Review
Immunology
Qiaonan Hong, Jian Liu, Yuquan Wei, Xiawei Wei
Summary: Vaccination is an effective strategy to control epidemics, and the baculovirus expression vector system (BEVS) is a promising technology for vaccine production due to its safety and rapid production. This review provides an overview of the development history of BEVS, the procedures for preparing recombinant protein vaccines using the BEVS platform, and the features and limitations of this platform. The progress of BEVS platform-related research, particularly in the field of vaccine, is highlighted, along with a future prospect for BEVS in vaccine manufacturing.
Article
Immunology
Mark R. Bruder, Marc G. Aucoin
Summary: The use of CRISPR-Cas9 technology to introduce indel mutations in the baculovirus expression vector system (BEVS) effectively blocked the release of virus particles and improved the production of virus-like particles (VLPs).
Review
Biochemical Research Methods
Carissa Grose, Zoe Putman, Dominic Esposito
Summary: Despite the success of generating recombinant proteins in insect cells using the Baculovirus Expression Vector System, challenges still exist, such as cytopathic effects, unpredictable co-infection and co-expression, and baculovirus genome instability. A better understanding of viral gene regulation and different promoter options is needed to advance the platform for expressing a greater variety of proteins and complexes in insect cells.
PROTEIN EXPRESSION AND PURIFICATION
(2021)
Article
Biochemical Research Methods
Zongtao Lin, Joanna Gongora, Xingyu Liu, Yixuan Xie, Chenfeng Zhao, Dongwen Lv, Benjamin A. Garcia
Summary: Chemical proteomics utilizes small-molecule probes to study protein interactions. The application of automated robotic systems in sample preparation can accelerate research progress and expand the future of chemical biology.
JOURNAL OF PROTEOME RESEARCH
(2023)
Article
Biochemical Research Methods
Ciaran N. Cronin
Summary: Genetic code expansion (GCE) allows for the production of recombinant proteins with unnatural amino acids, providing novel properties. The tRNApyl/PylRS pair from Methanosarcinaceae species is used to introduce new chemical functionalities, but its application in the baculovirus expression vector system (BEVS) is limited. This study develops novel transfer vectors and investigates their impact on protein production using the tRNApyl/PylRS pair in the more common Bac-to-Bac system.
PROTEIN EXPRESSION AND PURIFICATION
(2023)
Article
Biochemical Research Methods
Diana Santacruz, Francis O. Enane, Katrin Fundel-Clemens, Martin Giner, Gernot Wolf, Svenja Onstein, Christoph Klimek, Zachary Smith, Bhagya Wijayawardena, Coralie Viollet
Summary: RNA-seq has become a reliable method for unbiased assessment of gene expression, and integrating it with other omics datasets enhances our understanding of cell-specific regulatory patterns. However, the time-consuming library preparation process has been a bottleneck. To address this issue, an automated workflow was designed to increase efficiency.
Article
Biotechnology & Applied Microbiology
Lukas Kaesser, Julie Harnischfeger, Denise Salzig, Peter Czermak
Summary: The study found that different culture media have an impact on protein production during transient expression in Sf9 cells. Choosing the appropriate culture medium based on experimental objectives or production frameworks can improve the efficiency of transient expression.
ELECTRONIC JOURNAL OF BIOTECHNOLOGY
(2022)
Article
Medicine, Research & Experimental
Jiarui Wang, Jingyi Chen, Joey Studts, Gang Wang
Summary: The implementation of process analytical technologies is crucial in advancing biopharmaceutical manufacturing by addressing clinical, regulatory, and cost challenges. Raman spectroscopy is a key technology for in-line product quality monitoring, but its wide application is limited by laborious calibration and computational modeling efforts. In this study, we demonstrated new capabilities for real-time measurement of product aggregation and fragmentation in a clinical manufacturing bioprocess by using hardware automation and machine learning. Integration of existing workflows into a robotic system reduced the effort required to calibrate and validate multiple critical quality attribute models. Increased data throughput allowed for the training of calibration models that accurately measured product quality every 38 seconds. In-process analytics enable advanced process understanding in the short-term and ultimately result in controlled bioprocesses that ensure consistent product quality.
Review
Biotechnology & Applied Microbiology
Vijay Tejwani, Minal Chaudhari, Toyaj Rai, Susan T. Sharfstein
Summary: Mammalian cell line development is a multistep process involving single-cell cloning, monoclonality assurance, productivity and stability screening, which can be labor, time and resource intensive when performed manually. Introduction of automation and miniaturization has reduced manual labor, shortened timelines, and decreased resources required for developing manufacturing cell lines.
