous biomanufacturing for therapeutic proteins.
{"title":"Real‑Time Model Predictive Control of Monoclonal Antibody Capture in Continuous Manufacturing Using Physics‑Informed Neural Networks Accelerated Mechanistic Modeling","authors":"Si‐Yuan Tang, Yun‐Hao Yuan, Yan‐Na Sun, Wen‐Huang Pan, Shan‐Jing Yao, Dong‐Qiang Lin","doi":"10.1002/bit.70141","DOIUrl":"https://doi.org/10.1002/bit.70141","url":null,"abstract":"Continuous bioprocessing with Protein A affinity chromatography has demonstrated great potential to increase productivity and reduce the cost of goods in monoclonal antibody (mAb) production. However, maintaining process stability and responding to dynamic changes remains significant challenges, particularly in the real‐time optimization and control of multi‐column periodic counter‐current chromatography (PCC) for Protein A affinity chromatography, due to the computational complexity of rapidly solving mechanistic models. To address this challenge, this study developed distilled physics‐informed neural networks (PINNs) based on the general rate model (GRM) to accelerate and enhance the breakthrough curve fitting and four‐column PCC (4C‐PCC) process optimization. The distilled PINNs achieved a balance between prediction accuracy and computational speed. The 157k‐parameter distilled PINN enabled the breakthrough curve fitting and 4C‐PCC process optimization approximately 10 times faster than numerical methods while improving accuracy by about 40%. A smaller 2k‐parameter model achieved a 22‐fold acceleration with an acceptable trade‐off in accuracy, and the optimization time was reduced to 1.44 s. Explainability analyses confirmed the PINN's capability to capture nonlinear and interactive effects among key process parameters. The PINN‐accelerated GRM was then integrated with real‐time model predictive control (MPC) and applied to a lab‐scale continuous manufacturing process. PINN‐based MPC maintained robust control of binding capacity and yield, achieving a productivity of 35 g/L resin/h and resin capacity utilization of 90%, despite resin capacity decay and upstream variability. This work demonstrates that the PINNs can provide a computationally efficient and physically consistent framework for real‐time optimization and control of continuous processes. Integrating a mechanistic model with neural networks can enhance process understanding and robustness, supporting the implementation of continuous biomanufacturing for therapeutic proteins.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"1 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145847502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mariana Carvalho, Ana Cruz, Cees Haringa, Marcel Ottens, Marieke Klijn
The competition in the biopharmaceutical market is increasing due to the market entry of biosimilars and rising costs in research and development of new drugs. Hence, continuous manufacturing gained significant attention due to its potential in reducing production cycle times and costs, as well as the possibility of real‐time release testing. As a consequence, active monitoring and/or control systems are required for quantitative product quality measurements and in‐process control. Process analytical technology emerged as a robust strategy for the development and implementation of in situ real‐time testing, instead of the standard batch testing of end product. Through the evaluation of state‐of‐the‐art applications, this review highlights future opportunities in the field of quantitative real‐time analytical techniques for the characterization of monoclonal antibodies in continuous downstream biomanufacturing.
{"title":"A Review on Quantitative Process Analytical Technology for Continuous Downstream Processing of Monoclonal Antibodies","authors":"Mariana Carvalho, Ana Cruz, Cees Haringa, Marcel Ottens, Marieke Klijn","doi":"10.1002/bit.70139","DOIUrl":"https://doi.org/10.1002/bit.70139","url":null,"abstract":"The competition in the biopharmaceutical market is increasing due to the market entry of biosimilars and rising costs in research and development of new drugs. Hence, continuous manufacturing gained significant attention due to its potential in reducing production cycle times and costs, as well as the possibility of real‐time release testing. As a consequence, active monitoring and/or control systems are required for quantitative product quality measurements and in‐process control. Process analytical technology emerged as a robust strategy for the development and implementation of in situ real‐time testing, instead of the standard batch testing of end product. Through the evaluation of state‐of‐the‐art applications, this review highlights future opportunities in the field of quantitative real‐time analytical techniques for the characterization of monoclonal antibodies in continuous downstream biomanufacturing.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"91 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145829889","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Maisha M. Feroz, Seok‐Joon Kwon, Katherin Berman, Nicholas J. Mantis, Jonathan S. Dordick
As viral diseases like influenza, COVID‐19, and respiratory syncytial virus (RSV) become prevalent, demand for treatments and vaccinations is on the rise. Despite widespread vaccination, concerns remain that emerging virus strains may evade immunity. Monitoring these pathogens is crucial for developing effective treatments, but large‐scale population screening, while a public health need, is challenging in terms of speed, efficiency, and cost. Herein we demonstrate the use of a high‐throughput, chip‐based platform that is scalable to population‐wide immune surveillance using SARS‐CoV‐2 pseudovirus surrogates. We tested antibodies against pseudovirus surrogates bearing the spike protein of the Wuhan‐Hu‐1 (WT) SARS‐CoV‐2 on‐chip and screened patient‐derived sera and eluates from patient‐derived and contrived Dried Blood Spots (cDBS) for effectiveness against the WT strain. Interestingly, some sera and cDBS eluates inhibited infection by the Omicron BA.4/5 variant, which emerged a year after sample collection, suggesting cross‐reactivity against distinct SARS‐CoV‐2 spike proteins. This chip‐based platform may be used to screen populations for existing immunity and immune escape of emerging viruses and their variants from prior infection and/or vaccination.
