Profile Details

Create Project

★★★★★

☆☆☆☆☆

USD 200 /hr

★★★★★

☆☆☆☆☆

Hire Dr. Kapil G.

United Kingdom

USD 200 /hr

Expert scientist in biochemistry, biophysics, structural biology, process development. Science writer.

Profile Summary

I help biotech and pharma teams solve complex challenges in preclinical drug process development. With over 15 years of hands-on experience in protein science, I work as a trusted partner to de-risk development programs, accelerate timelines, and deliver practical, scalable solutions. My expertise includes process development, structural biology, and biochemistry, with extensive experience in multiprotein complexes, virus-like particles (VLPs), and other biologics. I also specialize in method development and optimization to ensure robust, reproducible processes that support successful preclinical advancement. If you’re looking for an experienced scientist who can quickly identify problems, design effective strategies, and move your program forward with confidence, I’d be happy to support your team.

Subject Matter Expertise

• ☆ Molecular Modeling
• ☆ Molecular Docking
• ☆ Analytical Chemistry
• ☆ Spectroscopy
• ☆ UV Spectroscopy
• ☆ NMR Spectroscopy
• ☆ Mass Spectrometry
• ☆ Molecular Imaging
• ☆ Microbiology
• ☆ Virology
• ☆ Coronaviruses
• ☆ Molecular Virology
• ☆ Biochemistry
• ☆ Molecular Biology
• ☆ Omics
• ☆ Proteomics
• ☆ Molecular Cloning
• ☆ Cell Biology
• ☆ Gene Regulation
• ☆ Biotechnology & Bioengineering
• ☆ Microscopy
• ☆ Biomolecular Engineering
• ☆ Science Outreach
• ☆ Science Education
• ☆ Infectious Disease
• ☆ Viral Diseases
• ☆ Respiratory Syncytial Virus (RSV)
• ☆ Medical Writing
• ☆ Healthcare Communications
• ☆ Technical & Scientific Writing
• ☆ Science Communication

Services

• Writing
• Research
• Consulting
• Data & AI
• Product Development

Writing Technical Writing

Research User Research, Technology Scouting, Scientific and Technical Research

Consulting Operations Consulting, Scientific and Technical Consulting

Data & AI Data Visualization, Big Data Analytics

Product Development Formulation, Recipe Development, Stability/Shelf Life Testing, Product Evaluation, Material Sourcing, Product Validation, Manufacturing, Quality Assurance & Control (QA/QC), Concept Development

Work Experience

Team Leader

Glen Clova Scientific limited

March 2025 - Present

Senior Protein Scientist

Imophoron Limited

January 2022 - March 2025

Senior Research Associate

University of Bristol

October 2019 - February 2022

Research Associate

University of Bristol

September 2016 - September 2019

Post Doctoral Fellow

European Molecular Biology Laboratory

October 2015 - June 2016

Pre Doctoral Fellow

European Molecular Biology Laboratory

May 2011 - September 2015

Education

PhD

EMBL Grenoble and University Grenoble Alps, France

May 2011 - September 2015

M.Sc.Biotechnology

Indian Institute of Technology, Roorkee-India

July 2008 - June 2010

B.Sc. Biotechnology

University of Rajasthan, India

July 2005 - May 2008

Certifications

• PhD

  EMBL

  September 2015 - Present

Publications

JOURNAL ARTICLE

Hybrid B- and T-Cell Immunity Associates With Protection Against Breakthrough Infection After Severe Acute Respiratory Syndrome Coronavirus 2 Vaccination in Avon Longitudinal Study of Parents and Children (ALSPAC) Participants @article{7b331346a2474e80bc29855336dd1589, title = "Hybrid B- and T-Cell Immunity Associates With Protection Against Breakthrough Infection After Severe Acute Respiratory Syndrome Coronavirus 2 Vaccination in Avon Longitudinal Study of Parents and Children (ALSPAC) Participants", abstract = "Background Immunological memory to vaccination and viral infection involves the coordinated action of B and T cells; thus, integrated analysis of these 2 components is critical for understanding their respective contributions to protection against breakthrough infections (BIs) after vaccination. Methods We investigated cellular and humoral immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and/or vaccination in 300 adult participants from the Avon Longitudinal Study of Parents and Children (ALSPAC). Participants were grouped by those with (cases) and without (controls) a history of SARS-CoV-2 infection. To provide a quantitative correlate for protection against BI in the 8-month period after the study, Youden index thresholds were calculated for all immune measures analyzed. Results The magnitude of antibody and T-cell responses following the second vaccine dose was associated with protection against BI in participants with a history of SARS-CoV-2 infection (cases), but not in infection-naive controls. Over 8 months of follow-up, 2 threshold combinations provided the best performance for protection against BI in cases: (i) anti-spike immunoglobulin G (IgG) (≥666.4 binding antibody units [BAU]/mL) combined with anti-nucleocapsid pan-immunoglobulin (pan-Ig) (≥0.1332 BAU/mL) and (ii) spike 1–specific T cells (≥195.6 spot-forming units/106 peripheral blood mononuclear cells) combined with anti-N pan-Ig (≥0.1332 BAU/mL). Both combinations offered 100\% specificity for detecting cases without BI, with sensitivities of 83.3\% and 72.2\%, respectively. Conclusions Collectively, these results suggest that hybrid B- and T-cell immunity offers superior protection from BI after coronavirus disease 2019 (COVID-19) vaccination, and this finding has implications for designing next-generation COVID-19 vaccines that are capable of eliciting immunity to a broader repertoire of SARS-CoV-2 proteins.", author = "Baum, \{Holly E\} and Marianna Santopaolo and Ore Francis and Milodowski, \{Emily J\} and Katrina Entwistle and Oliver, \{Elizabeth H\} and Hitchings, \{Benjamin E\} and Divya Diamond and Amy Thomas and Mitchell, \{Ruth E\} and Kibble, \{Milla M\} and Kapil Gupta and \{Di Bartolo\}, \{Natalie D\} and Paul Klenerman and Anthony Brown and Morales-Aza, \{M B\} and Oliver, \{Jennifer L\} and Imre Berger and Toye, \{Ash M\} and Finn, \{Adam H R\} and Anu Goenka and Davidson, \{Andrew D\} and Ring, \{Susan M\} and Molloy, \{Lynn C\} and Melanie Lewcock and Kate Northstone and Firona Roth and Timpson, \{Nicholas John\} and Linda Wooldridge and Alice Halliday and Laura Rivino", note = "Publisher Copyright: {\textcopyright} 2025 The Author(s). Published by Oxford University Press on behalf of Infectious Diseases Society of America.", year = "2025", month = may, day = "20", doi = "10.1093/infdis/jiaf246", language = "English", volume = "232", pages = "e327--e340", journal = "Journal of Infectious Diseases", issn = "0022-1899", publisher = "Oxford University Press", number = "2", } . Journal of Infectious Diseases.

Detection of SARS-CoV-2-specific mucosal antibodies in saliva following concomitant COVID-19 and influenza vaccination in the ComFluCOV trial @article{c67dabcfd92a49c9b3930585132285ea, title = "Detection of SARS-CoV-2-specific mucosal antibodies in saliva following concomitant COVID-19 and influenza vaccination in the ComFluCOV trial", abstract = "The ComFluCOV trial randomized 679 participants to receive an age-appropriate influenza vaccine, or placebo, alongside their second COVID-19 vaccine. Concomitant administration was shown to be safe, and to preserve systemic immune responses to both vaccines. Here we report on a secondary outcome of the trial investigating SARS-CoV-2-specific mucosal antibody responses. Anti-spike IgG and IgA levels in saliva were measured with in-house ELISAs. Concomitant administration of an influenza vaccine did not affect salivary anti-spike IgG positivity rates to Pfizer/BioNTech BNT162b2 (99.1 cf. 95.6\%), or AstraZeneca ChAdOx1 (67.8\% cf. 64.9\%), at 3-weeks post-vaccination relative to placebo. Furthermore, saliva IgG positively correlated with serum titres highlighting the potential utility of saliva for assessing differences in immunogenicity in future vaccine studies. Mucosal IgA was not detected in response to either COVID-19 vaccine, reinforcing the need for novel vaccines capable of inducing sterilising immunity or otherwise reducing transmission. The trial is registered as ISRCTN 14391248.", keywords = "Humans, Antibodies, Viral, BNT162 Vaccine, COVID-19/prevention \& control, COVID-19 Vaccines, Immunoglobulin G, Influenza Vaccines, Influenza, Human/prevention \& control, Saliva, SARS-CoV-2, Vaccination", author = "Baum, \{Holly E\} and Thirard, \{Russell C J\} and Alice Halliday and Baos, \{Sarah C\} and Amy Thomas and Harris, \{Rosie A\} and Oliver, \{Elizabeth H\} and Lucy Culliford and Hitchings, \{Benjamin E\} and Todd, \{Rachel L\} and Kapil Gupta and Anu Goenka and Finn, \{Adam H R\} and Rogers, \{Chris A\} and Rajeka Lazarus and \{ComfluCOV Trial Group\}", note = "Publisher Copyright: {\textcopyright} 2024.", year = "2024", month = apr, day = "30", doi = "10.1016/j.vaccine.2024.03.061", language = "English", volume = "42", pages = "2945--2950", journal = "Vaccine", issn = "0264-410X", publisher = "Elsevier Limited", number = "12", } . Vaccine.

Engineering the ADDobody protein scaffold for generation of high-avidity ADDomer super-binders @article{fdfc87fb63004ddf98d4581c653d4278, title = "Engineering the ADDobody protein scaffold for generation of high-avidity ADDomer super-binders", abstract = "Adenovirus-derived nanoparticles (ADDomer) comprise 60 copies of adenovirus penton base protein (PBP). ADDomer is thermostable, rendering the storage, transport, and deployment of ADDomer-based therapeutics independent of a cold chain. To expand the scope of ADDomers for new applications, we engineered ADDobodies, representing PBP crown domain, genetically separated from PBP multimerization domain. We inserted heterologous sequences into hyper-variable loops, resulting in monomeric, thermostable ADDobodies expressed at high yields in Escherichia coli. The X-ray structure of an ADDobody prototype validated our design. ADDobodies can be used in ribosome display experiments to select a specific binder against a target, with an enrichment factor of ∼104-fold per round. ADDobodies can be re-converted into ADDomers by genetically reconnecting the selected ADDobody with the PBP multimerization domain from a different species, giving rise to a multivalent nanoparticle, called Chimera, confirmed by a 2.2 {\AA} electron cryo-microscopy structure. Chimera comprises 60 binding sites, resulting in ultra-high, picomolar avidity to the target.", author = "Dora Buzas and Huan Sun and Christine Toelzer and Yadav, \{Sathish K N\} and Ufuk Borucu and Gunjan Gautam and Kapil Gupta and Bufton, \{Josh C\} and Capin, \{Julien M\} and Sessions, \{Richard B\} and Frederic Garzoni and Imre Berger and Berger-Schaffitzel, \{Christiane H\}", note = "Publisher Copyright: {\textcopyright} 2023 The Author(s)", year = "2024", month = jan, day = "9", doi = "10.1016/j.str.2023.12.010", language = "English", volume = "32", pages = "342--351.e6", journal = "Structure", issn = "0969-2126", publisher = "Cell Press", number = "3", } . Structure.

