AIBY meeting 21 May 2008

ICT, Prague – who we are? public university providing education and pursuing scientific, research, development and implementation activities,public university providing education and pursuing scientific, research, development and implementation activities, member of the family of 28 public and state universities in the Czech Republic.member of the family of 28 public and state universities in the Czech Republic.

ICT, Prague  Location  Praha 6 - Dejvice Technická 5 Czech Republic   Structure  Faculty of Chemical Technology  Faculty of Environmental Technology  Faculty of Food and Biochemical Technology  Faculty of Chemical Engineering

ICT, Prague - people  Staff(780)  40 professors  100 associate professors  280 assistant professors and assistants  Students (3100)  2500 Master and Bachelor students  600 post-graduate students

Faculty of Food and Biochemical Technology  The Faculty of Food Technology was established as part of the ICT, Prague in  The faculty took the name Faculty of Food and Biochemical Technology in the academic year

Faculty of Food and Biochemical Technology Department of Fermentation Chemistry and Bioengineering Department of Biochemistry and Microbiology Department of Carbohydrate Chemistry and Technology Department of Dairy and Fat Technology Department of Food Chemistry and Analysis Department of Food Preservation and Meat Technology Department of Chemistry of Natural Substances

Department of Fermentation Chemistry and Bioengineering Biotechnology GroupBrewery Group Prof. Karel Melzoch (Head of Department and Dean of Faculty)

Biotechnology group National and EU projects based on bioprocess optimization, microbial physiology, downstream processing, knowledge-based process control Training activities - biotechnology disciplines and physiology Research activities – cultivations, recombinant protein technology, bioprocess optimization, physiology (flow cytometry), downstream processing, analytics (HPLC, GC,…)

Biotechnology – teaching programme Combination of the principles of biological, chemical, engineering and economic studies role and behaviour of the biological element (microbial, plant and tissue cells, fragments and enzymes) regarding engineering and technological aspects of the bioprocess and connected up-stream and down-stream processing application of computers in bioprocess modelling, simulation studies and direct control of bioreactors students participate in regular research work in several research projects supported by Czech and European Grant Agencies (diploma thesis, PhD programme)

Biotechnology - research activities Optimization of bioprocesses employing microbial cells Optimization of heterologous protein production processes Study of physiological attributes of cells population under cultivation/stress condition Down-stream and Up-stream processes

Cooperation with Great Britain: University of Birmingham, University of Surrey, Brunel University of West London, Manchester Metropolitan University, University of Teeside Middlesbrough Portugal: INETI Lisboa, University of Lisbon Switzerland – Zurich University of Applied Sciences Poland: Akademia ekonomiczna, Wroclaw, Akademia rolnicza, Poznaň, Univesrsity of Lodz, Dept. Indust. Microbiol. Bulgary: Bulgarian Academy of Science, Inst. Microbiol., Sofia. Russia: Moscow State Academy of Chemical Engineering Holland: International Institute for Hydraulic and Environmental Engineering, Delft. Hungary: Biological Research Centre, Hungarian Acad. Sci., Biotechnol. Inst., Bay Z. Foundation Appl. Res. Techn. Szeged. Austria: Technische Universität Graz. Germany: Institut fü r Bioproceß und Analysenmeß technik Heiligenstadt, Forsungszentrum für Medizintechnik und Biotechnologie, Bad Langensalza. Italy: University of Naples "Federico II" Spain: Universidad de Valencia

Projets TEMPUS-PHARE COPERNICUS COST EUREKA ERASMUS/SOCRATES The Czech-American Program projects supported by the Czech Grant Agency, Czech Ministry of Agriculture National Agency for Agricultural Research, the Czech Ministry of the Environment, the Czech Academy of Sciences Grant Agency, etc.

Our contribution to AIBY Complete solution of heterologous protein production processes employing bacterial and yeast cells including cultivation, process optimization, down-stream and up-stream processes, process simulation, analytics including the analysis of selected physiological attributes of host strain

Cultivation of microorganisms Bacteria, yeasts, fungi B.Braun bioreactors (2 L, 5L, 7L) Possibility to scale-up to 1500 L (cooperation with Institute of Microbiology, Czech Academy of Sciences – Biotechnological Pilot Plant) 5-year experience with recombinant cultures (Eureka E!3415 succesfully finished in 2007)

Analysis of physiological attributes of cell population Flow cytometry (Flow cytometers Partec PAS III (Partec) and Quanta (Beckman-Coulter)) Ar-Ion laser for 488 nm excitation and HBO lamp for excitation in UVSSC, FSC, FL1( nm), FL2( nm), FL3 (>630 nm) Analysis of cell population on single cell level in rates of thousands in very short time period Staining with fluorescent probes – presence of active functions of cells Computer-aided picture analysis LUCIA G (CR)

ProbePhysiological parameter PI, EB, Sytox GreenViability, DNA AcriflavineGlycogen FITCProteins BOX, Rhodamine 123Membrane potential FDA, cFDAEsterase activity Pichia pastoris stained with BOXPichia pastoris stained with PI dead cells viable cells dead cells Cells stained with PI

Analytics HPLC analysis GC analysis Validation of methods Maldi-Toff analysis and protein sequencing (in cooperation with Department of Biochemistry and Microbiology) Mass spectrum of HPLC fraction determined by MALDI-TOF (Pichia pastoris)

Down-stream proceses Electrodialysis Micro-, ultra and nanofiltration Reverse osmosis Membrane units with ceramic membranes

Thank you for attention