A5049704234- Protein purification and stability
- Viral Infectious Diseases and Gene Expression in Insects
- Monoclonal and Polyclonal Antibodies Research
- Microfluidic and Bio-sensing Technologies
- Innovative Microfluidic and Catalytic Techniques Innovation
- Microfluidic and Capillary Electrophoresis Applications
- 3D Printing in Biomedical Research
- Analytical Chemistry and Chromatography
- SARS-CoV-2 detection and testing
- SARS-CoV-2 and COVID-19 Research
- Additive Manufacturing and 3D Printing Technologies
- Crystallization and Solubility Studies
- Transgenic Plants and Applications
- Chromatography in Natural Products
- Process Optimization and Integration
- Fault Detection and Control Systems
- Biomedical and Engineering Education
- Innovation Policy and R&D
- Glycosylation and Glycoproteins Research
- Microplastics and Plastic Pollution
- Biosensors and Analytical Detection
- Viral gastroenteritis research and epidemiology
- Scientific Computing and Data Management
- Protein Interaction Studies and Fluorescence Analysis
- Private Equity and Venture Capital
BOKU University
2014-2024
Austrian Centre of Industrial Biotechnology (Austria)
2020-2024
John Wiley & Sons (United States)
2019
Hudson Institute
2019
The use of nanomaterials in bioapplications demands a detailed understanding protein-nanoparticle interactions. Proteins can undergo conformational changes while adsorbing onto nanoparticles, but studies on the impact particle size are scarce. We have shown that happening upon adsorption myoglobin and BSA dependent nanoparticle they to. Out eight initially investigated model proteins, two (BSA myoglobin) showed changes, both cases this change was nanoparticle. Nanoparticle sizes ranged from...
The main objectives of bioprocesses are to reliably deliver drugs in a relatively short time frame with high quality within tight regulatory framework. Bioprocesses highly complex, the level automation is moderate, and there constant pressure improve efficiency costs. In addition, climate change resource scarcity mandate reduction environmental footprint production facilities. biopharmaceutical industry, two extreme scenarios applied: fully disposable factory characteristics full flexibility...
Currently, the golden standard for purification of recombinant humanized antibodies (rhAbs) from CHO cell culture is protein A chromatography. However, due to increasing rhAbs titers alternative methods have come into focus. new strategy human supernatant based on cold ethanol precipitation (CEP) and CaCl2 has been developed. This method precipitation, process used other components blood plasma. We proof applicability developed four different resulting in similar yield purity as a...
Continuous precipitation is a new unit operation for the continuous capture of antibodies. The step based on with PEG6000 and Zn++ in tubular reactor integrated two-stage tangential flow filtration unit. precipitate cannot be separated centrifugation, because highly compressed sediment results poor resolubilization. We developed microfiltration method, where part concentrated retentate first stage was directly fed to second stage, together wash buffer. Thus, washed process. obtained 97%...
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.
Downstream processing (DSP) of large bionanoparticles is still a challenge. The present study aims to systematically compare some the most commonly used DSP strategies for capture and purification enveloped viruses virus-like particles (eVLPs) by using same staring material analytical tools. As model, Human Immunodeficiency Virus-1 (HIV-1) gag VLPs produced in CHO cells were used. Four different tested. An anion-exchange monolith membrane adsorber, direct eVLPs, polymer-grafted resin...
The capture of recombinant antibodies from cell culture broth is the first critical step downstream processing. We were able to develop a precipitation-based method for and purification monoclonal based on divalent cations, namely ZnCl2. Traditional precipitation processes have deal with high dilution factors especially resolubilization higher viscosity due use PEG as or co-precipitation agent. By crosslinking nature cations without PEG, we kept supernatant very low. This beneficial...
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....
Integrated continuous production technology is of great interest in biopharmaceutical industry. Efficient, flexible and cost effective methods for cell removal have to be developed, before a fully integrated product train can realized. The paper describes the development testing such an disposable set-up separation by flocculation combined with depth filtration.Screening multiple agents, filters, conditions demonstrated that best performance was obtained 0.0375% polydiallyldimethylammonium...
