A5039371462- Viral Infectious Diseases and Gene Expression in Insects
- Protein purification and stability
- Monoclonal and Polyclonal Antibodies Research
- Bacterial Genetics and Biotechnology
- Bacteriophages and microbial interactions
- RNA and protein synthesis mechanisms
- Microbial Metabolic Engineering and Bioproduction
- Enzyme Production and Characterization
- CRISPR and Genetic Engineering
- SARS-CoV-2 and COVID-19 Research
- Enzyme Catalysis and Immobilization
- Innovative Microfluidic and Catalytic Techniques Innovation
- Enzyme Structure and Function
- Cancer Research and Treatments
- SARS-CoV-2 detection and testing
- Biofuel production and bioconversion
- Protein Structure and Dynamics
- Biosensors and Analytical Detection
- Glycosylation and Glycoproteins Research
- Peptidase Inhibition and Analysis
- Microplastics and Plastic Pollution
- Fault Detection and Control Systems
- thermodynamics and calorimetric analyses
- Fluid Dynamics and Mixing
- Process Optimization and Integration
Austrian Centre of Industrial Biotechnology (Austria)
2013-2024
BOKU University
2014-2024
Boehringer Ingelheim (Australia)
2022
Boehringer Ingelheim (Germany)
2019-2020
Boehringer Ingelheim (Austria)
2019-2020
Institute for Social Anthropology
2009
University of Agricultural Sciences, Dharwad
1999
Abstract Background In the biopharmaceutical industry, Escherichia coli ( E. ) strains are among most frequently used bacterial hosts for producing recombinant proteins because they allow a simple process set-up and Food Drug Administration (FDA)-approved human applications. Widespread use of in biotechnology has led to development many different strains, selecting an ideal host produce specific protein interest is important step developing production process. B K–12 employed large-scale...
The phage-derived T7 RNA polymerase is the most prominent orthogonal transcriptions system used in field of synthetic biology. However, gene expression driven by prone to read-through transcription due contextuality terminator. native terminator has a termination efficiency approximately 80% and therefore provides insufficient insulation unit. By using combination signal with two well-known transcriptional terminators (rrnBT1 T7), we have been able increase 99%. To characterize putative...
ABSTRACT Plasmid-based Escherichia coli BL21(DE3) expression systems are extensively used for the production of recombinant proteins. However, combination a high gene dosage with strong promoters exerts extremely stressful conditions on producing cells, resulting in multitude protective reactions and malfunctions host cell impact yield quality product. Here, we provide in-depth characterization plasmid-based perturbations protein production. A plasmid-free T7 system single copy interest...
Modulating resource allocation in bacteria to redirect metabolic building blocks the formation of recombinant proteins rather than biomass remains a grand challenge biotechnology. Here, we present novel approach for improved protein production (RPP) using Escherichia coli (E. coli) by decoupling synthesis from cell growth. We show that division and host mRNA transcription can be successfully inhibited coexpression bacteriophage-derived E. RNA polymerase (RNAP) inhibitor peptide genes...
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.
Tremendous advancements in cell and protein engineering methodologies bioinformatics have led to a vast increase bacterial production clones recombinant variants be screened evaluated. Consequently, an urgent need exists for efficient high-throughput (HTP) screening approaches improve the efficiency early process development as basis speed-up all subsequent steps course of design engineering. In this study, we selected BioLector micro-bioreactor (µ-bioreactor) system HTP cultivation platform...
Fusion protein technologies to facilitate soluble expression, detection, or subsequent affinity purification in
Abstract Background Escherichia coli is one of the most commonly used host organisms for production biopharmaceuticals, as it allows cost-efficient and fast recombinant protein expression. However, challenging proteins are often produced with low titres or inclusion bodies, manufacturing process needs to be developed individually each protein. Recently, we CASPON TM technology, a generic fusion tag-based platform high-titer soluble expression including standardized downstream processing...
Abstract Background Recombinant peptide production in Escherichia coli provides a sustainable alternative to environmentally harmful and size-limited chemical synthesis. However, in-vivo of disulfide-bonded peptides at high yields remains challenging, due degradation by host proteases/peptidases the necessity translocation into periplasmic space for disulfide bond formation. Results In this study, we established an expression system efficient soluble periplasm E. . We chose model with...
Recombinant protein production processes in Escherichia coli are usually operated fed-batch mode; therefore, the elaboration of a cultivation protocol microtiter plates that allows for screening under like conditions is particularly appealing. A highly reproducible plate E. micro-bioreactor system with advanced online monitoring capabilities was developed. synthetic enzymatic glucose release medium employed to provide carbon limited growth without external substrate feed and required buffer...