In vitro generated antibodies guide thermostable ADDomer nanoparticle design for nasal vaccination and passive immunization against SARS-CoV-2 @article{7f43378068d14beabe1ad42f4a04ca30, title = "In vitro generated antibodies guide thermostable ADDomer nanoparticle design for nasal vaccination and passive immunization against SARS-CoV-2", abstract = "Background Due to COVID-19, pandemic preparedness emerges as a key imperative, necessitating new approaches to accelerate development of reagents against infectious pathogens. Methods Here, we developed an integrated approach combining synthetic, computational and structural methods with in vitro antibody selection and in vivo immunization to design, produce and validate nature-inspired nanoparticle-based reagents against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Results Our approach resulted in two innovations: (i) a thermostable nasal vaccine called ADDoCoV, displaying multiple copies of a SARS-CoV-2 receptor binding motif derived epitope and (ii) a multivalent nanoparticle superbinder, called Gigabody, against SARS-CoV-2 including immune-evasive variants of concern (VOCs). In vitro generated neutralizing nanobodies and electron cryo-microscopy established authenticity and accessibility of epitopes displayed by ADDoCoV. Gigabody comprising multimerized nanobodies prevented SARS-CoV-2 virion attachment with picomolar EC50. Vaccinating mice resulted in antibodies cross-reacting with VOCs including Delta and Omicron. Conclusion Our study elucidates Adenovirus-derived dodecamer (ADDomer)-based nanoparticles for use in active and passive immunization and provides a blueprint for crafting reagents to combat respiratory viral infections.", author = "Dora Buzas and Bunzel, \{Adrian h\} and Oskar Staufer and Milodowski, \{Emily j\} and Edmunds, \{Grace l\} and Bufton, \{Joshua C\} and \{Vidana mateo\}, \{Beatriz V\} and Yadav, \{Sathish K N\} and Kapil Gupta and Charlotte Fletcher and Williamson, \{Maia K\} and Alexandra Harrison and Ufuk Borucu and Julien Capin and Ore Francis and Georgia Balchin and Sophie Hall and Vega, \{Mirella V\} and Fabien Durbesson and Srikanth Lingappa and Renaud Vincentelli and Joe Roe and Linda Wooldridge and Rachel Burt and Anderson, \{Ross j l\} and Mulholland, \{Adrian j\} and \{Bristol uncover group\}, \{West Midlands Paediatric High Dependency\} and Jonathan Hare and Mick Bailey and Davidson, \{Andrew d\} and Adam Finn and David Morgan and Jamie Mann and Joachim Spatz and Frederic Garzoni and Christiane Schaffitzel and Imre Berger", year = "2023", month = oct, day = "17", doi = "10.1093/abt/tbad024", language = "English", volume = "6", pages = "277--297", journal = "Antibody Therapeutics", issn = "2516-4236", publisher = "Oxford University Press", number = "4", } . Antibody Therapeutics.

Murine studies and expressional analyses of human cardiac pericytes reveal novel trajectories of SARS-CoV-2 Spike protein-induced microvascular damage @article{8bb75874e5f5472cb116ad6a7c2f8c54, title = "Murine studies and expressional analyses of human cardiac pericytes reveal novel trajectories of SARS-CoV-2 Spike protein-induced microvascular damage", keywords = "Cardiac pericyte, COVID-19, Spike protein, microangiopathy, Transcriptomics", author = "Elisa Avolio and Prashant Srivastava and Ji Jiahui and Michele Carrabba and Tsang, \{Christopher Tze Wei\} and Yue Gu and Thomas, \{Anita C\} and Kapil Gupta and Imre Berger and Costanza Emanueli and Madeddu, \{Paolo R\}", note = "Funding Information: The authors wish to acknowledge the members of the University of Bristol COVID-19 Emergency Research Group (UNCOVER) for their scientific support. Drawings were generated with BioRender.com. Funding Information: This work was supported by the British Heart Foundation (BHF) project grant “Targeting the SARS-CoV-2 S-protein binding to the ACE2 receptor to preserve human cardiac pericytes function in COVID-19” (PG/20/10285) (to P.M. and E.A.); European Commission H2020 CORDIS project COVIRNA (project/id/101016072) (to C.E. and P.K.S.) and BHF Chair award (CH/15/1/31199) (to C.E). In addition, it was supported by a grant from the National Institute for Health Research (NIHR) Biomedical Research Centre at University Hospitals Bristol NHS Foundation Trust and the University of Bristol. E.A. is a postdoctoral researcher supported by the Heart Research UK translational project grant “Targeting pericytes for halting pulmonary hypertension in infants with congenital heart disease” (RG2697/21/23) (to P.M. and E.A.). I.B. is an investigator of the Wellcome Trust (106115/Z/14/Z). ", year = "2023", month = jun, day = "2", doi = "10.1038/s41392-023-01489-2", language = "English", volume = "8", journal = "Signal Transduction and Targeted Therapy", issn = "2059-3635", publisher = "Springer Nature [academic journals on nature.com]", number = "1", } . Signal Transduction and Targeted Therapy.

Kapil Gupta, Elisa Avolio, Prashant K. Srivastava, Jiahui Ji, Michele Carrabba, Christopher T. W. Tsang, Yue Gu, Anita C. Thomas, Imre Berger, Costanza Emanueli, et al. (2023). Murine studies and expressional analyses of human cardiac pericytes reveal novel trajectories of SARS-CoV-2 Spike protein-induced microvascular damage . Signal Transduction and Targeted Therapy.

Evaluation and deployment of isotype-specific salivary antibody assays for detecting previous SARS-CoV-2 infection in children and adults @article{7e482718a0e34f25ae88a095a14ef032, title = "Evaluation and deployment of isotype-specific salivary antibody assays for detecting previous SARS-CoV-2 infection in children and adults", abstract = "Background Saliva is easily obtainable non-invasively and potentially suitable for detecting both current and previous SARS-CoV-2 infection, but there is limited evidence on the utility of salivary antibody testing for community surveillance. Methods We established 6 ELISAs detecting IgA and IgG antibodies to whole SARS-CoV-2 spike protein, to its receptor binding domain region and to nucleocapsid protein in saliva. We evaluated diagnostic performance, and using paired saliva and serum samples, correlated mucosal and systemic antibody responses. The best-performing assays were field-tested in 20 household outbreaks. Results We demonstrate in test accuracy (N = 320), spike IgG (ROC AUC: 95.0\%, 92.8–97.3\%) and spike IgA (ROC AUC: 89.9\%, 86.5–93.2\%) assays to discriminate best between pre-pandemic and post COVID-19 saliva samples. Specificity was 100\% in younger age groups (0–19 years) for spike IgA and IgG. However, sensitivity was low for the best-performing assay (spike IgG: 50.6\%, 39.8–61.4\%). Using machine learning, diagnostic performance was improved when a combination of tests was used. As expected, salivary IgA was poorly correlated with serum, indicating an oral mucosal response whereas salivary IgG responses were predictive of those in serum. When deployed to household outbreaks, antibody responses were heterogeneous but remained a reliable indicator of recent infection. Intriguingly, unvaccinated children without confirmed infection showed evidence of exposure almost exclusively through specific IgA responses. Conclusions Through robust standardisation, evaluation and field-testing, this work provides a platform for further studies investigating SARS-CoV-2 transmission and mucosal immunity with the potential for expanding salivo-surveillance to other respiratory infections in hard-to-reach settings.", author = "Amy Thomas and Oliver, \{Elizabeth H\} and Baum, \{Holly E\} and Kapil Gupta and Shelley, \{Kathryn L\} and Long, \{Anna E\} and Jones, \{Hayley E\} and Joyce Smith and Hitchings, \{Benjamin E\} and \{Di Bartolo\}, \{Natalie D\} and Kate Vasileiou and Rabi, \{Fruzsina A\} and Alamir, \{Hanin I A\} and Eghleilib, \{Malak A\} and Ore Francis and Oliver, \{Jennifer L\} and Begonia Morales-Aza and Ulrike Obst and Shattock, \{Debbie J\} and Barr, \{Rachael S\} and Lucy Collingwood and Kaltun Duale and Niall Grace and \{Gonnage Livera\}, Guillaume and Bishop, \{Lindsay C\} and Downing, \{Harriet E\} and Fernanda Rodrigues and Timpson, \{Nicholas John\} and Relton, \{Caroline L\} and Toye, \{Ash M\} and Woolfson, \{Dek N\} and Imre Berger and Anu Goenka and Davidson, \{Andrew D\} and Gillespie, \{Kathleen M\} and Williams, \{Alistair J K\} and Mick Bailey and \{Brooks Pollock\}, Ellen and Finn, \{Adam H R\} and Alice Halliday and \{Study Team\}, CoMMinS", year = "2023", month = mar, day = "15", doi = "10.1038/s43856-023-00264-2", language = "English", volume = "3", journal = "Communications Medicine", issn = "2730-664X", publisher = "Nature Research", number = "1", } . Communications Medicine.

Kapil Gupta, Amy C. Thomas, Elizabeth Oliver, Holly E. Baum, Kathryn L. Shelley, Anna E. Long, Hayley E. Jones, Joyce Smith, Benjamin Hitchings, Natalie di Bartolo, et al. (2023). Evaluation and deployment of isotype-specific salivary antibody assays for detecting previous SARS-CoV-2 infection in children and adults . Communications Medicine.

Cryo-EM reveals binding of linoleic acid to SARS-CoV-2 spike glycoprotein, suggesting an antiviral treatment strategy @article{665c8a597915491695b2295964082d94, title = "Cryo-EM reveals binding of linoleic acid to SARS-CoV-2 spike glycoprotein, suggesting an antiviral treatment strategy", abstract = "The COVID-19 pandemic and concomitant lockdowns presented a global health challenge and triggered unprecedented research efforts to elucidate the molecular mechanisms and pathogenicity of SARS-CoV-2. The spike glycoprotein decorating the surface of SARS-CoV-2 virions is a prime target for vaccine development, antibody therapy and serology as it binds the host cell receptor and is central for viral cell entry. The electron cryo-microscopy structure of the spike protein revealed a hydrophobic pocket in the receptor-binding domain that is occupied by an essential fatty acid, linoleic acid (LA). The LA-bound spike protein adopts a non-infectious locked conformation which is more stable than the infectious form and shields important immunogenic epitopes. Here, the impact of LA binding on viral infectivity and replication, and the evolutionary conservation of the pocket in other highly pathogenic coronaviruses, including SARS-CoV-2 variants of concern (VOCs), are reviewed. The importance of LA metabolic products, the eicosanoids, in regulating the human immune response and inflammation is highlighted. Lipid and fatty-acid binding to a hydrophobic pocket in proteins on the virion surface appears to be a broader strategy employed by viruses, including picornaviruses and Zika virus. Ligand binding stabilizes their protein structure and assembly, and downregulates infectivity. In the case of rhinoviruses, this has been exploited to develop small-molecule antiviral drugs that bind to the hydrophobic pocket. The results suggest a COVID-19 antiviral treatment based on the LA-binding pocket.", keywords = "synthetic biology", author = "Christine Toelzer and Kapil Gupta and Imre Berger and Berger-Schaffitzel, \{Christiane H\}", note = "Publisher Copyright: open access.", year = "2023", month = feb, day = "1", doi = "10.1107/S2059798323000049", language = "English", volume = "79", pages = "111--121", journal = "Acta Crystallographica Section D: Structural Biology", issn = "2059-7983", publisher = "John Wiley \& Sons, Inc", number = "2", } . Acta Crystallographica Section D: Structural Biology.

Kapil Gupta, Christine Toelzer, Imre Berger, Christiane Schaffitzel (2023). Cryo-EM reveals binding of linoleic acid to SARS-CoV-2 spike glycoprotein, suggesting an antiviral treatment strategy . Acta Crystallographica Section D Structural Biology.

The Free Fatty Acid-Binding Pocket is a Conserved Hallmark in Pathogenic β-Coronavirus Spike Proteins from SARS-CoV to Omicron @article{12cae0d5263047dfa3afc387ef11a527, title = "The Free Fatty Acid-Binding Pocket is a Conserved Hallmark in Pathogenic β-Coronavirus Spike Proteins from SARS-CoV to Omicron", abstract = "As COVID-19 persists, severe acquired respiratory syndrome coronavirus-2 (SARS-CoV-2) Variants of Concern (VOCs) emerge, accumulating spike (S) glycoprotein mutations. S receptor37 binding domain (RBD) comprises a free fatty acid (FFA)-binding pocket. FFA-binding stabilizes a locked S conformation, interfering with virus infectivity. We provide evidence that the pocket is conserved in pathogenic β-coronaviruses (β-CoVs) infecting humans. SARS-CoV, MERS-CoV, SARS-CoV-2 and VOCs bind the essential FFA linoleic acid (LA), while binding is abolished by one mutation in common cold-causing HCoV-HKU1. In the SARS-CoV S structure, LA stabilizes the locked conformation while the open, infectious conformation is devoid of LA. Electron tomography of SARS-CoV-2 infected cells reveals that LA treatment inhibits viral replication, resulting in fewer, deformed virions. Our results establish FFA-binding as a hallmark of pathogenic -CoV infection and replication, setting the stage for FFA-based antiviral strategies to overcome COVID-19.", keywords = "synthetic biology", author = "Christine Toelzer and Kapil Gupta and \{Yadav Kadapalakere\}, Sathish and Hodgson, \{Lorna R\} and \{Kavanagh Williamson\}, Maia and Dora Buzas and Ufuk Borucu and Powers, \{Kyle T\} and Stenner, \{Richard A\} and Kate Vasileiou and Frederic Garzoni and Fitzgerald, \{Daniel J\} and Christine Payr{\'e} and Gunjan Gautam and G{\'e}rard Lambeau and Davidson, \{Andrew D\} and Paul Verkade and Frank Martin and Imre Berger and Berger-Schaffitzel, \{Christiane H\}", year = "2022", month = nov, day = "25", doi = "10.1101/2022.04.22.489083", language = "English", volume = "8", journal = "Science Advances", issn = "2375-2548", publisher = "American Association for the Advancement of Science", number = "47", } . Science Advances.

