Novel calibration design improves knowledge transfer across products for the characterization of pharmaceutical bioprocesses
Wiley Online Library » Biotechnology Journal
by Laura M. Helleckes, Claus Wirnsperger, Jakub Polak, Gonzalo Guillén‐Gosálbez, Alessandro Butté, Moritz von Stosch
2d ago
Graphical Abstract and Lay Summary The development of pharmaceutical bioprocesses requires extensive screening of different process designs. In this work, we evaluate the use of transfer and meta learning on historical process data to predict the behaviour of new bioprocesses. Using a novel method that we coin calibration design, we suggest process designs for new campaigns with the help of knowledge transfer models. Consequently, this work demonstrates the potential to rethink process development by systematically leveraging knowledge transfer. Abstract Modern machine learning has the potent ..read more
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Streamlined screening of extracellularly expressed PETase libraries for improved polyethylene terephthalate degradation
Wiley Online Library » Biotechnology Journal
by Guy Orr, Yoav Niv, Maya Barakat, Alexandra Boginya, Moshe Dessau, Livnat Afriat‐Jurnou
2d ago
Graphical Abstract and Lay Summary This study presents the extracellular expression and screening of a genetic library based on a known PET-degrading enzyme (LCC-ICCG) aimed at enhancing its PETase activity towards untreated commercial PET. The process involved the creation of variant libraries followed by their extracellular expression and high-throughput screening performed on a 96 well-plate scale to identify the most efficient enzymes. The graphical abstract illustrates the cyclic workflow from library creation, expression, screening, and the identification of variants featuring improved a ..read more
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Engineering a 3‐dimensional tissue construct with adipose‐derived stem cells for healing bone defect: An ex vivo study with femur head
Wiley Online Library » Biotechnology Journal
by Aditi Mahajan, Siddhartha Sharma, Sanjay Kumar Bhadada, Aditya Aggarwal, Shalmoli Bhattacharyya
2d ago
Graphical Abstract and Lay Summary Mesenchymal stem cells (MSCs) were harvested from adipose tissue taken directly from patients undergoing orthopedic surgery. These MSCs were seeded on different commercially available bone graft substitutes such as hydroxyapatite, calcium sulfate, beta tricalcium phosphate and gelatin. Primary screening was done on the basis of the ability of these scaffolds to support MSCs adhesion and viability. The selected scaffold was then used in a patient derived ex vivo model of critical size bone defect. The study establishes for the first time the application of fem ..read more
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Biomass specific perfusion rate as a control lever for the continuous manufacturing of biosimilar monoclonal antibodies from CHO cell cultures
Wiley Online Library » Biotechnology Journal
by Jiayu Leong, Wen Qin Tang, Jake Chng, Wei Xuan Ler, Norhaizat Abdul Manan, Lyn Chiin Sim, Zi Ying Zheng, Wei Zhang, Ian Walsh, Gerben Zijlstra, Maarten Pennings, Say Kong Ng
2d ago
Graphical Abstract and Lay Summary Perfusion rate is a key process parameter in continuous manufacturing. This study determined the effects of using biomass specific perfusion rate (BSPR) which is a feed rate proportional to the biomass. This study demonstrated that BSPR influenced cell growth and productivity in cultures with variable cell volumes. Abstract Continuous manufacturing enables high volumetric productivities of biologics such as monoclonal antibodies. However, it is challenging to maintain both high viable cell densities and productivities at the same time for long culture durati ..read more
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Human platelet lysate supports SH‐SY5Y neuroblastoma cell proliferation and differentiation into a dopaminergic‐like neuronal phenotype under xenogeneic‐free culture conditions
Wiley Online Library » Biotechnology Journal
by Miguel Ribeiro, Jonas Campos, Tiffany S. Pinho, Belém Sampaio‐Marques, Sandra Barata‐Antunes, Jorge Ribeiro Cibrão, Ricardo Araújo, Sara Duarte‐Silva, Elsa Moreira, Rui A. Sousa, Pedro M. Costa, António J. Salgado
2d ago