Abstract The main goal of this work was to develop a strategy that enables tuning recombinant gene expression relative the metabolic capacity host cell synthesis machinery. In past, strong systems have been developed in order maximize expression. However, these exert an extremely high burden onto cell, which may even lead death. Hence, period is significantly reduced, and therefore, maximal yield cannot be attained. To extend production phase achieve optimal yields, adjustment by modulation...
Product quality assurance strategies in production of biopharmaceuticals currently undergo a transformation from empirical "quality by testing" to rational, knowledge-based design" approaches. The major challenges this context are the fragmentary understanding bioprocesses and severely limited real-time access process variables related product quantity. Data driven modeling combination with model predictive control concepts represent potential solution these problems. selection statistical...
Caspase-2 is the most specific protease of all caspases and therefore highly suitable as tag removal enzyme creating an authentic N-terminus overexpressed tagged proteins interest. The wild type human caspase-2 a dimer heterodimers generated by autocatalytic processing which required for its enzymatic activity. We designed circularly permuted (cpCasp2) to overcome drawback complex recombinant expression, purification activation, cpCasp2 was constitutively active expressed single chain...
Fusion protein technologies improve the expression and purification of recombinant proteins, but removal tags involved requires specific proteases. The circularly permuted caspase-2 (cpCasp2) with its cleavage site, efficiently generates untagged protein. While cpCasp2 is possible before all 20 proteinogenic amino acids, valine, leucine, isoleucine, aspartate glutamate suffers from slow, proline extremely turnover. To make platform fusion process even more general such that any an authentic...
Despite efforts to develop concepts for efficient antibody fragment (Fab) production in Escherichia coli (E. coli) and the high degree of similarity within this protein class, a generic platform technology is still not available. Indeed, feasible new Fab candidates remains challenging. In study, setup that enables direct characterization host cell response expression by utilizing genome-integrated (GI) systems established. Among multitude factors influence expression, variable domain,...
Abstract Background Glucosylglycerol (2- O -α- d -glucosyl- sn -glycerol; GG) is a natural osmolyte from bacteria and plants. It has promising applications as cosmetic food-and-feed ingredient. Due to its scarcity, GG must be prepared through dedicated synthesis, an industrial bioprocess for production been implemented. This process uses sucrose phosphorylase (SucP)-catalyzed glycosylation of glycerol sucrose, applying the isolated enzyme in immobilized form. A whole cell-based formulation...
The genome-integrated T7 expression system offers significant advantages, in terms of productivity and product quality, even when expressing the gene interest (GOI) from a single copy. Compared to plasmid-based systems, this does not incur plasmid-mediated metabolic load, it vary dosage GOI during production process. However, long-term with leads rapidly growing non-producing population, because RNA polymerase (RNAP) is prone mutations. present study aimed investigate whether two σ70...
Abstract Governance of the endogenous gene regulatory network enables navigation cells towards beneficial traits for recombinant protein production. CRISPRactivation and interference provides basis expression modulation but is primarily applied in eukaryotes. Particularly lack wide-ranging prokaryotic CRISPRa studies might be attributed to intrinsic limitations bacterial activators Cas9 proteins. While need accurate spatial orientation distancing target promoter functional, Cas9-based CRISPR...
Abstract Background Escherichia coli is a cost-effective expression system for production of antibody fragments like Fabs. Various yield improvement strategies have been applied, however, Fabs remain challenging to produce. This study aimed characterize the gene response commonly used E. strains BL21(DE3) and HMS174(DE3) periplasmic Fab using RNA sequencing (RNA-seq). Two Fabs, Fabx FTN2, fused post-translational translocation signal sequence, were produced in carbon-limited fed-batch...
Abstract In this work, we attempted to identify a method for the selective extraction of periplasmic endogenously expressed proteins, which is applicable at an industrial scale. For purpose, used expression model that allows coexpression two fluorescent each specifically targeted either cytoplasm or periplasm. We assessed number scalable lysis methods (high‐pressure homogenization, osmotic shock procedures, with ethylenediaminetetraacetic acid, and deoxycholate) ability selectively extract...
Abstract BACKGROUND Recombinant proteins produced for use as biopharmaceuticals need to harbor their native N‐terminus. A drawback in expression of recombinant fusion with an affinity fusion‐tag is that enzymatic or chemical processing required trim the artificial tag and release true protein interest. In many cases, however, this step generates incorrect RESULTS Human fibroblast growth factor 2 (FGF2) was expressed a Escherichia coli fed‐batch cultivations. The interest (POI) carried...
Abstract BACKGROUND A goal in the production of biopharmaceuticals is to replace cost‐intensive, empirical ‘quality by testing’ approach with rational, knowledge‐based design’ concepts. The major challenges this context are complexity bioprocesses and limited online access process variables related product quality. implementation advanced monitoring strategies combined chemometric‐based approaches represents a strategy overcome bottleneck. RESULTS series recombinant E. coli fed‐batch...