BIOTECHNOLOGY PROGRESS
(2021)
Article
Multidisciplinary Sciences
Kolesnikova Olga, Zachayus Amelie, Pichard Simon, Osz Judit, Rochel Natacha, Rossolillo Paola, Kolb-Cheynel Isabelle, Troffer-Charlier Nathalie, Compe Emmanuel, Bensaude Olivier, Berger Imrev, Poterszman Arnaud
Summary: The Baculovirus/insect cell expression system is a powerful technology for reconstituting eukaryotic macromolecular assemblies. We have established an open source multigene-expression toolbox using homologous recombination, which simplifies the generation of recombinant baculoviruses into a single-step procedure and shortens the time from cloning to protein production.
SCIENTIFIC REPORTS
(2022)
Article
Multidisciplinary Sciences
Charles J. Buchanan, Ben Gaunt, Peter J. Harrison, Yun Yang, Jiwei Liu, Aziz Khan, Andrew M. Giltrap, Audrey Le Bas, Philip N. Ward, Kapil Gupta, Maud Dumoux, Tiong Kit Tan, Lisa Schimaski, Sergio Daga, Nicola Picchiotti, Margherita Baldassarri, Elisa Benetti, Chiara Fallerini, Francesca Fava, Annarita Giliberti, Panagiotis Koukos, Matthew J. Davy, Abirami Lakshminarayanan, Xiaochao Xue, Georgios Papadakis, Lachlan P. Deimel, Virginia Casablancas-Antras, Timothy D. W. Claridge, Alexandre M. J. J. Bonvin, Quentin J. Sattentau, Simone Furini, Marco Gori, Jiandong Huo, Raymond J. Owens, Christiane Schaffitzel, Imre Berger, Alessandra Renieri, James H. Naismith, Andrew J. Baldwin, Benjamin G. Davis
Summary: This study utilizes universal saturation transfer analysis to reveal the interaction between the novel coronavirus and glycan compounds, and identifies protein mutations related to pathogenicity and zoonosis.
Article
Multidisciplinary Sciences
Alexander Domnick, Christian Winter, Lukas Susac, Leon Hennecke, Mario Hensen, Nicole Zitzmann, Simon Trowitzsch, Christoph Thomas, Robert Tampe
Summary: In this study, the authors determine the structure of the editing module of the peptide loading complex (PLC) and reveal the communication between peptide-MHC I assembly and glycan processing. These findings are important for understanding the initiation of adaptive immune responses and the coordination of events in endoplasmic reticulum quality control.
NATURE COMMUNICATIONS
(2022)
Article
Chemistry, Medicinal
Beatrice Noce, Elisabetta Di Bello, Clemens Zwergel, Rossella Fioravanti, Sergio Valente, Dante Rotili, Andrea Masotti, Mohammad Salik Zeya Ansari, Daniela Trisciuoglio, Alokta Chakrabarti, Christophe Romier, Dina Robaa, Wolfgang Sippl, Manfred Jung, Cecile Haeberli, Jennifer Keiser, Antonello Mai
Summary: Schistosoma mansoni HDAC8 is a reliable target for combating schistosomiasis, but most inhibitors lack selectivity over human deacetylases and have low or no activity against the parasite. In this study, a small library of HDAC inhibitors from the lab was tested for their in vitro enzyme and biological activity. These inhibitors showed submicromolar/nanomolar potency against smHDAC8 and varied selectivity over hHDAC1 and/or hHDAC6. Some compounds exhibited high activity against larval and adult stages of S. mansoni with moderate to no toxicity in human fibroblast cells.
Article
Multidisciplinary Sciences
Ines Katharina Mueller, Christian Winter, Christoph Thomas, Robbert M. Spaapen, Simon Trowitzsch, Robert Tampe
Summary: This study reveals the molecular details of client recognition by the tapasin-ERp57 complex and emphasizes the importance of peptide proofreading. These findings contribute to understanding the selector function of tapasin and the mechanism by which multiple MHC I allomorphs are chaperoned during peptide loading and editing.
NATURE COMMUNICATIONS
(2022)
Article
Multidisciplinary Sciences
Johannes F. Weijman, Sathish K. N. Yadav, Katherine J. Surridge, Jessica A. Cross, Ufuk Borucu, Judith Mantell, Derek N. Woolfson, Christiane Schaffitzel, Mark P. Dodding
Summary: By combining computational structure prediction with single-particle negative-stain electron microscopy, researchers have uncovered the coiled-coil architecture of heterotetrameric kinesin-1 in its compact state. The folding of the complex is enabled by an unusual flexion in the scaffold, bringing the kinesin heavy chain-light chain interface close to a tetrameric assembly. This framework provides insights into the activation mechanism of kinesin-1.