Precipitation has gained interest as alternative to the costly protein A chromatography for monoclonal antibody purification. Traditional precipitation processes are based on direct addition of precipitant in a single dose, with limited control co-precipitation impurities and not considering batch-to-batch variations. We propose gradual dosage polyethylene glycol prevent resulting floc size. used focused beam reflectance measurement demonstrate that PEG6000 time final concentration...
Staphylococcal protein A affinity chromatography is a well-established platform for purification of clinical-grade antibodies. The wild type ligand has been mutated to improve caustic stability, elution behavior, and/or increase binding capacity. Several modified ligands are nowadays commercially available, one them being the thermosensitive medium Byzen Pro from Nomadic Bioscience Co., Ltd. According manufacturer, ability release IgG upon change in temperature. It based on mutant which...
The economic benefits of continuous PEG precipitation antibodies compared to capture by protein A affinity chromatography have been always lowered the excess amount material needed for process. is added as a concentrated stock solution and therefore liquid handling increased during this step. To fully exploit precipitation, precipitant must be in solid form. We used an in-line feeding device with screw conveyor delivery system addition PEG6000 powder was connected tubular reactor where...
For scale up and efficient production of protein loaded nanoparticles continuous separation by size exclusion chromatography in simulated moving bed (SMB) mode helps do reduce unbound concentration increase yields for perfectly covered particles. Silica were with an excess beta casein or bovine serum albumin (BSA) the particles purified using Sephacryl300 as stationary phase a four zone SMB. We determined our working points SMB from batch separations triangle theory described Mazzotti et al....
3D printing represents a democratization of manufacturing processes, and inexpensive printed parts for cell culture have been tested as replacements single-use plastics currently unavailable due to worldwide supply chain issues. In addition, such distributed laboratory materials helps remote areas developing countries with limited resources. HEK293 cells were used test shake flasks applications their ease manufacture. Recorded growth curves showed that renewable biodegradable poly(lactic...
3D printing has become widespread for the manufacture of parts in various industries and enabled radically new designs. This trend not spread to bioprocess development yet, due a lack material suitable current workflow, including sterilization by autoclaving. work demonstrates that commercially available heat temperature stable poly-lactic acid (PLA) can be used easily novel bioreactor vessels with included features like harvest tubes printed spargers. Temperature responsiveness was tested...
Abstract The high‐throughput analysis of antibodies from processes can be enhanced when the linear range is expanded and sample preparation kept to a minimum. We developed fast chromatography method based on hexameric variant staphylococcal protein A immobilized Toyopearl matrix, TSK 5 PW using two wavelengths. protocol with min runtime single‐wavelength detection at 280 nm yielded an upper limit quantification 2.10 mg/mL lower 0.06 mg/mL. optimized 2 two‐wavelength 300 allowed us span valid...
The trend in the biopharmaceutical industry is changing from batch process to continuous process. For biomanufacturing, traceability of material required by regulatory authorities. recent ICH draft guideline Q13 on manufacturing drug substances and products requests an "understanding dynamics as a function input attributes (e.g., potency, flow properties), conditions mass rates) … One common approach characterization residence time distribution (RTD) for individual unit operations integrated...
Abstract Protein and in particular antibody precipitation by PEG is a cost‐effective alternative for the first capture step. The 3D structure of precipitates has large impact on process parameters recovery dissolution, but current technologies determination precipitate structures are either very time consuming (cryo‐TEM) or only generate an average fractal dimension (light scattering). We developed light microscopy based reconstruction individual particles with resolution 0.1–0.2 µm used...
Manufacturing of biopharmaceuticals requires high quality water. Overall conditions for production clean water vary across the globe due to availability and quality. We combined economic ecological modeling assess energy consumption from tap waste. defined a metric, WAter Related Impact ENergy (WARIEN) directly correlate amount CO2 emitted per kg biopharmaceutical included membrane- distillation-based methods production. Three scenarios antibodies with 500 L fed-batch fermentation stainless...
Abstract BACKGROUND A major improvement in biomanufacturing will arise with the transition from batch processing to continuous processing. Two important challenges address this change are definition and ability trace raw material through process. RESULTS We used an established simulation of a process train compare conventional based on fixed time new method greatest common divisor (GCD) period unit operations. successfully demonstrated that, by using concept GCD, we have constant periodic...