A5054323277- SARS-CoV-2 and COVID-19 Research
- SARS-CoV-2 detection and testing
- Influenza Virus Research Studies
- Viral Infectious Diseases and Gene Expression in Insects
- Respiratory viral infections research
- Transgenic Plants and Applications
- Protein purification and stability
- Viral gastroenteritis research and epidemiology
- Bacteriophages and microbial interactions
- COVID-19 Clinical Research Studies
- Advanced Fluorescence Microscopy Techniques
- Insect Resistance and Genetics
- vaccines and immunoinformatics approaches
- Animal Virus Infections Studies
- Advanced biosensing and bioanalysis techniques
- RNA Interference and Gene Delivery
- Advanced Electron Microscopy Techniques and Applications
- Monoclonal and Polyclonal Antibodies Research
- Lipid Membrane Structure and Behavior
- Immune Cell Function and Interaction
- Molecular Junctions and Nanostructures
- HIV Research and Treatment
- Viral Infections and Vectors
- Biosensors and Analytical Detection
- Photosynthetic Processes and Mechanisms
BOKU University
2012-2024
Institute of Molecular Biotechnology
2020-2024
Austrian Centre of Industrial Biotechnology (Austria)
2019
University of Lisbon
2008
Article23 August 2021Open Access Transparent process Identification of lectin receptors for conserved SARS-CoV-2 glycosylation sites David Hoffmann IMBA, Institute Molecular Biotechnology the Austrian Academy Sciences, Vienna, Austria These authors contributed equally to this work Search more papers by author Stefan Mereiter orcid.org/0000-0002-4832-3090 Yoo Jin Oh orcid.org/0000-0002-9636-3329 Biophysics, Johannes Kepler University Linz, Vanessa Monteil orcid.org/0000-0002-2652-5695...
Enveloped virus-like particles (VLPs) are increasingly used as vaccines and immunotherapeutics. Frequently, very time consuming density gradient centrifugation techniques for purification of VLPs. However, the progress towards optimized large-scale VLP production increased demand fast, cost efficient scale able processes. We developed a chromatographic procedure HIV-1 gag VLPs produced in CHO cells. The clarified filtered cell culture supernatant was directly processed on an anion-exchange...
The rapid quantification of enveloped virus-like particles (VLPs) requires orthogonal methods to obtain reliable results. Three methods—nanoparticle tracking analysis (NTA), size-exclusion HPLC (SE-HPLC) with UV detection, and detection multi-angle light scattering (MALS)—for VLPs have been compared, the lower upper limits evaluated. NTA directly counts VLPs, a particle number is obtained limit (LLOD) 1.7 × 107 part/mL (LLOQ) 3.4 108 part/mL. SE-HPLC was calibrated standards characterized by...
Nicotiana benthamiana is used worldwide as production host for recombinant proteins. Many proteins such monoclonal antibodies, growth factors or viral antigens require posttranslational modifications like glycosylation their function. Here, we transiently expressed different variants of the glycosylated receptor binding domain (RBD) from SARS-CoV-2 spike protein in N. benthamiana. We characterized impact variations RBD-length and on expression, yield functionality. found that a truncated RBD...
Abstract Recent waves of COVID-19 correlate with the emergence Delta and Omicron variant. We report that Spike trimer acts as a highly dynamic molecular caliper, thereby forming up to three tight bonds through its RBDs ACE2 expressed on cell surface. The both (B.1.1.529) Variant enhance markedly prolong viral attachment host receptor ACE2, opposed early Wuhan-1 isolate. shows rapid binding all different molecules considerably increased bond lifetime when compared reference strain,...
"Pan-coronavirus" antivirals targeting conserved viral components can be designed. Here, we show that the rationally engineered H84T-banana lectin (H84T-BanLec), which specifically recognizes high mannose found on proteins but seldom healthy human cells, potently inhibits Middle East respiratory syndrome coronavirus (MERS-CoV), severe acute 2 (SARS-CoV-2) (including Omicron), and other human-pathogenic coronaviruses at nanomolar concentrations. H84T-BanLec protects against MERS-CoV...
Chimeric virus-like particles formed by the co-expression of HIV-1 gag capsid protein and surface proteins from other viruses can be used as a platform for production wide range viral vaccines. These rapidly produced in an insect cell culture using baculovirus expression vector system. However, variety different process-related impurities such host proteins, double stranded DNA, chromatin, structurally similar bionanoparticles like extracellular vesicles are present harvested supernatant...
Purpose of work: A comparative analysis new and established insect cell lines, in regard to process relevant parameters, provide data that can be exploited for designing more robust effective protein production processes. The baculovirus-insect expression system has been efficiently used the heterologous proteins. Three different lines Tnao38, High Five Sf9 were compared terms virus susceptibility, baculovirus product yield an intra-cellularly (YFP) extra-cellularly (influenza...
Human infections with a novel influenza A H7N9 subtype virus were reported in China recently. The caused severe disease high mortality rates and it raised concerns over its pandemic potential. Here, we assessed the mouse model protective efficacy of single immunisations low vaccine doses insect cell-derived H7 virus-like particles, consisting hemagglutinin matrix protein. Vaccinated mice fully protected survived stringent lethal challenge (100 mLD50) H7N9, even after single, unadjuvanted,...