Article
Biochemistry & Molecular Biology
Samoil Sekulovski, Simon Trowitzsch
Summary: Aberrations in tRNA biogenesis can lead to neurological disorders; Mutations cause imbalances in immature and mature tRNA ratios; Further research and future directions are needed to understand the molecular mechanisms.
Article
Biochemical Research Methods
Christine Toelzer, Kapil Gupta, Imre Berger, Christiane Schaffitzel
Summary: The COVID-19 pandemic led to extensive research efforts to understand the molecular mechanisms of SARS-CoV-2. The spike glycoprotein on the virus surface, which is essential for viral entry into host cells, has a hydrophobic pocket that binds to a fatty acid called linoleic acid (LA). Binding of LA stabilizes the spike protein and reduces viral infectivity.
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
(2023)
Article
Multidisciplinary Sciences
Christine Toelzer, Kapil Gupta, Sathish K. N. Yadav, Lorna Hodgson, Maia Kavanagh Williamson, Dora Buzas, Ufuk Borucu, Kyle Powers, Richard Stenner, Kate Vasileiou, Frederic Garzoni, Daniel Fitzgerald, Christine Payre, Gunjan Gautam, Gerard Lambeau, Andrew D. Davidson, Paul Verkade, Martin Frank, Imre Berger, Christiane Schaffitzel
Summary: This study reveals the importance of fatty acid binding in pathogenic coronavirus infection and replication. The spike glycoprotein of SARS-CoV-2 and its variants bind to a fatty acid called linoleic acid, which stabilizes the locked conformation of the spike protein and interferes with virus infectivity. In contrast, a mutation in a common cold-causing coronavirus abolishes this binding. Treatment with linoleic acid inhibits viral replication and reduces the number of deformed virus particles. These findings suggest that targeting fatty acid binding could be a potential strategy for combating COVID-19.
Article
Biochemistry & Molecular Biology
Samoil Sekulovski, Lukas Susac, Lukas S. Stelzl, Robert Tampe, Simon Trowitzsch
Summary: The heterotetrameric human tRNA splicing endonuclease TSEN and its associated RNA kinase CLP1 are essential for intron excision from pre-tRNAs. This study presents cryo-EM reconstructions of human TSEN with intron-containing pre-tRNAs and reveals the molecular principles of pre-tRNA recognition and cleavage. Disease mutations are also rationalized, providing insights into the pathogenesis of pontocerebellar hypoplasia.
NATURE STRUCTURAL & MOLECULAR BIOLOGY
(2023)
Review
Biochemistry & Molecular Biology
Lingling Sun, Justine Mailliot, Christiane Schaffitzel
Summary: Nonsense-mediated mRNA decay (NMD) is a cellular surveillance mechanism that degrades mRNAs with a premature stop codon and downregulates the expression of endogenous transcripts. The core NMD factors are conserved from yeast to human, but mammals have diversified NMD pathways with additional factors. This review summarizes the molecular mechanisms and cellular roles of NMD and discusses its implications in neurodevelopmental diseases, cancer, and strategies used by RNA viruses to evade recognition by the NMD machinery.
Article
Cell Biology
A. Sofia F. Oliveira, Deborah K. Shoemark, Andrew D. Davidson, Imre Berger, Christiane Schaffitzel, Adrian J. Mulholland
Summary: The spike protein of SARS-CoV-2 contains a fatty acid binding site that is functionally important. Removing the fatty acid can significantly affect the conformation and functionality of the spike protein. Different variants of SARS-CoV-2 have varying responses to the removal of the fatty acid, indicating potential differences in transmissibility and virulence. Experimental comparison of the effects of the fatty acid on different variants is necessary.
JOURNAL OF MOLECULAR CELL BIOLOGY
(2023)
Correction
Multidisciplinary Sciences
Olga Kolesnikova, Amelie Zachayus, Simon Pichard, Judit Osz, Natacha Rochel, Paola Rossolillo, Isabelle Kolb-Cheynel, Nathalie Troffer-Charlier, Emmanuel Compe, Olivier Bensaude, Imre Berger, Arnaud Poterszman
SCIENTIFIC REPORTS
(2022)