Kapil Gupta, Christine Toelzer, Sathish K. N. Yadav, Lorna Hodgson, Maia Kavanagh Williamson, Dora Buzas, Ufuk Borucu, Kyle Powers, Richard Stenner, Kate Vasileiou, et al. (2022). The free fatty acid–binding pocket is a conserved hallmark in pathogenic β-coronavirus spike proteins from SARS-CoV to Omicron . Science Advances.

Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2 @article{379110c308e6432c91615b397d81d92d, title = "Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2", abstract = "Low-volume antibody assays can be used to track SARS-CoV-2 infection rates in settings where active testing for virus is limited and remote sampling is optimal. We developed 12 ELISAs detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid, spike protein or its receptor binding domain (RBD), 3 anti-RBD isotype specific luciferase immunoprecipitation system (LIPS) assays and a novel Spike-RBD bridging LIPS total-antibody assay. We utilized pre-pandemic (n=984) and confirmed/suspected recent COVID-19 sera taken pre-vaccination rollout in 2020 (n=269). Assays measuring total antibody discriminated best between pre-pandemic and COVID-19 sera and were selected for diagnostic evaluation. In the blind evaluation, two of these assays (Spike Pan ELISA and Spike-RBD Bridging LIPS assay) demonstrated >97\% specificity and >92\% sensitivity for samples from COVID-19 patients taken >21 days post symptom onset or PCR test. These assays offered better sensitivity for the detection of COVID-19 cases than a commercial assay which requires 100-fold larger serum volumes. This study demonstrates that low-volume in-house antibody assays can provide good diagnostic performance, and highlights the importance of using well-characterized samples and controls for all stages of assay development and evaluation. These cost-effective assays may be particularly useful for seroprevalence studies in low and middle-income countries.", keywords = "synthetic biology", author = "Alice Halliday and Long, \{Anna E\} and Baum, \{Holly E\} and Amy Thomas and Shelley, \{Kathryn L\} and Oliver, \{Elizabeth H\} and Kapil Gupta and Ore Francis and \{Kavanagh Williamson\}, Maia and \{Di Bartolo\}, \{Natalie D\} and Randell, \{Matthew J\} and \{Ben Khoud\}, Yassin and Ilana Kelland and Mortimer, \{Georgina L M\} and Ball, \{Olivia E\} and Charlie Plumptre and Chandler, \{M A\} and Ulrike Obst and Massimiliano Secchi and Lorenzo Piemonti and Vito Lampasona and Joyce Smith and Michaela Gregorova and Lea Knezevic and Jane Metz and Barr, \{Rachael S\} and Morales-Aza, \{M B\} and Oliver, \{Jennifer L\} and Lucy Collingwood and Hitchings, \{Benjamin E\} and Ring, \{Susan M\} and Linda Wooldridge and Laura Rivino and Timpson, \{Nicholas John\} and Jorgen McKernon and Peter Muir and Hamilton, \{Fergus W\} and Arnold, \{David T\} and Woolfson, \{Dek N\} and Anu Goenka and Davidson, \{Andrew D\} and Toye, \{Ash M\} and Imre Berger and Mick Bailey and Gillespie, \{Kathleen M\} and Williams, \{Alistair J K\} and Adam Finn", note = "Funding Information: This work was supported by multiple grants to AH, AL, OF, AMT, and IB awarded by the Elizabeth Blackwell Institute, and funded in part by the Wellcome Trust [Grant number 204813/Z/16/Z] with additional support from Bristol Alumni and Friends. AL is funded by a Diabetes UK/JDRF RD Lawrence Fellowship (18/0005778 and 3-APF-2018-591-A-N). The DISCOVER study was supported by donations to Southmead Hospital Charity (Registered Charity Number: 1055900). The LOGIC study was funded by The Grand Appeal (The Official Bristol Children{\textquoteright}s Hospital Charity; a registered charity in England and Wales (1043603)) through a grant awarded to AF and AG. ACT is supported by the Wellcome Trust (217509/Z/19/Z) and UKRI through the JUNIPER consortium MR/V038613/1 and CoMMinS study MR/V028545/1. The UK Medical Research Council and Wellcome (Grant ref: 217065/Z/19/Z) and the University of Bristol provide core support for ALSPAC. NT is a Wellcome Trust Investigator (202802/Z/16/Z), is the PI of the Avon Longitudinal Study of Parents and Children (MRC \& WT 217065/Z/19/Z), is supported by the University of Bristol NIHR Biomedical Research Centre (BRC-1215-2001), the MRC Integrative Epidemiology Unit (MC\_UU\_00011/1) and works within the CRUK Integrative Cancer Epidemiology Programme (C18281/A29019). LIPS assay development was supported by a joint grant from Diabetes UK/JDRF (20/0006217) to KMG. IB is supported by the Wellcome Trust (106115/Z/14/Z, 221708/Z/20/Z), the ERC (contr. nrs. 834631, 963992) and the EPSRC Impact Acceleration Account EP/R511663/1. We also acknowledge funding from BBSRC/EPSRC Synthetic Biology Research Centre (BB/L01386X/1, to DNW, NB and AMT), NHS Blood and Transplant (WP15-05, to NB and AMT), and the NIHR Blood and Transplant Research Unit in Red Cell Products (IS-BTU-1214-10032, to NB and AMT). This publication is the work of the authors and AH et al. will serve as guarantors for the contents of this paper. Funding Information: AF is a member of the Joint Committee on Vaccination and Immunisation, the UK National Immunisation Technical Advisory Group and is chair of the WHO European Regional Technical Advisory Group of Experts (ETAGE) on immunization and ex officio a member of the WHO SAGE working group on COVID vaccines. He is investigator COVID-19 vaccine on studies and trials funded by Pfizer, Sanofi, Valneva, the Gates Foundation and the UK government. This manuscript presents independent research funded in part by the National Institute for Health Research (NIHR). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health and Social Care. Publisher Copyright: Copyright {\textcopyright} 2022 Halliday, Long, Baum, Thomas, Shelley, Oliver, Gupta, Francis, Williamson, Di Bartolo, Randell, Ben-Khoud, Kelland, Mortimer, Ball, Plumptre, Chandler, Obst, Secchi, Piemonti, Lampasona, Smith, Gregorova, Knezevic, Metz, Barr, Morales-Aza, Oliver, Collingwood, Hitchings, Ring, Wooldridge, Rivino, Timpson, McKernon, Muir, Hamilton, Arnold, Woolfson, Goenka, Davidson, Toye, Berger, Bailey, Gillespie, Williams and Finn.", year = "2022", month = nov, day = "9", doi = "10.3389/fimmu.2022.968317", language = "English", volume = "13", journal = "Frontiers in Immunology", issn = "1664-3224", publisher = "Frontiers Media S.A.", } . Frontiers in Immunology.

Kapil Gupta, Charles J. Buchanan, Ben Gaunt, Peter J. Harrison, Yun Yang, Jiwei Liu, Aziz Khan, Andrew M. Giltrap, Audrey Le Bas, Philip N. Ward, et al. (2022). Pathogen-sugar interactions revealed by universal saturation transfer analysis . Science.

Synthetic virions reveal fatty acid-coupled adaptive immunogenicity of SARS-CoV-2 spike glycoprotein <head> <META HTTP-EQUIV="Refresh" CONTENT="0;URL=/servlet/useragent"> </head> . Nature Communications.

Synthetic virions reveal fatty acid-coupled adaptive immunogenicity of SARS-CoV-2 spike glycoprotein @article{470dcc47782a480aa32097dd1c4fa1be, title = "Synthetic virions reveal fatty acid-coupled adaptive immunogenicity of SARS-CoV-2 spike glycoprotein", abstract = "SARS-CoV-2 infection is a major global public health concern with incompletely understood pathogenesis. The SARS-CoV-2 spike (S) glycoprotein comprises a highly conserved free fatty acid binding pocket (FABP) with unknown function and evolutionary selection advantage1,2. Deciphering FABP impact on COVID-19 progression is challenged by the heterogenous nature and large molecular variability of live virus. Here we create synthetic minimal virions (MiniVs) of wild-type and mutant SARS-CoV-2 with precise molecular composition and programmable complexity by bottom-up assembly. MiniV-based systematic assessment of S free fatty acid (FFA) binding reveals that FABP functions as an allosteric regulatory site enabling adaptation of SARS-CoV-2 immunogenicity to inflammation states via binding of pro-inflammatory FFAs. This is achieved by regulation of the S open-to-close equilibrium and the exposure of both, the receptor binding domain (RBD) and the SARS-CoV-2 RGD motif that is responsible for integrin co-receptor engagement. We find that the FDA-approved drugs vitamin K and dexamethasone modulate S-based cell binding in an FABP-like manner. In inflammatory FFA environments, neutralizing immunoglobulins from human convalescent COVID-19 donors lose neutralization activity. Empowered by our MiniV technology, we suggest a conserved mechanism by which SARS-CoV-2 dynamically couples its immunogenicity to the host immune response.", keywords = "synthetic biology", author = "Oskar Staufer and Kapil Gupta and Gunjita Singh and Kate Vasileiou and Berger-Schaffitzel, \{Christiane H\} and Ilia Platzman and Imre Berger and Spatz, \{Joachim Pius\}", note = "Funding Information: We would like to thank Sabine Gr{\"u}newald for cell culture support, Isabelle Kajzar for critical discussion of the data, Ulrike Mersdorf for support in MiniV negative staining and TEM imaging and Mirko Cortese and Christopher J. Neufeldt (Molecular Virology Heidelberg) for sharing their expertise with SARS-CoV-2 infection. Some elements in the figures were created with BioRender.com. Support from the Heidelberg Bioscience International Graduate School and the Max Planck School Matter to Life is acknowledged by O.S. J.P.S. is the Weston Visiting Professor at the Weizmann Institute of Science and part of the excellence cluster CellNetworks at the University of Heidelberg. O.S. is the Meurer Visiting Professor at the University of Bristol. The Max Planck Society is appreciated for its general support by O.S., J.E.H.B., A.Y.R., M.M., S.F., E.A.C.A., I.P., J.P.S. J.P.S. and I.P. acknowledge funding from the Federal Ministry of Education and Research of Germany, Grant Agreement no. 13XP5073A, PolyAntiBak and the MaxSynBio Consortium, which is jointly funded by the Federal Ministry of Education and Research of Germany and the Max Planck Society. J.P.S. and I.P. also acknowledge the support from the Volkswagen Stiftung (priority call {\textquoteleft}Life?{\textquoteright}). The German Science Foundation SFB1129 (project nr. 240245600-SFB1129 P15 to A.E.C.A and J.P.S. and P13 to A.R.) is acknowledged by E.A.C.A., A.R., and J.P.S. J.P.S acknowledges the support from Germany{\textquoteright}s Excellence Strategy via the Excellence Cluster 3D Matter Made to Order (EXC-2082/1–390761711). This research received support from the Elizabeth Blackwell Institute for Health Research and the EPSRC Impact Acceleration Account EP/R511663/1 University of Bristol to C.S. and I.B., the Deutsche Forschungsgemeinschaft via the Gottfried-Wilhelm-Leibniz Program to H.D. and J.P.S., a European Research Council Consolidator Grant (\#724261) to H.D., the Max Planck School Matter to Life (a joint program of BMBF and Max Planck Society) to O.S., M.M., H.D., and J.P.S., and the EU FET Open Project Virofight (\#899619) to H.D. I.B. acknowledges support from UK Research and Innovation (UKRI) through the Bristol Synthetic Biology Centre BrisSynBio (BB/L01386X/1). C.S. and I.B. are Investigators of the Wellcome Trust (210701/Z/18/Z; 106115/Z/14/Z). Funding Information: We would like to thank Sabine Gr{\"u}newald for cell culture support, Isabelle Kajzar for critical discussion of the data, Ulrike Mersdorf for support in MiniV negative staining and TEM imaging and Mirko Cortese and Christopher J. Neufeldt (Molecular Virology Heidelberg) for sharing their expertise with SARS-CoV-2 infection. Some elements in the figures were created with BioRender.com. Support from the Heidelberg Bioscience International Graduate School and the Max Planck School Matter to Life is acknowledged by O.S. J.P.S. is the Weston Visiting Professor at the Weizmann Institute of Science and part of the excellence cluster CellNetworks at the University of Heidelberg. O.S. is the Meurer Visiting Professor at the University of Bristol. The Max Planck Society is appreciated for its general support by O.S., J.E.H.B., A.Y.R., M.M., S.F., E.A.C.A., I.P., J.P.S. J.P.S. and I.P. acknowledge funding from the Federal Ministry of Education and Research of Germany, Grant Agreement no. 13XP5073A, PolyAntiBak and the MaxSynBio Consortium, which is jointly funded by the Federal Ministry of Education and Research of Germany and the Max Planck Society. J.P.S. and I.P. also acknowledge the support from the Volkswagen Stiftung (priority call {\textquoteleft}Life?{\textquoteright}). The German Science Foundation SFB1129 (project nr. 240245600-SFB1129 P15 to A.E.C.A and J.P.S. and P13 to A.R.) is acknowledged by E.A.C.A., A.R., and J.P.S. J.P.S acknowledges the support from Germany{\textquoteright}s Excellence Strategy via the Excellence Cluster 3D Matter Made to Order (EXC-2082/1–390761711). This research received support from the Elizabeth Blackwell Institute for Health Research and the EPSRC Impact Acceleration Account EP/R511663/1 University of Bristol to C.S. and I.B., the Deutsche Forschungsgemeinschaft via the Gottfried-Wilhelm-Leibniz Program to H.D. and J.P.S., a European Research Council Consolidator Grant (\#724261) to H.D., the Max Planck School Matter to Life (a joint program of BMBF and Max Planck Society) to O.S., M.M., H.D., and J.P.S., and the EU FET Open Project Virofight (\#899619) to H.D. I.B. acknowledges support from UK Research and Innovation (UKRI) through the Bristol Synthetic Biology Centre BrisSynBio (BB/L01386X/1). C.S. and I.B. are Investigators of the Wellcome Trust (210701/Z/18/Z; 106115/Z/14/Z). Publisher Copyright: {\textcopyright} 2022, The Author(s).", year = "2022", month = feb, day = "14", doi = "10.1038/s41467-022-28446-x", language = "English", volume = "13", journal = "Nature Communications", issn = "2041-1723", publisher = "Nature Research", number = "1", } . Nature Communications.