Graphical Abstract and Lay Summary SH-SY5Y cells are a widely used in vitro model for early-stage development of novel therapies for neurodegenerative conditions such as Parkinson's disease. Most studies so far use Fetal Bovine Serum as a growth medium supplement for SH-SY5Y cell culture, but current guidelines strongly recommend to avoid the use of animal-derived products during early-development of novel therapies. This work presents the possibility of differentiating SH-SY5Y cells into a neuronal phenotype with dopaminergic-like features under animal-free culture conditions, using hPL and F ..read more
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Efficient CRISPR‐mediated C‐to‐T base editing in Komagataella phaffii
Wiley Online Library » Biotechnology Journal
by Ling‐Yu Wu, Yan Xu, Xiao‐Wei Yu
2d ago
Graphical Abstract and Lay Summary The methylotrophic yeast Komagataella phaffii is widely applied in the production of industrial enzymes, and pharmaceutical proteins. The development of robust and versatile genome editing tools for K. phaffii is crucial for the design of increasingly advanced cell factories. This study developed a base editing method based on the CRISPR-nCas9 system, which efficiently performed C-to-T editing with single-, and double-locus editing efficiencies of up to 96.0%, and 65.0%, respectively, within a 7-nucleotide window. This study also expanded the targetable genom ..read more
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Toehold region triggered CRISPR/Cas12a trans‐cleavage for detection of uracil‐DNA glycosylase activity
Wiley Online Library » Biotechnology Journal
by Chenyu Cui, Guihuan Guo, Ting‐Hsuan Chen
2d ago
Graphical Abstract and Lay Summary Abnormal experssion of uracil-DNA glycosylase (UDG) has been proved to be associated with different diseases such as cancer, making it a potential biomaker and therapeutic target. Here, we developed a simple method to detect UDG activity based on toehold region triggered CRISPR/Cas12a trans-cleavage. Abstract DNA glycosylases are a group of enzymes that play a crucial role in the DNA repair process by recognizing and removing damaged or incorrect bases from DNA molecules, which maintains the integrity of the genetic information. The abnormal expression of ur ..read more
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Fast‐track adaptive laboratory evolution of Cupriavidus necator H16 with divalent metal cations
Wiley Online Library » Biotechnology Journal
by Sepwin Nosten Sitompul, Laura Andrea Diaz Garcia, Joseph Price, Kang Lan Tee, Tuck Seng Wong
2d ago
Graphical Abstract and Lay Summary Accelerated Adaptive Laboratory Evolution (aALE) is a novel tool designed to optimize microbial strains. This method enhances genetic diversity by utilizing divalent metal cations, which reduce the fidelity of genome replication, thereby speeding up the evolution process. Abstract Microbial strain improvement through adaptive laboratory evolution (ALE) has been a key strategy in biotechnology for enhancing desired phenotypic traits. In this Biotech Method paper, we present an accelerated ALE (aALE) workflow and its successful implementation in evolving Cupri ..read more
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Issue Information: Biotechnology Journal 7/2024
Wiley Online Library » Biotechnology Journal
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1w ago
Biotechnology Journal, Volume 19, Issue 7, July 2024 ..read more
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Unlocking the formate utilization of wild‐type Yarrowia lipolytica through adaptive laboratory evolution
Wiley Online Library » Biotechnology Journal
by Qian Chen, Yunhong Chen, Zeming Hou, Yuyue Ma, Jianfeng Huang, Zhidan Zhang, Yefu Chen, Xue Yang, Yanfei Zhang, Guoping Zhao
3w ago
Graphical Abstract and Lay Summary In this study, we utilized gene editing tools to promote adaptive evolution, enabling wild-type Yarrowia lipolytica to achieve a formate tolerance of 1 M within 2 months, resulting in strains with enhanced formate utilization. Importantly, this study highlights the presence of a complete natural formate metabolic pathway and a highly efficient formate-based energy production module in wild-type Y. lipolytica. Abstract Synthetic biology is contributing to the advancement of the global net-negative carbon economy, with emphasis on formate as a member of t ..read more
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