Antibody tests are essential tools to investigate humoral immunity following SARS-CoV-2 infection or vaccination. While first-generation antibody have primarily provided qualitative results, accurate seroprevalence studies and tracking of levels over time require highly specific, sensitive quantitative test setups.
Vaccination against influenza virus can reduce the risk of by 40% to 60%, they rely on production neutralizing antibodies specific hemagglutinin (HA) ignoring neuraminidase (NA) as an important surface target. with standardized NA concentration may offer broader and longer-lasting protection infection. In this regard, we aimed compare potency a displayed VLP soluble NA. The baculovirus expression system (BEVS) novel virus-free Tnms42 insect cell line were used express N2 gag-based VLPs. To...
Abstract Background FAD dependent glucose dehydrogenase (GDH) currently raises enormous interest in the field of biosensors. Due to its superior properties such as high turnover rate, substrate specificity and oxygen independence, GDH makes way into biosensing. The recently discovered from ascomycete Glomerella cingulata is a novel candidate for an electrochemical application, but also study plant-pathogen interaction family wide-spread, crop destroying fungi. Heterologous expression...
The plant-pathogenic fungus Glomerella cingulata (anamorph Colletotrichum gloeosporoides ) secretes high levels of an FAD-dependent glucose dehydrogenase (GDH) when grown on tomato juice-supplemented media. To elucidate its molecular and catalytic properties, GDH was produced in submerged culture. highest volumetric activity obtained shaking flasks after 6 days cultivation (3400 U l −1 , 4.2 % total extracellular protein). is a monomeric protein with isoelectric point 5.6. masses the...
The aim of this study was the development a scalable production process for high titer (108 pfu/mL and above) recombinant baculovirus stocks with low cell line-derived impurities virus-like particles (VLP). To achieve this, we developed density (HCD) culture footprint proliferation, compared different infection strategies at multiplicity (MOI) 0.05 0.005, validated generally applicable harvest criteria viability ≤ 80%. We also investigated online measurable parameters to observe production....
The downstream processing of enveloped virus-like particles is very challenging because the biophysical and structural similarity between correctly assembled contaminating vesicular present in feedstock. We used hydroxyl-functionalized polymethacrylate monoliths, providing hydrophobic electrostatic binding contributions, for purification HIV-1 gag particles. clarified culture supernatant was conditioned with ammonium sulfate after membrane filtration loaded onto a 1 mL monolith. capacity 2 ×...
Significant genetic variability in the head region of influenza A hemagglutinin, main target current vaccines, makes it challenging to develop a long-lived seasonal prophylaxis. Vaccines based on conserved hemagglutinin stalk domain might provide broader cross-reactive immunity. However, this is immunosubdominant region. Peptide-based vaccines have gained much interest as they allow immune system focus relevant but less immunogenic epitopes. We developed novel hemagglutinin-based display...
The receptor binding domain (RBD) of the SARS-CoV-2 spike (S)-protein is a prime target virus-neutralizing antibodies present in convalescent sera COVID-19 patients and thus considered key antigen for immunosurveillance studies vaccine development. Although recombinant expression RBD has been achieved several eukaryotic systems, mammalian cells have proven particularly useful. authors aimed to optimize produced HEK293-6E towards stable homogeneous preparation addressed its O-glycosylation as...
The receptor binding domain (RBD) of the SARS-CoV-2 spike protein plays a key role in virus-host cell interaction, and viral infection. RBD is major target for neutralizing antibodies, whilst recombinant commonly used as an antigen serological assays. Such assays are essential tools to gain control over pandemic detect extent durability immune response infected or vaccinated populations. Transient expression plants can contribute fast production antigens, which required by industry high...
Clinical and historical data underscore the ability of influenza viruses to ally with Staphylococcus aureus predispose host for secondary bacterial pneumonia, which is a leading cause influenza-associated mortality. This fundamental because no vaccine S. available number antibiotic-resistant strains alarmingly rising. Hence, this leaves vaccination only strategy prevent postinfluenza staphylococcal infections. In present work, we assessed off-target effects Tnms42 insect cell-expressed...
The nucleocapsid protein (NP) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for several steps the viral life cycle, and abundantly expressed during infection, making it an ideal diagnostic target protein. This has a strong tendency dimerization interaction with nucleic acids. For first time, high titers NP were in E. coli CASPON tag, using growth-decoupled expression system. Purification was accomplished by nuclease treatment cell homogenate sequence downstream...
Current influenza virus vaccines poorly display key neuraminidase (NA) epitopes and do not robustly induce NA-reactive antibodies; instead, they focus on the induction of hemagglutinin (HA)-reactive antibodies. Next-generation should be optimized in order to activate B cells a broadly cross-reactive protective antibody response. We aimed at enhancing immunogenicity NA by two strategies: (i) modifying HA:NA ratio vaccine preparation (ii) exposing lateral surface or beneath head extending...