Structural insights in cell-type specific evolution of intra-host diversity by SARS-CoV-2 <head> <META HTTP-EQUIV="Refresh" CONTENT="0;URL=/servlet/useragent"> </head> . Nature Communications.

Structural insights in cell-type specific evolution of intra-host diversity by SARS-CoV-2 @article{fd11d5cd6b174e38a2aecebae1a95962, title = "Structural insights in cell-type specific evolution of intra-host diversity by SARS-CoV-2", abstract = "As the global burden of SARS-CoV-2 infections escalates, so does the evolution of viral variants with increased transmissibility and pathology. In addition to this entrenched diversity, RNA viruses can also display genetic diversity within single infected hosts with co-existing viral variants evolving differently in distinct cell types. The BriSΔ variant, originally identified as a viral subpopulation from SARS-CoV-2 isolate hCoV-19/England/02/2020, comprises in the spike an eight amino-acid deletion encompassing a furin recognition motif and S1/S2 cleavage site. We elucidate the structure, function and molecular dynamics of this spike providing mechanistic insight into how the deletion correlates to viral cell tropism, ACE2 receptor binding and infectivity of this SARS-CoV-2 variant. Our results reveal long-range allosteric communication between functional domains that differ in the wild-type and the deletion variant and support a view of SARS-CoV-2 probing multiple evolutionary trajectories in distinct cell types within the same infected host. ", keywords = "synthetic biology", author = "Kapil Gupta and Christine Toelzer and \{Kavanagh Williamson\}, Maia and Shoemark, \{Deborah K\} and A Oliveira and Davidson, \{Andrew D\} and Almuqrin, \{Abdulaziz M\} and Oskar Staufer and \{Yadav Kadapalakere\}, Sathish and Ufuk Borucu and Frederic Garzoni and Fitzgerald, \{Daniel J\} and Spatz, \{Joachim Pius\} and Mulholland, \{Adrian J\} and Davidson, \{Andrew D\} and Berger-Schaffitzel, \{Christiane H\} and Imre Berger", note = "Funding Information: We thank all members of the Berger and Schaffitzel teams as well as Robin Shattock (Imperial College, UK) and Adam Finn (Bristol UNCOVER Group and Children{\textquoteright}s Vaccine Centre, Bristol Medical School) for their assistance and advice. We thank Simon Burbidge, Thomas Batstone and Matt Williams for computation infrastructure support. We would like to thank the Advanced Computing Research Centre (ACRC) at the University of Bristol for access to BlueCryo, BlueCrystal Phase 4 and BlueGEM, and the UK HECBioSim for access to the UK supercomputer, ARCHER. We are particularly grateful to Thiru Thangarajah (Genscript) for early access to Genscript{\textquoteright}s cPass{\texttrademark} SARS-CoV-2 Neutralization Antibody Detection/Surrogate Virus Neutralization Test Kit (L00847). We thank Sebastian Fabritz and the Core Facility for Mass Spectrometry at the Max Planck Institute for Medical Research for their support on MS measurements. For the purpose of Open Access, the authors have applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. This research received support from the Elizabeth Blackwell Institute for Health Research and the EPSRC Impact Acceleration Account EP/R511663/1, University of Bristol, from Bris-SynBio a BBSRC/EPSRC Research Centre for synthetic biology at the University of Bristol (BB/L01386X/1) (I.B., C.S., A.J.M., D.K.S., and A.S.F.O.) and from the BBSRC (BB/P000940/1) (C.S. and I.B.). This work received generous support from the Oracle Higher Education and Research program to enable cryo-EM data processing using Oracle{\textquoteright}s high-performance public cloud infrastructure (https://cloud.oracle.com/en\_US/ cloud-infrastructure) and the EPSRC through a COVID-19 project award via HECBioSim to access ARCHER (A.J.M.). We acknowledge support and assistance by the Wolfson Bioimaging Facility and the GW4 Facility for High-Resolution Electron Cryo-Microscopy funded by the Wellcome Trust (202904/Z/16/Z and 206181/Z/17/Z) and BBSRC (BB/R000484/1). The authors are grateful to University of Bristol{\textquoteright}s Alumni and Friends, which funded the ImageXpress Pico Imaging System. O.S. acknowledges support from the Elisabeth Muerer Foundation, the Max Planck School Matter to Life and the Heidelberg Biosciences International Graduate School. J.S. is the Weston Visiting Professor at the Weizmann Institute of Science, part of the excellence cluster CellNetworks at Heidelberg University and acknowledges funding from the European Research Council (ERC, contract no. 294852), SynAd and the MaxSynBio Consortium, funded by the Federal Ministry of Education and Research of Germany and the Max Planck Society, from the SFB 1129 and Project 240245660-SFB1129 P15 of the German Research Foundation (DFG) and from the Volkswagen Stiftung (priority call “Life?”). A.D.D. and D.A.M. are supported by the United States Food and Drug Administration (HHSF223201510104C) and UK Research and Innovation/Medical Research Council (MRC) (MR/V027506/1). M.K.W is supported by MRC grants MR/R020566/1 and MR/ V027506/1 (awarded to A.D.D). A.J.M. is supported by the British Society for Antimicrobial Chemotherapy (BSAC-COVID-30) and the EPSRC (EP/M022609/1, CCP-BioSim). I.B. acknowledges support from the EPSRC Innovative Future Vaccine Manufacturing and Research Hub (EP/R013764/1). C.S. and I.B. are Investigators of the Wellcome Trust (210701/Z/18/Z; 106115/Z/14/Z). Publisher Copyright: {\textcopyright} 2022, The Author(s).", year = "2022", month = jan, day = "11", doi = "10.1038/s41467-021-27881-6", language = "English", volume = "13", journal = "Nature Communications", issn = "2041-1723", publisher = "Nature Research", number = "1", } . Nature Communications.

The SARS-CoV-2 Spike protein disrupts human cardiac pericytes function through CD147-receptor-mediated signalling @article{9019d2446a2f40fd8461c33ec130e93f, title = "The SARS-CoV-2 Spike protein disrupts human cardiac pericytes function through CD147-receptor-mediated signalling: a potential non-infective mechanism of COVID-19 microvascular disease", abstract = "The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes a broad range of clinical responses including prominent microvascular damage. The capacity of SARS-CoV-2 to infect vascular cells is still debated. Additionally, the SARS-CoV-2 Spike (S) protein may act as a ligand to induce non-infective cellular stress. We tested this hypothesis in pericytes (PCs), which are reportedly reduced in the heart of patients with severe coronavirus disease-2019 (COVID-19). Here we newly show that the in vitro exposure of primary human cardiac PCs to the SARS-CoV-2 wild type strain or the Alpha and Delta variants caused rare infection events. Exposure to the recombinant S protein alone elicited signalling and functional alterations, including: (1) increased migration, (2) reduced ability to support endothelial cell (EC) network formation on Matrigel, (3) secretion of pro-inflammatory molecules typically involved in the cytokine storm, and (4) production of pro-apoptotic factors causing EC death. Next, adopting a blocking strategy against the S protein receptors angiotensin-converting enzyme 2 (ACE2) and CD147, we discovered that the S protein stimulates the phosphorylation/activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) through the CD147 receptor, but not ACE2, in PCs. The neutralisation of CD147, either using a blocking antibody or mRNA silencing, reduced ERK1/2 activation, and rescued PC function in the presence of the S protein. Immunoreactive S protein was detected in the peripheral blood of infected patients. In conclusion, our findings suggest that the S protein may prompt PC dysfunction, potentially contributing to microvascular injury. This mechanism may have clinical and therapeutic implications.", keywords = "pericyte, Microvascular disease, COVID-19, Spike protein, CD147, angiotensin converting enzyme 2", author = "Elisa Avolio and Michele Carrabba and Rachel Milligan and \{Kavanagh Williamson\}, Maia and Beltrami, \{Antonio Paolo\} and Kapil Gupta and Elvers, \{Karen T\} and Monica Gamez and Foster, \{R R\} and Gillespie, \{Kathleen M\} and Hamilton, \{Fergus W\} and Arnold, \{David T\} and Imre Berger and Davidson, \{Andrew D\} and Hill, \{Darryl J\} and Massimo Caputo and Madeddu, \{Paolo R\}", note = "Publisher Copyright: {\textcopyright} 2021 The Author(s).", year = "2021", month = nov, day = "22", doi = "10.1042/CS20210735", language = "English", volume = "135", pages = "2667--2689", journal = "Clinical Science", issn = "0143-5221", publisher = "Portland Press", number = "24", } . Clinical Science.

TAF8 regions important for TFIID lobe B assembly or for TAF2 interactions are required for embryonic stem cell survival @article{d691f7fd8ba740a789ee826300bab0f5, title = "TAF8 regions important for TFIID lobe B assembly or for TAF2 interactions are required for embryonic stem cell survival", abstract = "The human general transcription factor TFIID is composed of the TATA-binding protein (TBP) and 13 TBP-associated factors (TAFs). In eukaryotic cells, TFIID is thought to nucleate RNA polymerase II (Pol II) preinitiation complex formation on all protein coding gene promoters and thus, be crucial for Pol II transcription. TFIID is composed of three lobes, named A, B, and C. A 5TAF core complex can be assembled in vitro constituting a building block for the further assembly of either lobe A or B in TFIID. Structural studies showed that TAF8 forms a histone fold pair with TAF10 in lobe B and participates in connecting lobe B to lobe C. To better understand the role of TAF8 in TFIID, we have investigated the requirement of the different regions of TAF8 for the in vitro assembly of lobe B and C and the importance of certain TAF8 regions for mouse embryonic stem cell (ESC) viability. We have identified a region of TAF8 distinct from the histone fold domain important for assembling with the 5TAF core complex in lobe B. We also delineated four more regions of TAF8 each individually required for interacting with TAF2 in lobe C. Moreover, CRISPR/Cas9-mediated gene editing indicated that the 5TAF core-interacting TAF8 domain and the proline-rich domain of TAF8 that interacts with TAF2 are both required for mouse embryonic stem cell survival. Thus, our study defines distinct TAF8 regions involved in connecting TFIID lobe B to lobe C that appear crucial for TFIID function and consequent ESC survival.", keywords = "synthetic biology", author = "Elisabeth Scheer and Jie Luo and Andrea Bernardini and Frank Ruffenach and Jean-Marie Garnier and Isabelle Kolb-Cheynel and Kapil Gupta and Imre Berger and Jeff Ranish and L{\'a}szl{\'o} Tora", note = "Funding Information: We are grateful to the IGBMC cell culture and flow cytometry services for assistance. We thank D. Schmit for starting the initial experiments on this project, F. El-Saafin and the Tora lab members for helpful discussions, N. Jung and B. Reina San Martin for help with the CRISPR/Cas9 experiments, and L. Negroni with MS experiments and data submission. This study was supported by grants from Agence Nationale de la Recherche (ANR): ANR-19-CE11-0003-02, ANR-PRCI-19-CE12-0029-01, ANR-20-CE12-0017-03, NIH R01 GM131626 and NSF (Award Number: 1933344) grants (to L. T.); NIH R01 GM110064 and GM136974 (to J. R.) and supported by funds from CNRS, INSERM, Strasbourg University, and Investissements d?Avenir grants (ANR-10-IDEX-0002-02 and ANR-10-LABX-0030-INRT). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or other granting agencies. Funding Information: Funding and additional information—This study was supported by grants from Agence Nationale de la Recherche (ANR): ANR-19-CE11-0003-02, ANR-PRCI-19-CE12-0029-01, ANR-20-CE12-0017-03, NIH R01 GM131626 and NSF (Award Number: 1933344) grants (to L. T.); NIH R01 GM110064 and GM136974 (to J. R.) and supported by funds from CNRS, INSERM, Strasbourg University, and Investissements d{\textquoteright}Avenir grants (ANR-10-IDEX-0002-02 and ANR-10-LABX-0030-INRT). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or other granting agencies. Publisher Copyright: {\textcopyright} 2021 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology.", year = "2021", month = nov, day = "1", doi = "10.1016/j.jbc.2021.101288", language = "English", volume = "297", journal = "Journal of Biological Chemistry", issn = "0021-9258", publisher = "American Society for Biochemistry and Molecular Biology Inc.", number = "5", } . Journal of Biological Chemistry.

