1

schedule – AIBY 21 Mai 2008

08:30

• complementary competences (groups presentations)

• proposal of joint projects

• collaborators/competitors outside of AIBY

• arguments to EU 12:15

• vision

• AIBY focus (expression systems, products, technologies)

• next steps (schedule, To Do)

end 15:00

2

resources

http://www.biotech2008.ch

aiby2009

http://cms.biotechlab.net/information/materials_aiby/

3

Wädenswil

Bioprocess Technology

Institute of Biotechnology (IBT)

School of Life Sciences and Facility Management

Zurich University of Applied Sciences (ZHAW)

Waedenswil

4

Zürich University of Applied Sciences

the largest multidisciplinary UAS, > 6000 students

8 departments (Wädenswil, Winterthur, Zurich)

Masters’ Programme in Pharmaceutical Biotechnology

5

bioprocess technology

processtechnology

"software" forbiotechnology

high-productivity processesmicrobial glycosylation (E. coli, P. pastoris)

dynamics of physiologyfedbatch, flowcytometry

process control and simulation(software sensors, predictive control)

computer-intensive methodssignal extraction, clustering, etc.

eLearning www.biotechLAB.netknowledge managementadvanced training

6

high-productivity processes

1

7

methanol:glucose mixtures – trypsinogen

5

4

3

2

1

0

CD

W

glu

cose

m

eth

ano

l (

g L

-1)

121086420

time (h)

5

4

3

2

1

0

met

abo

lite

form

iate

(g

L-1

)

2.5

2.0

1.5

1.0

0.5

0.0

pro

du

ct (

U L

-1)

A B DC E

8

HCD-fedbatch Mut+ P. pastoris – trypsinogen

120

100

80

60

40

20

0

CD

W

met

han

ol

(g

L-1

)

3020100-10-20

production time (h)

300

250

200

150

100

50

0

pro

du

ct (

U L

-1)

A B C

open symbols – 100% methanolclosed symbols – 40%:60% mixture of glucose and methanol

9

10

product forms – Pichia pastoris

* product forms

rP productivity

rL rate of proteolytic degradation

rA rate of autocatalytic degradation

methanolbiomass (CDW)

metabolites

product 1product 2*product 3*...product i *

heat

O2

P. pastoris, Mut+

CO2

OH-/H+

methanol accumulation

proteases

rL

rA

rPdegradation product adegradation product bdegradation product c*...degradation product j *

11

microbial glycosylation

2

12

US$ 5000 per mg – mGM-CSF

16

14

12

10

8

6

4

2

0

f

eed

rat

e (g

CH

3O

H L

-1 h

-1)

100806040200

time (h)

400

300

200

100

0

mG

M-C

SF

(m

g L

-1)

120

100

80

60

40

20

0

CD

W (

g L

-1)

13

yield and glycosylation pattern = f (physiology)

14

yield and glycosylation pattern = f (physiology)

start of induction

end of cultivation

reference glycans

malto-dextrose reference

start of induction

end of cultivation

reference glycans

malto-dextrose reference

Man5GlcNAc2

15

16

single cell analysis

3

17

single cell analysis

averaged population subpopulations

18

100

80

60

40

20

0

50403020100

production time (h)

100

80

60

40

20

0

PI

stai

ned

(%

)

10 %

porcine trypsinogen, pH 4, (methanol accumulation < 80 g L-1

, Figure 1) porcine trypsinogen, pH 4, stepwise decreasing methanol addition porcine trypsinogen, pH 5 horseradish peroxidase, pH 4 horseradish peroxidase, pH 5 wilde type X33 strain, pH 4

vitality by FCM (Mut+ P. pastoris)– single cell analysis

100

80

60

40

20

0

50403020100

production time (h)

100

80

60

40

20

0

PI

stai

ned

(%

)

10 %

porcine trypsinogen, pH 4, (methanol accumulation < 80 g L-1

, Figure 1) porcine trypsinogen, pH 4, stepwise decreasing methanol addition porcine trypsinogen, pH 5 horseradish peroxidase, pH 4 horseradish peroxidase, pH 5 wilde type X33 strain, pH 4

19

20

complex functionalised biomolecules

1+2 = 4collagene III(high titre,

proper structure)

growth factor(e.g. PDGF )+ = dental implant

21

complex functionalised biomolecules

1+3 = 5novel

promotorsphysiology+ = knowledge

/advice base

http://www.biotechLAB.net

22

> 1000 proteins – Pichia pastoris

biomaterials(polymers)

pharmaceuticals

enzymes

peptides

http://faculty.kgi.edu/cregg/Pichia2004.htm

23

24

5 + 1 high-value product categories – yeasts (Pichia pastoris)

• therapeutic proteins

• (catalytic proteins)

• enzymes for the synthesis/modification of pharmaceuticals

• new (functional) biomaterials for (e.g. health care)

• drug target proteins

• proteins for diagnostics