{"title":"A Chip‐Based Surveillance Platform for Detecting Antibody‐Based Immunity and Immune Escape Using SARS‐CoV‐2 Pseudovirus Surrogates","authors":"Maisha M. Feroz, Seok‐Joon Kwon, Katherin Berman, Nicholas J. Mantis, Jonathan S. Dordick","doi":"10.1002/bit.70143","DOIUrl":"https://doi.org/10.1002/bit.70143","url":null,"abstract":"As viral diseases like influenza, COVID‐19, and respiratory syncytial virus (RSV) become prevalent, demand for treatments and vaccinations is on the rise. Despite widespread vaccination, concerns remain that emerging virus strains may evade immunity. Monitoring these pathogens is crucial for developing effective treatments, but large‐scale population screening, while a public health need, is challenging in terms of speed, efficiency, and cost. Herein we demonstrate the use of a high‐throughput, chip‐based platform that is scalable to population‐wide immune surveillance using SARS‐CoV‐2 pseudovirus surrogates. We tested antibodies against pseudovirus surrogates bearing the spike protein of the Wuhan‐Hu‐1 (WT) SARS‐CoV‐2 on‐chip and screened patient‐derived sera and eluates from patient‐derived and contrived Dried Blood Spots (cDBS) for effectiveness against the WT strain. Interestingly, some sera and cDBS eluates inhibited infection by the Omicron BA.4/5 variant, which emerged a year after sample collection, suggesting cross‐reactivity against distinct SARS‐CoV‐2 spike proteins. This chip‐based platform may be used to screen populations for existing immunity and immune escape of emerging viruses and their variants from prior infection and/or vaccination.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"9 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145822888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jessica L. King, Christopher Spencer, Richard Youngblood, Kelly Crumley, Elizabeth Bealer, Peter D. Rios, Ira Joshi, Sofia Ghani, Douglas Isa, James J. McGarrigle, David Cook, Conor Locke, Adam Abraham, Andrea Clark, José Oberholzer, Lonnie D. Shea
Clinical islet transplantation has long been investigated as a potential cure for type 1 diabetes (T1D), yet standard intrahepatic delivery leaves islets prone to an instant blood‐mediated inflammatory response. Herein, we investigated the design of microporous poly( D , l ‐lactide‐co‐glycolide) (PLG) scaffolds for extrahepatic islet transplantation in mouse and nonhuman primate (NHP) models. Acellular scaffolds elicited only a mild inflammatory response following implantation into the omentum. On scaffold islet transplantation had extensive insulin staining at 4 weeks yet modest insulin requirement reductions in diabetic NHP recipients. Scaffolds were sterilized by irradiation and exhibited fragility during seeding and implantation, motivating an increase in the manufacturing ratio of PLG:NaCl from 1:30 to 1.25:30 w/w. These scaffolds exhibited no differences in porosity or interior geometry between sterilization conditions, and transplants in mice restored normoglycemia. We piloted a modified scaffold study in a fourth NHP, and although scaffold integrity was improved, the transplant outcome was similar. We subsequently tested intermediate PLG:NaCl ratios in mice, finding that a 1.15:30 ratio achieved a balance of mechanical stability and islet compatibility. Overall, these studies identify that scaffold porosity can be adjusted to account for the impact of sterilization on transplantation.