Young infants exhibit robust functional antibody responses and restrained IFN-γ production to SARS-CoV-2 @article{a2a5b26147214953894a25f29d198540, title = "Young infants exhibit robust functional antibody responses and restrained IFN-γ production to SARS-CoV-2", abstract = "Severe COVID-19 appears rare in children. This is unexpected, especially in young infants, who are vulnerable to severe disease caused by other respiratory viruses. We evaluate convalescent immune responses in 4 infants under 3 months old with confirmed COVID-19 who presented with mild febrile illness, alongside their parents, and adult controls recovered from confirmed COVID-19. Although not statistically significant, compared to seropositive adults, infants have high serum levels of IgG and IgA to SARS-CoV-2 spike protein, with a corresponding functional ability to block SARS-CoV-2 cellular entry. Infants also exhibit robust saliva anti-spike IgG and IgA responses. Spike-specific IFN-γ production by infant peripheral blood mononuclear cells appears restrained, but the frequency of spike-specific IFN-γ- and/or TNF-α-producing T cells is comparable between infants and adults. On principal-component analysis, infant immune responses appear distinct from their parents. Robust functional antibody responses alongside restrained IFN-γ production may help protect infants from severe COVID-19.", keywords = "synthetic biology, immunity, COVID-19, antibody, infant, T cell", author = "Anu Goenka and Alice Halliday and Michaela Gregorova and Milodowski, \{Emily J\} and Amy Thomas and \{Kavanagh Williamson\}, Maia and Baum, \{Holly E\} and Oliver, \{Elizabeth H\} and Long, \{Anna E\} and Lea Knezevic and Kapil Gupta and \{Di Bartolo\}, \{Natalie D\} and Imre Berger and Toye, \{Ash M\} and Jolanta Bernatoniene and Mick Bailey and Gillespie, \{Kathleen M\} and Davidson, \{Andrew D\} and Linda Wooldridge and Laura Rivino and Adam Finn", year = "2021", month = jul, day = "20", doi = "10.1016/j.xcrm.2021.100327", language = "English", volume = "2", journal = "Cell Reports Medicine", issn = "2666-3791", publisher = "Cell Press", number = "7", } . Cell Reports Medicine.

Molecular Simulations suggest Vitamins, Retinoids and Steroids as Ligands of the Free Fatty Acid Pocket of the SARS-CoV-2 Spike Protein* @article{a6b7637eed3346908046466435208570, title = "Molecular Simulations suggest Vitamins, Retinoids and Steroids as Ligands of the Free Fatty Acid Pocket of the SARS-CoV-2 Spike Protein*", abstract = "Following our recent identification of a fatty acid binding site in the SARS-CoV-2 spike protein (Toelzer et al., Science eabd3255 (2020)), we investigate the binding of linoleate and other potential ligands at this site using molecular dynamics simulations. The results support the hypothesis that linoleate stabilises the locked form of the spike, in which its interaction interface for the ACE2 receptor is occluded. The simulations indicate weaker binding of linoleate to the partially open conformation. Simulations of dexamethasone bound at this site indicate that it binds similarly to linoleate, and thus may also stabilize a locked spike conformation. In contrast, simulations suggest that cholesterol bound at this site may destabilize the locked conformation, and in the open conformation, may preferentially bind at an alternative site in the hinge region between the receptor binding domain and the domain below, which could have functional relevance. We also use molecular docking to identify potential ligands that may bind at the fatty acid binding site, using the Bristol University Docking Engine (BUDE). BUDE docking successfully reproduces the linoleate complex and also supports binding of dexamethasone at the spike fatty acid site. Virtual screening of a library of approved drugs identifies vitamins D, K and A, as well as retinoid ligands with experimentally demonstrated activity against SARS-CoV-2 replication in vitro, as also potentially able to bind at this site. Our data suggest that the fatty acid binding site of the SARS-CoV-2 spike protein may bind a diverse array of candidate ligands. Targeting this site with small molecules, including dietary components such as vitamins, which may stabilise its locked conformation and represents a potential avenue for novel therapeutics or prophylaxis for COVID-19.", keywords = "synthetic biology", author = "Shoemark, \{Deborah K\} and Colenso, \{Charlotte K.\} and Christine Toelzer and Kapil Gupta and Sessions, \{Richard B\} and Sessions, \{Richard B\} and Davidson, \{Andrew D\} and Imre Berger and Christiane Schaffitzel and James Spencer and Mulholland, \{Adrian J\}", year = "2021", month = mar, day = "22", doi = "10.26434/chemrxiv.13143761.v1", language = "English", volume = "60", journal = "Angewandte Chemie", issn = "0044-8249", publisher = "John Wiley \& Sons, Ltd.", number = "13", } . Angewandte Chemie.

VLP-factory™ and ADDomer © @article{3dd9f0bdea564d799d0b103fb437b6a4, title = "VLP-factory{\texttrademark} and ADDomer {\textcopyright}: Self-assembling Virus-Like Particle (VLP) technologies for multiple protein and peptide epitope display", abstract = "Virus‐like particles (VLPs) play a prominent role in vaccination as safe and highly versatile alternatives to attenuated or inactivated viruses or subunit vaccines. We present here two innovations, VLP‐factory{\texttrademark} and ADDomer{\textcopyright}, for creating VLPs displaying entire proteins or peptide epitopes as antigens, respectively, to enable efficient vaccination. For producing these VLPs, we use MultiBac, a baculovirus expression vector system (BEVS) that we developed for producing complex protein biologics in insect cells transfected with an engineered baculovirus. VLPs are protein assemblies that share features with viruses but are devoid of genetic material, and thus considered safe. VLP‐factory{\texttrademark} represents a customized MultiBac baculovirus tailored to produce enveloped VLPs based on the M1 capsid protein of influenza virus. We apply VLP‐factory{\texttrademark} to create an array of influenza‐derived VLPs presenting functional mutant influenza hemagglutinin (HA) glycoprotein variants. Moreover, we describe MultiBac‐based production of ADDomer{\textcopyright}, a synthetic self‐assembling adenovirus‐derived protein‐based VLP platform designed to display multiple copies of pathogenic epitopes at the same time on one particle for highly efficient vaccination. {\textcopyright} 2021 The Authors.", keywords = "synthetic biology, protein and peptide display, virus‐like particle (VLP), antigenic epitope, immunization, baculovirus expression vector system (BEVS), MultiBac, vaccine", author = "Duygu Sari-Ak and Bufton, \{Josh C\} and Kapil Gupta and Frederic Garzoni and Fitzgerald, \{Daniel J\} and Berger-Schaffitzel, \{Christiane H\} and Imre Berger", year = "2021", month = mar, day = "17", doi = "10.1002/cpz1.55", language = "English", volume = "1", journal = "Current Protocols", issn = "2691-1299", publisher = "Wiley", number = "3", } . Current Protocols.

TBPL2/TFIIA complex establishes the maternal transcriptome through oocyte-specific promoter usage <head> <META HTTP-EQUIV="Refresh" CONTENT="0;URL=/servlet/useragent"> </head> . Nature Communications.

Kapil Gupta, Christine Toelzer, Sathish K. N. Yadav, Ufuk Borucu, Andrew D. Davidson, Maia Kavanagh Williamson, Deborah K. Shoemark, Frederic Garzoni, Oskar Staufer, Rachel Milligan, et al.(2020). Free fatty acid binding pocket in the locked structure of SARS-CoV-2 spike protein . Science. 370. (6517). p. 725--730. American Association for the Advancement of Science ({AAAS})

Free fatty acid binding pocket in the locked structure of SARS-CoV-2 spike protein @article{c70a4d1122ec417db1a84ad0d6c32766, title = "Free fatty acid binding pocket in the locked structure of SARS-CoV-2 spike protein", abstract = "COVID-19, caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2), represents a global crisis. Key to SARS-CoV-2 therapeutic development is unraveling the mechanisms driving high infectivity, broad tissue tropism and severe pathology. Our 2.85 {\AA} cryo-EM structure of SARS-CoV-2 spike (S) glycoprotein reveals that the receptor binding domains (RBDs) tightly bind the essential free fatty acid (FFA) linoleic acid (LA) in three composite binding pockets. The pocket also appears to be present in the highly pathogenic coronaviruses SARS-CoV and MERS-CoV. LA binding stabilizes a locked S conformation giving rise to reduced ACE2 interaction in vitro. In human cells, LA supplementation synergizes with the COVID-19 drug remdesivir, suppressing SARS-CoV-2 replication. Our structure directly links LA and S, setting the stage for intervention strategies targeting LA binding by SARS-CoV-2.", keywords = "synthetic biology", author = "Christine Toelzer and Kapil Gupta and Yadav, \{Sathish K N\} and Ufuk Borucu and Davidson, \{Andrew D\} and \{Kavanagh Williamson\}, Maia and Shoemark, \{Deborah K\} and Frederic Garzoni and Oskar Staufer and Rachel Milligan and Julien Capin and Mulholland, \{Adrian J\} and Imre Berger and Christiane Schaffitzel", year = "2020", month = nov, day = "6", doi = "10.1126/science.abd3255", language = "English", volume = "370", journal = "Science", issn = "0036-8075", publisher = "American Association for the Advancement of Science", number = "6517", } . Science.

DNA origami-based single-molecule force spectroscopy elucidates RNA Polymerase III pre-initiation complex stability <head> <META HTTP-EQUIV="Refresh" CONTENT="0;URL=/servlet/useragent"> </head> . Nature Communications.

DNA origami-based single-molecule force spectroscopy elucidates RNA Polymerase III pre-initiation complex stability @article{67c460ecd7e8471380777961cd4d4480, title = "DNA origami-based single-molecule force spectroscopy elucidates RNA Polymerase III pre-initiation complex stability", abstract = "The TATA-binding protein (TBP) and a transcription factor (TF) IIB-like factor are important constituents of all eukaryotic initiation complexes. The reason for the emergence and strict requirement of the additional initiation factor Bdp1 in the RNA polymerase (RNAP) III system, however, remained elusive. A poorly studied aspect in this context is the effect of DNA strain arising from DNA compaction and transcriptional activity on initiation complex formation. We made use of a DNA origami-based force clamp to follow the assembly of human initiation complexes in the RNAP II and RNAP III systems at the single-molecule level under piconewton forces. We demonstrate that TBP-DNA complexes are force-sensitive and TFIIB is sufficient to stabilise TBP on a strained promoter. In contrast, Bdp1 is the pivotal component that ensures stable anchoring of initiation factors, and thus the polymerase itself, in the RNAP III system. Thereby, we offer an explanation for the crucial role of Bdp1 for the high transcriptional output of RNAP III.", keywords = "TFIIB, transcription, synthetic biology, Bdp1, single molecule force measurements, single-molecule FRET, DNA origami, RNA polymerase, TBP", author = "Kapil Gupta and Imre Berger", year = "2020", month = jun, day = "5", doi = "10.1038/s41467-020-16702-x", language = "English", journal = "Nature Communications", issn = "2041-1723", publisher = "Nature Research", } . Nature Communications.

