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:
ÄKTA Pure 25 and 150, Cytiva (formerly GE Healthcare Life Sciences) coupled to
MALS Detector (DAWN® Heleos, Wyatt)
- 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.
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.
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.
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
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
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.
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.
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.
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.
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.
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-
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