临床胰岛移植作为治疗1型糖尿病（T1D）的潜在方法已经被研究了很长时间，然而标准的肝内移植使胰岛容易发生即时血液介导的炎症反应。在此，我们研究了用于小鼠和非人灵长类动物（NHP）模型肝外胰岛移植的微孔聚（D， l -丙交酯- co -乙醇酸酯）（PLG）支架的设计。无细胞支架在植入网膜后仅引起轻度炎症反应。支架胰岛移植在4周时有广泛的胰岛素染色，但糖尿病NHP受体的胰岛素需求略有下降。支架经过辐照灭菌，在播种和植入过程中表现出脆性，促使PLG:NaCl的制造比从1:30增加到1.25:30 w/w。这些支架在灭菌条件下的孔隙度和内部几何形状没有差异，小鼠移植后恢复了正常血糖。我们在第四个NHP中进行了改良支架研究，尽管支架的完整性得到了改善，但移植结果相似。随后，我们在小鼠中测试了中间PLG:NaCl比例，发现1.15:30的比例达到了机械稳定性和胰岛相容性的平衡。总的来说，这些研究确定支架孔隙度可以调整，以考虑灭菌对移植的影响。
{"title":"Gamma Irradiation of Poly(lactide‐co‐glycolide) Scaffolds Reduces the Mechanical Stability and Function of Islet Grafts in Diabetic Nonhuman Primates","authors":"Jessica L. King, Christopher Spencer, Richard Youngblood, Kelly Crumley, Elizabeth Bealer, Peter D. Rios, Ira Joshi, Sofia Ghani, Douglas Isa, James J. McGarrigle, David Cook, Conor Locke, Adam Abraham, Andrea Clark, José Oberholzer, Lonnie D. Shea","doi":"10.1002/bit.70134","DOIUrl":"https://doi.org/10.1002/bit.70134","url":null,"abstract":"Clinical islet transplantation has long been investigated as a potential cure for type 1 diabetes (T1D), yet standard intrahepatic delivery leaves islets prone to an instant blood‐mediated inflammatory response. Herein, we investigated the design of microporous poly( <jats:sc>D</jats:sc> , <jats:sc>l</jats:sc> ‐lactide‐co‐glycolide) (PLG) scaffolds for extrahepatic islet transplantation in mouse and nonhuman primate (NHP) models. Acellular scaffolds elicited only a mild inflammatory response following implantation into the omentum. On scaffold islet transplantation had extensive insulin staining at 4 weeks yet modest insulin requirement reductions in diabetic NHP recipients. Scaffolds were sterilized by irradiation and exhibited fragility during seeding and implantation, motivating an increase in the manufacturing ratio of PLG:NaCl from 1:30 to 1.25:30 w/w. These scaffolds exhibited no differences in porosity or interior geometry between sterilization conditions, and transplants in mice restored normoglycemia. We piloted a modified scaffold study in a fourth NHP, and although scaffold integrity was improved, the transplant outcome was similar. We subsequently tested intermediate PLG:NaCl ratios in mice, finding that a 1.15:30 ratio achieved a balance of mechanical stability and islet compatibility. Overall, these studies identify that scaffold porosity can be adjusted to account for the impact of sterilization on transplantation.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"251 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145813175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Halimatun Sakdiah Zainuddin, Sanjeeva Kumar Murali, Thomas J. Mansell
The application of engineered live biotherapeutic products (LBPs) to secrete small molecules, peptides, or proteins to benefit a human or animal host, relies on heterologous protein expression. Key challenges in this area include expressing protein in a targeted location, the use of antibiotic‐free platforms, and expressing recombinant proteins at titers capable of the desired therapeutic effect. In this study, we sought to engineer the promising candidate probiotic chassis Escherichia coli Nissle 1917 (EcN) as an in situ drug delivery platform. Despite its long history of safe human use and general probiotic characteristics, wild‐type EcN is not optimal for routine protein expression. In this work, we present several approaches to improve protein production in this host. First, we enable stable antibiotic‐free protein expression system via native cryptic plasmids. Next, we integrate the T7 RNA polymerase for high level protein expression. Finally, we knock out OmpT protease activity, enabling expression levels comparable to the industry standard E. coli BL21 (DE3). To demonstrate its application, the above system was adapted to express antimicrobial peptide microcin L (MccL) from EcN, which can potentially reduce gut related pathogens and enhance fitness of the probiotic in the competitive niche of the gut. Overall, this study establishes an antibiotic free and high level protein expression platform in EcN, expandable for in situ delivery of therapeutic proteins.
{"title":"Engineered Stable, Antibiotic‐Free, High‐Level Protein Expression in the Probiotic Chassis Escherichia coli Nissle 1917","authors":"Halimatun Sakdiah Zainuddin, Sanjeeva Kumar Murali, Thomas J. Mansell","doi":"10.1002/bit.70130","DOIUrl":"https://doi.org/10.1002/bit.70130","url":null,"abstract":"The application of engineered live biotherapeutic products (LBPs) to secrete small molecules, peptides, or proteins to benefit a human or animal host, relies on heterologous protein expression. Key challenges in this area include expressing protein in a targeted location, the use of antibiotic‐free platforms, and expressing recombinant proteins at titers capable of the desired therapeutic effect. In this study, we sought to engineer the promising candidate probiotic chassis <jats:italic>Escherichia coli</jats:italic> Nissle 1917 (EcN) as an <jats:italic>in situ</jats:italic> drug delivery platform. Despite its long history of safe human use and general probiotic characteristics, wild‐type EcN is not optimal for routine protein expression. In this work, we present several approaches to improve protein production in this host. First, we enable stable antibiotic‐free protein expression system via native cryptic plasmids. Next, we integrate the T7 RNA polymerase for high level protein expression. Finally, we knock out OmpT protease activity, enabling expression levels comparable to the industry standard <jats:italic>E. coli</jats:italic> BL21 (DE3). To demonstrate its application, the above system was adapted to express antimicrobial peptide microcin L (MccL) from EcN, which can potentially reduce gut related pathogens and enhance fitness of the probiotic in the competitive niche of the gut. Overall, this study establishes an antibiotic free and high level protein expression platform in EcN, expandable for <jats:italic>in situ</jats:italic> delivery of therapeutic proteins.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"23 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145813201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Biotechnology and Bioengineering Welcomes New Early Career Researcher Editorial Board","authors":"Paul Trevorrow","doi":"10.1002/bit.70136","DOIUrl":"https://doi.org/10.1002/bit.70136","url":null,"abstract":"","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"45 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145807418","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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