(2019). Synthetic energy sensor AMPfret deciphers adenylate-dependent AMPK activation mechanism . Nature Communications.

(2019). The MultiBac system: a perspective . Emerging Topics in Life Sciences.

The MultiBac System @article{6c20bbab69be46f8ab8aec2224913334, title = "The MultiBac System: A Perspective", abstract = "Baculovirus expression is a time-tested technique to produce proteins in insect cells, in high quality and quantity for a range of applications. MultiBac is a baculovirus expression system we developed originally for producing multiprotein complexes comprising many subunits, for structural and mechanistic studies. First introduced in 2004, MultiBac is now in use in many laboratories worldwide, accelerating research programmes in academia and industry. We have continuously optimized our MultiBac system, providing customized reagents and standard operating protocols to facilitate its use also by non-specialists. More recently, we have generated MultiBac genomes tailored for specific purposes, for example, to produce humanized glycoproteins, high-value pharmaceutical targets including kinases, viral polymerases, and virus-like particles (VLPs) as promising vaccine candidates. By altering the host tropism of the baculovirion, we created MultiBacMam, a heterologous DNA delivery toolkit to target mammalian cells, tissues and organisms. Introducing CRISPR/Cas modalities, we set the stage for large-scale genomic engineering applications utilizing this high-capacity DNA delivery tool. Exploiting synthetic biology approaches and bottom-up design, we engage in optimizing the properties of our baculoviral genome, also to improve manufacturing at scale. Here we provide a perspective of our MultiBac system and its developments, past, present and future.", keywords = "synthetic biology, Synthetic biology", author = "Imre Berger and Christine Toelzer and Kapil Gupta", year = "2019", month = sep, day = "3", doi = "10.1042/ETLS20190084", language = "English", journal = "Emerging Topics in Life Sciences", issn = "2397-8554", publisher = "Portland Press", } . Emerging Topics in Life Sciences.

(2019). Sequential Digestion with Trypsin and Elastase in Cross-Linking Mass Spectrometry . Analytical Chemistry.

Sequential Digestion with Trypsin and Elastase in Cross-Linking Mass Spectrometry @article{d0a4a8965c2c471cb9f39ea9f21841cc, title = "Sequential Digestion with Trypsin and Elastase in Cross-Linking Mass Spectrometry", abstract = "Cross-linking mass spectrometry has become an important approach for studying protein structures and protein-protein interactions. The amino acid compositions of some protein regions impede the detection of cross-linked residues, although it would yield invaluable information for protein modeling. Here, we report on a sequential-digestion strategy with trypsin and elastase to penetrate regions with a low density of trypsin-cleavage sites. We exploited intrinsic substrate-recognition properties of elastase to specifically target larger tryptic peptides. Our application of this protocol to the TAF4-12 complex allowed us to identify cross-links in previously inaccessible regions.", keywords = "synthetic biology, Synthetic biology", author = "Therese Dau and Kapil Gupta and Imre Berger and Juri Rappsilber", year = "2019", month = apr, day = "2", doi = "10.1021/acs.analchem.8b05222", language = "English", volume = "91", pages = "4472--4478", journal = "Analytical Chemistry", issn = "0003-2700", publisher = "American Chemical Society", number = "7", } . Analytical Chemistry.

Synthetic energy sensor AMPfret deciphers adenylate-dependent AMPK activation mechanism @article{3d2c10226842433181bdb88e74cb8c5a, title = "Synthetic energy sensor AMPfret deciphers adenylate-dependent AMPK activation mechanism", abstract = " AMP-activated protein kinase AMPK senses and regulates cellular energy state. AMPK activation by increasing AMP and ADP concentrations involves a conformational switch within the heterotrimeric complex. This is exploited here for the construction of a synthetic sensor of cellular energetics and allosteric AMPK activation, AMPfret. Based on engineered AMPK fused to fluorescent proteins, the sensor allows direct, real-time readout of the AMPK conformational state by fluorescence resonance energy transfer (FRET). AMPfret faithfully and dynamically reports the binding of AMP and ADP to AMPK γ-CBS sites, competed by Mg 2+ -free ATP. FRET signals correlate with activation of AMPK by allosteric mechanisms and protection from dephosphorylation, attributed here to specific CBS sites, but does not require activation loop phosphorylation. Moreover, AMPfret detects binding of pharmacological compounds to the AMPK α/β-ADaM site enabling activator screening. Cellular assays demonstrate that AMPfret is applicable in vivo for spatiotemporal analysis of energy state and allosteric AMPK activation. ", keywords = "synthetic biology, cell biology, biochemistry, SYNTHETIC BIOLOGY, biophysics, drug discovery, biological techniques", author = "Martin Pelosse and C{\'e}cile Cottet-Roussellea and C{\'e}cile Bidand and Aur{\'e}lie Dupont and Kapil Gupta and Imre Berger and Uwe Schlattner", year = "2019", month = mar, day = "4", doi = "10.1038/s41467-019-08938-z", language = "English", volume = "10", journal = "Nature Communications", issn = "2041-1723", publisher = "Nature Research", } . Nature Communications.

MultiBac @article{24704f629ce0479ab7e533fbe919446d, title = "MultiBac: Baculovirus-Mediated Multigene DNA Cargo Delivery in Insect and Mammalian Cells", abstract = "The baculovirus/insect cell system (BICS) is widely used in academia and industry to produce eukaryotic proteins for many applications, ranging from structure analysis to drug screening and the provision of protein biologics and therapeutics. Multiprotein complexes emerged as vital catalysts of cellular function. To unlock the structure and mechanism of these essential molecular machines and decipher their function, we developed MultiBac, a BICS particularly tailored for heterologous multigene transfer and multiprotein complex production. Baculovirus is unique among common viral vectors in its capacity to accommodate very large quantities of heterologous DNA and to faithfully deliver this cargo to a host cell of choice. We exploited this beneficial feature to outfit insect cells with synthetic DNA circuitry conferring new functionality during heterologous protein expression, developing customized MultiBac baculovirus variants in the process. By altering its tropism, recombinant baculovirions can be used for highly efficient delivery of customized DNA cargo in mammalian cells and tissues. Current advances in synthetic biology greatly facilitates the construction or recombinant baculoviral genomes for gene editing and genome engineering, mediated by MultiBac baculovirus tailored to this purpose. Here, recent developments and exploits of the MultiBac system are presented and discussed.", keywords = "Multiprotein Complex, synthetic biology, GPCR, Transduction, Drug Screening, Virus-like Particle VLP, Heterologous Expression, Genome Engineering, Synthetic Biology, Human TFIID, Baculovirus", author = "Kapil Gupta and Christine Toelzer and Duygu Sari-Ak and Fitzgerald, \{Daniel J\} and Christiane Berger-Schaffitzel and Imre Berger", year = "2019", month = mar, doi = "10.3390/v11030198", language = "English", volume = "11", journal = "Viruses", issn = "1999-4915", publisher = "MDPI AG", number = "3", } . Viruses.

Kapil Gupta, Christine T&#246;lzer, Duygu Sari-Ak, Daniel J. Fitzgerald, Christiane Schaffitzel, Imre Berger (2019). MultiBac: Baculovirus-Mediated Multigene DNA Cargo Delivery in Insect and Mammalian Cells . Viruses.

Kapil Gupta, Christine T&#246;lzer, Duygu Sari-Ak, Daniel J. Fitzgerald, Christiane Schaffitzel, Imre Berger (2019). MultiBac: Baculovirus-Mediated Multigene DNA Cargo Delivery in Insect and Mammalian Cells . Viruses.

Gupta, K., T&#246;lzer, C., Sari-Ak, D., Fitzgerald, D.J., Schaffitzel, C., Berger, I.(2019). Multibac: Baculovirus-mediated multigene DNA cargo delivery in insect and mammalian cells . Viruses. 11. (3).

Dau, T., Gupta, K., Berger, I., Rappsilber, J.(2019). Sequential Digestion with Trypsin and Elastase in Cross-Linking Mass Spectrometry . Analytical Chemistry. 91. (7). p. 4472-4478.

Pelosse, M., Cottet-Rousselle, C., Bidan, C.M., Dupont, A., Gupta, K., Berger, I., Schlattner, U.(2019). Synthetic energy sensor AMPfret deciphers adenylate-dependent AMPK activation mechanism . Nature Communications. 10. (1).

Chaperonin CCT Checkpoint Function in Basal Transcription Factor TFIID Assembly @article{7cb0b29ba4ee473fb61c56492e05ee3f, title = "Chaperonin CCT Checkpoint Function in Basal Transcription Factor TFIID Assembly", abstract = "TFIID is a cornerstone of eukaryotic gene regulation. Distinct TFIID complexes with unique subunit composition exist and several TFIID subunits are shared with other complexes, conveying intricate cellular decision making to control subunit allocation and functional assembly of this essential transcription factor. However, the underlying molecular mechanisms remain poorly understood. Here, we used quantitative proteomics to examine TFIID submodules and assembly mechanisms in human cells. Structural and mutational analysis of the cytoplasmic TAF5-TAF6-TAF9 submodule identified novel interactions crucial for TFIID integrity, and for allocating TAF9 to TFIID or the SAGA co-activator complex. We discover a key checkpoint function for the chaperonin CCT, which specifically associates with nascent TAF5 for subsequent handover to TAF6-TAF9 and ultimate holo-TFIID formation. Our findings illustrate at the molecular level how multisubunit complexes are crafted in the cell, involving checkpoint decisions facilitated by a chaperone machine.", keywords = "synthetic biology", author = "Antonova, \{Simona V\} and Matthias Haffke and Eleanora Corradini and Mykolas Mikuciunas and Low, \{Teck Y\} and Luca Signor and Van-Es, \{Robert M\} and Kapil Gupta and Elisabeth Scheer and Vos, \{Harmjan R\} and L{\'a}szl{\'o} Tora and Heck, \{Albert J R\} and Timmers, \{H T Marc\} and Imre Berger", year = "2018", month = dec, doi = "10.1038/s41594-018-0156-z", language = "English", volume = "25", pages = "1119--1127", journal = "Nature Structural and Molecular Biology", issn = "1545-9993", publisher = "Nature Research", number = "12", } . Nature Structural and Molecular Biology.

Kapil Gupta, Simona V. Antonova, Matthias Haffke, Eleonora Corradini, Mykolas Mikuciunas, Teck Y. Low, Luca Signor, Robert M. van Es, Elisabeth Scheer, Harmjan R. Vos, et al.(2018). Chaperonin CCT checkpoint function in basal transcription factor TFIID assembly . Nature Structural & Molecular Biology. 25. (12). p. 1119--1127. Springer Science and Business Media {LLC}

Antonova, S.V., Haffke, M., Corradini, E., Mikuciunas, M., Low, T.Y., Signor, L., van Es, R.M., Gupta, K., Scheer, E., Vos, H.R., et al.(2018). Chaperonin CCT checkpoint function in basal transcription factor TFIID assembly . Nature Structural and Molecular Biology. 25. (12). p. 1119-1127.

Architecture of TAF11/TAF13/TBP complex suggests novel regulation properties of general transcription factor TFIID @article{9d2cc4222ff3400d8c8e2e37316fa5bb, title = "Architecture of TAF11/TAF13/TBP complex suggests novel regulation properties of general transcription factor TFIID", abstract = "General transcription factor TFIID is a key component of RNA polymerase II transcription initiation. Human TFIID is a megadalton-sized complex comprising TATA-binding protein (TBP) and 13 TBP-associated factors (TAFs). TBP binds to core promoter DNA, recognizing the TATA-box. We identified a ternary complex formed by TBP and the histone fold (HF) domain-containing TFIID subunits TAF11 and TAF13. We demonstrate that TAF11/TAF13 competes for TBP binding with TATA-box DNA, and also with the N-terminal domain of TAF1 previously implicated in TATA-box mimicry. In an integrative approach combining crystal coordinates, biochemical analyses and data from cross-linking mass-spectrometry (CLMS), we determine the architecture of the TAF11/TAF13/TBP complex, revealing TAF11/TAF13 interaction with the DNA binding surface of TBP. We identify a highly conserved C-terminal TBP-interaction domain (CTID) in TAF13, which is essential for supporting cell growth. Our results thus have implications for cellular TFIID assembly and suggest a novel regulatory state for TFIID function.", keywords = "SYNTHETIC BIOLOGY, synthetic biology", author = "Kapil Gupta and Watson, \{Aleksandra A\} and Tiago Baptista and Elisabeth Scheer and Chambers, \{Anna L\} and Christine Koehler and Juan Zou and Ima Obong-Ebong and Eaazhisai Kandiah and Arturo Temblador and Adam Round and Eric Forest and Petr Man and Christoph Bieniossek and Laue, \{Ernest D\} and Lemke, \{Edward A\} and Juri Rappsilber and Robinson, \{Carol V\} and Didier Devys and L{\`a}szl{\`o} Tora and Imre Berger", year = "2017", month = nov, day = "16", doi = "10.7554/eLife.30395", language = "English", volume = "6", journal = "eLife", issn = "2050-084X", publisher = "eLife Sciences Publications Ltd", } . eLife.

