AIBY meeting

AIBY meeting

21 May 2008

ICT, PragueICT, Prague – who we are? – who we are?

• public university providing education and public university providing education and pursuing scientific, research, development and pursuing scientific, research, development and implementation activitiesimplementation activities,,

• member of the family of 28 public and state member of the family of 28 public and state universities in the Czech Republicuniversities in the Czech Republic..

ICT, PragueICT, Prague LocationLocation

Praha 6Praha 6 - Dejvice - Dejvice Technická 5Technická 5Czech RepublicCzech Republic

www.vscht.cz

StructureStructure Faculty of Chemical Technology Faculty of Chemical Technology Faculty of Environmental TechnologyFaculty of Environmental Technology Faculty of Food and Biochemical TechnologyFaculty of Food and Biochemical Technology Faculty of Chemical EngineeringFaculty of Chemical Engineering

ICT, Prague - peopleICT, Prague - people StaffStaff (780)(780)

40 professors40 professors 100 100 associate professors associate professors 280 assistant professors 280 assistant professors

and assistants and assistants

Students Students (3100)(3100) 2500 Master and Bachelor students2500 Master and Bachelor students 600 600 post-graduate post-graduate studentsstudents

Faculty of Food and Biochemical Faculty of Food and Biochemical TechnologyTechnology

The Faculty of Food Technology was The Faculty of Food Technology was established as part of the ICT, Prague in established as part of the ICT, Prague in 1952. 1952.

The faculty took the name The faculty took the name Faculty of Food Faculty of Food and Biochemical Technologyand Biochemical Technology in the in the academic year 1969-1970. academic year 1969-1970.

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 Group Brewery 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(512-545 nm), FL2(575-610 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)

Probe Physiological parameter

PI, EB, Sytox Green Viability, DNA

Acriflavine Glycogen

FITC Proteins

BOX, Rhodamine 123 Membrane potential

FDA, cFDA Esterase activity

Pichia pastoris stained with BOX Pichia pastoris stained with PI

dead cells

viable 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