Biotechnological Infrastructure
Upstream process (USP) development
Multibioreactorsystem with four parallel reactors:
DASGIP® system for cell culture, Eppendorf
- Bioreactor system in a scalable benchtop format configured for cell culture and virus applications
- Parallel processing and precise control of all relevant process parameters;
- Highly suitable for process development and characterisation studies.
Lab-scale bioreactor systems:
BioFlo® 320, Eppendorf
- Lab-scale bioreactor system for cell culture and virus application for scales of up to 10 Litres;
- Single-use bioreactor options available;
- Test for upscale consistency and feed stock supply for downstream development studies.
Alternating Tangential Flow system:
XCell™ ATF 2, Repligen
- Scalable cell retention system delivering high cell concentration and process intensification during cell culture;
- Connects to any bioreactor and can be used in a variety of applications.
Downstream process (DSP) development
Preparative Chromatography:
ÄKTA Pure 25 and 150, Cytiva
- On-line detection of particles during chromatographic experiments;
- Accurate tracking of particle elution which is not possible by using only UV signals;
- Faster and more efficient DSP development and optimization.
Continuous Ultracentrifuge:
Pilot scale (Alfa Wassermann pKII) with fluid handling system (Alfa Wassermann AFH)
- Capture, concentration and primary purification of bionanoparticles directly from clarified harvest material in a single step;
- Fully automated filling and fractionation of ultracentrifuge density gradient processes.
Analytics
Field flow fractionation coupled to UV-MALS-DLS-RI detectors (Wyatt) & Analytic HPLCs (Agilent and Thermo Fisher) coupled to UV-MALS-FLD-RI detectors
- FFF: high-resolution separation of bionanoparticles based on hydrodynamic radius in analytical and semi-preparative manner;
- HPLC: SEC, AEX, HIC and affinity based analytical chromatography;
- MALS and DLS: direct determination of particle number, size and shape by multi-angle and dynamic light scattering in the same flow cell;
- RI: universal concentration measurement, determination of solvent absolute refractive index.
- FLD: fluorescence detection
Nanoparticle Tracking Analysis: NanoSight NS300 incl. Sample Assistant (Malvern Panalytical)
- Analysis of the size and concentration of particles in solution by combining the properties of both Brownian motion and light scattering;
- Particles are individually tracked, resulting in a size distribution of hydrodynamic diameters.
Microflow Cytometer (Apogee)
- Flow Cytometer tuned for extremely small particle applications including extracellular vesicles (EVs), virus and protein aggregates;
- Highest sensitivity and resolution from multiple light scattering and fluorescence detectors;
- Able to measure biological particles down to about 100nm diameter by light scatter.
Publications
Publications (USP)
Strobl, F., Ghorbanpour, S.M., Palmberger, D. et al. Evaluation of screening platforms for virus-like particle production with the baculovirus expression vector system in insect cells (2020) Scientific Reports 10, 1065
https://doi.org/10.1038/s41598-020-57761-w
Puente-Massaguer, E., Strobl, F., Grabherr, R., Striedner, G., Lecina, M., Gòdia, F., PEI-Mediated Transient Transfection of High Five Cells at Bioreactor Scale for HIV-1 VLP Production (2020) Nanomaterials 10(8), 1580
https://doi.org/10.3390/nano10081580
https://doi.org/10.1038/s41598-020-57761-w
Puente-Massaguer, E., Strobl, F., Grabherr, R., Striedner, G., Lecina, M., Gòdia, F., PEI-Mediated Transient Transfection of High Five Cells at Bioreactor Scale for HIV-1 VLP Production (2020) Nanomaterials 10(8), 1580
https://doi.org/10.3390/nano10081580
Publications (DSP)
Strategies for the purification of enveloped bionanoparticles have been developed based on chromatographic methods including conventional ion exchange resins, core shell beads with multimodal functionality, ion exchange membrane adsorbers and affinity resins. Additionally, particle detection and characterization methods are a main focus of our research. Recent data can be found in the following references:
Pereira Aguilar, P., Reiter, K., Wetter, V., Steppert, P., Maresch, D., Ling, W.L., Satzer, P., Jungbauer, A. “Capture and purification of Human Immunodeficiency Virus-1 virus-like particles: Convective media vs porous beads” (2020) Journal of Chromatography A.