Gupta K, Watson AA, Baptista T, Scheer E, Chambers AL, Koehler C, Zou J, Obong-Ebong I, Kandiah E, Temblador A, et al.(2017). Architecture of TAF11/TAF13/TBP complex suggests novel regulation properties of general transcription factor TFIID . eLife.

Gupta, K., Watson, A.A., Baptista, T., Scheer, E., Chambers, A.L., Koehler, C., Zou, J., Obong-Ebong, I., Kandiah, E., Temblador, A., et al.(2017). Architecture of TAF11/TAF13/TBP complex suggests novel regulation properties of general transcription factor TFIID . eLife. 6.

Genetic code expansion for multiprotein complex engineering @article{2be0611e9d7540ab92cc92ea97d67290, title = "Genetic code expansion for multiprotein complex engineering", abstract = "We present a baculovirus-based protein engineering method that enables site-specific introduction of unique functionalities in a eukaryotic protein complex recombinantly produced in insect cells. We demonstrate the versatility of this efficient and robust protein production platform, 'MultiBacTAG', (i) for the fluorescent labeling of target proteins and biologics using click chemistries, (ii) for glycoengineering of antibodies, and (iii) for structure–function studies of novel eukaryotic complexes using single-molecule F{\"o}rster resonance energy transfer as well as site-specific crosslinking strategies.", keywords = "Biophysical chemistry, Chemical tools, Genetic engineering, Protein design, Proteins", author = "Christine Koehler and Sauter, \{Paul F.\} and Mirella Wawryszyn and Girona, \{Gemma Estrada\} and Kapil Gupta and Landry, \{Jonathan J M\} and Fritz, \{Markus Hsi Yang\} and Ksenija Radic and Hoffmann, \{Jan Erik\} and Chen, \{Zhuo A.\} and Juan Zou and Tan, \{Piau Siong\} and Bence Galik and Sini Junttila and Peggy Stolt-Bergner and Giancarlo Pruneri and Attila Gyenesei and Carsten Schultz and Biskup, \{Moritz Bosse\} and Hueseyin Besir and Vladimir Benes and Juri Rappsilber and Martin Jechlinger and Korbel, \{Jan O.\} and Imre Berger and Stefan Braese and Lemke, \{Edward A.\}", year = "2016", month = dec, doi = "10.1038/nmeth.4032", language = "English", volume = "13", pages = "997–1000", journal = "Nature Methods", issn = "1548-7091", publisher = "Nature Research", number = "12", } . Nature Methods.

Kapil Gupta, Christine Koehler, Paul F Sauter, Mirella Wawryszyn, Gemma Estrada Girona, Jonathan J M Landry, Markus Hsi-Yang Fritz, Ksenija Radic, Jan-Erik Hoffmann, Zhuo A Chen, et al.(2016). Genetic code expansion for multiprotein complex engineering . Nature Methods. Springer Nature

Gupta K, Sari-Ak D, Haffke M, Trowitzsch S, Berger I, Journal of molecular biology, 2016, vol. 428, no. 12, pp. 2581-2591, 2016 .

Gupta, K., Sari-Ak, D., Haffke, M., Trowitzsch, S., Berger, I.(2016). Zooming in on Transcription Preinitiation . Journal of Molecular Biology. 428. (12). p. 2581-2591.

Koehler, C., Sauter, P.F., Wawryszyn, M., Girona, G.E., Gupta, K., Landry, J.J.M., Fritz, M.H.-Y., Radic, K., Hoffmann, J.-E., Chen, Z.A., et al.(2016). Genetic code expansion for multiprotein complex engineering . Nature Methods. 13. (12). p. 997-1000.

Sari D, Gupta K, Thimiri Govinda Raj DB, Aubert A, Drncová P, Garzoni F, Fitzgerald D, Berger I, Advances in experimental medicine and biology, 2016, vol. 896, pp. 199-215, 2016 .

Kapil Gupta, Wassim Abdulrahman, Laura Radu, Frederic Garzoni, Olga Kolesnikova, Judit Osz-Papai, Imre Berger, Arnaud Poterszman(2015). The production of multiprotein complexes in insect cells using the baculovirus expression system . Methods in Molecular Biology. 1261. p. 91--114. Humana Press Inc.

Abdulrahman W, Radu L, Garzoni F, Kolesnikova O, Gupta K, Osz-Papai J, Berger I, Poterszman A, Methods in molecular biology (Clifton, N.J.), 2015, vol. 1261, pp. 91-114, 2015 .

More pieces to the puzzle @article{e131a3749ba043fab99e67e7528a785b, title = "More pieces to the puzzle: recent structural insights into class II transcription initiation", abstract = "Class II transcription initiation is a highly regulated process and requires the assembly of a pre-initiation complex (PIC) containing DNA template, RNA polymerase II (RNAPII), general transcription factors (GTFs) TFIIA, TFIIB, TFIID, TFIIE, TFIIF, TFIIH and Mediator. RNAPII, TFIID, TFIIH and Mediator are large multiprotein complexes, each containing 10 and more subunits. Altogether, the PIC is made up of about 60 polypeptides with a combined molecular weight of close to 4MDa. Recent structural studies of key PIC components have significantly advanced our understanding of transcription initiation. TFIID was shown to bind promoter DNA in a reorganized state. The architecture of a core-TFIID complex was elucidated. Crystal structures of the TATA-binding protein (TBP) bound to TBP-associated factor 1 (TAF1), RNAPII-TFIIB complexes and the Mediator head module were solved. The overall architectures of large PIC assemblies from human and yeast have been determined by electron microscopy (EM). Here we review these latest structural insights into the architecture and assembly of the PIC, which reveal exciting new mechanistic details of transcription initiation.", keywords = "Animals, Humans, Mediator Complex, Models, Molecular, Protein Conformation, RNA Polymerase II, Transcription Factors, General, Transcriptional Activation", author = "Eaazhisai Kandiah and Simon Trowitzsch and Kapil Gupta and Matthias Haffke and Imre Berger", note = "Copyright {\textcopyright} 2014 Elsevier Ltd. All rights reserved.", year = "2014", month = feb, doi = "10.1016/j.sbi.2013.12.005", language = "English", volume = "24", pages = "91--7", journal = "Current Opinion in Structural Biology", issn = "0959-440X", publisher = "Elsevier Limited", } . Current Opinion in Structural Biology.

Gupta, Kapil, Kandiah, Eaazhisai, Trowitzsch, Simon, Haffke, Matthias, Berger, Imre(2014). More pieces to the puzzle: recent structural insights into class II transcription initiation . Current opinion in structural biology. 24. p. 91-97.

Kandiah, E., Trowitzsch, S., Gupta, K., Haffke, M., Berger, I.(2014). More pieces to the puzzle: Recent structural insights into class II transcription initiation . Current Opinion in Structural Biology. 24. (1). p. 91-97.

Gene gymnastics @article{355c1b0a5cae4f21af1c4a55fa035f85, title = "Gene gymnastics: Synthetic biology for baculovirus expression vector system engineering", abstract = "Most essential activities in eukaryotic cells are catalyzed by large multiprotein assemblies containing up to ten or more interlocking subunits. The vast majority of these protein complexes are not easily accessible for high resolution studies aimed at unlocking their mechanisms, due to their low cellular abundance and high heterogeneity. Recombinant overproduction can resolve this bottleneck and baculovirus expression vector systems (BEVS) have emerged as particularly powerful tools for the provision of eukaryotic multiprotein complexes in high quality and quantity. Recently, synthetic biology approaches have begun to make their mark in improving existing BEVS reagents by de novo design of streamlined transfer plasmids and by engineering the baculovirus genome. Here we present OmniBac, comprising new custom designed reagents that further facilitate the integration of heterologous genes into the baculovirus genome for multiprotein expression. Based on comparative genome analysis and data mining, we herein present a blueprint to custom design and engineer the entire baculovirus genome for optimized production properties using a bottom-up synthetic biology approach.", keywords = "Baculoviridae, Gene Expression, Data Mining, Plasmids, synthetic biology, Genome, Viral, Chromosome Mapping, Genetic Engineering, Eukaryotic Cells, Multiprotein Complexes, Synthetic Biology, Cloning, Molecular, Genetic Vectors, Gene Transfer Techniques", author = "Vijayachandran, \{Lakshmi S\} and \{Thimiri Govinda Raj\}, \{Deepak B\} and Evelina Edelweiss and Kapil Gupta and Josef Maier and Valentin Gordeliy and Fitzgerald, \{Daniel J\} and Imre Berger", year = "2013", month = jan, day = "19", doi = "10.4161/bioe.22966", language = "English", volume = "4", pages = "279--87", journal = "Bioengineered Online", issn = "2165-5979", publisher = "Taylor \& Francis Group", number = "5", } . Bioengineered Online.

Gupta, Kapil, Vijayachandran, Lakshmi S, Thimiri Govinda Raj, Deepak B, Edelweiss, Evelina, Maier, Josef, Gordeliy, Valentin, Fitzgerald, Daniel J, Berger, Imre(2013). Gene gymnastics: Synthetic biology for baculovirus expression vector system engineering . Bioengineered. 4. (5). p. 279-287.

Vijayachandran, L.S., Thimiri Govinda Raj, D.B., Edelweiss, E., Gupta, K., Maier, J., Gordeliy, V., Fitzgerald, D.J., Berger, I.(2013). Gene gymnastics synthetic biology for baculovirus expression vector system engineering . Bioengineered. 4. (5).

Gupta, Kapil, Berger, Imre, Garzoni, Frederic, Chaillet, Maxime, Haffke, Matthias, Aubert, Alice(2013). The multiBac protein complex production platform at the EMBL . Journal of visualized experiments : JoVE.

Berger, I., Garzoni, F., Chaillet, M., Haffke, M., Gupta, K., Aubert, A.(2013). The MultiBac protein complex production platform at the EMBL . Journal of Visualized Experiments. (77).

Kapil Gupta, Imre Berger, Frederic Garzoni, Maxime Chaillet, Matthias Haffke, Alice Aubert(2013). The MultiBac protein complex production platform at the EMBL . Journal of Visualized Experiments. (77). p. e50159. MyJoVE Corporation

PREPRINT

Dora Buzas, H Adrian Bunzel, Oskar Staufer, Emily J Milodowski, Grace L Edmunds, Joshua C Bufton, Beatriz V Vidana Mateo, Sathish K N Yadav, Charlotte Fletcher, et al., Kapil Gupta (2023). Antibodies generatedin vitroandin vivoelucidate design of a thermostable ADDomer COVID-19 nasal nanoparticle vaccine .