https://doi.org/10.1016/j.chroma.2020.461378
Pereira Aguilar, P., Schneider, T.A., Wetter, V., Maresch, D., Ling, W.L., Tover, A., Steppert, P., Jungbauer, A. “Polymer-grafted chromatography media for the purification of enveloped virus-like particles, exemplified with HIV-1 gag VLP” (2019) Vaccine.
https://doi.org/10.1016/j.vaccine.2019.07.001
Pereira Aguilar, P., González-Domínguez, I., Schneider, T.A., Gòdia, F., Cervera, L., Jungbauer, A. “At-line multi-angle light scattering detector for faster process development in enveloped virus-like particle purification” (2019) Journal of Separation Science. https://doi.org/10.1002/jssc.201900441
Steppert, P., Burgstaller, D., Klausberger, M., Tover, A., Berger, E., Jungbauer, A. Quantification and characterization of virus-like particles by size-exclusion chromatography and nanoparticle tracking analysis(2017) Journal of Chromatography A, 1487, pp. 89-99. https://DOI: 10.1016/j.chroma.2016.12.085
Steppert, P., Burgstaller, D., Klausberger, M., Kramberger, P., Tover, A., Berger, E., Nöbauer, K., Razzazi-Fazeli, E., Jungbauer, A. Separation of HIV-1 gag virus-like particles from vesicular particles impurities by hydroxyl-functionalized monoliths (2017) Journal of Separation Science, 40 (4), pp. 979-990. https://DOI: 10.1002/jssc.201600765
Steppert, P., Burgstaller, D., Klausberger, M., Berger, E., Aguilar, P.P., Schneider, T.A., Kramberger, P., Tover, A., Nöbauer, K., Razzazi-Fazeli, E., Jungbauer, A. Purification of HIV-1 gag virus-like particles and separation of other extracellular particles (2016) Journal of Chromatography A, 1455, pp. 93- https://DOI: 10.1016/j.chroma.2016.05.053
Pereira Aguilar, P., Reiter, K., Wetter, V., Steppert, P., Maresch, D., Ling, W.L., Satzer, P., Jungbauer, A. “Capture and purification of Human Immunodeficiency Virus-1 virus-like particles: Convective media vs porous beads” (2020) Journal of Chromatography A.
https://doi.org/10.1016/j.chroma.2020.461378
Pereira Aguilar, P., Schneider, T.A., Wetter, V., Maresch, D., Ling, W.L., Tover, A., Steppert, P., Jungbauer, A. “Polymer-grafted chromatography media for the purification of enveloped virus-like particles, exemplified with HIV-1 gag VLP” (2019) Vaccine.
https://doi.org/10.1016/j.vaccine.2019.07.001
Pereira Aguilar, P., González-Domínguez, I., Schneider, T.A., Gòdia, F., Cervera, L., Jungbauer, A. “At-line multi-angle light scattering detector for faster process development in enveloped virus-like particle purification” (2019) Journal of Separation Science. https://doi.org/10.1002/jssc.201900441
Steppert, P., Burgstaller, D., Klausberger, M., Tover, A., Berger, E., Jungbauer, A. Quantification and characterization of virus-like particles by size-exclusion chromatography and nanoparticle tracking analysis(2017) Journal of Chromatography A, 1487, pp. 89-99. https://DOI: 10.1016/j.chroma.2016.12.085
Steppert, P., Burgstaller, D., Klausberger, M., Kramberger, P., Tover, A., Berger, E., Nöbauer, K., Razzazi-Fazeli, E., Jungbauer, A. Separation of HIV-1 gag virus-like particles from vesicular particles impurities by hydroxyl-functionalized monoliths (2017) Journal of Separation Science, 40 (4), pp. 979-990. https://DOI: 10.1002/jssc.201600765
Steppert, P., Burgstaller, D., Klausberger, M., Berger, E., Aguilar, P.P., Schneider, T.A., Kramberger, P., Tover, A., Nöbauer, K., Razzazi-Fazeli, E., Jungbauer, A. Purification of HIV-1 gag virus-like particles and separation of other extracellular particles (2016) Journal of Chromatography A, 1455, pp. 93- https://DOI: 10.1016/j.chroma.2016.05.053
Partners & Contact
Funding for the infrastructure was received by the Austrian Research Promotion Agency (FFG) through the “F&E Infrastrukturförderung”