Kapil Gupta, Alice Halliday, Anna E Long, Holly E Baum, Amy C Thomas, Kathryn L Shelley, Elizabeth Oliver, Ore Francis, Maia Kavanagh Williamson, Natalie di Bartolo, et al.(2022). Development and evaluation of low-volume tests to detect and characterise antibodies to SARS-CoV-2 . Cold Spring Harbor Laboratory

OTHER

Antibodies generated<i>in vitro</i>and<i>in vivo</i>elucidate design of a thermostable ADDomer COVID-19 nasal nanoparticle vaccine @misc{b766b0365dcb40d1ac6d40aeee98b362, title = "Antibodies generatedin vitroandin vivoelucidate design of a thermostable ADDomer COVID-19 nasal nanoparticle vaccine", abstract = "COVID-19 continues to damage populations, communities and economies worldwide. Vaccines have reduced COVID-19-related hospitalisations and deaths, primarily in developed countries. Persisting infection rates, and highly transmissible SARS-CoV-2 Variants of Concern (VOCs) causing repeat and breakthrough infections, underscore the ongoing need for new treatments to achieve a global solution. Based on ADDomer, a self-assembling protein nanoparticle scaffold, we created ADDoCoV, a thermostable COVID-19 candidate vaccine displaying multiple copies of a SARS-CoV-2 receptor binding motif (RBM)-derived epitope.In vitrogenerated neutralising nanobodies combined with molecular dynamics (MD) simulations and electron cryo-microscopy (cryo-EM) established authenticity and accessibility of the epitopes displayed. A Gigabody comprising multimerized nanobodies prevented SARS-CoV-2 virion attachment with picomolar EC50. Antibodies generated by immunising mice cross-reacted with VOCs including Delta and Omicron. Our study elucidates nasal administration of ADDomer-based nanoparticles for active and passive immunisation against SARS-CoV-2 and provides a blueprint for designing nanoparticle reagents to combat respiratory viral infections.", author = "Dora Buzas and Bunzel, \{Hans Adrian\} and Oskar Staufer and Emily Milodowski and Grace Edmonds and Joshua Bufton and Beatriz Vidana and sathish Yadav and Kapil Gupta and Charlotte Fletcher and Williamson, \{Maia Kavanagh\} and Alexandra Harrison and Ufuk Borucu and Julien Capin and Ore Francis and Georgia Balchin and Sophie Hall and Vega, \{Mirella Vivoli\} and DURBESSON Fabien and Renaud Vincentelli and Joe Roe and Linda Wooldridge and Rachel Burt and Ross Anderson and Adrian Mulholland and Jonathan Hare and Mick Bailey and Andrew Davidson and Adam Finn and David Morgan and Jamie Mann and Joachim Spatz and Frederic Garzoni and Berger-Schaffitzel, \{Christiane Helene\} and Imre Berger and Group, \{Bristol UNCOVER\}", year = "2023", month = mar, day = "17", doi = "10.1101/2023.03.17.533092", language = "Undefined/Unknown", type = "Other", } .

Kapil Gupta, Christine Toelzer, Maia Kavanagh Williamson, Deborah K. Shoemark, A. Sofia F. Oliveira, David A. Matthews, Abdulaziz Almuqrin, Oskar Staufer, Sathish K.N. Yadav, Ufuk Borucu, et al.(2021). Structural basis for cell-type specific evolution of viral fitness by SARS-CoV-2 . Cold Spring Harbor Laboratory

Frontispiz @article{ad37e5781737430bb3a7fa3c09a7f48e, title = "Frontispiz: Molecular Simulations suggest Vitamins, Retinoids and Steroids as Ligands of the Free Fatty Acid Pocket of the SARS-CoV-2 Spike Protein", author = "Shoemark, \{Deborah K\} and Colenso, \{Charlotte K\} and Christine Toelzer and Kapil Gupta and Sessions, \{Richard B\} and Davidson, \{Andrew D\} and Imre Berger and Christiane Schaffitzel and James Spencer and Mulholland, \{Adrian J\}", year = "2021", month = mar, day = "22", doi = "10.1002/ange.202181362", language = "English", volume = "133", journal = "Angewandte Chemie", issn = "0044-8249", publisher = "John Wiley \& Sons, Ltd.", number = "13", } . Angewandte Chemie.

Kapil Gupta, Christine Toelzer, Sathish K.N. Yadav, Ufuk Borucu, Frederic Garzoni, Oskar Staufer, Julien Capin, Joachim Spatz, Daniel Fitzgerald, Imre Berger, et al.(2020). Unexpected free fatty acid binding pocket in the cryo-EM structure of SARS-CoV-2 spike protein . Cold Spring Harbor Laboratory

Kapil Gupta, Changwei Yu, Nevena Cvetesic, Vincent Hisler, Tao Ye, Emese Gazdag, Luc Negroni, Petra Hajkova, Imre Berger, Boris Lenhard, et al.(2020). TBPL2/TFIIA complex establishes the maternal transcriptome by an oocyte-specific promoter usage . Cold Spring Harbor Laboratory

TFIID Enables RNA Polymerase II Promoter-Proximal Pausing @article{b68e339f8df9451eb9e56baccbab3b5c, title = "TFIID Enables RNA Polymerase II Promoter-Proximal Pausing", abstract = "RNA polymerase II (RNAPII) transcription is governed by the pre-initiation complex (PIC), which contains TFIIA, TFIIB, TFIID, TFIIE, TFIIF, TFIIH, RNAPII, and Mediator. After initiation, RNAPII enzymes pause after transcribing less than 100 bases; precisely how RNAPII pausing is enforced and regulated remains unclear. To address specific mechanistic questions, we reconstituted human RNAPII promoter-proximal pausing in vitro, entirely with purified factors (no extracts). As expected, NELF and DSIF increased pausing, and P-TEFb promoted pause release. Unexpectedly, the PIC alone was sufficient to reconstitute pausing, suggesting RNAPII pausing is an inherent PIC function. In agreement, pausing was lost upon replacement of the TFIID complex with TATA-binding protein (TBP), and PRO-seq experiments revealed widespread disruption of RNAPII pausing upon acute depletion (t = 60 min) of TFIID subunits in human or Drosophila cells. These results establish a TFIID requirement for RNAPII pausing and suggest pause regulatory factors may function directly or indirectly through TFIID", keywords = "synthetic biology, DSIF, TRIM-Away, TAF1, RNA polymerase II, NELF, P-TEFb, TFIID, PRO-seq, TBP, pausing", author = "Fant, \{Charli B\} and Levandowski, \{Cecilia B\} and Kapil Gupta and Maas, \{Zachary L\} and John Moir and Rubin, \{Jonathan D\} and Andrew Sawyer and Esbin, \{Meagan N\} and Rimel, \{Jenna K\} and Olivia Luyties and Marr, \{Michael T\} and Imre Berger and Dowell, \{Robin D\} and Taatjes, \{Dylan J\}", year = "2020", month = may, day = "21", doi = "10.1016/j.molcel.2020.03.008", language = "English", volume = "78", pages = "785--793.e8", journal = "Molecular Cell", issn = "1097-2765", publisher = "Cell Press", number = "4", } . Molecular Cell.

JOURNAL ISSUE

The SARS-CoV-2 Spike protein alters human cardiac pericyte function and interaction with endothelial cells through a non-infective mechanism involving activation of CD147 receptor signalling. @article{f143d405c5bd4a498b90406734348544, title = "The SARS-CoV-2 Spike protein alters human cardiac pericyte function and interaction with endothelial cells through a non-infective mechanism involving activation of CD147 receptor signalling.", abstract = "Background: Human cardiac pericytes (PC) were proposed as the main cellular target for SARS-CoV-2 in the heart due to high transcriptional levels of the angiotensin-converting enzyme 2 (ACE2) receptor. Emerging reports indicate CD147/Basigin (BSG), highly expressed in endothelial cells (EC), is an alternative SARS-CoV-2 receptor. To date, the mechanism by which the virus infects and disrupts the heart vascular cells was not identified yet. Moreover, cleaved Spike (S) protein molecules could be released into the bloodstream from the leaking pulmonary epithelial-endothelial barrier in patients with severe COVID-19, opening to the possibility of non-infective diseases in organs distant from the primary site of infection. Purposes: (1) to confirm that human primary cardiac PC express ACE2 and CD147; (2) to verify if PC are permissible to SARS-CoV-2 infection; (3) to investigate if the recombinant SARS-CoV-2 S protein alone, without the other viral elements, can trigger molecular signalling and induce functional alterations in PC; (4) to explore which viral receptor is responsible for the observed events. Methods and results: Cardiac PC express both the ACE2 and CD147 receptors at mRNA and protein level. Incubation of PC for up to 5 days with SARS-CoV-2 expressing the green fluorescent protein (GFP) did not show any evidence of cell infection or viral replication. Next, we exposed the PC to the recombinant S protein (5.8 nM) and confirmed that the protein engaged with cellular receptors (western blot analysis of S protein in treated and control PC). Incubation with the S protein increased PC migration (wound closure assay, P<0.01 vs ctrl) and reduced the formation of tubular structures between PC and EC in a Matrigel assay (P<0.01 vs ctrl). Moreover, the S protein promoted the production of pro-inflammatory factors typical of the cytokine storm in PC (ELISA measurement of MCP1, IL-6, IL-1β, TNFα, P<0.05 vs ctrl), and induced the secretion of pro-apoptotic factors responsible for EC death (Caspase 3/7 assay, P<0.05 vs ctrl). Signalling studies revealed that the S protein triggers the phosphorylation/activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) through the CD147 receptor, but not ACE2, in cardiac PC. The neutralization of CD147, using a blocking antibody, prevented ERK1/2 activation in PC, and was reflected into a partial rescue of the cell functional behaviour (migration and pro-angiogenic capacity). In contrast, blockage of CD147 failed to prevent the pro-inflammatory response in PC. Conclusions: We propose the novel hypothesis that COVID-19 associated heart{\textquoteright}s microvascular dysfunction is prompted by circulating S protein molecules rather than by the direct coronavirus infection of PC. Besides, we propose CD147, and not ACE2, as the leading receptor mediating S protein signalling in cardiac PC.", keywords = "covid-19, pericyte, microvascular disease, cardiac pericytes, Endothelial dysfunction, CD147, SPIKE", author = "Elisa Avolio and Michele Carrabba and Rachel Milligan and \{Kavanagh Williamson\}, Maia and Kapil Gupta and Monica Gamez and Rebecca Foster and Imre Berger and Massimo Caputo and Davidson, \{Andrew D\} and Hill, \{Darryl J\} and Madeddu, \{Paolo R\}", year = "2021", month = oct, day = "14", doi = "10.1093/eurheartj/ehab724.3383", language = "English", volume = "42", journal = "European Heart Journal", issn = "0195-668X", publisher = "Oxford University Press", number = "1", note = "ESC Congress 2021 ; Conference date: 27-08-2021 Through 30-08-2021", url = "https://www.escardio.org/Congresses-\&-Events/ESC-Congress", } . European Heart Journal.

Zooming in on Transcription Preinitiation @article{e91a1a2fdc764aab8ac2b67caefb4458, title = "Zooming in on Transcription Preinitiation", abstract = "Class II gene transcription commences with the assembly of the Preinitiation Complex (PIC) from a plethora of proteins and protein assemblies in the nucleus, including the General Transcription Factors (GTFs), RNA polymerase II (RNA pol II), co-activators, co-repressors and more. TFIID, a megadalton-sized multiprotein complex comprising 20 subunits is among the first GTFs to bind the core promoter. TFIID assists in nucleating PIC formation, completed by binding of further factors in a highly regulated step-wise fashion. Recent results indicate that TFIID itself is built from distinct pre-formed submodules, which reside in the nucleus but also in the cytosol of cells. Here we highlight recent insights in transcription factor assembly and the regulation of transcription preinitiation.", keywords = "Preinitiation complex (PIC), General Transcription Factor TFIID, Transcription Initiation, RNA polymerase II (RNA pol II), Multiprotein Complex, Structural Biology", author = "Kapil Gupta and Duygu Sari-Ak and Matthias Haffke and Simon Trowitzsch and Imre Berger", note = "Special Issue: Macromolecular Complexes in Transcription and Co-transcriptional RNA Processing", year = "2016", month = jun, day = "19", doi = "10.1016/j.jmb.2016.04.003", language = "English", volume = "428", pages = "2581--2591", journal = "Journal of Molecular Biology", issn = "0022-2836", publisher = "Academic Press", number = "12", } . Journal of Molecular Biology.

BOOK CHAPTER

Kapil Gupta, Imre Berger(2016). The MultiBac Baculovirus/Insect Cell Expression Vector System for Producing Complex Protein Biologics . Advanced Technologies for Protein Complex Production and Characterization. IV. p. 199--215. Springer

Sari, D., Gupta, K., Raj, D.B.T.G., Aubert, A., Drncov&#225;, P., Garzoni, F., Fitzgerald, D., Berger, I.(2016). The multibac baculovirus/insect cell expression vector system for producing complex protein biologics . Advances in Experimental Medicine and Biology. 896. p. 199-215.

Abdulrahman, W., Radu, L., Garzoni, F., Kolesnikova, O., Gupta, K., Osz-Papai, J., Berger, I., Poterszman, A.(2014). The production of multiprotein complexes in insect cells using the baculovirus expression system . Structural Proteomics: High-Throughput Methods: Second Edition. p. 91-114.

BOOK

Abdulrahman, W., Radu, L., Garzoni, F., Kolesnikova, O., Gupta, K., Osz-Papai, J., Berger, I., Poterszman, A.(2015). The production of multiprotein complexes in insect cells using the baculovirus expression system . Methods in Molecular Biology. 1261. p. 91-114.